用于降解错误折叠的蛋白质的组合物和方法与流程

文档序号:14915481发布日期:2018-07-11 00:36

本申请要求享有2015年5月29日提交的美国临时专利申请号62/168,309的优先权,其内容通过引证以其全部并入本文。

有关联邦政府资助的研究或开发的声明

本发明是伴随国立卫生研究院授予的CA088868、GM060911、CA182675、CA184867和P30AI045008下的政府支持做出的。政府对本发明具有一定的权利。



背景技术:

蛋白质是细胞的最丰富的大分子并且对几乎所有生理学过程是重要的。为了发挥它们的生物学功能,大部分蛋白质需要折叠成并维持它们的天然构象。尽管通过其氨基酸序列确定了蛋白质的天然构象,但是折叠过程异常复杂并且高度易于出错,并且在遗传突变、生物来源的错误和翻译后损伤等情况下可以进一步限制其应用(Dobson,2003,Nature,426:884-890;Goldberg,2003,Nature,426:895-899)。采取异常构象的蛋白质和由它们所形成的聚集体对细胞活力和功能具有持续的威胁。无法消除这些蛋白质与各种衰竭性人类疾病的病理发生密切相关(Selkoe,2003,Nature,426:900-904;Taylor et al.,2002,Science,296:1991-1995)。

为了对抗蛋白质错误折叠,细胞采用两套广泛的蛋白质质量控制(PQC)系统:辅助蛋白质实现它们天然构象的系统,和一旦形成错误折叠的蛋白质,消除它们的系统。前者主要由大量的分子伴侣和它们的协同分子伴侣组成,其以ATP依赖性的方式保护处于它们的非天然状态的蛋白质并减少错误折叠和聚集体。著名的实例包括(1)热休克蛋白70(Hsp70),其辅助各种蛋白质的折叠;(2)Hsp60/伴侣蛋白,其形成大分子笼以包封相对小的蛋白质用于不间断的折叠;和(3)HSP90,其最通常作用于参与细胞信号传导和转录的蛋白(Hartl et al.,2011,Nature,475:324-332)。

去除错误折叠的蛋白质的系统包括蛋白质解聚酶。例如,原核细胞或低等真核细胞中的Hsp100蛋白(例如,细菌中的ClpB和酵母中的Hsp104)可以再溶解蛋白质聚集体,从而与Hsp70及其协同分子伴侣Hsp40共同作用(Glover and Linquist,1998,Cell,94:73-82)。尽管如此,考虑到蛋白质错误折叠是不可避免的并且由于突变、生物来源的错误或不可修复的损伤而通常无法逆转,因此细胞最终依赖于降解系统来维持蛋白质量。然而,对这些系统的理解仍较差。尽管泛素-蛋白酶体途径以及自噬一定是这些系统的重要部分,但是它们如何选择性识别错误折叠的蛋白质并靶向它们来降解的关键问题仍是难以理解的(Goldberg,2003,Nature,426:895-899;Tyedmers et al.,2010,Nat Rev Mol Cell Biol,11:777-788)。

此外,与其它细胞隔室,如内质网相比(Buchberger et al.,2010,Mol Cell,40:238-252),细胞核中的PQC系统是非常不清楚的。细胞核中的错误折叠的蛋白质对分裂期后的哺乳动物细胞(例如,神经元和心肌细胞)可以是特别有害的,不能在有丝分裂期间通过破环核膜来除去这些蛋白。通过神经元包涵体的形成强调了理解该细胞隔室中PQC的重要性,神经元包涵体的形成与多种显性遗传的神经退行性疾病,包括亨廷顿舞蹈病(HD)和几种类型的脊髓小脑性共济失调(SCA)有关。这些疾病是由编码polyQ的CAG重复的相关基因内的扩增引起的。它们在polyQ延伸超过疾病特异的阈值长度时出现,并且随其长度增加而逐渐变得更加严重(Orr and Zoghbi,2007,Annu Rev Neurosci,30:575-621)。

因此,本领域存在对于消除错误折叠的蛋白质的组合物和方法的需要。本发明满足了该尚未满足的需要。



技术实现要素:

在一个方面,本发明提供了用于治疗或预防与错误折叠的蛋白质或者蛋白质聚集体有关的疾病或病症的组合物,其中组合物包含一种或多种TRIM蛋白的调节剂。在一个实施方式中,调节剂增加一种或多种TRIM蛋白的表达或活性。在一个实施方式中,调节剂为以下中的至少一种:化合物、蛋白质、肽、拟肽、抗体、核糖酶、小分子化合物、核酸、载体和反义核酸。

在一个实施方式中,调节剂增加人TRIM3、TRIM4、TRIM5、TRIM6、TRIM7、TRIM9、TRIM11、TRIM13、TRIM14、TRIM15、TRIM16、TRIM17、TRIM19(在本文中还称为“PML”)、TRIM20、TRIM21、TRIM24、TRIM25、TRIM27、TRIM28、TRIM29、TRIM32、TRIM34、TRIM39、TRIM43、TRIM44、TRIM45、TRIM46、TRIM49、TRIM50、TRIM52、TRIM58、TRIM59、TRIM65、TRIM67、TRIM69、TRIM70、TRIM74和TRIM75;和小鼠TRIM30中至少一种的表达或活性。

在一个实施方式中,组合物包含含有一种或多种TRIM蛋白的分离的肽。在一个实施方式中,分离的肽还包含细胞穿透肽(CPP)以使得分离的肽进入细胞。在一个实施方式中,CPP包含HIV tat的蛋白转导域。

在一个实施方式中,组合物包含编码一种或多种TRIM蛋白的分离的核酸分子。

在一个实施方式中,疾病或病症为polyQ病症。在一个实施方式中,疾病或病症是神经退行性疾病或病症,其选自由脊髓小脑性共济失调(SCA)1型(SCA1)、SCA2、SCA3、SCA6、SCA7、SCA17、亨廷顿舞蹈病、齿状核红核苍白球路易体萎缩症(DRPLA)、阿尔茨海默氏病、帕金森氏症、肌萎缩性侧索硬化(ALS)、传染性海绵状脑病(朊病毒疾病)、滔蛋白病变和额颞叶脑退行性病变(FTLD)。在一个实施方式中,疾病或病症选自AL淀粉样变性、AA淀粉样变性、家族性地中海热、老年系统性淀粉样变性、家族性淀粉样多神经病、透析相关性淀粉样变性、ApoAI淀粉样变性、ApoAII淀粉样变性、ApoAIV淀粉样变性、芬兰型遗传性淀粉样变性、溶菌酶淀粉样变性、纤维蛋白原淀粉样变性、冰岛型遗传性脑淀粉样血管病、II型糖尿病、甲状腺髓样癌、心房性淀粉样变性、遗传性脑出血伴淀粉样变性、垂体泌乳素瘤、注射定位的淀粉样变性、主动脉内侧淀粉样变性、遗传性格子状角膜变性、与倒睫相关的角膜淀粉样变性、白内障、牙源性钙化上皮肿瘤、肺泡蛋白沉积症、包涵体肌炎和苔藓型皮肤淀粉样变性。在一个实施方式中,疾病或病症是与p53突变聚集体有关的癌症。

在一个方面,本发明提供了治疗或预防对其有需要的受试者的与错误折叠的蛋白质或者蛋白质聚集体有关的疾病或病症的方法,其中方法包括向受试者给予包含一种或多种TRIM蛋白的调节剂的组合物。在一个实施方式中,调节剂增加一种或多种TRIM蛋白的表达或活性。在一个实施方式中,调节剂为以下中的至少一种:化合物、蛋白质、肽、拟肽、抗体、核糖酶、小分子化合物、核酸、载体和反义核酸。

在一个实施方式中,调节剂增加人TRIM3、TRIM4、TRIM5、TRIM6、TRIM7、TRIM9、TRIM11、TRIM13、TRIM14、TRIM15、TRIM16、TRIM17、TRIM19(在本文中还称为“PML”)、TRIM20、TRIM21、TRIM24、TRIM25、TRIM27、TRIM28、TRIM29、TRIM32、TRIM34、TRIM39、TRIM43、TRIM44、TRIM45、TRIM46、TRIM49、TRIM50、TRIM52、TRIM58、TRIM59、TRIM65、TRIM67、TRIM69、TRIM70、TRIM74和TRIM75;和小鼠TRIM30中至少一种的表达或活性。

在一个实施方式中,组合物包含含有一种或多种TRIM蛋白的分离的肽。在一个实施方式中,分离的肽还包含细胞穿透肽(CPP)以使得分离的肽进入细胞。在一个实施方式中,CPP包含HIV tat的蛋白转导域。

在一个实施方式中,组合物包含编码一种或多种TRIM蛋白的分离的核酸分子。

在一个实施方式中,疾病或病症为polyQ病症。在一个实施方式中,疾病或病症是神经退行性疾病或病症,其选自脊髓小脑性共济失调(SCA)1型(SCA1)、SCA2、SCA3、SCA6、SCA7、SCA17、亨廷顿舞蹈病、齿状核红核苍白球路易体萎缩症(DRPLA)、阿尔茨海默氏病、帕金森氏症、肌萎缩性侧索硬化(ALS)、传染性海绵状脑病(朊病毒疾病)、滔蛋白病变和额颞叶脑退行性病变(FTLD)。在一个实施方式中,疾病或病症选自AL淀粉样变性、AA淀粉样变性、家族性地中海热、老年系统性淀粉样变性、家族性淀粉样多神经病、透析相关的淀粉样变性、ApoAI淀粉样变性、ApoAII淀粉样变性、ApoAIV淀粉样变性、芬兰型遗传性淀粉样变性、溶菌酶淀粉样变性、纤维蛋白原淀粉样变性、冰岛型遗传性脑淀粉样血管病、II型糖尿病、甲状腺髓样癌、心房性淀粉样变性、遗传性脑出血伴淀粉样变性、垂体泌乳素瘤、注射定位的淀粉样变性、主动脉内侧淀粉样变性、遗传性格子状角膜变性、与倒睫相关的角膜淀粉样变性、白内障、牙源性钙化上皮肿瘤、肺泡蛋白沉积症、包涵体肌炎和苔藓型皮肤淀粉样变性。在一个实施方式中,疾病或病症是与p53突变聚集体有关的癌症。

在一个实施方式中,方法包括向受试者的至少一种神经细胞给予组合物。

在一个方面,本发明提供了用于治疗或预防与功能突变蛋白的降解有关的疾病或病症的组合物,其中组合物包含一种或多种TRIM蛋白的调节剂。在一个实施方式中,调节剂增加一种或多种TRIM蛋白的表达或活性。在一个实施方式中,调节剂为以下中的至少一种:化合物、蛋白质、肽、拟肽、抗体、核糖酶、小分子化合物、核酸、载体和反义核酸。在一个实施方式中,疾病或病症为囊性纤维化。

在一个方面,本发明提供了用于治疗或预防对其有需要的受试者中与功能突变蛋白的降解有关的疾病或病症的方法,其中方法包括向受试者给予包含一种或多种TRIM蛋白的调节剂的组合物。在一个实施方式中,调节剂增加一种或多种TRIM蛋白的表达或活性。在一个实施方式中,调节剂为以下中的至少一种:化合物、蛋白质、肽、拟肽、抗体、核糖酶、小分子化合物、核酸、载体和反义核酸。在一个实施方式中,疾病或病症为囊性纤维化。

在一个方面,本发明提供了治疗或预防与错误折叠的蛋白质或者蛋白质聚集体有关的疾病或病症的组合物,其中组合物包含一种或多种SUMO靶向的泛素连接酶(STUbl)的调节剂。在一个实施方式中,调节剂增加一种或多种STUbL的表达或活性。在一个实施方式中,调节剂为以下中的至少一种:化合物、蛋白质、肽、拟肽、抗体、核糖酶、小分子化合物、核酸、载体和反义核酸。在一个实施方式中,调节剂增加RNF4的表达或活性。

在一个方面,本发明提供了治疗或预防对其有需要的受试者中与错误折叠的蛋白质或者蛋白质聚集体有关的疾病或病症的方法,其中方法包括向受试者给予包含一种或多种STUbL的调节剂的组合物。在一个实施方式中,调节剂增加一种或多种STUbL的表达或活性。在一个实施方式中,调节剂为以下中的至少一种:化合物、蛋白质、肽、拟肽、抗体、核糖酶、小分子化合物、核酸、载体和反义核酸。在一个实施方式中,调节剂增加RNF4的表达或活性。

在一个方面,本发明提供了用于产生重组蛋白的方法,其中方法包括向修饰以表达重组蛋白的细胞给予一种或多种TRIM蛋白的调节剂。在一个实施方式中,调节剂包含含有一种或多种TRIM蛋白的分离的肽。在一个实施方式中,调节剂包含编码一种或多种TRIM蛋白的分离的核酸分子。

附图说明

当结合附图阅读时,将更好地理解本发明优选实施方案的下列详细说明。出于说明本发明的目的,在附图中显示了本发明优选的实施方式。然而,应理解本发明不限于附图中所示实施方式的特定布置和方法。

图1包含图1A至图1L,其显示了表明PML促进Atxn1 82Q及其它细胞核错误折叠的蛋白质的降解的实例实验结果。(图1A)用抗PML抗体(红色)和DAPI(蓝色)对Atxn1 82Q-GFP转染的HeLa细胞染色。显示了单个和合并的图像。比例尺10μm。(图1B)在HeLa细胞中单独或与PML一起表达Atxn1 82Q-GFP。左图,细胞的代表性荧光图像。比例尺20μm。右图,基于Atxn1 82Q-GFP包涵体尺寸的细胞定量。(图1C)在HeLa细胞中单独或与PML一起表达Atxn1 82Q-GFP(左图),或者在先前用对照(-)或PML siRNA处理的HeLa细胞中单独表达Atxn182Q-GFP。通过过滤器阻滞测定(对于SR部分)或免疫印迹分析(WB;对于其它部分),分析细胞裂解液部分(当指明时)和全细胞裂解液(WCL)。指出了分子量标准(以kDa计)和SS或SR Atxn1相对于肌动蛋白的相对比率。(图1D)通过WB分析,当在HeLa细胞中单独或与PML一起表达时(左图),或者在用对照或PML siRNA处理的HeLa细胞中单独表达时(右图),FLAG-Atxn1 82Q或30Q的稳态水平。(图1E)通过脉冲追踪测定和放射自显影分析,PML对总FLAG-Atxn1 82Q蛋白稳定性的影响。显示了35S标记的Atxn1 82Q的相对量。(图1F和图1G)通过CHX处理和WB分析,PML过表达(图1F)和降低(图1G)对Atxn1 82Q-GFP稳定性的影响。(图1H)在不存在或存在MG132的情况下,PML对Atxn182Q-GFP水平的影响。(图1I)顶部图,在不存在或存在PML的情况下,具有细胞质(左图)和细胞核(右图)包涵体的表达Httex1p 97QP细胞的相对百分比(平均值+SD,n=3)。底部图,用抗Htt抗体免疫染色的转染细胞的代表性荧光图像。箭头表示Httex1p 97QP聚集体。(图1J和图1K)在有和没有PML过表达的细胞中,HA-Httex1p 97QP和HA-Httex1p 97QP(KR)(图1J)和GFP-TDP-43(图1K)的水平。几乎所有Htt聚集体处于SR部分中。(图1L)通过CHX处理和WB分析,对照和PML-耗尽的细胞中nFucDM-GFP的稳定性。

图2包含图2A至图2E,显示了实例实验的结果,其表明了PML对错误折叠的蛋白质的识别。(图2A)通过体外牵出试验,然后通过WB(顶部和底部图)和丽春红S染色(中间图)分析,GST-Htt 25Q和GST-Htt 103Q对固定的FLAG-PML和FLAG-GFP(阴性对照)的结合。*IgG重链。(图2B)如(图2A)中分析的,GST-PML(右图所示)和对照GST蛋白对固定在Ni-NTA珠上的天然(N)和脲变性的(D)荧光素酶(luc)的结合。*结合至对照珠的来自BL21细菌裂解液的非特异性蛋白质。(图2C)通过WB(顶部和中间图)和考马斯亮蓝染色(底部图)分析,所指示的GST-Htt融合蛋白对缀合在抗Flag M2珠或对照珠上的FLAG-PML CC的结合。(图2D)PML F12/SRS2(右图所示)对来源于荧光素酶的肽文库的结合。显示了每行中斑点印迹的第一个肽的N末端氨基酸和最后一个肽的数目。(图2E)相对于其在荧光素酶肽阵列中的出现,PML SRS2-结合肽中每个氨基酸的出现。

图3包含图3A至图3F,显示了表明SUMO2/3参与泛素化和PML介导的Atxn1 82Q降解的实例实验结果。(图3A)不使用或使用MG132处理的HeLa细胞中Atxn1 82Q的SUMO1和SUMO2/3修饰。为了更好地比较修饰的Atxn1 82Q,通过WB分析具有类似的未修饰的Atxn1 82Q水平的变性免疫沉淀(d-IP)产物。*非特异性条带。(图3B)使用或不使用MG132处理的表达GFP-SUMO2-或GFP-SUMO3的U2OS细胞中Atxn182Q的定位(通过抗FLAG抗体检测,红色)。比例尺,10μm。(图3C)通过d-IP,随后通过WB(顶部图)和丽春红S染色(底部图)分析的HeLa细胞中Atxn1 82Q和30Q的SUMO2/3修饰。(图3D)在先前用指明的siRNA转染的或者未转染的(-)HeLa细胞中,单独或者与SUMO1或HA-SUMO2KR一起表达Atxn1 82Q。细胞用或不用MG132处理。通过d-IP和WB分析FLAG-Atxn1 82Q的SUMO化和泛素化。(图3E)在用指出的siRNA预处理的HeLa细胞中单独或者与增加量的PML一起表达的Atxn1 82Q-GFP的水平。(图3F)在存在或不存在SUMO1或SUMO2KR的情况下,PML对Atxn1 82Q-GFP水平的影响。

图4包含图4A至图4F,显示了表明PML促进Atxn1 82Q的SUMO化的实例实验的结果。(图4A)在不存在或存在HA-PML细胞并且不使用或使用MG132处理的情况下,HeLa细胞中FLAG-Atxn1 82Q的SUMO化。调节用于转染的Atxn1 82Q DNA的量以获得相当水平的未修饰的蛋白质。(图4B和图4C)在不使用或使用MG132处理的对照和PML siRNA转染的HeLa细胞中FLAG-Atxn1 82Q(图4B)和FLAG-nFlucDM-GFP(图4C)的SUMO化。(图4D和图4E)在存在如指出的重组FLAG-PML、FLAG-PML M6和SUMO2的情况下,进行纯化的HA-Atxn1 82Q-FLAG的SUMO化。在(图4D)中,调节不同d-IP产物的量以获得类似水平的未修饰的Atxn1 82Q(中间)。(图4F)在不存在或存在增加量的PML或PML M6的情况下,HeLa细胞中Atxn1 82Q-GFP的水平。

图5包含图5A至图5I,显示了表明RNF4在Atxn1 82Q降解中的作用的实例实验结果。(图5A)不具有和具有RNF4过表达的HeLa细胞中Atxn1 82Q-GFP的水平。(图5B和图5C)通过CHX处理和WB分析,对照和消除PML的HeLa细胞中RNF4过表达对Atxn1 82Q-GFP稳定性的影响。(图5C)中显示了相对SS Atxn1 82Q/肌动蛋白比率。(图5D)用对照siRNA(-)或者三种RNF4siRNA的组合预处理的HeLa细胞中FLAG-Atxn1 82Q的水平。(图5E)对照和RNF4降低的HeLa细胞中Atxn182Q-GFP的代表性荧光图像。比例尺,20μm。(图5F)用载体(DMSO)或者MG132处理的HeLa细胞中Atxn1 82Q-GFP和内源RNF4的定位。比例尺,10μm。(图5G和图5H)在不具有和具有RNF4过表达的HeLa细胞中FLAG-Atxn1 82Q和FLAG-Atxn1 30Q(图5G)或者HA-Httex1p97QP和HA-Httex1p 97QP(KR)(图5H)的水平。(图5I)稳定表达shCtrl和shRNF4的HeLa细胞中nFlucDM-GFP的稳定性(左图)和这些细胞中RNF4的水平(右图)。

图6包含图6A至图6I,显示了表明RNF4促进SUMO2/3-修饰的Atxn182Q的泛素化和降解的实例实验结果。(图6A和图6B)在使用或不使用MG132处理的HeLa细胞中,在不存在或存在RNF4的情况下,SUMO化的FLAG-Atxn1 82Q(图6A)和FLAG-nFlucDM-GFP(图6B)的水平。(图6C)如(图6A)中所分析,在用对照siRNA或RNF4siRNA的组合预处理的HeLa细胞中SUMO化的FLAG-Atxn1 82Q的水平。(图6D和图6E)在不存在或存在GST-RNF4的情况下,将结合在M2珠(+)上的未修饰和SUMO2-修饰的FLAG-Atxn1 82Q蛋白或者对照M2珠(-)与泛素化反应混合物一起温育。(图6D)实验设计的示意图。(图6E)FLAG-Atxn182Q(左图)和GST-RNF4(右图)的WB分析。(图6F和图6G)HeLa中Atxn1 82Q-GFP和RNF4蛋白(通过抗FLAG抗体检测)的定位。比例尺,10μm。(图6H)HeLa细胞中指出的RNF4蛋白对Atxn1 82Q-GFP水平的影响。(图6I)用对照或RNF4siRNA预处理的HeLa细胞中PML过表达对Atxn1 82Q-GFP水平的影响。

图7包含图7A至图7I,显示了表明PML缺陷加剧SCA1小鼠模型的行为和病理表型的实例实验结果。(图7A和图7B)在7(图7A)和11(图7B)周龄时,在加速转棒上的停留时间(平均值+SEM),括号中显示了动物的数目。(图7C和图7D)用苏木精染色的12周大动物的小脑切片。(图7C)分子层厚度的定量(平均值±SEM,n=3只小鼠/基因型)。(图7D)染色的代表性图像。比例尺,200μm。(图7E和图7F)用抗钙结合蛋白抗体染色的1岁大动物的小脑切片。(E)浦肯野细胞的定量,绘图为每1mm长度的体细胞平均数目(平均值±SEM,n=4只小鼠/基因型)。(F)染色的代表性图像。比例尺,200μm。(图7G和图7H)用抗泛素抗体染色并用苏木精复染的12周大小鼠的小脑皮层切片。(图7G)具有聚集体的浦肯野细胞的百分比(平均值±SEM,n=3只小鼠/基因型)。(图7H)免疫组织化学染色的代表性图像。箭头表示浦肯野细胞体中的泛素阳性的聚集体。比例尺,50μm。在无Atxn1tg/-的小鼠中的浦肯野细胞中未观察到泛素阳性的聚集体(参见图14D)。(图7I)通过PML-RNF4系统的PQC的模型。PML通过SRS识别错误折叠的蛋白质并通过其SUMO E3连接酶活性将它们与聚SUMO2/3链结合(a)。RNF4使SUMO化-错误折叠的蛋白质泛素化(b)并将它们靶向蛋白酶体降解(c)。

图8包含图8A至图8G,显示了表明PML与Atxn1 82Q聚集体共定位并且降低不溶性Atxn1 82Q的实例实验结果。(图8A)在PML缺乏的(PML-/-)小鼠胚胎成纤维细胞(MEF)中与每种指出的PML同种型一起表达Atxn1 82Q-GFP。用抗PML抗体(红色)和DAPI(蓝色)对细胞染色。(图8B)用对照siRNA(-)、PML siRNA#4或PML siRNA#9处理的HeLa细胞中FLAG-Atxn1 82Q的水平。通过免疫印迹和过滤器阻滞测定分析细胞裂解液。显示了归一化至对照的SS部分中的Atxn1 82Q相对于肌动蛋白的比率。(图8C)用对照siRNA、PML siRNA#4(其靶向PMLmRNA的5'UTR)或PML siRNA#4加表达PML的开放阅读框的质粒转染HeLa细胞(因此耐受siRNA)。分析各个部分中Atxn1 82Q-GFP的水平。(图8D)用对照siRNA、PML siRNA#4和PML siRNA#9处理,然后用FLAG-Atxn1 82Q转染HeLa细胞。通过归一化至18S rRNA的水平的定量RT-PCR确定FLAG-Atxn1 82Q转录的水平。(图8E)在不存在或存在PML的情况下,在HeLa细胞中表达Atxn1 30Q-GFP。然后,用CHX将细胞处理指定的时间。(图8F)用nFlucDM-GFP和相应野生型荧光素酶(WT)蛋白转染HeLa细胞。通过抗PML抗体(红色)检测内源PML,通过DAPI(蓝色)检测DNA。比例尺:10μm。(图8G)PML降低导致不溶性突变体荧光素酶的积累。用对照或PML siRNA,然后用nFluc-GFP或nFlucDM-GFP转染HeLa细胞。SR部分含有极少量的nFlucDM-GFP。

图9包含图9A至图9E,显示了表明PML与致病性Htt蛋白的相互作用的实例实验结果。(图9A)PML优先与致病性Htt相互作用。将表达FLAG-GFP或FLAG-PML的293T细胞的裂解液与固定在谷胱甘肽珠上的GST-Htt 25Q或者GST-Htt 103Q(泳道1-3和5-7)或者与对照谷胱甘肽珠(泳道4和8)温育。通过免疫印迹分析输入和拉下部分。(图9B)野生型PML同种型IV(aa 1-633)(在本发明中称为PML)和多个缺失片段(F1-F10)的示意图。标记了RING域(R)、B1盒、B2盒和卷曲螺旋区(CC)。由线指示底物识别位点SRS1和SRS2。显示了PML与致病性Htt蛋白(Htt 103Q或者52Q)以及变性荧光素酶的相互作用。ND:未进行。(图9C)全长PML和PML缺失突变体与Htt的相互作用。在存在[35S]Met的情况下通过结合的体外转录/翻译产生PML蛋白,并且将PML蛋白与纯化的GST-Htt 103Q、GST-Htt 25Q或固定在谷胱甘肽珠上的GST温育。通过放射自显影分析输入样品和拉下样品中[35S]Met-标记的蛋白,并且通过考马斯亮蓝染色分析拉下样品中的GST蛋白。同时并且以相同方式(包括样品的量和放射自显影的暴露时间)分析了三个拉下样品组。将输入样品暴露较短的时间段。(图9D)Htt 52Q和CC-去稳定(cc-)突变体的序列。用红色显示了改变为Htt 52Q cc-中的Pro的Htt 52Q中的氨基酸。(图9E)如图2D所示,测定了纯化的PML CC区(在右侧显示)与纤维素-结合的荧光素酶肽扫描的相互作用。*TEV蛋白酶。

图10包含图10A至图10C,显示了表明PML与变性的荧光素酶的相互作用的实例实验结果。(图10A)PML片段与变性的荧光素酶的相互作用。将体外翻译的[35S]标记的全长FLAG-PML和FLAG-PML缺失突变体与固定在珠上的天然或变性的荧光素酶或与对照珠温育。通过放射自显影分析输入和与珠结合的PML蛋白,并通过考马斯亮蓝染色分析荧光素酶。*,非特异性条带。同时并且以相同方式(包括样品的量和放射自显影的暴露时间)分析了三个拉下样品组。将输入样品暴露较短的时间段。(图10B和图10C)PML上第二底物识别位点(SRS2)的确认。(图10B)涵盖C末端的氨基酸的PML缺失突变以及它们与变性的荧光素酶相互作用的总结。(图10C)如(图10A)中描述的,测试了体外翻译的[35S]标记的PML片段的GST融合蛋白或者GST与荧光素酶的相互作用。

图11包含图11A至图11G,显示了表明SUMO2/3对错误折叠的蛋白质的修饰的实例实验结果。(图11A)用FLAG-Atxn1 82Q或FLAG-Atxn182Q(5KR)转染并使用或不使用MG132处理HeLa细胞。通过d-IP分离FLAG-Atxn1 82Q和FLAG-Atxn1 82Q(5KR),并且通过免疫印迹分析它们的SUMO2/3修饰。(图11B)不使用或使用GFP-TDP-43转染并使用载体(DMSO)(-)或MG132(+)处理HeLa细胞。使用抗GFP抗体或对照抗体进行d-IP。通过免疫印迹分析d-IP产物。(图11C)用nFluc-GFP、nFlucSM-GFP和nFlucSM-GFP(每个还使用FLAG表位N末端标签)转染并使用或不使用MG132处理HeLa细胞。通过d-IP分离nFluc蛋白。通过免疫印迹分析全细胞裂解液(WCL)和IP产物。注意WT荧光素酶相对于SM/DM荧光素酶(顶部图,泳道1-3)的SUMO2/3修饰之间的差异不是由于WCL中整体SUMO2/3结合中的改变造成的(底部图)。(图11D)在用对照(Ctrl)、SUMO2/3或SUMO1siRNA预处理的HeLa细胞中表达Atxn1 82Q-GFP。通过免疫印迹分析细胞裂解液。(图11E和图11F)用对照(Ctrl)siRNA、SUMO2/3siRNA或SUMO1siRNA(图11E)处理或者用这些siRNA处理并然后用GFP转染的HeLa细胞(图11F)。通过免疫印迹分析细胞裂解液。(图11G)将PML和PML M6蛋白用于体外SUMO化测定。在293T细胞中表达FLAG-PML和FLAG-PML M6并通过抗FLAG(M2)珠纯化。通过考马斯亮蓝染色以及BSA标准(左图)和通过免疫印迹(右图)分析蛋白质。基于免疫印迹(右图)和通过质谱分析,确定PML和PML M6泳道(左图)两者中存在的两条其它条带(箭头)是PML片段。在底部显示了M6突变体的示意图。*:在本文其它处描述了点突变。

图12包含图12A至图12I,显示了表明RNF4促进错误折叠的蛋白质降解的实例实验结果。(图12A)单独或与RNF4一起表达Atxn1 82Q-GFP的HeLa细胞的代表性荧光图像。比例尺:20μm。图像来自与图1B中所示的相同的实验。(图12B)具有和不具有RNF4过表达的HeLa细胞中Atxn1 82Q-GFP的半衰期。样品来自与图1F中所示那些相同的实验。(图12C)用对照siRNA(-)、单独的RNF4siRNA或RNF4siRNA加siRNA-耐受性RNF4转染HeLa细胞。通过免疫印迹分析细胞裂解液。(图12D)在用对照siRNA和指出的RNF4siRNA预处理的HeLa细胞中表达FLAG-Atxn1 82Q。用抗FLAG M2珠,通过d-IP分离FLAG-Atxn1 82Q。使用指出的抗体,通过免疫印迹分析WCL和IP产物。(图12E)用载体(DMSO)或MG132处理表达Atxn1 82Q-GFP和FLAG-RNF4两者的HeLa细胞。通过抗FLAG抗体检测外源(Exo)RNF4。在用DMSO处理的对照细胞中,外源RNF4显示出与Atxn1 82Q-GFP聚集体的部分共定位。在MG132处理时,在100%的细胞中观察到外源RNF4与Atxn1 82Q聚集体的完全共定位。比例尺:10μm。(图12F)单独或与增加量的RNF4一起使用GFP-TDP-43转染的HeLa细胞的免疫印迹分析。(图12G)在用对照siRNA(-)或指出的RNF4siRNA预处理的细胞中表达GFP-TDP-43。在每个实验中对500个细胞计数。显示了具有GFP-TDP-43灶点的细胞百分比。(图12H)用GFP-TDP-43转染用RNF4siRNA处理的HeLa细胞。用抗RNF4抗体(红色)和DAPI(蓝色)对细胞染色。注意在其中RNF4降低的细胞中TDP-43形成聚集体(实心箭头),但是在对照细胞中扩散(空心箭头)。(图12I)用GFP-TDP-43转染并用MG132处理HeLa细胞。用抗RNF4抗体(红色)对内源RNF4免疫染色。注意RNF4还在没有TDP-43的细胞中形成细胞核灶点(图12E-图12H)。

图13包含图13A至图13F,显示了表明SUMO2/3参与RNF4介导的Atxn1 82Q降解的实例实验结果。(图13A)在用对照siRNA、SUMO2/3siRNA和SUMO1siRNA预处理的HeLa细胞中表达Atxn1 82Q-GFP和/或RNF4。通过免疫印迹分析细胞裂解液。(图13B)首先用指出的RNF4siRNA或对照siRNA(-),然后用FLAG-Atxn1 82Q转染稳定表达GFP-SUMO2的U2OS细胞。显示了具有GFP-SUMO2阳性的Atxn1 82Q聚集体的转染的细胞的百分比(平均+SD,n=3)。在每个实验中,计数200个细胞。在右图中显示了转染的细胞的代表性图像。在无Atxn1 82Q表达的细胞中GFP-SUMO2不形成聚集结构。(图13C)野生型和突变体RNF4蛋白的示意图。显示了SUMO相互作用基序(SIM)1至4和RING域。*:在本文其它处描述了点突变。(图13D和图13E)将纯化的鼠(r)RNF4和RNF4CS1(图13D)或人RNF4和RNF4SIMm(图13E)的重组GST融合蛋白与泛素E1、E2(UbcH5a)和指出的泛素(Ub)加上Mg2+-ATP一起温育。使用抗GST抗体通过免疫印迹分析反应混合物。(图13F)Atxn182Q-GFP在用所指明的对照、PML和RNF4siRNA的所指明的组合处理的细胞中的表达。通过免疫印迹分析细胞裂解液。

图14包含图14A至图14D,显示了表明PML缺乏降低浦肯野细胞树突的分枝但不导致在浦肯野细胞中的聚集体的实验结果。(图14A)用抗浦肯野细胞特异性蛋白质钙结合蛋白的抗体对12周大小鼠的中矢状面小脑切片染色。由前顶裂的矩形区绘制荧光强度图(对于PML+/+,n=2只小鼠,并且对于所有其它基因型,n=3只小鼠)。与PML+/+小鼠相比,PML-/-小鼠显示出树突分枝的显著损失(ANOVA,p=0.031)。(图14B)钙结合蛋白免疫荧光的代表性共聚焦成像。比例尺:100μm。(图14C)具有和不具有Atxn1tg/-的12周的PML+/+、PML+/-和PML-/-小鼠中浦肯野细胞密度的定量,作为每1mm长度的体细胞平均数目作图(平均值±SEM,n=3只小鼠/基因型)。(图14D)来自不具有Atxn1tg/-的12周大的PML+/+和PML-/-小鼠的小脑皮层切片的免疫组织化学染色。对切片用抗泛素抗体染色并用苏木精复染。比例尺:50μm。注意,在那些切片中未检测到泛素阳性聚集体。图7G显示了PML+/+:Atxn1tg/-和PML-/-:Atxn1tg/-小鼠的染色切片。

图15包含图15A至图15D,显示了表明TRIM27、TRIM32和TRIM5δ与EGFP-Atxn1 82Q和Httex1p 97QP共定位的实施例实验结果。(图15A)在HeLa细胞中与指出的FLAG标签的TRIM蛋白一起表达Atxn182Q-GFP。用抗FLAG抗体(红色)和DAPI(蓝色)对细胞染色。比例尺:10μm。(图15B和图15C)在HeLa细胞中与FLAG标签的TRIM5δ、TRIM27(图15B)和TRIM32(图15C)一起表达Httex1p 97QP。用抗亨廷顿蛋白(绿色)和抗FLAG(红色)抗体对细胞免疫染色。注意不考虑Httex1p 97QP的细胞定位,TRIM32与Httex1p 97QP共定位。(图15D)用Atxn1 82Q-GFP转染HeLa细胞。通过抗TRIM27抗体(红色)检测内源TRIM27。箭头表示与Atxn1 82Q聚集体共定位的内源TRIM27体。比例尺:10μm。

图16包含图16A至图16D,显示了表明通过TRIM27、TRIM32和TRIM5δ降低聚集的Atxn1 82Q的实例实验结果。(图16A和图16B)当单独(-)或与指出的FLAG标签的TRIM蛋白一起在HeLa细胞中表达时Atxn1 82Q-GFP的水平。通过免疫印迹分析细胞裂解液。在(图16A)的左图中,提供了SS部分中的Atxn1 82Q相对于肌动蛋白的比率,并且可以在长时间暴露后检测到TRIM11的表达。(图16C)用对照(-)或TRIM27siRNA处理的HeLa细胞中Atxn1 82Q的表达水平。*,非特异性条带。(图16D)PIASy不抑制Atxn1 82Q蛋白的水平。使用指出的Atxn1 82Q-GFP、PIASy和PML转染的HeLa细胞的免疫印迹分析。

图17包含图17A至图17C,显示了表明TRIM27和TRIM32不依赖PML降低聚集的Atxn1 82Q的实例实验结果。(图17A)TRIM27与PML的部分共定位。用抗FLAG(绿色)和抗PML(红色)抗体对用FLAG-TRIM27转染的HeLa细胞免疫染色。(图17B)TRIM27与PML+/+和PML-/-MEF中Atxn1 82Q-GFP聚集体的共定位。通过抗FLAG(红色)抗体对FLAG-TRIM 27染色。(图17C)单独或与PML+/+和PML-/-MEF中指示的TRIM蛋白一起表达Atxn1 82Q-GFP。通过过滤器阻滞测定分析提取物。为了更好地比较TRIM蛋白对聚集体的影响,在右图显示了较轻的暴露。

图18包含图18A至图18G,显示了表明TRIM蛋白依赖于SUMO2/3和蛋白酶体来除去不溶性Atxn1 82Q的实例实验结果。(图18A-图18F)在不使用或使用MG132处理的细胞(图18A-图18C)或者在对照或SUMO2/3降低的细胞(图18D-图18F)中单独或与TRIM5δ、TRIM27和TRIM32一起表达Atxn1 82Q-GFP。通过免疫印迹分析细胞裂解液。(图18G)在存在指示的重组FLAG-TRIM11和SUMO2的情况下,进行纯化的HA-Atxn1 82Q-FLAG的SUMO化。使用变性免疫沉淀分离HA-Atxn182Q-FLAG。通过免疫印迹分析反应混合物和IP样品。

图19包含图19A至图19F,显示了表明TRIM蛋白相对于Atxn1 82Q的定位的实例实验结果。在HeLa细胞中与指示的HA标签的TRIM蛋白一起表达Atxn1 82Q-GFP(绿色)。用抗HA抗体(红色)对细胞免疫染色。显示了每种TRIM蛋白的代表性定位模式。(图19A)TRIM1-TRIM 16的代表性定位模式。(图19B)TRIM17-TRIM32的代表性定位模式。(图19C)TRIM33-TRIM41和TRIM44-TRIM 41的代表性定位模式。(图19D)TRIM52、TRIM54-TRIM58、TRIM62-TRIM66、TRIM73-TRIM74和TRIM76的代表性定位模式。用黄色表示在大多数细胞中显示与Atxn1 82Q-GFP共定位的TRIM蛋白。比例尺:10μm。

图20包含图20A和图20B,显示了TRIM蛋白对Atxn1 82Q和Httex1p97QP的系统分析结果。Atxn1 82Q-GFP(图20A)或Httex1p 97QP(图20B)在HeLa细胞中与指出的TRIM蛋白共表达。通过免疫印迹分析细胞裂解液。TRIM蛋白标记的红色和绿色分别是降低和增加的polyQ蛋白水平的那些,而标记为黑色的TRIM蛋白没有可观察的影响。注意TRIM蛋白的影响可以受它们的表达水平的影响,如在本文其它地方描述的。

图21包含图21A至图21C,显示了表明重组TAT-TRIM11降低错误折叠的蛋白质水平的实例实验结果。用Atxn1 82Q-GFP、Atxn1 30Q或Htt97Q-GFP转染HeLa细胞,并且随后与重组TRIM11或SUMO2蛋白一起温育。(图21A)用TRIM11处理HeLa细胞导致Atxn1 82Q水平强烈降低。(图21B)用TRIM11处理HeLa细胞也导致Htt 97Q水平强烈降低。(图21C)SUMO2对Atxn1 82Q水平的影响最小。

图22显示了在实例实验中使用的Tat-TRIM11的氨基酸序列。

图23包含图23A至图23F,显示了表明人脑样品的RNF4染色的实验结果。HD脑组织的抗RNF4(绿色)免疫染色。RNF4显示出扩散的细胞核定位(图23A-图23C)或形成的灶点(图23D-图23F)(通过箭头指示)。比例尺:10μm。

图24包含图24A至图24H,显示了表明RNF4与SCA1患者的神经元包涵体共定位的实验结果。SCA1脑组织使用抗polyQ(1C2)和抗RNF4抗体的免疫染色。比例尺:10μm。

图25包含图25A至图25L,显示了表明RNF4与SCA1患者的神经元包涵体共定位的实验结果。图25显示了HD脑组织使用抗Htt、抗泛素和两种单独的RNF4抗体的免疫染色。注意Htt和泛素形成了环状结构(图25A-图25H)或者均匀分布的包涵体(图25I-图25L),其中RNF4信号位于中心。

图26包含图26A至图26H,显示了表明RNF4与HD患者的神经元中的包涵体共定位的实验结果。显示了HD脑组织的Htt和RNF4免疫染色。使用了两种不同的RNF4抗体(#1和#2)。比例尺:10μm。

图27包含图27A至图27D,显示了表明Atxn1 82Q、p53和α-突触核蛋白的SUMO化的实验结果。图27A显示了其中当TRIM11WT、TRIM11MUT或PML共表达时,分析Atxn1 82Q的SUMO化的实验。在溶胞前,加入10μM MG132 4小时。图27B显示了在存在E1、E2和SUMO2的情况下,与TRIM11WT或TRIM11MUT温育的纯化的Atxn1 82Q的体外SUMO化。图27C显示了在存在E1、E2和SUMO2的情况下,与TRIM11或PML温育的纯化的Flag-p53的体外SUMO化。图27D显示了在存在E1、E2和SUMO2的情况下,与TRIM11WT温育的α-突触核蛋白的体外SUMO化。

图28包含图28A至图28G,显示了表明TRIM11补充至Atxn1 82Q聚集体的实验结果。图28A显示了293T细胞中转染的GFP-Hsp70的免疫荧光分析。图28B显示了293T细胞中转染的GFP-TRIM11的免疫荧光分析。图28C显示了免疫荧光分析,其示出可以将Hsp70补充到Atxn1 82Q的聚集体。图28D显示了免疫荧光分析,其示出可以将TRIM11补充到Atxn1 82Q的聚集体。图28E显示了用Atxn1 82Q、TRIM11或Hsp70转染的细胞的去污剂可溶性和不溶性部分的免疫印迹分析。在指示处加入10μM MG132 3小时。图28F显示了用Atxn1 82Q、TRIM11WT(野生型)或TRIM11MUT(突变)转染的细胞的去污剂可溶性和不溶性部分的免疫印迹分析。在指示处加入10μM MG132 3小时。图28G显示了其中用指示的质粒转染的HCT116细胞的免疫印迹。48小时后,将细胞裂解,然后用20μM硫黄素T(ThT)染色。

图29包含图29A至图29C,显示了表明TRIM11结合至Atxn1 82Q的实验结果。图29A显示了其中固定在珠上的FLAG-Atxn1 82Q与GST或GST-TRIM11温育的纯化实验。图29B是其中显示固定在珠上的纯化的FLAG-Atxn1 82Q或Flag-Atxn1 30Q与GST或GST-TRIM11温育的实验。图29C显示了GST-TRIM11和GST蛋白与固定在Ni-NTA珠上的天然(N)和脲变性的(D)荧光素酶(luc)的结合。

图30包含图30A至图30D,显示了表明TRIM11降低细胞聚集体的实验结果。图30A显示了其中使稳定表达GFP-Atxn1 82Q的HCT116细胞裂解,然后用20μM硫黄素T(ThT)染色的实验。图30B显示了用TRIM11转染的GFP-Atxn1 82Q-HCT116细胞的沉降分析。图30C显示了其中用TRIM11WT或TRIM11MUT转染稳定表达GFP-Atxn1 82Q的HCT116细胞的实验。48小时后,使细胞裂解并用20μM ThT染色。图30D显示了用TRIM11WT或TRIM11MUT转染GFP-Atxn1 82Q-HCT116的细胞的沉降分析。

图31包含图31A至图31H,显示了表明TRIM11作用为分子伴侣以防止聚集体形成的实验结果。图31A显示了其中将荧光素酶(10nM)与200nM GST、200nM GST-TRIM11或者200nM Hsp70在45℃温育指定时间的实验。将天然荧光素酶活性设置为100%。N=3。图31B显示了其中将GFP(0.45uM)与200nM GST、200nM GST-TRIM11或者200nM Hsp70在45℃温育指定时间的实验。将天然GFP荧光设置为100%。N=3。图31C显示了在无热冲击的情况下作为对照测量的HCT116中转染的荧光素酶的活性。在45℃热冲击30分钟后或者在温育箱中恢复3小时后,将荧光素酶活性相对于对照表示。图31D显示了在无热冲击的情况下作为对照测量的HCT116中转染的荧光素酶的活性。在45℃热冲击60分钟后或者在温育箱中恢复1.5小时或3小时后,将荧光素酶活性相对于对照表示。图31E显示了稳定表达对照载体或Flag-TRIM11的HCT116细胞的免疫印迹分析。图31F显示了ThT分析,示出通过GST、TRIM11或Hsp70防止β-淀粉样蛋白原纤维形成。图31G显示了沉降测定,显示了通过溶菌酶、GST或TRIM11防止Atxn1 82Q聚集体形成。通过免疫印迹和点-印迹测定显示了结果。图31H显示了沉降测定,示出通过L TRIM11防止p53聚集体形成。在指示处应用E1、E2、SUMO2或ATP。通过免疫印迹和点-印迹测定显示了结果。

图32包含图32A至图32G,显示了表明TRIM11转录的调节不需要HSF1的实验结果。图32A显示了其中用或不用热冲击(42℃)处理细胞1小时,然后恢复不同的时间的实验。使用指示的抗体对总细胞裂解液进行免疫印迹。图32B显示了其中用或不用热冲击(42℃)处理稳定表达Flag-TRIM11的HCT116细胞1小时,然后恢复不同时间的实验。使用指示的抗体对总细胞裂解液进行免疫印迹。图32C显示了其中用或不用热冲击(42℃)处理HeLa细胞1小时,然后恢复不同时间的实验。使用指示的抗体对总细胞裂解液进行免疫印迹。图32D显示了其中用或不用As2O3处理A549细胞30分钟,然后恢复不同时间的实验。使用指示的抗体对总细胞裂解液进行免疫印迹。图32E显示了其中用或不用H2O2处理A549细胞100分钟,然后恢复不同时间的实验。使用指示的抗体对总细胞裂解液进行免疫印迹。图32F显示了对热冲击响应的TRIM11、HSP70、HSP90和GAPDH的半定量PCR分析。图32G显示了其中用或不用热冲击处理稳定表达载体或HSF1的A549细胞并恢复3小时的实验。分析免疫印迹和半定量PCR。

图33包含图33A至图33F,显示了表明p53是热冲击响应中上调TRIM11的因子的实验结果。图33A显示了用热冲击处理并恢复的HCT116p53野生型或无p53细胞的免疫印迹。图33B显示了用热冲击处理并恢复的HCT116p53野生型或无p53细胞中TRIM11mRNA水平的qPCR分析。图33C显示了用或不用热冲击处理并恢复3小时的稳定表达对照(Ctrl)或p53shRNA的A549细胞的免疫印迹和半定量PCR分析。图33D显示了用或不用热冲击处理并恢复3小时的Ctrl或p53siRNA转染的HCT116细胞的免疫印迹和半定量PCR分析。图33E显示了加热并恢复24小时的HCT116细胞的存活的结晶紫分析。在指示处加入KRIBB11。图33F显示了通过OD490分析的图33E中存在的相对细胞数目结果。

图34包含图34A至图34G,显示了表明TRIM11作用为解聚酶以溶解预形成的聚集体的实验结果。图34A显示了使用提高浓度的溶菌酶、GST或GST-TRIM11的预形成的荧光素酶聚集体的解聚和再激活(n=3)。图34B显示了沉降测定,示出通过GST或GST-TRIM11溶解的热聚集的荧光素酶。通过免疫印迹显示结果。图34C显示了使用增加浓度的溶菌酶、GST或GST-TRIM11预形成的GFP聚集体的解聚和再激活(n=3)。图34D显示了沉降测定,示出通过GST或GST-TRIM11溶解的热聚集的GFP。通过免疫印迹显示结果。图34E显示了沉降测定,示出通过溶菌酶、GST或TRIM11溶解的预形成的Atxn1 82Q聚集体。通过免疫印迹显示结果。图34F显示了沉降测定,示出通过1μM Hsp70和0.5μM Hsp40解聚的预形成的Atxn1 82Q聚集体(左图)和p53聚集体(右图)。图34G显示了沉降测定,示出通过0.5μM GST、0.5μM TRIM11、1μM Hsp70、0.5μM Hsp40或1μM Hsp104解聚的预形成的Atxn1 82Q聚集体。

图35包含图35A至图35D,其显示了表明再折叠活性需要全长TRIM11的实验结果。图35A显示了TRIM11结构的示意图。图35B显示了使用增加浓度的指示的蛋白质对预形成的荧光素酶聚集体的解聚和再激活(n=3)。图35C显示了沉降测定,示出通过GST、TRIM11、RBC或B30.2解聚的加热的荧光素酶聚集体。图35D显示了使用指示的蛋白对预形成的荧光素酶聚集体的解聚和再激活(n=3)。

图36包含图36A和图36B,其显示了表明TRIM11的解聚功能需要TRIM11结合至底物的实验结果。图36A显示固定在珠上的纯化的FLAG-Atxn1 82Q与GST、GST-TRIM11、GST-RBC或者GST-B30.2温育。图36B显示固定在珠上的纯化的FLAG-Atxn1 82Q与GST、GST-TRIM11或其它TRIM11片段温育。

图37包含图37A至图37E,显示了表明TRIM11不依赖于其SUMO E3连接酶活性进行解聚的实验结果。图37A显示了其中将荧光素酶(10nM)与200nM GST、200nM GST-TRIM11WT或MUT在45℃下温育指定时间的实验。将天然荧光素酶活性设置为100%。N=3。图37B显示了200nMGST、200nM GST-TRIM11WT或MUT对预形成的荧光素酶聚集体的解聚和再激活。图37C显示了GST、GST-TRIM11WT或MUT与固定在Ni-NTA珠上的天然的(N)和脲变性的(D)的荧光素酶(luc)的结合。图37D显示了293T细胞中转染的GFP-TRIM11MUT的免疫荧光分析。图37E显示了显示可以将TRIM11MUT补充至Atxn1 82Q聚集体的免疫荧光分析。

图38包含图38A至图38E,显示了表明TRIM11还可以预形成α-突触核蛋白淀粉样蛋白原纤维形成并解聚预形成的α-突触核蛋白纤维的实验结果。图38A显示了ThT分析,示出通过GST、TRIM11、Hsp70、Hsp40或Hsp104防止α-Syn原纤维形成。图38B显示了ThT分析,示出TRIM11以剂量依赖性方式防止α-Syn原纤维形成。图38C显示了与GST或GST-TRIM11温育的α-Syn单体的纤维形成的EM图像。图38D显示了沉降测定,示出通过TRIM11和Hsp104A503S解聚的预形成的α-Syn纤维。图38E显示了ThT分析,示出GST、TRIM11或Hsp104对预先形成的α-Syn纤丝的解聚。

图39包含图39A至图39D,显示了表明TRIM21具有与TRIM11类似的解聚功能的实验结果。图39A显示GST、GST-TRIM11WT或MUT与固定在Ni-NTA珠上的天然的(N)和脲变性的(D)荧光素酶(luc)的结合。图39B显示了沉降测定,示出通过GST或GST-TRIM21溶解的热-聚集的荧光素酶。图39C显示了使用增加浓度的GST或GST-TRIM21对预形成的荧光素酶聚集体的解聚和再激活(n=3)。图39D显示了其中将荧光素酶(10nM)与200nM GST或200nM GST-TRIM21在45℃温育1分钟的荧光素酶测定。将天然荧光素酶活性设置为100%。N=3。

图40包含图40A至图40E,显示了表明PML和Atxn1 82Q解聚的实验结果。图40A显示了从293T细胞纯化的Flag-PML6的考马斯亮蓝染色。图40B显示了使用增加浓度的溶菌酶或Flag-PML6对预形成的GFP聚集体的解聚和再激活(n=3)。图40C显示了使用溶菌酶、Flag-PML6或GST-TRIM11对预形成的GFP聚集体的解聚和再激活(n=3)。图40D显示了从293T细胞纯化的Flag-PML4片段的考马斯亮蓝染色。图40E显示了使用不同的PML4片段对预形成的GFP聚集体的解聚和再激活(n=3)。

图41包含图41A至图41B,显示了表明小鼠原代海马神经元中TRIM11的实验结果。图41A显示了MTT分析,示出GST或TRIM11温育的α-Syn诱导的细胞死亡。图41B显示了α-Syn纤维处理的海马神经元细胞中p-α-Syn或者p62的免疫荧光分析。

图42包含图42A至图42C,其显示了表明在皮层和海马神经元中对热冲击响应上调TRIM11的实验结果。图42A显示了在42℃热冲击处理30分钟并恢复3小时的小鼠原代皮层神经元的免疫印迹。图42B显示了在42℃热冲击处理30分钟并恢复3小时的小鼠原代海马神经元的免疫印迹。图42C显示了海马神经元中对热冲击响应的TRIM11、HSP70、HSP90和GAPDH的半定量PCR分析。

具体实施方式

本发明涉及对三结构域(TRIM)蛋白家族成员和SUMO依赖性泛素连接酶RNF4在错误折叠的蛋白质的识别和降解中的作用的发现,它们在多种神经退行性病症的病理中起作用。

在一个方面,本发明提供了治疗或预防与错误折叠的蛋白质或者蛋白质聚集体有关的疾病或病症的组合物和方法。本文表明TRIM蛋白在作为伴侣蛋白靶向错误折叠的蛋白质用于蛋白酶体降解和在使蛋白质聚集体或包涵体解聚中起作用。因此,在某些方面,本发明可以用于消除胞内或胞外错误折叠的蛋白质、蛋白质聚集体或蛋白质包涵体。

例如,在某些实施方式中,本发明提供了在对其有需要的受试者中治疗或预防神经退行性病症的组合物和方法。例如,在某些实施方式中,本发明提供了治疗或预防神经退行性病症的组合物和方法,神经退行性病症是多聚谷氨酰胺(polyQ)病症,其中CAG密码子的重复编码了具有多聚谷氨酰胺片段的蛋白质,其可以导致产生错误折叠的蛋白质聚集体。示例性的polyQ病症包括(但不限于)脊髓小脑性共济失调(SCA)1型(SCA1)、SCA2、SCA3、SCA6、SCA7、SCA17、亨廷顿舞蹈病和齿状核红核苍白球路易体萎缩症(DRPLA)。在某些实施方式中,本发明提供了治疗与错误折叠的蛋白质或者蛋白质聚集体有关的神经退行性病症的组合物和方法,神经退行性病症包括(但不限于)阿尔茨海默氏病、帕金森氏症、肌萎缩性侧索硬化(ALS)、传染性海绵状脑病(朊病毒疾病)、滔蛋白病变和额颞叶脑退行性病变(FTLD)。然而,本发明不局限于神经退行性病症的治疗或预防。而是,本发明涵盖了与错误折叠的蛋白质或蛋白质聚集体有关的任何疾病或病症的治疗或预防。其它该类疾病和病症包括(但不限于)AL淀粉样变性、AA淀粉样变性、家族性地中海热、老年系统性淀粉样变性、家族性淀粉样多神经病、透析相关的淀粉样变性、ApoAI淀粉样变性、ApoAII淀粉样变性、ApoAIV淀粉样变性、芬兰型遗传性淀粉样变性、溶菌酶淀粉样变性、纤维蛋白原淀粉样变性、冰岛型遗传性脑淀粉样血管病、II型糖尿病、甲状腺髓样癌、心房性淀粉样变性、遗传性脑出血伴淀粉样变性、垂体泌乳素瘤、注射局部化淀粉样变性、主动脉内侧淀粉样变性、遗传性格子状角膜变性、与倒睫相关的角膜淀粉样变性、白内障、牙源性钙化上皮肿瘤、肺泡蛋白沉积症、包涵体肌炎和苔藓型皮肤淀粉样变性。在某些实施方式中,本发明涵盖了与p53突变聚集体有关的癌症的治疗或预防,其包括(但不限于)膀胱癌、星形细胞瘤、咽癌、淋巴瘤和腺癌。

在一个方面,本发明涵盖了一种或多种TRIM蛋白稳定错误折叠的蛋白质的用途。在某些方面,通过本文描述的一种或多种TRIM蛋白稳定化功能性错误折叠的蛋白质可以治疗或预防与错误折叠的蛋白质有关的疾病或病症。例如,在一个实施方式中,通过本文所述的一种或多种TRIM蛋白稳定化突变囊性纤维化跨膜传导调节剂(CFTR)将允许突变的CFTR发挥功能而不是被降解。设想使用TRIM蛋白稳定错误折叠的蛋白质可以用于治疗与部分功能蛋白降解有关的囊性纤维化及其它疾病。通过本文所述的一种或多种TRIM蛋白对蛋白质的稳定可以用于治疗与功能突变蛋白降解有关的任何疾病或病症,其包括(但不限于)囊性纤维化和溶酶体贮积症,如戈谢病和法布里病。

在一个方面,本发明提供了增加TRIM蛋白的表达、活性或两者的组合物和方法。在某些实施方式中,组合物包含核酸分子、表达载体、蛋白质、肽、小分子等,其增加一种或多种TRIM蛋白的表达、活性或两者。

在一个方面,本发明提供了增加一种或多种SUMO靶向的泛素连接酶(STUbL)的表达、活性或两者的组合物和方法。在某些实施方式中,组合物包含核酸分子、表达载体、蛋白质、肽、小分子等,其增加一种或多种STUbL的表达、活性或两者。

定义

除非另外定义,本文所使用的所有技术和科学名词的含义与本发明所属领域中技术人员通常理解的含义相同。尽管可以在本发明的实践或测试中使用与本文的那些类似或等价的任何方法和材料,但是描述了优选的方法和材料。

如本文所使用的,下列术语中的每一个具有在本部分中与其相关的含义。

本文所使用的冠词“一个”和“一种”表示一个或多于一个(即至少一个)冠词的语法对象。举例来说,“一个元素”表示一个元素或多于一个元素。

如本文所使用的,当表示可测量的值,如量、时距等时,“约”表示涵盖了指定值的±20%、±10%、±5%、±1%或±0.1%的变化,这些改变适合实施所公开的方法。

当在生物、组织、细胞或其组分的背景中使用时,术语“异常”是指那些生物、组织、细胞或其组分在至少一个可观察或可检测的特征(例如,年龄、治疗、当日时间等)中不同于显示“正常”(预期的)各个特征的那些生物、组织、细胞或其组分。对一种细胞或组织类型正常或预期的特征可以对不同的细胞或组织类型是异常的。

“疾病”是其中动物不能维持体内平衡并且其中如果疾病不能得到改善,那么动物的健康持续变差的动物健康状况。

相反,在动物中“病症”是其中动物能够维持体内平衡,但是其中动物的健康状况相比没有病症时较差的健康状况。保持不治疗,病症不一定导致动物健康状况的进一步降低。

如果疾病或病症的病征或症状的严重程度、患者经受这种病征或症状的频率或两者降低,则疾病或病症“减轻”。

化合物的“有效量”或“治疗有效量”是足以对给予化合物的受试者提供有益作用的化合物的量。递送载体的“有效量”是足以有效结合或递送化合物的量。

如本文所使用的,“说明材料”包括出版物、录像、图表或任何其它表达媒介,其可以用于传达试剂盒中本发明的化合物、组合物、载体或递送系统对于引起本文列举的多种疾病或病症的有效减轻的有用性。可选地或可替换地,说明材料可以描述减轻哺乳动物细胞或组织中疾病或病症的一种或多种方法。本发明的试剂盒的说明材料可以(例如)附在包含本发明指示的化合物、组合物、载体或递送系统的盒上,或者与含有指示的化合物、组合物、载体或递送系统的盒一起提供。可替换地,说明材料可以与盒分开提供,旨在接受者协同使用说明材料和化合物。

术语“患者”、“受试者”、“个体”等在本文中是可互换使用的并且表示无论体外或体内,受本文的方法作用的任何动物或其细胞。在某些非限制性实施方式中,患者、受试者或个体是人。

“治疗性”治疗是出于减轻或消除那些病征或症状的目的,给予于显示出疾病或病症的病征或症状的受试者的治疗。

如本文所使用的,“治疗疾病或病症”表示降低患者经受的疾病或病症的病征或症状的严重程度和/或频率。

如本文所使用的短语“生物样品”旨在包括包含其中存在或者可以检测到核酸的表达或多肽的细胞、组织或体液的任何样品。性质上为液体的样品在本文中称为“体液”。可以通过多种技术从患者获得生物样品,包括(例如)通过刮拭或擦拭受试者的区域或者通过使用针头获得体液。采集多种身体样品的方法在本领域中是熟知的。

如本文所使用的,“免疫测定”是指使用能够特异性结合至靶标分子的抗体来检测和定量靶标分子的任何结合测定。

如本文所使用的,相对于抗体,术语“特异性结合”是指识别特异性抗原,但是基本不识别或结合样品中其它分子的抗体。例如,特异性结合至来自一个物种的抗原的抗体还可以结合至来自一种或多种物种的抗原。但是,这种种间反应性本身不会改变抗体作为特异性的分类。在另一个实例中,特异性结合至抗原的抗体还可以结合至抗原的不同等位形式。然而,这种种间反应性本身不会改变抗体作为特异性的分类。

在一些情况下,术语“特异性结合”可以涉及抗体、蛋白质或肽与第二化学物质的相互作用,用于表示相互作用取决于化学物质上特定结构(例如,抗原决定簇或表位)的存在;例如,抗体识别并结合至具体的蛋白结构,而不是一般地结合至蛋白质。如果抗体对表位“A”特异,则在含有标记的“A”和抗体的反应中,含有表位A(或者不含,未标记的A)的分子的存在将降低结合至抗体的标记的A的量。

基因的“编码区”由基因的编码链的核苷酸残基和基因的非编码链的核苷酸组成,其分别与通过基因转录产生的mRNA分子的编码区同源或互补。

mRNA分子的“编码区”由与mRNA分子翻译期间的转运核糖核酸分子的反密码子区匹配或编码终止密码子的mRNA分子的核苷酸残基组成。因此,编码区可以包括包含在mRNA分子编码的成熟蛋白质中不存在的氨基酸残基(例如,蛋白质输出信号序列中的氨基酸残基)的密码子的核苷酸残基。

如本文提及核酸时所使用的“互补”是指两条核酸链的区域之间或者相同核酸链的两个区域之间序列互补性的广泛概念。已知第一核酸区的腺嘌呤残基能够与和第一区反向平行的第二核酸区的残基(如果残基是胸腺嘧啶或尿嘧啶)形成特异性氢键(“碱基配对”)。类似地,已知第一核酸链的胞嘧啶残基能够与和第一链反向平行的第二核酸链的残基(如果残基是鸟嘌呤)碱基配对。如果当两个区域以反向平行方式布置时,第一区的至少一个核苷酸残基能够与第二区的残基碱基配对,则第一核酸区与具有相同或不同核酸的第二区互补。优选地,第一区包含第一部分并且第二区包含第二部分,其中当第一和第二部分以反向平行方式布置时,至少约50%,并且优选地至少约75%,至少约90%或至少约95%的第一部分的核苷酸残基能够与第二部分的核苷酸残基碱基配对。更优选地,第一部分的所有核苷酸残基能够与第二部分中的核苷酸残基碱基配对。

“分离的”表示从天然状态改变或移除的。例如,在活的动物中在其正常背景中天然存在的核酸或肽不是“分离的”,但是与其天然背景中共存的材料部分或完全分离的相同核酸或肽是“分离的”。分离的核酸或蛋白质可以以基本纯化的形式存在,或者可以在非天然环境中存在,如(例如)宿主细胞。

“分离的核酸”是指与序列分离的核酸部分或片段,序列以天然存在的状态与其侧接,即从通常邻近于片段的序列中移除的DNA片段,即在其天然存在的基因组中邻近于片段的序列。该术语还适用于基本从天然伴随核酸的其它组分,即在细胞中天然伴随核酸的RNA或DNA或蛋白质中纯化的核酸。因此,该术语包括(例如)引入载体中、引入自主复制质粒或病毒或者引入原核生物或真核生物的基因组DNA,或者作为独立于其它序列的单独的分子(即,作为通过PCR或者限制酶消化产生的cDNA或者基因组或cDNA片段)存在的重组DNA。还包括作为编码其它多肽序列的杂合基因的一部分的重组DNA。

在本发明的上下文中,使用了以下用于常见核酸碱基的缩写。“A”是指腺苷酸,“C”是指胞嘧啶,“G”是指鸟嘌呤,“T”是指胸腺嘧啶,并且“U”是指尿嘧啶。

本文所使用的术语“多核苷酸”定义为核苷酸的链。此外,核酸是核苷酸的聚合物。因此,如本文所使用的核酸和多核苷酸是可互换的。本领域技术人员具有核酸是多核苷酸,并且其可以水解成单体“核苷酸”的一般知识。单体核苷酸可以水解成核苷。如本文所使用的多核苷酸包括(但不限于)通过本领域中可用的任何方式获得的所有核酸序列,该方式无限制地包括重组方式,即使用常规克隆技术和PCR等从重组文库或细胞基因组克隆核酸序列,和合成方式。

如本文所使用的,术语“肽”、“多肽”和“蛋白质”是可互换使用的,并且表示由通过肽键共价连接的氨基酸残基组成的化合物。蛋白质或肽必须含有至少两个氨基酸,并且对蛋白质或肽序列可以包含的最大氨基酸个数无限制。多肽包括包含通过肽键彼此连接在一起的两个或更多个氨基酸的任何肽或蛋白。如本文所使用的,该术语是指短链,其在本领域中还通常称为(例如)肽、寡肽和寡聚物,和长链,其在本领域中通常称为蛋白质,其存在多种类型。“多肽”包括(例如)生物学活性片段、基本上同源的多肽、寡肽、同型二聚体、异质二聚物、多肽变体、修饰的多肽、衍生物、类拟物、融合蛋白等。多肽包括天然肽、重组肽、合成肽或它们的组合。

如本文所使用的,“结合的”是指一个分子与第二分子的共价连接。

作为在本文中使用的术语“变体”是分别与参比核酸序列或肽序列在序列方面存在差异,但保留了参比分子的必需生物学性质的核酸序列或肽序列。核酸变体序列的变化可以不改变参比核酸所编码的肽的氨基酸序列,或者可以导致氨基酸取代、添加、缺失、融合和截短。肽变体序列的改变通常是有限或保守的,从而参比肽和变体的序列在整体上非常类似并且在多个区域中相同。变体和参比肽可以在氨基酸序列方面以任意组合相差一个或多个取代、添加、缺失。核酸或肽的变体可以是天然存在的,如等位变体,或者可以是未知或非天然存在的变体。可以通过突变技术或通过直接合成制备核酸和肽的非天然存在的变体。

如本文所使用的,“一个或多个TRIM蛋白的调节剂”是与不存在调节剂的情况下的TRIM蛋白的表达、活性或生物学功能相比,改变TRIM蛋白的表达、活性或生物学功能的化合物。

如本文所使用的,“RNF4的调节剂”是与不存在调节剂的情况下的RNF4的表达、活性或生物学功能相比,改变RNF4的表达、活性或生物学功能的化合物。

范围:在本发明公开中,本发明的多个方面可以以范围形式存在。应理解以范围形式进行说明仅是为了方便和简洁,并不应将其视作对本发明范围刻板的限制。因此,范围的说明应认为具有具体公开的所有可能的子范围以及该范围内的单个数值。例如,范围的说明(如1至6)应认为具有具体公开的子范围,如1至3、1至4、1至5、2至4、2至6、3至6等,以及该范围内的单个数值,例如,1、2、2.7、3、4、5、5.3和6。不考虑范围的宽度,这都是适用的。

描述

在一个方面,本发明提供了治疗或预防与错误折叠的蛋白质或者蛋白质聚集体有关的疾病或病症的组合物和方法。例如,本发明提供了增加错误折叠的蛋白质的识别和消除的组合物和方法。在某些实施方式中,本发明提供了错误折叠的蛋白质的SUMO介导的泛素化和最终的降解。在某些实施方式中,本发明提供了蛋白质聚集体或包涵体的解聚。因此,本发明可以用于胞内或胞外消除错误折叠的蛋白质、蛋白质聚集体或蛋白质包涵体。

本发明涉及对三结构域(TRIM)蛋白家族成员和SUMO靶向的泛素连接酶(STUbL)RNF4在错误折叠的蛋白质的识别和降解中的作用的发现,它们在多种疾病和病症,包括多种神经退行性疾病和病症的病理中起作用。

本文所提供的数据显示TRIM蛋白家族成员与错误折叠的蛋白质共定位并且介导错误折叠的蛋白质的降解。此外,本文中描述RNF4是泛素连接酶,其介导错误折叠的蛋白质的泛素化和降解。因此,本发明提供了增加一种或多种TRIM蛋白、一种或多种STUbL或它们的组合的表达、活性或两者的组合物。例如,证明编码TRIM蛋白和重组TRIM蛋白的核酸分子促进错误折叠的蛋白质的降解。

在一个实施方式中,组合物包含一种或多种TRIM蛋白的表达或活性的调节剂。例如,在一个实施方式中,调节剂增加一种或多种TRIM蛋白的表达或活性。一种或多种TRIM蛋白包括TRIM蛋白家族的任何成员,包括哺乳动物和非哺乳动物成员。在某些实施方式中,调节剂增加以下中的一种或多种的表达或活性:人TRIM3、TRIM4、TRIM5、TRIM6、TRIM7、TRIM9、TRIM11、TRIM13、TRIM14、TRIM15、TRIM16、TRIM17、TRIM19(在本文中还称为“PML”)、TRIM20、TRIM21、TRIM24、TRIM25、TRIM27、TRIM28、TRIM29、TRIM32、TRIM34、TRIM39、TRIM43、TRIM44、TRIM45、TRIM46、TRIM49、TRIM50、TRIM52、TRIM58、TRIM59、TRIM65、TRIM67、TRIM69、TRIM70、TRIM74和TRIM75;和小鼠TRIM30。

在一个实施方式中,组合物包含一种或多种STUbL的调节剂。例如,在一个实施方式中,调节剂增加一种或多种STUbL的表达或活性。示例性的STUbL包括(但不限于)RNF4和RNF111(也称为Arkadia)。

在一个实施方式中,组合物包含一种或多种TRIM蛋白的调节剂和一种或多种STUbL的调节剂。

本发明提供了治疗或预防对其有需要的受试者中与错误折叠的蛋白质或者蛋白质聚集体有关的疾病或病症的方法。在本文中发现增加TRIM蛋白或STUbL的表达或活性水平可以促进错误折叠的蛋白质降解,由此治疗受试者中的疾病或病症。

可以通过本发明的组合物和方法治疗或预防的神经退行性疾病或病症的实例包括(但不限于)polyQ病症,如SCA1、SCA2、SCA3、SCA6、SCA7、SCA17、亨廷顿舞蹈病、齿状核红核苍白球路易体萎缩症(DRPLA)、阿尔茨海默氏病、帕金森氏症、肌萎缩性侧索硬化(ALS)、传染性海绵状脑病(朊病毒疾病)、滔蛋白病变和额颞叶脑退行性病变(FTLD)。然而,本发明不局限于神经退行性病症的治疗或预防。而是,本发明涵盖了与错误折叠的蛋白质或蛋白质聚集体有关的任何疾病或病症的治疗或预防。其它该类疾病和病症包括(但不限于)AL淀粉样变性、AA淀粉样变性、家族性地中海热、老年系统性淀粉样变性、家族性淀粉样多神经病、透析相关的淀粉样变性、ApoAI淀粉样变性、ApoAII淀粉样变性、ApoAIV淀粉样变性、芬兰型遗传性淀粉样变性、溶菌酶淀粉样变性、纤维蛋白原淀粉样变性、冰岛型遗传性脑淀粉样血管病、II型糖尿病、甲状腺髓样癌、心房性淀粉样变性、遗传性脑出血伴淀粉样变性、垂体泌乳素瘤、注射局部化淀粉样变性、主动脉内侧淀粉样变性、遗传性格子状角膜变性、与倒睫相关的角膜淀粉样变性、白内障、牙源性钙化上皮肿瘤、肺泡蛋白沉积症、包涵体肌炎和苔藓型皮肤淀粉样变性。在某些实施方式中,本发明涵盖了与p53突变聚集体有关的癌症的治疗或预防,其包括(但不限于)膀胱癌、星形细胞瘤、咽癌、淋巴瘤和腺癌。

在一个方面,本发明涵盖了一种或多种TRIM蛋白稳定错误折叠的蛋白质的用途。在某些方面,通过本文的一种或多种TRIM蛋白对功能性错误折叠的蛋白质的稳定化可以治疗或预防与错误折叠的蛋白质有关的疾病或病症,其包括(但不限于)囊性纤维化和溶酶体贮积症,如戈谢病和法布里病。

在一个方面,本发明提供了诊断与错误折叠的蛋白质或蛋白质聚集体有关的疾病或病症的方法。例如,在一个实施方式中,将一种或多种TRIM蛋白或者一种或多种STUbL用作诊断标志物。

在一个方面,本发明涉及生产所关心的重组蛋白的组合物和方法。例如,在某些实施方式中,一种或多种TRIM蛋白和一种或多种STUbL可以用于使感兴趣的重组蛋白的蛋白质聚集体解聚。

组合物

在各个实施方式中,本发明包括预防和治疗与错误折叠的蛋白质或者蛋白质聚集体有关的疾病或病症的调节剂组合物和方法。在各个实施方式中,本发明的预防或治疗的调节剂组合物和方法调节基因或基因产物的水平或活性。在一些实施方式中,本发明的调节剂组合物是增加基因或基因产物的水平或活性的激活因子。

基于本文所提供的公开,本领域技术人员将理解调节基因或基因产物涵盖了调节基因或基因产物的水平或活性,其包括(但不限于)调节转录、翻译、拼接、降解、酶活力、结合活力或它们的组合。因此,调节基因或基因产物的水平或活性包括(但不限于)调节核酸的转录、翻译、降解、拼接或它们的组合;并且还包括调节多肽基因产物的任何活性。

在一个实施方式中,调节剂通过增加基因或基因产物的产生,例如,通过调节基因的转录或者基因产物的翻译来增加基因或基因产物的表达或活性。在一个实施方式中,调节剂通过提供外源基因或基因产物增加基因或基因产物的表达或活性。例如,在某些实施方式中,调节剂包含编码一种或多种TRIM蛋白或者一种或多种STUbL的分离的核酸。在一个实施方式中,调节剂包含编码一种或多种TRIM蛋白和一种或多种STUbL的分离的核酸。在某些实施方式中,调节剂包含含有一种或多种TRIM蛋白或者一种或多种STUbL的分离的肽。在一个实施方式中,调节剂包含含有一种或多种TRIM蛋白和一种或多种STUbL的分离的肽。在一个实施方式中,调节剂通过抑制基因或基因产物的降解来增加基因或基因产物的表达或活性。例如,在一个实施方式中,调节剂降低了一种或多种TRIM蛋白或一种或多种STUbL的泛素化、蛋白酶体降解或蛋白水解。在一个实施方式中,调节剂增加基因产物的稳定性或半衰期。

在各个实施方式中,调节的基因或基因产物是一种或多种TRIM蛋白。例如,本文描述了TRIM蛋白识别错误折叠的蛋白质并且介导错误折叠的蛋白质和蛋白质聚集体的降解。在一个实施方式中,基因或基因产物是一种或多种STUbL。例如,本文描述了STUbl蛋白质家族的成员RNF4介导错误折叠的蛋白质和蛋白质聚集体的降解。

可以使用多种方法评估基因或基因产物的调节,其包括本文所公开的那些和本领域中已知的或者将来开发的方法。也就是说,基于本文所提供的公开,技术人员将理解可以使用评估编码基因产物的核酸(例如,mRNA)的水平、生物样品中存在的多肽基因产物的水平、生物样品中存在的多肽基因产物的活性或它们的组合的方法,容易地评估对基因或基因产物的水平或活性的调节。

本发明的调节基因或基因产物的水平或活性的调节剂组合物和方法包括(但不应解释为限于)化合物、蛋白质、肽、拟肽、抗体、核糖酶、小分子化合物、核酸、载体、反义核酸分子(例如,siRNA、miRNA等)或它们的组合。基于本文所提供的公开,本领域技术人员将容易地理解调节剂组合物包含调节基因或基因产物的水平或活性的化合物。另外,调节剂组合物包含化学修饰的化合物和衍生物,如化学领域中技术人员所熟知的。

在一个实施方式中,本发明的调节剂组合物是激动剂,其增加基因或基因产物的表达、活性或生物学功能。例如,在某些实施方式中,本发明的调节剂是一种或多种TRIM蛋白或一种或多种STUbL的激动剂。

此外,当与本公开和本文中举例说明的方法一起提供时,本领域技术人员将理解调节剂包括将来发现的那些调节剂,如可以通过药学领域中熟知的标准,如基因和基因产物调节的生理学结果,如本文中详细描述的和/或如本领域中已知的。因此,本发明不以任何方式受限于本文所举例说明或公开的任何特定调节剂组合物;而是,本发明包含了常规人员将理解在本领域中已知并且将来会发现有用的那些调节剂组合物。

鉴别和产生调节剂组合物的其它方法是本领域那些技术人员熟知的。可替换地,可以化学合成调节剂。此外,基于本文所提供的教导,技术人员将理解调节剂组合物可以获得自重组生物。用于化学合成调节剂和用于从天然来源获得它们的组合物和方法在本领域中是熟知的并且在本领域已描述。

本领域技术人员将理解可以作为小分子化合物、多肽、肽、抗体、编码蛋白的核酸构建体、反义核酸、编码反义核酸的核酸构建体或它们的组合给予调节剂。已知多种载体及其它组合物和方法用于将蛋白质或编码蛋白质的核酸构建体给予至细胞或组织来说。因此,本发明包括作为基因或基因产物的调节剂的肽或编码肽的核酸。例如,本发明包括包含一种或多种TRIM蛋白、一种或多种STUbL或它们的组合的肽或编码肽的核酸。(Sambrook et al.,2001,Molecular Cloning:A Laboratory Manual,Cold Spring Harbor Laboratory,New York;Ausubel et al.,1997,Current Protocols in Molecular Biology,John Wiley&Sons,New York)。

在一个实施方式中,本发明的组合物包含一种或多种肽。例如,在一个实施方式中,组合物的肽包含一种或多种TRIM蛋白的氨基酸序列。例如,在一个实施方式中,肽包含TRIM5δ、TRIM 11、TRIM19、TRIM 21、TRIM27和TRIM32中的一种或多种。在某些实施方式中,肽包含一种或多种STUbL的氨基酸序列。例如,在一个实施方式中,肽包含RNF4和RNF111(Arkadia)中的一种或多种。

下表1中提供了TRIM蛋白的示例性氨基酸序列和编码TRIM蛋白的cDNA核苷酸序列。

还应将本发明解释为包含与本文所公开的肽具有显著同源性的任何形式的肽。优选地,“显著同源的”肽与本文所公开的肽的氨基酸序列约50%同源、更优选地约70%同源、更优选地约80%同源、更优选地约90%同源、更优选地约95%同源、并且更优选地约99%同源。

在一个实施方式中,本发明的组合物包含肽、肽的片段、本文描述的肽的同源物、变体、衍生物或盐。例如,在某些实施方式中,组合物包含含有一种或多种TRIM蛋白、一种或多种TRIM蛋白的片段、一种或多种TRIM蛋白的同源物、一种或多种TRIM蛋白的变体、一种或多种TRIM蛋白的衍生物或一种或多种TRIM蛋白的盐的肽。在某些实施方式中,组合物包含含有一种或多种STUbL、一种或多种STUbL的片段、一种或多种STUbL的同源物、一种或多种STUbL的变体、一种或多种STUbL的衍生物或一种或多种STUbL的盐的肽。

在一个实施方式中,组合物包含本文描述的肽的组合。例如,在某些实施方式中,组合物包含含有一种或多种TRIM蛋白的肽和含有一种或多种STUbL的肽。在一个实施方式中,组合物包含含有一种或多种TRIM蛋白和一种或多种STUbL的肽。

在某些实施方式中,肽包含靶向域,其将肽靶向期望的位置。例如,在某些实施方式中,靶向域结合至靶细胞、蛋白质或蛋白质聚集体,从而将治疗肽递送至期望的位置。例如,在一个实施方式中,导向靶向域以结合至与疾病或病症有关的蛋白质或蛋白质聚集体,其包括(但不限于)淀粉样蛋白-β、α-突触核蛋白、τ蛋白、朊病毒、SOD1、TDP-43、FUS、p53突变体的蛋白质和蛋白质聚集体以及与多聚谷氨酰胺重复有关的蛋白质,如亨廷顿蛋白、脊髓小脑共济失调蛋白。

在某些实施方式中,靶向域包含肽、核酸、小分子等,其具有结合至靶细胞、蛋白质或蛋白质聚集体的能力。例如,在一个实施方式中,靶向域包含结合至靶细胞、蛋白质或蛋白质聚集体的抗体或抗体片段。

可以使用化学方法制备本发明的肽。例如,可以通过固相技术(Roberge J Y et al.(1995)Science 269:202-204)合成,从树脂剪切和通过制备型高效液相色谱法纯化肽。可以例如根据生产商所提供的说明书使用ABI 431A肽合成仪(Perkin Elmer)实现自动合成。

可替换地,可以通过重组方式或通过从较长的多肽剪切来制备肽。可以通过氨基酸分析或测序确认肽的组成。

根据本发明的肽的变体可以是(i)其中一个或多个氨基酸残基被保守或非保守氨基酸残基(优选保守氨基酸残基)取代的肽并且该取代的氨基酸残基可以或可以不是遗传密码所编码的,(ii)其中存在一个或多个修饰的氨基酸残基,例如,通过连接取代基团而修饰的残基的肽,(iii)其中肽是本发明的肽的替代性拼接变体的肽,(iv)肽的片段和/或(v)其中肽与另一种肽,如前导序列或分泌序列或用于纯化(例如,His标签)或检测(例如,Sv5表位标签)的序列融合的肽。片段包括通过原始序列的蛋白水解切割(包括多位点蛋白水解)所产生的肽。变体可以是翻译后或化学修饰的。根据本文的教导,这些变体被认为在本领域技术人员的范围内的。

本发明的肽可以是翻译后修饰的。例如,本发明范围内的翻译后修饰包括信号肽切割、糖基化、乙酰化、异戊二烯化、蛋白水解、豆蔻酰化、蛋白质折叠和蛋白水解处理等。一些修饰或处理事件需要引入额外的生物机制。例如,通过将狗微粒体膜或者非洲蟾蜍卵提取物(美国专利号6,103,489)添加至标准翻译反应来检验处理事件,如信号肽切割和核心糖基化。

本发明的肽可以包括通过翻译后修饰或通过在翻译期间引入非天然氨基酸所形成的非天然氨基酸。多种方法可用于在蛋白质翻译期间引入非天然氨基酸。举例来说,在定点非天然氨基酸替代(SNAAR)过程中,使用特殊的tRNA,如具有抑制剂性质的tRNA、抑制剂tRNA。在SNAAR中,在mRNA和抑制剂tRNA上需要唯一的密码子,其作用为在蛋白质合成期间将非天然氨基酸靶向唯一的位点(WO90/05785中描述)。然而,抑制剂tRNA必需是蛋白质翻译系统中存在的氨酰TRNA合成酶不可识别的。在某些情况下,可以在使用特异性修饰天然氨基酸并且不显著改变氨酰化tRNA的功能活性的化学反应使tRNA分子氨酰化后形成非天然氨基酸。这些反应被认为是氨酰化后修饰。例如,可以用胺特异性光亲和标记修饰与其同源tRNA(tRNALYS)连接的赖氨酸的ε-氨基。

本发明的肽可以与其它分子,如蛋白质结合以制备融合蛋白。这可以通过例如N末端或C末端融合蛋白的合成来完成,条件是得到的融合蛋白保留了本发明的肽的功能性。

本发明的肽的环状衍生物也是本发明的一部分。环化可以使肽呈现出对于与其它分子结合更有利的构象。可以使用本领域中已知的技术实现环化。例如,可以在具有自由巯基基团的两个适当间隔的组分之间形成二硫键,或者可以在一个组分的氨基和另一个组分的羧基之间形成酰胺键。还可以使用含偶氮苯的氨基酸实现环化,如Ulysse,L.et al.,J.Am.Chem.Soc.1995,117,8466-8467描述的。形成键的组分可以是氨基酸的侧链、非氨基酸组分或两者的组合。在本发明的一个实施方式中,环状肽可以包含处于右侧位置的β-转角。可以通过在右侧位置添加氨基酸Pro-Gly将β-转角引入本发明的肽。

可以期望产生比如上描述的含有肽键的环肽更柔性的环肽。可以通过在肽的右侧和左侧位置引入半胱氨酸并在两个半胱氨酸之间形成二硫键来制备更柔性的肽。两个半胱氨酸布置为不会使β-折叠和转角变形。由于β-折叠部分中二硫键的长度以及较少数目的氢键,肽更柔性。可以通过分子动力学模拟确定环肽的相对柔性。

可以通过与无机酸,如盐酸、硫酸、氢溴酸、磷酸等,或有机酸,如甲酸、乙酸、丙酸、乙醇酸、乳酸、丙酮酸、草酸、丁二酸、苹果酸、酒石酸、柠檬酸、苯甲酸、水杨酸、苯磺酸和甲苯磺酸反应,将本发明的肽转化为药物盐。

本发明的肽还可以具有修饰。修饰(其通常不改变一级序列)包括多肽的体内或体外化学衍生化,例如,乙酰化或羧化。还包括糖基化修饰,例如,在其合成和处理期间或在其它处理步骤中通过修饰多肽的糖基化型式所产生的那些;例如,通过将多肽暴露于影响糖基化的酶,例如,哺乳动物的糖基化或去糖基化酶。还包含具有磷酸化氨基酸残基,例如,磷酸酪氨酸、磷酸丝氨酸或磷酸苏氨酸的序列。

还包括已使用常规分子生物技术修饰以改善它们对蛋白水解降解的耐受性或优化溶解度性质或使它们更适合用作治疗剂的肽。这些变体包括含有天然存在的L-氨基酸,例如,D-氨基酸或非天然存在的合成氨基酸以外的残基的那些。本发明的肽可以进一步结合至在它们的治疗应用中有用的非氨基酸部分。具体地,改善肽的稳定性、生物半衰期、水溶性和/或免疫学特征的部分是有用的。这种部分的非限制性实例是聚乙二醇(PEG)。

生物学活性化合物与水溶性聚合物的共价连接是用于改变和控制这些化合物的生物分布、药物动力学并且通常地,毒性的一种方法(Duncan et al.,1984,Adv.Polym.Sci.57:53-101)。多种水溶性聚合物已用于实现这些作用,如聚(唾液酸)、葡聚糖、聚(N-(2-羟丙基)甲基丙烯酰胺)(PHPMA)、聚(N-乙烯基吡咯烷酮)(PVP)、聚(乙烯醇)(PVA)、聚(乙二醇-共-丙二醇)、聚(N-丙烯酰基吗啉)(PAcM)和聚(乙二醇)(PEG)(Powell,1980,Polyethylene glycol.In R.L.Davidson(Ed.)Handbook of Water Soluble Gums and Resins.McGraw-Hill,New York,chapter 18)。PEG具有一组理想的性质:极低的毒性(Pang,1993,J.Am.Coll.Toxicol.12:429-456)、在水溶液中优异的溶解度(Powell,如上)、低免疫原性和抗原性(Dreborg et al.,1990,Crit.Rev.Ther.Drug Carrier Syst.6:315-365)。已在科学文献中描述了在蛋白质上含有单个或多个聚乙二醇链的PEG结合的或“PEG化的”蛋白质治疗剂(Clark et al.,1996,J.Biol.Chem.271:21969-21977;Hershfield,1997,Biochemistry and immunology of poly(ethylene glycol)-modified adenosine deaminase(PEG-ADA).In J.M.Harris and S.Zalipsky(Ed.)Poly(ethylene glycol):Chemistry and Biological Applications.American Chemical Society,Washington,D.C.,p 145-154;Olson et al.,1997,Preparation and characterization of poly(ethylene glycol)ylated human growth hormone antagonist.In J.M.Harris and S.Zalipsky(Ed.)Poly(ethylene glycol):Chemistry and Biological Applications.American Chemical Society,Washington,D.C.,p 170-181)。

本发明的肽可以通过常规方法合成。例如,可以使用固相肽合成通过化学合成来合成本发明的肽。这些方法使用了固相或液相合成法(参见,例如,对于固相合成技术,J.M.Stewart,and J.D.Young,Solid Phase Peptide Synthesis,2nd Ed,Pierce Chemical Co.,Rockford Ill.(1984)and G.Barany and R.B.Merrifield,The Peptides:Analysis Synthesis,Biology editors E.Gross and J.Meienhofer Vol.2Academic Press,New York,1980,pp.3-254;和对于经典溶液合成,M Bodansky,Principles of Peptide Synthesis,Springer-Verlag,Berlin 1984,and E.Gross and J.Meienhofer,Eds.,The Peptides:Analysis,Synthesis,Biology,suprs,Vol 1)。

可以通过Merrifield型固相肽合成来化学合成肽。可以常规地实施该方法以获得长度至多达约60-70个残基的肽,并且在一些情况下该方法可以用于制备长达约100个氨基酸的肽。还可以通过片段缩聚或天然化学连接来合成产生较大的肽(Dawson et al.,2000,Ann.Rev.Biochem.69:923-960)。使用合成肽途径的优势在于能够产生大量的肽,甚至是天然很少存在的那些,同时具有相对高的纯度,即足以用于研究、诊断或治疗目的的纯度。

以下文献中描述了固相肽合成:Stewart et al.,in Solid Phase Peptide Synthesis,2nd Edition,1984,Pierce Chemical Company,Rockford,Ill.;和Bodanszky and Bodanszky in The Practice of Peptide Synthesis,1984,Springer-Verlag,New York。开始,将适当保护的氨基酸残基通过羧基基团连接至不溶性聚合物载体,如交联的聚苯乙烯或聚酰胺树脂。“适当保护的”是指在氨基酸的α-氨基和任何侧链官能团上保护基团的存在。侧链保护基团通常对整个合成中使用的溶剂、试剂和反应条件稳定,并且是在不影响最终肽产物的条件下可除去的。寡肽的逐步合成通过从初始氨基酸除去N-保护基团并将其连接至期望的肽序列中下一个氨基酸的羧基端来进行。该氨基酸也是适当保护的。可以激活即将加入的氨基酸的羧基,从而通过形成反应基团,如形成碳二亚胺、对称酸酐或“活性酯”基团,如羟基苯并三唑或五氟苯基酯与载体-结合的氨基酸的N-末端反应。

固相肽合成法的实例包括BOC法,其使用叔丁氧羰基作为α-氨基保护基团,和FMOC法,其使用9-芴甲氧羰基保护氨基酸残基的α-氨基,两种方法均是本领域技术人员熟知的。

还可以使用固相肽合成法的常规流程实现N-和/或C-封端基团的并入。对于C端封端基团的并入,例如,通常使用化学修饰的载体树脂作为固相来进行所期望的肽的合成,从而树脂的剪切导致产生了具有期望的C末端封端基团的肽。为了提供其中C端具有伯氨基封端基团的肽,例如,使用对甲基二苯甲基胺(MBHA)树脂进行合成,从而当肽合成完成时,用氢氟酸的处理释放了期望的C端酰胺化的肽。类似地,使用N-甲基氨基乙基衍生化的DVB树脂实现了在C端处的N-甲胺封端基团的并入,其在HF处理时释放具有N-甲基酰胺化的C端的肽。还可以使用常规程序实现通过酯化对C端的封端。这需要使用允许侧链肽从树脂释放的树脂/封端基团组合,从而使其与所期望的醇进行后续反应以形成酯官能团。可以将FMOC保护基团与甲氧基烷氧基苄醇或等同的连接基衍生化的DVB树脂结合用于该目的,其中载体的剪切受二氯甲烷中TFA的影响。然后,可以通过加入期望的醇,然后去保护并分离酯化肽产物来进行用(例如)DCC适当激活的羧基官能团的酯化。

可以通过肽合成的标准化学或生物方式来制备本发明的肽。生物学方法无限制地包括宿主细胞或体外翻译系统中编码肽的核酸的表达。

本发明包括编码本发明的肽的核酸序列。在一个实施方式中,本发明包括编码一种或多种TRIM蛋白或一种或多种STUbL的氨基酸序列的核酸序列。因此,可以使用常规的对亚克隆基因片段的分子遗传操作来产生编码本发明的肽的核酸序列的亚克隆,如Sambrook et al.,Molecular Cloning:A Laboratory Manual,Cold Springs Laboratory,Cold Springs Harbor,New York(2012)和Ausubel et al.(ed.),Current Protocols in Molecular Biology,John Wiley&Sons(New York,NY)(1999以及之前版本)描述的,其各自通过引证以其全部内容并入本文。然后,在细菌细胞中体外或体内表达亚克隆以获得可以测试特定活性的较小的蛋白质或多肽。

与特定制剂组合,这些肽可以是有效的胞内试剂。然而,为了增加这些肽的效力,本发明的一种或多种肽可以与促进“转胞吞作用”,例如通过细胞的肽的吸收的第二肽一起提供融合肽。例如,在一个实施方式中,肽可以包含细胞穿透域,例如,细胞穿透肽(CPP)以允许肽进入细胞。在一个实施方式中,CPP来源于HIV Tat。

为了说明,可以将本发明的一种或多种肽作为具有可以促进转胞吞作用的HIV蛋白Tat的N端域的全部或片段,例如,Tat的残基1-72或其更小的片段的融合多肽的一部分提供。在一个实施方式中,肽包含HIV Tat的蛋白转导域(YGRKKRRQRRR;(SEQ ID NO:163))。在其它实施方式中,一种或多种肽可以提供为具有触角足蛋白III(antenopedia III protein)的全部或部分的融合多肽。调节肽的吸收的其它细胞穿膜域在本领域中是已知的,并且同样适合在本发明的融合肽中使用。

核酸

在一个实施方式中,本发明的组合物包含一种或多种分离的核酸。例如,在一个实施方式中,一种或多种分离的核酸编码一种或多种TRIM蛋白。例如,在一个实施方式中,一种或多种分离的核酸编码以下中的一种或多种:人TRIM3、TRIM4、TRIM5、TRIM6、TRIM7、TRIM9、TRIM11、TRIM13、TRIM14、TRIM15、TRIM16、TRIM17、TRIM19(在本文中还称为“PML”)、TRIM20、TRIM21、TRIM24、TRIM25、TRIM27、TRIM28、TRIM29、TRIM32、TRIM34、TRIM39、TRIM43、TRIM44、TRIM45、TRIM46、TRIM49、TRIM50、TRIM52、TRIM58、TRIM59、TRIM65、TRIM67、TRIM69、TRIM70、TRIM74和TRIM75;和鼠TRIM30。在某些实施方式中,一种或多种分离的核酸编码一种或多种STUbL。例如,在一个实施方式中,一种或多种分离的核酸编码RNF4和RNF111(Arkadia)中的一种或多种。

表1中提供了编码TRIM蛋白的示例性核苷酸序列。

在某些实施方式中,使用已知的技术在细胞中由一种或多种核酸体内或体外表达对应于一种或多种TRIM蛋白或一种或多种STUbL的肽。

分离的核酸的核苷酸序列包括转录为RNA的DNA序列和翻译成多肽的RNA序列两者。根据其它实施方式,从本发明的肽的氨基酸序列推定核苷酸序列。如本领域中已知的,由于冗余密码子,一些替代核苷酸序列是可能的,同时保留了翻译的肽的生物活性。

此外,本发明涵盖了包含与编码本文公开的肽的核苷酸序列具有基本上的同源性的核苷酸序列的分离的核酸。优选地,分离的核酸的核苷酸序列是“基本同源的”,即与编码本发明的肽的分离的核酸的核苷酸序列约60%同源、更优选地约70%同源、更优选地约80%同源、更优选地约90%同源、更优选地约95%同源、并且甚至更优选地约99%同源。

在一个实施方式中,组合物包含本文描述的核酸分子的组合。例如,在某些实施方式中,组合物包含编码一种或多种TRIM蛋白的分离的核酸分子和编码一种或多种STUbL的分离的核酸分子。在一个实施方式中,组合物包含编码一种或多种TRIM蛋白和一种或多种STUbL的分离的核酸分子。

因此,本发明涵盖了将外源DNA引入细胞并具有外源DNA在细胞中的伴随表达的表达载体和方法,如在例如Sambrook et al.(2012,Molecular Cloning:A Laboratory Manual,Cold Spring Harbor Laboratory,New York)和Ausubel et al.(1997,Current Protocols in Molecular Biology,John Wiley&Sons,New York)中描述的那些。

可以将编码一种或多种TRIM蛋白或一种或多种STUbL的期望的核酸克隆到多种类型的载体中。然而,本发明不应解释为受限于任何特定载体。相反,本发明应视为涵盖了易于获得和/或在本领域熟知的各种载体。例如,可以将期望的本发明的多核苷酸克隆到载体中,其包括(但不限于)质粒、噬菌粒、噬菌体衍生物、动物病毒和粘粒。特别感兴趣的载体包括表达载体、复制载体、探针产生载体和测序载体。

在具体的实施方式中,表达载体选自由病毒载体、细菌载体和哺乳动物细胞载体组成的组。存在多种表达载体系统,其包含以上所讨论的组合物的至少一部分或全部。可以将基于原核生物载体和/或真核生物载体的系统用于本发明以产生多核苷酸或它们的同源多肽。多种这些系统是可用的并且是广泛获得的。

此外,可以以病毒载体的形式向细胞提供表达载体。病毒载体技术在本领域中是熟知的并且在例如Sambrook et al.(2012)和在Ausubel et al.(1997)和在其它病毒学和分子生物学手册中描述。用作载体的病毒包括但不限于反转录病毒、腺病毒、腺相关病毒、疱疹病毒和慢病毒。通常,适合的载体含有在至少一种生物体中的复制功能的起点、启动子序列、方便的限制性核酸内切酶位点和一种或多种可选择的标志物。(参见,例如,WO 01/96584;WO 01/29058;和美国专利号6,326,193)。

开发了一些基于病毒系统用于将基因转移至哺乳动物细胞。例如,反转录病毒提供了用于基因递送系统的方便的平台。可以使用本领域中已知的技术将选择的基因插入载体并装包在反转录病毒颗粒中。然后,可以将重组病毒分离并递送至体内或离体的受试者细胞。一些反转录病毒系统在本领域中是已知的。在一些实施方式中,使用了腺病毒载体。一些腺病毒载体在本领域中是已知的。在一个实施方式中,使用了慢病毒载体。

例如,来源于反转录病毒,如慢病毒的载体是实现长期基因转移的适合工具,因为它们允许长期、稳定地整合转基因及其在子代细胞中的繁殖。与来源于其中它们可以转导非增殖细胞,如肝细胞的致癌反转录病毒,如鼠白血病毒相比,慢病毒载体具有额外的优势。它们还具有低免疫原性的额外优势。在优选的实施方式中,组合物包括来源于腺相关病毒(AAV)的载体。腺相关病毒(AAV)载体已成为治疗多种病症的强大基因递送工具。AAV载体具有使它们非常适合于基因疗法的一些特征,包括缺少病原性、极低的免疫原性和以稳定且有效的方式转导有丝分裂期后细胞的能力。通过选择AAV血清型、启动子和递送方法的适当组合,可以将包含在AAV载体内的特定基因的表达特异性靶向一种或多种类型的细胞

在一个实施方式中,编码序列包含在AAV载体中。有超过30种天然存在的AAV血清型可用。AAV衣壳中存在多种天然变体,允许鉴别和使用具有特别适合于骨骼肌的性质的AAV。可以使用常规分子生物学技术工程化AAV病毒,从而使得可以优化这些颗粒,用于核酸序列的细胞特异性递送、最大程度降低免疫原性、调节稳定性和颗粒寿命、有效降解、向核的准确递送等。

因此,可以通过递送含有一种或多种编码序列的重组工程化的AAV或人工AAV实现一种或多种TRIM蛋白或一种或多种STUbL的表达。由于其相对无毒,AAV的使用是DNA外源递送的常用模式,其提供了有效的基因转移并且可以对特定目的容易地优化。示例性的AAV血清型包括但不限于AAV1、AAV2、AAV3、AAV4、AAV5、AAV6、AAV7、AAV8和AAV9。

对于组装为载体期望的AAV片段包括cap蛋白,其包括vp1、vp2、vp3和高变区、rep蛋白,其包括rep 78、rep 68、rep 52和rep 40,和编码这些蛋白的序列。这些片段可以在多种载体系统和宿主细胞中容易地使用。这些片段可以单独使用,结合其它AAV血清型序列或片段使用,或者结合来自其它AAV或非AAV病毒序列的元件使用。如本文所使用的,人工AAV血清型无限制地包括具有非天然存在的衣壳蛋白的AAV。使用所选的AAV序列(例如,vp1衣壳蛋白的片段)结合可以获得自不同的选择的AAV血清型、相同的AAV血清型的非邻接部分、获得自非AAV病毒来源或者获得自非病毒来源的异源序列,可以通过任何适合的技术产生这种人工衣壳。人工AAV血清型可以无限制地为嵌合AAV衣壳、重组AAV衣壳或“人源化的”AAV衣壳。因此,适合于一种或多种TRIM蛋白或一种或多种STUbL的表达的示例性AAV或人工AAV包括AAV2/8(参见美国专利号7,282,199)、AAV2/5(获得自国立卫生研究院)、AAV2/9(国际专利公开号WO2005/033321)、AAV2/6(美国专利号6,156,303)和AAVrh8(国际专利公开号WO2003/042397)等。

对于期望的多核苷酸的表达,每个启动子中的至少一个模块作用为定位RNA合成的起始位点。其最熟知的实例是TATA盒,但是在缺少TATA盒的一些启动子中,如哺乳动物末端脱氧核苷酰基转移酶基因的启动子和SV40基因的启动子,覆盖起始位点本身的独立元件帮助固定起始的位置。

其它启动子元件,即增强子调节转录起始的频率。通常,这些位于起始位点上游30-110bp的区域,尽管最近显示一些启动子还含有起始位点下游的功能元件。启动子元件之间的距离通常是可变的,从而当元件相对彼此反转或移动时,保持启动子功能。在胸苷激酶(tk)启动子中,在活性开始降低之前,启动子元件之间的间距可以增加至相距50bp。基于启动子,似乎单个元件可以协同或独立起作用以激活转录。

启动子可以是与基因或多核苷酸序列天然结合的,如可以通过分离位于编码段和/或外显子上游的5'非编码序列获得的。这种启动子可以称为“内源的”。类似地,增强子可以与位于该序列的下游或上游的多核苷酸序列天然结合。可替换地,通过定位受重组或异源启动子控制的编码多核苷酸段将获得某些优势,重组或异源启动子是指在其自然环境中通常不与多核苷酸序列结合的启动子。重组或异源增强子也表示在其自然环境中通常不与多核苷酸序列结合的增强子。这些启动子或增强子可以包括其它基因的启动子或增强子,和分离自任何其它原核细胞、病毒或真核细胞的启动子或增强子,和非“天然存在”的启动子或增强子,即含有不同转录调控区的不同元件和/或改变表达的突变。除通过合成产生启动子和增强子的核酸序列之外,可以使用重组克隆和/或核酸扩增技术,包括PCRTM,结合本文所公开的组合物产生序列(美国专利4,683,202、美国专利5,928,906)。此外,考虑还可以使用指导序列在非核细胞器,如线粒体、叶绿体等中转录和/或表达的控制序列。

天然地,使用有效引导DNA片段在选择用于表达的细胞类型、细胞器和生物体中表达的启动子和/或增强子将是重要的。分子生物学领域技术人员通常知道如何使用启动子、增强子和细胞类型组合来用于蛋白质表达,例如,参见Sambrook et al.(2012)。所使用的启动子可以是组成型的、组织特异的、诱导型的和/或在引导引入的DNA片段的高水平表达的适当条件下有用的,如在重组蛋白和/或肽的大规模生产中是有利的。启动子可以是异源的或内源的。

在一个实施方式中,启动子或增强子特异地引导一种或多种TRIM蛋白或一种或多种STUbL在神经组织中在肠上皮细胞中表达。例如,在某些实施方式中,启动子或增强子特异地引导一种或多种TRIM蛋白或一种或多种STUbL在神经元、星形胶质细胞、少突胶质细胞、浦肯野细胞、锥体细胞等中的表达。

为了评估期望的多核苷酸的表达,待引入细胞的表达载体还可以含有可选择的标志物基因或报告基因或两者以有利于从试图转染或通过病毒载体感染的细胞群体中鉴别和选择表达细胞。在其它实施方式中,可选择的标志物可以位于单独的DNA片段上并在共转染程序中使用。可选择标志物和报告基因两者可以侧接适当的调控序列以使得能够在宿主细胞中表达。有用的可选择标志物在本领域中是已知的并且包括(例如)抗生素抗性基因,如neo等。

报告基因用于鉴别潜在转染的细胞并用于评价调控序列的功能性。编码易于测定的蛋白质的报告基因在本领域中是熟知的。通常,报告基因是受体生物或组织中不存在或不表达并且编码通过一些易于检测的性质,例如,酶活力来显示其表达的蛋白质的基因。在DNA引入受体细胞后,在适合的时间测定报告基因的表达。

适合的报告基因可以包括编码荧光素酶、β-半乳糖苷酶、氯霉素乙酰转移酶、分泌型碱性磷酸酶的基因或绿色荧光蛋白基因(参见,例如,Ui-Tei et al.,2000FEBS Lett.479:79-82)。适合的表达系统是熟知的并且可以使用熟知的技术制备或商购获得。可以使用唯一的内部限制位点或者通过非唯一限制位点的部分消化产生内部缺失的构建体。然后,可以将构建体转染到显示出高水平siRNA多核苷酸和/或多肽表达的细胞中。通常,将具有最小5'侧接区并显示出最高报告基因表达水平的构建体确定为启动子。这些启动子区可以连接至报告基因并用于评价试剂调节启动子驱动的转录的能力。

在表达载体的背景中,可以通过本领域中的任何方法容易地将载体引入宿主细胞,例如,哺乳动物、细菌、酵母或昆虫细胞。例如,表达载体可以通过物理、化学或生物方式转移到宿主细胞中。

将多核苷酸引入宿主细胞的物理方法包括磷酸钙沉淀、脂质转染、粒子轰击、微注射、电穿孔等。产生包含载体和/或外源核酸的细胞的方法在本领域中是熟知的。参见,例如,Sambrook et al.(2012,Molecular Cloning:A Laboratory Manual,Cold Spring Harbor Laboratory,New York)和Ausubel et al.(1997,Current Protocols in Molecular Biology,John Wiley&Sons,New York)。

将感兴趣的多核苷酸引入宿主细胞的生物学方法包括DNA和RNA载体的使用。病毒载体,并且特别是反转录病毒载体已成为将基因插入哺乳动物,例如,人细胞的最广泛使用的方法。其它病毒载体可以来源于慢病毒、痘病毒、单纯疱疹病毒I型、腺病毒和腺相关病毒等。参见,例如,美国专利号5,350,674和5,585,362。

将多核苷酸引入宿主细胞的化学方法包括胶体分散系统,如大分子复合物、纳米胶囊、微球、珠和基于脂质的系统,包括水包油乳化剂、胶束、混合胶束和脂质体。用作体外和体内递送载体的优选胶体系统是脂质体(即,人工膜囊泡)。这些系统的制备和使用在本领域中是熟知的。

无论用于将外源核酸引入宿主细胞的方法,为了确认宿主细胞中重组DNA序列的存在,可以进行多种测定。这些测定包括例如本领域技术人员熟知的“分子生物学”测定,如DNA和RNA印迹、RT-PCR和PCR;“生物化学”测定,如检测特定肽是否存在,例如,通过免疫学方式(ELISA和免疫印迹)或者通过本文的测定以鉴别属于本发明范围的试剂。

可以使用任何DNA载体或递送载体将期望的多核苷酸体外或体内递送至细胞。在其中使用非病毒递送系统的情况下,优选的递送载体是脂质体。因此,上述递送系统和流程可见于Gene Targeting Protocols,2ed,pp1-35(2002)和Gene Transfer and Expression Protocols,Vol.7,Murray ed.,pp81-89(1991)。

“脂质体”是涵盖了通过产生封闭的脂质双层或聚集体所形成的各种单层和多层脂质载体的一般术语。脂质体可以特征为具有囊泡结构,其具有磷脂双分子层膜和内部水性介质。多层脂质体具有通过水介质分开的多个脂质层。当将磷脂混悬在过量水溶液中时,它们自发形成。在形成封闭结构前,脂质组分经历自重排并在脂质双分子层之间捕获水和溶解的溶质。然而,本发明还涵盖了在溶液中具有与正常囊泡结构不同的结构的组合物。例如,脂质可以呈现出胶束结构或者仅作为脂质分子的非均一聚集体存在。还设想了脂质体-核酸复合物。

在一个实施方式中,本发明的组合物包含编码一种或多种TRIM蛋白或一种或多种STUbL的一种或多种组分的体外转录的(IVT)RNA。在一个实施方式中,可以将IVT RNA作为瞬时转染的形式引入细胞。使用通过合成产生的质粒DNA模板体外转录产生RNA。可以使用适当的引物和RNA聚合酶,通过PCR将来自任何来源的感兴趣的DNA直接转化为用于体外mRNA合成的模板。DNA的来源可以是例如基因组DNA、质粒DNA、噬菌体DNA、cDNA、合成DNA序列或任何其它适合来源的DNA。对于体外转录期望的模板是一种或多种TRIM蛋白或一种或多种STUbL。

在一个实施方式中,用于PCR的DNA含有开放阅读框。DNA可以来自生物的基因组的天然存在的DNA序列。在一个实施方式中,DNA是感兴趣的全长基因或基因的一部分。基因可以包括5'和/或3'未翻译区(UTR)的一些或全部。基因可以包括外显子和内含子。在一个实施方式中,用于PCR的DNA是人基因。在另一个实施方式中,用于PCR的DNA是包含5'和3'UTR的人基因。可替换地,DNA可以是通常不在天然存在的生物中表达的人工DNA序列。示例性的人工DNA序列是含有连接在一起形成编码融合蛋白的开放阅读框的基因部分的DNA序列。连接在一起的DNA部分可以来自单一生物体或者来自多于一种生物体。

在一个实施方式中,本发明的组合物包含编码本文描述的一种或多种TRIM蛋白或RNF4的修饰的核酸。例如,在一个实施方式中,组合物包含核苷修饰的RNA。在一个实施方式中,组合物包含核苷修饰的mRNA。与未修饰的mRNA相比,核苷修饰的mRNA具有特别的优势,包括例如增加的稳定性、低免疫原性和增强的翻译。在美国专利号8,278,036中进一步描述了在本发明中有用的核苷修饰的mRNA,以其全部内容通过引证并入本文。

修饰的细胞

本发明包括包含细胞的组合物,细胞包含一种或多种TRIM蛋白、一种或多种STUbL、编码一种或多种TRIM蛋白的核酸、编码一种或多种STUbL的核酸或它们的组合。在一个实施方式中,细胞遗传修饰以表达本发明的蛋白质和/或核酸。在某些实施方式中,遗传修饰的细胞与使用本发明的组合物治疗的受试者是自体同源的。可替换地,细胞相对于受试者可以是同种异体的、同基因的或异基因的。在某些实施方式中,细胞能够将所表达的蛋白质分泌或释放到细胞间隙中以将肽递送至一种或多种其它细胞。

可以使用本领域中的标准技术,体内或离体修饰遗传修饰的细胞。可以使用表达载体或者使用裸露的分离的核酸构建体进行细胞的遗传修饰。

在一个实施方式中,使用编码本文描述的一种或多种蛋白质的分离的核酸离体获得和修饰细胞。在一个实施方式中,细胞获得自受试者,遗传修饰以表达蛋白质和/或核酸并再次给予于受试者。在某些实施方式中,将细胞离体或体外扩增以产生细胞群体,其中将群体中的至少一部分给予于对其有需要的受试者。

在一个实施方式中,将细胞遗传修饰以稳定表达蛋白质。在另一个实施方式中,将细胞遗传修饰以瞬时表达蛋白质。

基底

本发明提供了包含本发明的蛋白质、本发明的分离的核酸、表达本发明的蛋白质的细胞或它们的组合的支架或基底组合物。例如,在一个实施方式中,将本发明的蛋白质、本发明的分离的核酸、表达本发明的蛋白质的细胞或它们的组合并入支架内。在另一个实施方式中,将本发明的蛋白质、本发明的分离的核酸、表达本发明的蛋白质的细胞或它们的组合施加于支架表面。本发明的支架可以是本领域中已知的任何类型。这种支架的非限制性实例包括水凝胶、静电纺丝支架、泡沫、网丝、片材、贴片和海绵。

治疗方法

本发明还提供了用于与蛋白质错误折叠、蛋白质聚集体或它们的组合有关的疾病或病症的治疗方法。

在各种实施方式中,通过本发明的方法可以治疗的疾病和病症包括但不限于:polyQ病症,如SCA1、SCA2、SCA3、SCA6、SCA7、SCA17、亨廷顿舞蹈病、齿状核红核苍白球路易体萎缩症(DRPLA)、阿尔茨海默氏病、帕金森氏症、肌萎缩性侧索硬化(ALS)、传染性海绵状脑病(朊病毒疾病)、滔蛋白病变和额颞叶脑退行性病变(FTLD)、AL淀粉样变性、AA淀粉样变性、家族性地中海热、老年系统性淀粉样变性、家族性淀粉样多神经病、透析相关的淀粉样变性、ApoAI淀粉样变性、ApoAII淀粉样变性、ApoAIV淀粉样变性、芬兰型遗传性淀粉样变性、溶菌酶淀粉样变性、纤维蛋白原淀粉样变性、冰岛型遗传性脑淀粉样血管病、II型糖尿病、甲状腺髓样癌、心房性淀粉样变性、遗传性脑出血伴淀粉样变性、垂体泌乳素瘤、注射局部淀粉样变性、主动脉内侧淀粉样变性、遗传性格子状角膜变性、与倒睫相关的角膜淀粉样变性、白内障、牙源性钙化上皮肿瘤、肺泡蛋白沉积症、包涵体肌炎和苔藓型皮肤淀粉样变性。在某些实施方式中,方法包括与p53突变聚集体有关的癌症的治疗或预防,其包括但不限于膀胱癌、星形细胞瘤、咽癌、淋巴瘤和腺癌。

当提供有包括本文详细描述的方法的本公开时,本领域技术人员将理解本发明不局限于已建立的与蛋白质错误折叠或蛋白质聚集体有关的疾病的治疗。具体地,疾病或病症不需要显示出达到损害受试者的程度;事实上,在给予治疗前,不需要在受试者中检测疾病或病症。也就是说,在本发明可以提供益处前,不必发生疾病或病症的明显病征或症状。因此,本发明包括预防与蛋白质错误折叠或蛋白质聚集体有关的疾病或病症的方法,其中如以上在本文其它处所讨论的,可以在疾病或病症发生前将调节剂组合物给予于受试者,由此预防疾病或病症。

当提供有本文中的公开时,本领域技术人员将理解与蛋白质错误折叠或蛋白质聚集体有关的疾病的预防涵盖了作为对与蛋白质错误折叠或蛋白质聚集体有关的疾病的发展或进展的预防措施向受试者给予调节剂组合物。如本文中其它处更充分讨论的,调节基因或基因产物的水平或活性的方法涵盖了不但用于调节多肽基因产物的水平和活性,而且还用于调节核酸表达,包括转录、翻译或两者的多种技术。

另外,如本文中其它处所公开的,一旦提供有本文所提供的教导,本领域技术人员将理解本发明涵盖了治疗或预防与蛋白质错误折叠或蛋白质聚集体有关的多种疾病的方法,其中调节基因或基因产物的水平或活性治疗或预防疾病。评价疾病是否与蛋白质错误折叠或蛋白质聚集体有关的多种方法在本领域中是已知的。此外,本发明涵盖了未来将发现的这类疾病的治疗或预防。

在一个方面,方法包括使用一种或多种TRIM蛋白稳定化错误折叠的蛋白质。在某些方面,通过本文描述的一种或多种TRIM蛋白稳定化功能性错误折叠的蛋白质可以治疗或预防与错误折叠的蛋白质有关的疾病或病症。例如,在一个实施方式中,通过本文描述的一种或多种TRIM蛋白对突变囊性纤维化跨膜传导调节因子(CFTR)的稳定化将允许突变CFTR发挥功能而不是被降解。设想使用TRIM蛋白稳定化错误折叠的蛋白质可以用于治疗与部分功能蛋白降解有关的囊性纤维化及其它疾病。通过本文描述的一种或多种TRIM蛋白对蛋白质的稳定化可以用于治疗与功能突变蛋白降解有关的任何疾病或病症,其包括但不限于囊性纤维化和溶酶体贮积症,如戈谢病和法布里病。

本发明涵盖了基因或基因产物的调节剂的给予。为了实践本发明的方法,基于本文所提供的公开,技术人员将理解如何配制和给予适当的调节剂组合物至受试者。本发明不局限于任何具体给予方法或治疗方案。

在一个实施方式中,方法包括向对其有需要的受试者给予有效量的增加一种或多种TRIM蛋白或一种或多种STUbL的表达或活性的组合物。

例如,在一个实施方式中,方法包括向对其有需要的受试者给予有效量的组合物,组合物增加以下中的一种或多种的表达或活性:人TRIM3、TRIM4、TRIM5、TRIM6、TRIM7、TRIM9、TRIM11、TRIM13、TRIM14、TRIM15、TRIM16、TRIM17、TRIM19(在本文中还称为“PML”)、TRIM20、TRIM21、TRIM24、TRIM25、TRIM27、TRIM28、TRIM29、TRIM32、TRIM34、TRIM39、TRIM43、TRIM44、TRIM45、TRIM46、TRIM49、TRIM50、TRIM52、TRIM58、TRIM59、TRIM65、TRIM67、TRIM69、TRIM70、TRIM74和TRIM75;和鼠TRIM30。

在一个实施方式中,方法包括向对其有需要的受试者给予有效量的增加RNF4和RNF111(Arkadia)中的一种或多种的表达或活性的组合物。

在一个实施方式中,方法包括向受试者给予有效量的增加一种和多种TRIM蛋白和一种和多种STUbL的表达和活性的组合物。

在一个实施方式中,方法包括在受试者的至少一种神经细胞中增加一种或多种TRIM蛋白或一种或多种STUbL的表达或活性。例如,在某些实施方式中,方法包括在至少一种神经元、星形胶质细胞、少突神经胶质细胞、浦肯野细胞、锥体细胞等中增加一种或多种TRIM蛋白或一种或多种STUbL的表达或活性。

在一个实施方式中,方法包括将受试者的神经组织与有效量的增加一种或多种TRIM蛋白或一种或多种STUbL的一种或多种组分的表达或活性的组合物接触。例如,在某些实施方式中,方法包括将受试者的神经元、星形胶质细胞、少突神经胶质细胞、浦肯野细胞、锥体细胞等与有效量的增加一种或多种TRIM蛋白或一种或多种STUbL的表达或活性的组合物接触。

本领域技术人员将理解本发明的调节剂可以单独或以任意组合给予。此外,本发明的调节剂可以在瞬时意义上单独或以任意组合给予,其中它们可以彼此同时或之前和/或之后给予。基于本文所提供的发明公开,本领域的技术人员将理解本发明的调节剂组合物可以用于预防或治疗与错误折叠的蛋白质或蛋白质聚集体有关的疾病或病症,并且调节剂组合物可以单独使用或与另一种调节剂以任意组合使用以引起预防或治疗结果。

在多个实施方式中,本文的本发明的任何调节剂可以单独给予或者和与蛋白质错误折叠或蛋白质聚集体有关的疾病相关的其它分子的其它调节剂组合给予。在多个实施方式中,本文的本发明的任何调节剂可以单独给予或者与其它治疗剂或预防剂组合给予,治疗剂或预防剂可以用于治疗或预防与蛋白质错误折叠或蛋白质聚集体有关的疾病。可以与本发明的调节剂组合使用的示例性治疗剂包括(但不限于)抗淀粉样蛋白-β抗体和抗τ抗体。

基因疗法

将受试者中的细胞与编码增加一种或多种TRIM蛋白或一种或多种STUbL的表达或活性的蛋白质的核酸组合物接触可以抑制或延缓与蛋白质错误折叠或蛋白质聚集体有关的疾病或病症的一种或多种症状的发生。

在一个实施方式中,本发明的核酸组合物编码一种或多种肽。例如,在一个实施方式中,核酸组合物可以编码包含一种或多种TRIM蛋白的氨基酸序列的肽。例如,在一个实施方式中,核酸组合物编码包含以下中的一种或多种的肽:人TRIM3、TRIM4、TRIM5、TRIM6、TRIM7、TRIM9、TRIM11、TRIM13、TRIM14、TRIM15、TRIM16、TRIM17、TRIM19(在本文中还称为“PML”)、TRIM20、TRIM21、TRIM24、TRIM25、TRIM27、TRIM28、TRIM29、TRIM32、TRIM34、TRIM39、TRIM43、TRIM44、TRIM45、TRIM46、TRIM49、TRIM50、TRIM52、TRIM58、TRIM59、TRIM65、TRIM67、TRIM69、TRIM70、TRIM74和TRIM75;和鼠TRIM30。在某些实施方式中,核酸组合物编码包含一种或多种STUbL的氨基酸序列的肽。例如,在一个实施方式中,核酸组合物编码包含RNF4和RNF111(Arkadia)中的一种或多种的肽。

还应将本发明视为包含编码与本文所公开的肽具有显著同源性的肽的任何形式的核酸。优选地,“显著同源的”肽与本文所公开的肽的氨基酸序列约50%同源、更优选地约70%同源、更优选地约80%同源、更优选地约90%同源、更优选地约95%同源、并且更优选地约99%同源。

在一个实施方式中,本发明的组合物包含编码肽、肽片段、本文的肽的同源物、变体、衍生物或盐的核酸。例如,在某些实施方式中,组合物包含编码肽的核酸,肽包含一种或多种TRIM蛋白、一种或多种TRIM蛋白的片段、一种或多种TRIM蛋白的同源物、一种或多种TRIM蛋白的变体或一种或多种TRIM蛋白的衍生物。在某些实施方式中,组合物包含编码肽的核酸,肽包含一种或多种STUbL、一种或多种STUbL的片段、一种或多种STUbL的同源物、一种或多种STUbL的变体或一种或多种STUbL的衍生物。

根据本发明,还提供了向细胞提供蛋白质的方法,细胞带有与一种或多种TRIM蛋白或一种或多种STUbL的减弱或不足的活性有关的正常或突变的基因。向具有突变基因的细胞提供蛋白质应允许受体细胞正常作用。可以将编码肽的核酸在载体中引入细胞,从而使核酸保持在染色体外。在这种情况下,细胞将从染色体外的位置表达核酸。更优选的是其中将核酸或其部分以使得其整合至细胞的基因组或与细胞中存在的内源突变基因重组的方式引入细胞的情况。对于重组、对于整合和对于染色体外维持,用于基因引入的载体在本领域中是已知的,并且可以使用任何适合的载体。将DNA引入细胞的方法,如电穿孔、磷酸钙共沉淀和病毒转导在本领域中是已知的,并且方法的选择在开业医生的能力范围内。

如以上一般地讨论的,在适用的情况下,核酸可以在基因疗法中采用以增加本发明的肽的水平或活性,甚至可以在其中野生型基因以“正常”水平表达,但基因产物功能不足的那些人中使用。

“基因疗法”包括其中通过单一治疗实现持久作用的常规基因疗法和包括一次或重复给予治疗有效的DNA或mRNA的基因治疗剂的给予两者。可以修饰寡核苷酸以增加它们的吸收,例如,通过不带电的基团取代它们带负电荷的磷酸二酯基团。可以使用基因疗法递送本发明的一种或多种TRIM蛋白或一种或多种STUbL,例如,在神经细胞或组织中局部递送或者全身递送(例如,通过选择性靶向特异性组织类型的载体,例如,组织特异性腺相关病毒载体)。在一些实施方式中,可以用编码本发明的任何肽的核酸离体转染从个体收获的原代细胞,并且然后将转染的细胞返回至个体的身体。

基因疗法在本领域中是熟知的。参见,例如,WO96/07321,其公开了使用基因疗法产生胞内抗体。还成功在人患者中证实了基因疗法。参见,例如,Baumgartner et al.,Circulation 97:12,1114-1123(1998);Fatham,C.G.‘A gene therapy approach to treatment of autoimmune diseases’,Immun.Res.18:15-26(2007);和美国专利号7,378089,两者通过引证并入本文。还参见Bainbridge JWB et al.,“Effect of gene therapy on visual function in Leber’s congenital Amaurosis”.N Engl J Med 358:2231-2239,2008;和Maguire AM et al.“Safety and efficacy of gene transfer for Leber’s Congenital Amaurosis”.N Engl J Med 358:2240-8,2008。

存在两种将编码肽或蛋白质的核酸(可选地包含在载体中)体内和离体引入患者细胞的主要方法。对于体内递送,在某些情况下,将核酸直接注入患者,有时在最需要蛋白质的位点。对于离体治疗,取出患者细胞,将核酸引入这些分离的细胞并将修饰的细胞直接或例如包封在将植入患者的多孔膜内给予于患者(参见,例如,美国专利号4,892,538和5,283,187)。存在多种可用于将核酸引入活细胞的技术。取决于核酸是体外转移到培养的细胞中还是体内在预期宿主的细胞中,技术是不同的。适合于将核酸体外转移至哺乳动物细胞的技术包括脂质体、电穿孔、微注射、细胞融合、DEAE-葡聚糖、磷酸钙沉淀法等的使用。用于基因离体递送的常用载体是反转录病毒和慢病毒载体。

将根据公认的方法进行基因疗法,例如,如通过Friedman et al.,1991,Cell 66:799-806或Culver,1996,Bone Marrow Transplant 3:S6-9;Culver,1996,Mol.Med.Today 2:234-236描述的。在一个实施方式中,首先通过本领域中已知的诊断方法分析来自患者的细胞以确定一种或多种TRIM蛋白或一种或多种STUbL的表达或活性。制备含有与表达控制元件连接并且能够在细胞内部复制的基因的拷贝或其功能等价物的病毒或质粒载体。载体能够在细胞内部复制。可替换地,载体可以是复制缺陷型的并且在辅助细胞中复制以用于在基因疗法中使用。适合的载体是已知的,如在美国专利号5,252,479和PCT公开的专利申请WO 93/07282和美国专利号5,691,198;5,747,469;5,436,146和5,753,500中所公开的。然后,将载体注入患者。如果转染的基因不是永久引入到每个靶细胞的基因组,则可能必须定期重复治疗。

本领域中已知的基因转移系统可以在本发明的基因疗法的实践中是有用的。这些包括病毒和非病毒转移方法。一些病毒已用作基因转移载体或者用作修复基因转移载体的基础,包括乳多空病毒(例如,SV40,Madzak et al.,1992,J.Gen.Virol.73:1533-1536)、腺病毒(Berkner,1992;Curr.Topics Microbiol.Immunol.158:39-66)、牛痘病毒(Moss,1992,Current Opin.Biotechnol.3:518-522;Moss,1996,PNAS 93:11341-11348)、腺相关病毒(Russell and Hirata,1998,Mol.Genetics 18:325-330)、疱疹病毒,包括HSV和EBV(Fink et al.,1996,Ann.Rev.Neurosci.19:265-287)、慢病毒(Naldini et al.,1996,PNAS 93:11382-11388)、辛德毕斯和生里基森林病毒(Berglund et al.,1993,Biotechnol.11:916-920)和鸟类(Petropoulos et al.,1992,J.Virol.66:3391-3397)、鼠(Miller,1992,Hum.Gene Ther.3:619-624)以及人来源的反转录病毒(Shimada et al.,1991;Helseth et al.,1990;Page et al.,1990;Buchschacher and Panganiban,1992,J.Virol.66:2731-2739)。大部分人基因疗法流程基于失去功能的鼠反转录病毒,尽管也在使用腺病毒和腺相关病毒。

本领域中已知的非病毒基因转移法包括化学技术,如磷酸钙共沉淀;机械技术,例如,微注射;通过脂质体的膜融合介导的转移;以及直接DNA吸收和受体介导的DNA转移(Curiel et al.,1992,Am.J.Respir.Cell.Mol.Biol 6:247-252)。病毒介导的基因转移可以与使用脂质体递送的直接体外基因转移组合,使得将病毒载体导向肿瘤细胞而不是周围未分化的细胞。然后,生产细胞的注射将提供连续的载体颗粒源。该技术已批准用于具有不能手术的脑肿瘤的人。

在组合了生物和物理基因转移方法的方法中,将任何尺寸的质粒DNA与对腺病毒六邻体蛋白特异的聚赖氨酸结合的抗体组合,并且将得到的复合物结合至腺病毒载体。然后,将三分子复合物用于感染细胞。在连接的DNA受损前,腺病毒载体允许内涵体有效结合、内化和降解。对于腺病毒基载体递送的其它技术,参见美国专利号5,691,198;5,747,469;5,436,146和5,753,500。

已显示脂质体/DNA复合物能够介导直接的体内基因转移。尽管在标准脂质体制剂中,基因转移过程是非特异性的,但是例如,在直接原位给予后,在肿瘤种植处已报道了局部化的体内吸收和表达。

基因疗法背景中的表达载体表示包括含有足以表达克隆到其中的多核苷酸的序列的那些构建体。在病毒表达载体中,构建体含有足以支持构建体的包装的病毒序列。如果多核苷酸编码蛋白质,则表达将产生蛋白质。如果多核苷酸编码反义多核苷酸或核糖酶,则表达将产生反义多核苷酸或核糖酶。因此,在该背景中,表达不要求合成蛋白产物。除克隆到表达载体中的多核苷酸之外,载体还含有在真核细胞中起作用的启动子。克隆的多核苷酸序列受该启动子的控制。适合的真核启动子包括如上描述的那些。表达载体还可以包含序列,如可选择的标志物及本文的其它序列。

在某些实施方式中,方法包括基因转移技术的使用,其将分离的核酸直接靶向神经组织。例如,受体介导的基因转移伴随着核酸分子(通常处于共价键封闭的超螺旋质粒形式)与蛋白配体通过聚赖氨酸的结合。根据靶细胞/组织类型的细胞表面上的相应配体受体的存在,选择配体。如果需要,可以将这些配体-DNA结合物直接注射到血液中,并导向在此发生受体结合和DNA-蛋白质复合物内化的靶组织。为了克服DNA胞内破坏的问题,可以包括使用腺病毒的共感染以破坏内涵体功能。

药物组合物和制剂

本发明还涵盖了本发明的药物组合物或其盐实践本发明的方法的用途。这种药物组合物可以包括处于适合于向受试者给予的形式的本发明的至少一种调节剂组合物或其盐,或者药物组合物可以包含本发明的至少一种调节剂组合物或其盐,和一种或多种药用载体、一种或多种其它成分或这些成分的一些组合。本发明的化合物或结合物可以以生理学上可接受的盐的形式(如与生理学上可接受的的阳离子或阴离子结合,如本领域中熟知的)存在于药物组合物中。

在一个实施方式中,可以给予用于实践本发明方法的药物组合物以递送1ng/kg/天至100mg/kg/天的剂量。在另一个实施方式中,可以给予用于实践本发明的药物组合物以递送1ng/kg/天至500mg/kg/天的剂量。

本发明的药物组合物中活性成分、药用载体和任何其它成分的相对量将根据待治疗的受试者的特征、身材和病况并且还根据组合物给予的途径而变化。举例来说,组合物可以包含0.1%至100%(w/w)的活性成分。

可以适当地开发在本发明的方法中有用的药物组合物以用于口服、直肠、阴道、肠胃外、局部、肺、鼻内、口含、眼部给予途径或另一种给予途径。在本发明的方法内有用的组合物可以直接给予至哺乳动物的皮肤、阴道或任何其它组织。其它所考虑的制剂包括脂质体制剂、含有活性成分的重新包封的红细胞和基于免疫学的制剂。给予途径对于技术人员将是显而易见的,并且将基于多种因素,包括待治疗的疾病的类型和严重性、待治疗的动物或人受试者的类型和年龄等。

可以通过任何已知的或此后在药学领域中发展出的方法制备本文的药物组合物的制剂。通常,这些制备方法包括以下步骤:使活性成分与载体或一种或多种其它辅助成分结合,然后,如有必要或期望,将产物成型或包装成所需的单剂量或多剂量单元。

如本文所使用的,“单位剂量”是包含预定量的活性成分的药物组合物的单个的量。活性成分的量通常等于将给予于受试者的活性成分的剂量或该剂量的方便的分数,如例如该剂量的二分之一或三分之一。单位剂量形式可以用于单次每日给药或多次每日给药(例如,每天约1至4次或更多次)。当使用多次每日给药时,对于每次给药,单位剂量形式可以是相同或不同的。

尽管对本文所提供的药物组合物的描述主要涉及在伦理上适合于向人给予的药物组合物,但本领域技术人员应理解这些组合物通常还适合于向所有种类的动物给予。改变适合于向人给予的药物组合物进行以使组合物适合于向多种动物给予的方法是众所周知的,并且兽医药理学领域的普通技术人员可以仅通过常规实验(如果需要)来设计和做出这种改变。考虑对其给予本发明的药物组合物的受试者包括但不限于人及其它灵长类、哺乳动物,包括商业相关的哺乳动物,如牛、猪、马、绵羊、猫和狗。

在一个实施方式中,使用一种或多种药用赋形剂或载体配制本发明的组合物。在一个实施方式中,本发明的药物组合物包含治疗有效量的本发明的化合物或结合物和药用载体。有用的药用载体包括但不限于甘油、水、盐水、乙醇及其它药用盐溶液,如磷酸盐和有机酸盐。在Remington's Pharmaceutical Sciences(1991,Mack Publication Co.,New Jersey)中描述了这些及其它药用载体的实例。

载体可以是溶剂或分散介质,其含有例如水、乙醇、多元醇(例如,甘油、丙二醇和液体聚乙二醇等)、它们适合的混合物和植物油。可以通过例如使用覆层如卵磷脂,在分散体系的情况下通过保持所需的粒度以及通过使用表面活性剂来保持合适的流动性。可以通过各种抗菌和抗真菌剂例如,尼泊金、氯代丁醇、苯酚、抗坏血酸、硫柳汞等防止微生物的作用。在许多情况下,优选在组合物中包括等渗剂,例如糖、氯化钠或多元醇,如甘露糖醇和山梨糖醇。可注射组合物的延长吸收可以通过在组合物中包括使吸收延迟的试剂例如,单硬脂酸铝或明胶来实现。在一个实施方式中,药用载体不是单独的DMSO。

可以与常规赋形剂,即本领域已知的适合于口服、阴道、肠胃外、鼻部、静脉内、皮下、肠内或任何其它适合的给予形式的药用有机或无机载体物质混合使用制剂。药物制剂可以灭菌,并且如果需要,可以与助剂,例如润滑剂、防腐剂、稳定剂、润湿剂、乳化剂、影响渗透压缓冲的盐、着色剂、调味剂和/或芳香物等混合。需要时,它们还可以与其它活性剂,例如,其它止痛剂混合。

如本文所使用的,“其它成分”包括但不限于以下中的一种或多种:赋形剂;表面活性剂;分散剂;惰性稀释剂;成粒剂和崩解剂;粘结剂;润滑剂;甜味剂;调味剂;着色剂;防腐剂;生理学可降解的组合物,如明胶;水性载体和溶剂;油性载体和溶剂;助悬剂;分散剂或润湿剂;乳化剂,镇痛剂;缓冲剂;盐;增稠剂;填充剂;乳化剂;抗氧化剂;抗生素;抗真菌剂;稳定剂;和药用聚合物或疏水性材料。可以包含在本发明的药物组合物中的其它“其它成分”是本领域已知的,并且在例如Genaro ed.(1985,Remington's Pharmaceutical Sciences,Mack Publishing Co.,Easton,PA)中描述,其通过引证并入本文。

本发明的组合物可以包含按组合物总重量计约0.005%至2.0%的防腐剂。防腐剂用于在暴露于环境中的污染物的情况下防止腐败。根据本发明,有用的防腐剂的实例包括但不限于选自由苯甲醇、山梨酸、对羟苯甲酸、咪唑烷基脲及它们的组合组成的组。特别优选的防腐剂是约0.5%至2.0%的苯甲醇和0.05%至0.5%的山梨酸的组合。

组合物优选地包含抑制化合物降解的抗氧化剂和螯合剂。对于一些化合物,优选的抗氧化剂是按组合物总重量计约0.01wt%至0.3wt%的优选范围内的BHT、BHA、α-生育酚和抗坏血酸,并且更优选地,0.03wt%至0.1wt%的范围内的BHT。优选地,螯合剂以按组合物总重量按重量计0.01%至0.5%的量存在。特别优选的螯合剂包括按组合物总重量计约0.01%至0.20%的重量范围内,并且更优选地,0.02%至0.10%的范围内的乙二胺四乙酸盐(例如,乙二胺四乙酸二钠)和柠檬酸。螯合剂对螯合组合物中对制剂寿命可能有害的金属离子是有用的。尽管对于一些化合物,BHT和乙二胺四乙酸二钠分别是特别优选的抗氧化剂和螯合剂,但是如本领域技术人员所知的,其它适合的和等价的抗氧化剂和螯合剂可以被取代。

可以使用常规方法制备液体混悬液以实现活性成分在水性或油性媒介中的混悬。水性载体包括例如水和等渗盐水。油性载体包括例如杏仁油、油酯、乙醇、植物油,如花生油、橄榄油、芝麻油或椰子油、分馏植物油,和矿物油,如液体石蜡。液体混悬剂还可以包含一种或多种其它成分,其包括但不限于助悬剂、分散剂或润湿剂、乳化剂、镇痛剂、防腐剂、缓冲剂、盐、调味剂、着色剂和甜味剂。油性混悬剂还可以包含增稠剂。已知的助悬剂包括但不限于葡萄糖醇糖浆、氢化可食用脂肪、海藻酸钠、聚乙烯基吡咯烷酮、黄芪胶、阿拉伯树胶和纤维素衍生物,如羧甲基纤维素钠、甲基纤维素、羟丙基甲基纤维素。已知的分散剂或润湿剂包括但不限于天然存在的磷脂,如卵磷脂、氧化烯烃与脂肪酸、与长链脂肪醇、与来源于脂肪酸和己糖醇的偏酯或者与来源于脂肪酸和已糖醇酐的偏酯的缩合产物(例如,分别为聚氧化乙烯硬脂酸酯、十七乙烯氧基鲸蜡醇、聚氧乙烯单油酸山梨糖醇酯和聚氧乙烯单油酸脱水山梨糖醇酯)。已知的乳化剂包括但不限于卵磷脂和阿拉伯胶。已知的防腐剂包括但不限于对羟基苯甲酸甲酯、对羟基苯甲酸乙酯或对羟基苯甲酸正丙酯、抗坏血酸和山梨酸。已知的甜味剂包括例如甘油、丙二醇、山梨糖醇、蔗糖和糖精。已知用于油性混悬液的增稠剂包括例如蜂蜡、固体石蜡和鲸蜡醇。

可以通过与液体混悬剂基本相同的方式在水性或油性溶剂中制备活性成分的液体溶液,其主要差别在于活性成分是溶解的,而不是在溶剂中悬浮的。如本文所使用的,“油性”液体是包含含碳液体分子并且显示出低于水的极性的液体。本发明的药物组合物的液体溶液可以包含液体混悬液的每种成分,应理解助悬剂不必需辅助活性成分在溶剂中的溶解。水性溶剂包括例如水和等渗盐水。油性溶剂包括例如杏仁油、油酯、乙醇、植物油,如花生油、橄榄油、芝麻油或椰子油、分馏植物油,和矿物油,如液体石蜡。

可以使用已知的方法制备本发明的药物制剂的粉末和颗粒制剂。可以将这些制剂直接给予于受试者,或者将其用于例如形成片剂、填充胶囊或通过向其中添加水性或油性载体制备水性或油性混悬液。这些制剂中的每种还可以包含一种或多种分散剂或润湿剂、助悬剂和防腐剂。这些制剂中还可以包含其它赋形剂,如填充剂和甜味剂、调味剂或着色剂。

还可以以水包油乳剂或油包水乳剂形式制备、包装或销售本发明的药物组合物。油相可以是植物油,如橄榄油或花生油、矿物油,如液体石蜡或者这些的组合。这些组合物还可以包含一种或多种乳化剂,如天然存在的胶,如阿拉伯胶或黄芪胶、天然存在的磷脂,如大豆磷脂或卵磷脂、来源于脂肪酸和已糖醇酐的组合的酯或偏酯,如单油酸脱水山梨糖醇酯,和这些偏酯与氧化乙烯的缩合产物,如聚氧化乙烯单油酸脱水山梨糖醇酯。这些乳剂还可以含有其它成分,包括例如甜味剂或调味剂。

用化学组合物浸渍或涂覆材料的方法在本领域中是已知的,并且包括但不限于使用或不使用后续干燥,将化合组合物沉积或结合在表面上的方法,在材料(即,如使用生理学可降解材料)合成期间将化合组合物引入到材料结构中的方法,和将水性或油性溶液或混悬液吸附到吸附性材料中的方法。

给予方案可以影响有效量的构成。可以在疾病诊断之前或之后,向受试者给予治疗制剂。另外,可以每天或顺序给予一些独立的剂量和错开的剂量,或者可以连续输注剂量,或者可以是弹丸注射。另外,可以根据治疗或预防情况的紧急需要的指示,按比例增加或减少治疗制剂的剂量。

可以使用已知的程序,以对疾病的预防或治疗有效的剂量和时间段进行对受试者,优选哺乳动物,更优选人的本发明的组合物的给予。可以根据以下因素改变实现治疗效果所必需的治疗化合物的有效量,如所使用的特定化合物的活性;给予时间;化合物外排速率;治疗持续时间;与化合物结合使用的其它药物、化合物或材料;待治疗受试者的疾病或病症状况、年龄、性别、体重、病况、一般健康状况和先前病史,以及医学领域中熟知的类似因素。可以调节给予方案以提供最佳的治疗反应。例如,可以每天给予一些单独的剂量,或者可以如治疗情况紧急需要所指示的,按比例降低剂量。本发明的治疗化合物的有效量范围的非限制性实例为约1至5000mg/kg体重/天。本领域的技术人员将能够研究相关因素并做出与治疗化合物的有效量有关的决定而无需过多实验。

可以向受试者以频繁至每天几次的频率给予化合物,或者可以较不频繁地给予,如每天一次、每周一次、每两周一次、每月一次或甚至更不频繁,如每几个月一次或甚至每年一次或更少。应理解在非限制性实例中,可以每天、每隔一天、每2天、每3天、每4天或每5天给予每天剂量给予的化合物的量。例如,通过每隔一天给予,可以在周一开始5mg每天的剂量,并在周三给予第一次后续的5mg每天的剂量,在周五给予第二次后续的5mg每天的剂量。剂量的频率对于技术人员将是显而易见的,并且将基于多种因素,如但不限于待治疗疾病的类型和严重性、动物的类型和年龄等。

可以改变本发明药物组合物中活性成分的真实剂量水平,从而对特定受试者、组合物和给予形式获得实现所需治疗反应有效的活性成分的量,而不会对受试者有毒。

具有本领域常规技术的医生,例如,医师或兽医可以容易地确定并开具所需的药物组合物的有效量。例如,医师或兽医可以以低于实现所需治疗效果所需的水平开始在药物组合物中使用的本发明的化合物的剂量,并逐渐增加剂量直至实现所需效果。

在具体的实施方式中,将化合物配制成给药单元的形式是尤其有利的,以便于给予和剂量均匀性。如本文所使用的给药单元的形式是指适用于待治疗受试者的单一剂量的物理上分离的单元;每个单元含有计算为产生与所需药物媒介相关的所需治疗效果的预定量的治疗化合物。本发明的给药单元的形式由(a)治疗化合物的独有特性和待实现的具体治疗效果,以及(b)制备/配置这种治疗化合物用于受试者中疾病的治疗的领域中固有的限制所决定,并且直接取决于它们。

在一个实施方式中,将本发明的组合物以每天1至5次或以上的范围内的剂量给予于受试者。在另一个实施方式中,将本发明的组合物以以下剂量范围给予于受试者,包括但不限于每天一次、每两天一次、每三天一次至每周一次和每两周一次。对于本领域技术人员显而易见的是对于不同受试者,本发明组合物的各自组合的给予频率将是不同的,这取决于多种因素,包括但不限于年龄、待治疗的疾病或病症、性别、整体健康状况及其它因素。因此,本发明不应视为仅限于任何特定给药方案,并且给予任何受试者的精确剂量和组合物将由主治医生考虑受试者有关的所有其它因素来决定。

用于给予的本发明的化合物可以在约1mg至约10000mg、约20mg至约9500mg、约40mg至约9000mg、约75mg至约8500mg、约150mg至约7500mg、约200mg至约7000mg、约3050mg至约6000mg、约500mg至约5000mg、约750mg至约4000mg、约mg至约3000mg、约10mg至约2500mg、约20mg至约2000mg、约25mg至约1500mg、约50mg至约1000mg、约75mg至约900mg、约100mg至约800mg、约250mg至约750mg、约300mg至约600mg、约400mg至约500mg的范围内以及它们之间的任何或全部的全增量或偏增量。

在一些实施方式中,本发明化合物的剂量为约1mg至约2500mg。在一些实施方式中,在本文的组合物中使用的本发明的化合物的剂量小于约10000mg、或小于约8000mg、或小于约6000mg、或小于约5000mg、或小于约3000mg、或小于约2000mg、或小于约1000mg、或小于约500mg、或小于约200mg、或小于约50mg。类似地,在一些实施方式中,如本文的第二化合物(即,用于治疗如本发明组合物所治疗的相同或另一种疾病的药物)的剂量小于约1000mg、或小于约800mg、或小于约600mg、或小于约500mg、或小于约400mg、或小于约300mg、或小于约200mg、或小于约100mg、或小于约50mg、或小于约40mg、或小于约30mg、或小于约25mg、或小于约20mg、或小于约15mg、或小于约10mg、或小于约5mg、或小于约2mg、或小于约1mg、或小于约0.5mg以及它们的任何或全部的全增量或偏增量。

在一个实施方式中,本发明涉及包装的药物组合物,其包括容纳单独的或与第二药剂结合的治疗有效量的本发明的化合物或结合物的容器;和使用化合物或结合物治疗、预防或减轻受试者中一种或多种疾病症状的说明书。

术语“容器”包括用于容纳药物组合物的任何容器。例如,在一个实施方式中,容器为含有药物组合物的包装。在其它实施方式中,容器不是含有药物组合物的包装,即容器是含有包装的药物组合物或未包装的药物组合物以及药物组合物的使用说明书的的容器,如盒或小瓶。此外,包装技术在本领域中是熟知的。应理解药物组合物的使用说明书可以包含在含有药物组合物的包装上,并且由此说明书与包装产品形成了增加的功能关系。然而,应理解说明书可以包含有关化合物发挥其预定功能的能力的信息,例如,治疗或预防受试者中的疾病或向受试者递送图像或诊断试剂。

本发明的任何组合物的给予途径包括口服、鼻部、直肠、肠胃外、舌下、经皮、经粘膜(例如,舌下、舌、口含(透颊)、(经)尿道、阴道(例如,经阴道和阴道周)、鼻(内)和(经)直肠)、膀胱内、肺内、脑内、硬膜外、脑室内、十二指肠内、胃内、鞘内、皮下、肌内、皮内、动脉内、静脉内、支气管内、吸入和局部给予。

适合的组合物和给药形式包括例如片剂、胶囊、囊片、丸剂、胶丸、锭剂、分散剂、混悬剂、溶液、糖浆、颗粒剂、球剂、透皮贴片、凝胶剂、粉剂、小粒、乳浆剂、糖锭、乳膏、糊剂、硬膏剂、洗剂、盘剂、栓剂、用于鼻部或口服给予的液体喷雾、用于吸入的干粉或气雾制剂、用于膀胱内给予的组合物和制剂等。应理解在本发明中有用的制剂和组合物不限于本文的具体制剂和组合物。

诊断方法

本发明提供了诊断受试者患有与蛋白质错误折叠或蛋白质聚集体有关的疾病或病症或者有发展该疾病或病症的风险的方法。例如,在一个实施方式中,方法包括使用一种或多种TRIM蛋白或一种或多种STUbL的表达或活性水平作为诊断标志物。在一个实施方式中,方法包括检测编码一种或多种TRIM蛋白或一种或多种STUbL的核酸中基因突变的存在。

在一个实施方式中,方法用于诊断受试者患有与蛋白质错误折叠或蛋白质聚集体有关的疾病或病症。在一个实施方式中,方法用于诊断受试者有发展与蛋白质错误折叠或蛋白质聚集体有关的疾病或病症的风险。在一个实施方式中,方法用于评价与蛋白质错误折叠或蛋白质聚集体有关的神经退行性疾病或病症的疗法的有效性。

在一个实施方式中,方法包括从受试者采集生物样品。示例性的样品包括但不限于血液、尿、粪便、汗、胆汁、血清、血浆、组织活检等。例如,在一个实施方式中,样品包含神经组织的至少一种细胞。在一个实施方式中,样品包含神经元、星形胶质细胞、少突神经胶质细胞、浦肯野细胞、锥体细胞等。

用于检测一种或多种TRIM蛋白或一种或多种STUbL的表达或活性降低的方法包括在核酸或蛋白水平上检测基因或其产物的任何方法。这些方法在本领域中是熟知的并且包括但不限于核酸杂交技术、核酸反转录方法和核酸扩增方法、免疫印迹、RNA印迹、DNA印迹、ELISA、免疫沉淀、免疫荧光、流式细胞术、免疫细胞化学。在具体的实施方式中,使用例如对特定蛋白的抗体在蛋白水平上检测破坏的基因转录。这些抗体可以用于多种方法,如免疫印迹、ELISA、免疫沉淀、流式细胞术或免疫细胞化学技术。

制造重组蛋白的方法

在某些实施方式中,本发明提供了在生产感兴趣的重组蛋白中使用一种或多种TRIM蛋白、一种或多种STUbL或它们的组合的方法。例如,一种或多种TRIM蛋白、一种或多种STUbL或它们的组合可以用于使感兴趣的重组蛋白的蛋白质聚集体解聚,从而允许生产并收集感兴趣的重组蛋白。

在某些实施方式中,方法包括向细胞给予一种或多种TRIM蛋白、一种或多种STUbL、编码一种或多种TRIM蛋白的核酸分子、编码一种或多种STUbL的核酸分子或它们的组合。在某些实施方式中,修饰细胞以表达感兴趣的重组蛋白。细胞可以是任何表达系统,包括但不限于酵母表达系统、细菌表达系统、昆虫表达系统或哺乳动物表达系统。

实验实施例

通过参考以下实验实施例,进一步详细描述了本发明。这些实施例仅以说明的目的提供,并且除非另外说明,否则不旨在是限制性的。因此,本发明决不应视为受限于以下实施例,而应视为由于本文所提供的教导内容,涵盖了显而易见的任何和全部变化。

无需进一步描述,据信使用以上说明以及以下说明性实施例,本领域的技术人员可以制备和使用本发明并实践所主张的方法。因此,以下工作实施例具体指出了本发明优选的实施方式,并且不应将其视为以任何方式限制本发明公开的其它内容。

实施例1:降解错误折叠的蛋白质并且保护不受神经变性的细胞系统

错误折叠的蛋白质损害细胞功能并导致疾病。尚未良好理解如何检测并降解这些蛋白质。本文所提供的实验显示PML(也称为TRIM19)和SUMO依赖性泛素连接酶RNF4一起作用以促进哺乳动物细胞核中错误折叠的蛋白质的降解。PML通过不同底物识别位点选择性地与错误折叠的蛋白质相互作用并通过其SUMO连接酶活性将这些蛋白质与小类泛素修饰因子(SUMO)结合。然后,通过RNF4识别并泛素化SUMO化的错误折叠的蛋白质,并且随后靶向错误折叠的蛋白质用于蛋白酶体降解。此外,本文表明PML缺陷加速了脊髓小脑性共济失调1型(SCA1)的小鼠模型中的多聚谷氨酰胺(polyQ)疾病。这些发现显示了通过顺序SUMO化和泛素化除去错误折叠的蛋白质的哺乳动物系统并且定义了它在保护抵抗蛋白质错误折叠疾病中的作用。

早幼粒细胞性白血病蛋白质(PML;也称为TRIM19)是三结构域(TRIM)蛋白家族的成员,其含有N端TRIM/BRCC区,包括RING域、一个或两个B盒和螺旋卷曲(CC)基序,随后是可变C端区。PML主要是细胞核蛋白并且是PML核体的同名组件。其涉及多种细胞过程,包括细胞凋亡、转录、DNA损伤信号传导和抗病毒反应(Bernardi and Pandolfi,2007,Nat Rev Mol Cell Biol,8:1006-1016)。值得注意地,PML还与通过与SCA有关的polyQ蛋白所形成的聚集体共定位(Skinner et al.,1997,Nature,389,971-974;Takahashi et al.,2003,Neurobiol Dis,13:230-237)并且,一旦过表达,促进它们中的至少一种(突变体脊髓小脑共济失调蛋白-7)的降解(Janer et al.,2006,J Cell Biol,174:65-76)。尽管这些观察结果具有潜在重要性,但PML在除去错误折叠的蛋白质中的作用尚未完全清楚。具体地,不清楚PML是否在细胞核错误折叠的蛋白质的除去中具有广泛作用。PML除去错误折叠的蛋白质的分子机制的关键问题尚未解决。此外,PML对错误折叠的蛋白质的影响的生理学相关性仍是未知的。

现将描述在这些实验中使用的材料和方法。

质粒

除非另外说明,否则所有蛋白质是人源的。在哺乳动物细胞中表达以下蛋白质的质粒是通过PCR在pRK5中制备的,并且如以下所示,各自在NH2-或COOH-末端与HA、FLAG或6×His标签或者GST或GFP蛋白质融合:FLAG-PML突变体(除非另外说明,否则为同种型IV);GST-PML;Atxn1 82Q-GFP、HA-Atxn1 82Q-FLAG、FLAG-Atxn1 82Q;HA-Httex1p97QP和HA-Httex1p 97QP(KR);FLAG-nFluc-GFP、FLAG-nFlucSM-GFP和FLAG-nFlucDM-GFP;HA-RNF4、HA-RNF4SIMm、HA-RNF4-FLAG和HA-RNF4SIMm-FLAG;和HA-SUMO2KR。Atxn1 82Q质粒是基于H.Orr所提供的FLAG-Atxn1 82Q/pcDNA质粒所制备的(Riley et al.,2005,J Biol Chem,280:21942-21948);Httex1p 97QP和Httex1p 97QP(KR)(其中K6、K9和K15改变为Arg)基于Steffan et al.,2004,Science,304:100-104;并且nFluc质粒基于Gupta et al.,2011,Nat Methods,8:879-884。将每种nFluc蛋白在NH2-末端融合至SV40核定位信号(PKKKRKV)(SEQ ID NO:147)并在COOH-末端融合至GFP。在FlucDM中,R188和R261改变为Glu;在FlucSM中,R188改变为Glu(Gupta et al.,2011,Nat Methods,8:879-884)。RNF4的PCR扩增的模板购自Open Biosystems(基因登录号:NM002938)。在RNF4SIMm中,将SIM内的下列位点改变为Ala:I36、L38和V39(SIM1);I46、V47和L49(SIM2);V57、V58和V59(SIM3);和V67、V68、I69和V70(SIM4)。在SUMO2KR中,内部SUMO化共有位点Lys11突变为Arg。

对于细菌表达,在pGEX-1ZT(具有额外的克隆位点的pGEX-1λT的衍生物)中构建Htt 25Q、Htt 103Q、Htt 52Q、Htt 52Q cc-、PML CC-FLAG、RNF4和RNF4SIMm的GST融合。Htt 25Q、Htt 52Q和Htt 103Q含有Htt氨基酸1-17,并且随后是指定长度的polyQ延伸(Krobitsch and Lindquist,2000,Proc Natl Acad Sci USA,97:1589-1594)。通过连接合成的寡核苷酸组装Htt 52Q和Htt 52Q cc-cDNAs。在pET28a中构建FLAG-PML F12(571-633)-6×His。通过DNA测序确认对本研究所产生的全部质粒。

先前描述了以下质粒:FLAG-PML、FLAG-PML M6(其具有C57S、C60S、C129A、C132A、C189A和H194A突变)、6×His-SUMO1和6×His-SUMO2(Chu and Yang,2011,Oncogene,30:1108-1116);FLAG-Atxn182Q和FLAGAtxn1 30Q(Riley et al.,2005,J Biol Chem,280:21942-21948);荧光素酶-6×his(北美萤火虫(Photinus pyralis)荧光素酶变体)(Sharma et al.,2010,Nat Chem Biol,6:914-920);GST-rRNF4(其中“r”表示鼠来源,下同)、GSTrRNF4CS1(其中C136和C139改变为Ser)、FLAG-rRNF4和FLAGrRNF4CS(其中C136、C139、C177和C180改变为Ser)(Hakli et al.,2004,FEBS Lett,560:56-62);和PML同种型I、II、III、IV和VI(图8A中所使用的)(Xu et al.,2005,Mol Cel,17:721-732)。

siRNA

PML和RNF4siRNA购自Qiagen,并且有义链序列为:PML#4,CTCCAAGATCTAAACCGAGAA(SEQ ID NO:148);PML#9,CACCCGCAAGACCAACAACAT(SEQ ID NO:149);RNF4#5,CCCTGTTTCCTAAGAACGAAA(SEQ ID NO:150);RNF4#6,TAGGCCGAGCTTTGCGGGAAA(SEQ ID NO:151);RNF4#8,AAGACTGTTTCGAAACCAACA(SEQ ID NO:152)。使用单独的siRNA或等摩尔比组合降低RNF4。SUMO1siRNA(Thermo Scientific,siGENOME SMARTpool M-016005-03-0005)是4种靶标特异性siRNA双螺旋的混合。有义链序列为:TCAAGAAACUCAAAGAATC(SEQ ID NO:153)、GACAGGGTGTTCCAATGAA(SEQ ID NO:154)、GGTTTCTCTTTGAGGGTCA(SEQ ID NO:155)和GAATAAATGGGCATGCCAA(SEQ ID NO:156)。SUMO2/3siRNA(Santa Cruz sc-37167)是三种不同的siRNA双螺旋的混合,并且有义链序列为CCCAUUCCUUUAUUGUACA(SEQ ID NO:157)、CAGAGAAUGACCACAUCAA(SEQ ID NO:158)和CAGUUAUGUUGUCGUGUAU(SEQ ID NO:159)。

细胞培养和转染

将HeLa细胞(来自ATCC)和表达GFP-SUMO2或者GFP-SUMO3的U2OS细胞(Mukhopadhyay et al.,2006,J Cell Biol,174:939-949)维持在标准培养条件下。使用Lipofectamine 2000将DNA质粒转染到细胞中。当PML和Atxn1共转染时,使用FLAG-PML加Atxn1 82Q/30Q-GFP或者HA-PML加FLAG-Atxn1 82Q/30Q。分别将HA-RNF4和HA-RNF4-FLAG质粒用于测试蛋白表达和细胞定位。

根据生产商的说明,使用Lipofectamine 2000或RNAiMAX(Invitrogen)的siRNA。对于降低实验,连续两天进行两轮siRNA转染。当转染DNA和siRNA两者时,处理后4-6小时用组合的RNF4siRNA转染DNA,并且在处理后1天用其它siRNA转染。在上次转染后24小时,以7.5-10μM(最终浓度)加入MG132(Sigma)4-5小时。

RNF4shRNA稳定细胞系的产生

克隆到pLKO.1的抗人RNF4的shRNA得自Thermo Scientific。shRNF4的反义序列为TGGCGTTTCTGGGAGTATGGG(SEQ ID NO:160)(TRCN0000017054)。对于慢病毒产生,用慢病毒载体、Gag辅助质粒、Rev辅助质粒和VSVG辅助质粒转染293T细胞。在48小时和72小时收集含有病毒的培养基并以100g离心5分钟。使用含有病毒的上清液通过聚凝胺转导HeLa细胞并通过嘌罗霉素进行选择。pLKO.1载体用于产生对照稳定细胞。

细胞裂解液分级、过滤器阻滞测定和免疫印迹

在含有NP-40的缓冲液中制备细胞裂解液并通过离心分级为上清液(NS)和颗粒。将两部分在含有2%SDS的缓冲液中煮沸并通过免疫印迹分析。对于SR物质,通过过滤器阻滞测定分析一部分颗粒。

如描述的(有修改)制备样品(Janer et al.,2006,J Cell Biol,174:65-76)。收获细胞并在冰上在含有50mM Tris,pH 8.8、100mM NaCl、5mM MgCl2、0.5%NP-40、2mM DTT、250IU/ml核酸酶(Sigma)、1mM PMSF、1×完全蛋白酶混合物(Roche)和20mM N-乙基马来酰亚胺(NEM;Sigma)的缓冲液中裂解30分钟。通过Bradford测定(Bio-Rad Labs)确定蛋白浓度。将全细胞裂解液以13,000rpm在4℃下离心15分钟。通过SDS-PAGE分析含有NP-40可溶(NS)蛋白的上清液。将颗粒在颗粒缓冲液(20mM Tris,pH 8.0、15mM MgCl2、2mM DTT、250IU/ml核酸酶、1mM PMSF、1×完全蛋白酶混合物和20mM NEM)中再悬浮并在冰上温育30分钟。将颗粒部分在2%SDS,50mM DTT中煮沸。通过SDS-PAGE溶解一部分煮沸的颗粒部分,并通过免疫印迹检测进入凝胶的蛋白(SDS-可溶的,SS)。如前所述,将另一部分施加于0.2μm孔径的膜过滤器(Wanker et al.,1999,Methods Enzymol,309:375-386),并通过免疫印迹法分析保留在过滤器上的SDS-耐受性(SR)聚集体。

将抗以下蛋白的第一抗体用于免疫印迹,其中指明了产品信息和稀释液:PML(兔,H-238,1:1,000和山羊,N-19,1:500)、泛素(小鼠,P4D1,1:10,000)和HA(兔,Y-11,1:500)Santa Cruz Biotechnology);FLAG(小鼠,M2,1:7,500),肌动蛋白(兔,1:10,000)和β-微管蛋白(小鼠,1:5,000)(Sigma);GFP(小鼠,1:4,000)(Clonetech);GST(山羊,1:1000,GEHealthcare Life Sciences);SUMO1(小鼠,1:500,Invitrogen);SUMO2/3(兔,1:250,Abgent);HA(用于转染的HA-RNF4)(大鼠,3F10,辣根过氧化物酶或者HRP结合的,1:10,000)(Roche);RNF4(小鼠,1:500,Abnova和使用抗原肽DLTHNDSVVI(SEQ ID NO:161)Abmart开发的小鼠单克隆抗体,1:1,000)。通过抗FLAG抗体检测转染的FLAG-PML,通过HA抗体检测转染的HA-PML和HA-RNF4。

将第二抗体结合至HRP(Santa Cruz Biotechnology),或用IRD Fluor 800或IRD Fluor 680(LI-COR,Inc.)标记。使用ECL试剂进行免疫印迹并使用ImageJ分析,或者使用Odyssey红外成像系统扫描并使用Image Studio Lite(LI-COR,Inc.)分析。

培养的细胞的免疫荧光

将在盖玻片上培养的细胞用4%多聚甲醛固定15分钟,用0.2%Triton X-100透性化15分钟,用1%BSA封闭并如所说明的用抗体温育。用含有DAPI(用于DNA检测)的培养基(Vector Labs)装载细胞,并用Nikon Eclipse E800或者Olympus IX81显微镜采集图像。以指明的产品信息和浓度使用以下第一抗体:PML(兔,H-238和小鼠,PG-M3,1:100)、RNF4(山羊,C-15,1:25)(Santa Cruz Biotechnology)和FLAG(用于转染的FLAG-PML和HA-RNF4-FLAG)(小鼠,M2,1:2,000)(Sigma)。第二抗体为FITC结合的抗小鼠、抗兔(Zymed)和抗山羊(Invitrogen)IgG;德克萨斯红结合的抗小鼠和抗兔IgG(Vector labs);和罗丹明红-X结合的抗山羊(Jackson ImmunoResearch Labs)。

对于Atxn1 82Q、Httex1p 97QP和SUMO2阳性Atxn1 82Q聚集体的定量,检验分别来自10个或更多随机选择区域的约400、500和200个细胞。使用ImageJ测量Atxn1 82Q包涵体的尺寸,并基于细胞中最大的包涵体对细胞归类。使用卡方检验计算存在和不存在PML时具有不同尺寸的聚集体的细胞比例的P值。对于用Httex1p 97QP转染的细胞,它们中约30%具有胞质或核聚集体。

蛋白质半衰期的测定

在补充有[35S]Met和[35S]Cys的不含Met-和Cys-的DMEM培养基中脉冲标记细胞,然后在常规DMEM中培养。可替换地,用CHX处理细胞。通过放射自显影或免疫印迹分析细胞裂解液中免疫沉淀的[35S]Atxn1 82Q或未标记的Atxn1 82Q。

对于脉冲追踪分析,单独或者与适量的PML一起用FLAG-Atxn1 82Q转染HeLa细胞。转染后17h,在不含Met和Cys的DMEM培养基中培养细胞30分钟,然后用[35S]Met和[35S]Cys(各100μCi/ml)脉冲标记30min。之后,用PBS漂洗细胞两次并在具有10%FBS的DMEM中追踪0-18h。在含有2%SDS和50mM DTT的IP-裂解缓冲液(50mM HEPES,pH 7.5、150mM NaCl、0.5%NP-40和2mM DTT)中裂解细胞,并在95℃下煮沸10分钟。将全细胞裂解液以13,000rpm离心15分钟。将上清液在IP-裂解缓冲液中稀释20倍,并与抗FLAG M2珠在4℃下温育过夜。用具有另外的0、0.5M和1M KCl的IP裂解缓冲液依次清洗珠,并在2%SDS样品缓冲液中煮沸。通过SDS-PAGE分离样品并通过放射自显影分析。为了在不同条件下更好地比较Atxn1 82Q的半衰期,显示在0小时以类似的信号强度下的暴露。

对于Atxn1 82Q转染的细胞的环己酰亚胺(CHX)处理,在转染后4-5h将150μg/mL CHX加入至细胞培养基。收获细胞,并在指定的时间点在干冰上快速冷冻、裂解和分级用于免疫印迹分析。对于nFlucDM转染的细胞的CHX处理,在转染后17小时将50μg/mL CHX加入至细胞培养基。在指定时间点收获细胞,并将全细胞裂解液用于免疫印迹分析。

定量RT-PCR分析

使用TRIzol(Invitrogen)提取总RNA。使用第一链cDNA合成试剂盒(Marligen Biosciences)通过总RNA的反转录进行cDNA合成。将使用人Atxn1(Hs00165656_m1)和18s rRNA(4333760F)引物/探针组的Taqman基因表达测定(Applied Biosystems)用于qPCR分析。

蛋白质纯化

如前描述的(Tang et al.,2006,Nat Cell Biol,8:855-862;Tang et al.,2004,J Biol Chem,279:20369-20377)(有修改),在293T细胞中表达FLAG-PML、FLAG-PML M6和FLAG-Atxn1 82Q-HA并通过抗FLAG M2珠(Sigma)纯化。在补充有1mM PMSF和1×完全蛋白酶混合物的IP-裂解缓冲液(50mM Tris,pH 7.5、150mM NaCl、0.5%Triton X-100、0.5%NP-40和2mM DTT)中裂解细胞。对于PML纯化,IP-裂解缓冲液还补充有20μM ZnCl2。将裂解液以13,000rpm离心15分钟。将上清液与抗-FLAG M2珠在4℃下温育4小时至过夜。用含有0、0.5和1M KCl的IP裂解缓冲液,并用洗脱缓冲液(50mM Tris,pH 7.5、150mM NaCl和2mM DTT)依次清洗M2珠。在含有0.1-0.3mg/ml 3×FLAG肽(Sigma)的洗脱缓冲液中洗脱结合的蛋白质。基于免疫印迹和质谱分析两者,在FLAG-PML和FLAG-PML M6制备物中观察到的主要的其它条带源自PML(图11G)。

在293T细胞中表达PML的GST融合物,并通过与以上描述的类似的溶胞和清洗条件使用谷胱甘肽-SepharoseTM 4B珠(GE Healthcare Life Sciences)纯化。在大肠杆菌(Escherichia coli)BL21DE3或Rosetta 2(EMD Chemicals)中表达rRNF4、rRNF4CS1、RNF4和RNF4SIMm的GST融合并如前描述的纯化(Hakli et al.,2001,J Biol Chem,276:23653-23660)。将细菌在37℃下生长至A 600nm=0.6-0.8,并在30℃下用0.3mM IPTG诱导蛋白表达3小时。用谷胱甘肽珠纯化GST-标签的蛋白。除了将0.1mM IPTG用于诱导蛋白表达外,类似地纯化了Htt 25Q、Htt 103Q的GST和GST融合。在含有30mM谷胱甘肽(Sigma)的洗脱缓冲液中洗脱结合的蛋白质。

如前所述,在BL21 DE3中表达荧光素酶-6xHis(Sharma et al.,2010,Nat Chem Biol,6:914-920)。为了产生固定化的天然荧光素酶,根据生产商的说明,将Luc(N)、Ni-NTA珠(Qiagen)与细菌裂解液温育并清洗。通过用8M脲处理固定化的Luc(N)5分钟产生了变性的荧光素酶,Luc(D)。荧光素酶活力测定显示在脲处理后仅保留0.2%的酶活力。对于对照珠,同时将来自不表达荧光素酶的细菌的裂解液与Ni-NTA珠温育。

为了产生用于荧光素酶肽扫描的PML突变体,在室温下用0.1mM IPTG分别诱导3小时和1小时,在BL21DE细胞中表达FLAG-PML F12(571-633)-6xHis和GST-PML CC-FLAG。首先用M2珠和FLAG肽洗脱纯化Flag-PML F12(571-633)-6xHis,然后根据生产商的说明,使用Ni-NTA珠进行第二纯化。为了产生PML CC域,在GST和PML CC之间引入TEV蛋白酶切割位点。根据生产商的说明,将GST-PML CC-FLAG结合的谷胱甘肽珠与TEV蛋白酶(Sigma)温育,从而从GST部分(和珠)释放PML CC-FLAG。如上所述,通过将纯化的PML CC-FLAG蛋白与M2珠温育并清洗,产生了用于拉下试验(Pull-down assay)的PML CC-FLAG

拉下试验

对于FLAG拉下试验,如上文描述的制备结合至抗FLAG M2珠的FLAG-PML、FLAG-GFP和PML CC-FLAG。将纯化的GST-Htt 25Q、GST-Htt 103Q、GST-Htt 52Q或GST-Htt 52Q cc以13,000rpm离心15分钟以除去任何聚集的蛋白。将摩尔浓度相当的FLAG-PML(2.5μg)或FLAG-GFP(1.1μg)在4℃下在最终体积200μl的测定缓冲液(50mM Tris,pH 7.5、150mM NaCl和2mM DTT、0.5%NP-40)中在不存在或存在Hsp70(2.5μg)和Hsp40(1.4μg)(EnzoLife Sciences)的情况下与GST-Htt 25Q或GST-Htt 103Q(分别为2.5μg)温育2小时。在小型反应柱(Affymetrix/USB)中用IP-裂解缓冲液清洗珠三次并在2%SDS样品缓冲液中煮沸。通过免疫印迹分析样品。除将含有1.6μg PML CC-FLAG的M2珠或对照M2珠与5μg GST或GST-Htt蛋白温育外,类似地进行PML CC拉下试验。

对于GST拉下试验,将各自为2μg的结合至谷胱甘肽-SepharoseTM 4B珠的GST、GST-Htt 25Q和GST-Htt 103Q蛋白与来自表达FLAG-PML的293T细胞的400μg裂解液在4℃下温育4小时。在小型反应柱中用IP-裂解缓冲液清洗珠三次并在2%SDS样品缓冲液中煮沸。通过免疫印迹分析样品。对于PML突变体和Htt之间相互作用的检测,使用SP6连接的转录/翻译系统(Promega)产生[35S]Met-标记的全长和突变体PML蛋白并与结合至珠的GST、GST-25Q和GST-Htt 103Q在4℃下在150μl IP-裂解缓冲液中温育过夜。如上所述清洗并煮沸珠。通过放射自显影和考马斯亮蓝染色分析输入样品和下拉样品。对于放射自显影,将在不同凝胶上分离的来自相同实验的下拉样品进行相同的曝露时间。

对于荧光素酶拉下试验,将从等量细菌裂解液制备的3μg Luc(N)或Luc(D)结合的Ni-NTA珠或对照珠在不存在或存在Hsp70(3μg)和Hsp40(1.7μg)的情况下与纯化的GST(1μg)、GST-PML(3μg)或者与[35S]Met标记的全长和突变PML蛋白在200μl的含有15mM咪唑、1mM DTT、0.5%Triton X-100和0.5%NP-40的PBS中在4℃下温育4小时。用含有20mM咪唑、1mM DTT、0.5%Triton X-100和0.5%NP-40的PBS清洗珠三次并煮沸。如上所述分析输入样品和下拉样品。

筛选结合至PML域的纤维素结合的肽

通过自动化的点合成(spot synthesis)(JPT peptide Technologies)制备了北美萤火虫荧光素酶的肽文库(重叠10个氨基酸的13聚体)。用纯化的PML SRS1和SRS2片段探针检测肽阵列膜。用Odyssey封闭缓冲液(LI-COR,Inc)封闭肽阵列膜,并将其与FLAG-PML F12(571-633)-Hisx6或者PML F4/CC-FLAG(分别为150nM)在TBS-T(50mM TRIS,pH 8.0、137mM NaCl、2.7mM KCl、0.05%Tween和1mM DTT)中在4℃下温育2小时。清洗膜并按照生产商的说明用小鼠抗Flag和抗小鼠-HRP抗体印迹。使用ECL试剂使印迹显像。即使长期暴露,单独用抗Flag和抗小鼠-HRP抗体印迹的背景信号最小。根据生产商的规程,再生肽阵列膜。

SUMO化和泛素化分析

将细胞或体外反应混合物在含有2%SDS的缓冲液中煮沸,然后在无SDS的缓冲液中稀释并通过Bio-Spin色谱柱以降低SDS浓度。将蛋白质免疫沉淀(变性IP或d-IP)并通过免疫印迹分析。

体内SUMO化和泛素化测定

如指示的,用FLAG-Atxn1、FLAG-nFluc-GFP及其它表达质粒转染细胞。对于图4A中所示的实验,还使用了SUMO2-表达质粒。转染后24小时,用7.5μM MG132或DMSO处理细胞5小时或者保持不处理,并在补充有2%SDS和50mM DTT的IP-裂解缓冲液中收获。对于变性免疫沉淀(d-IP),将细胞裂解液在95℃下煮沸10分钟。保存1等份用于免疫印迹分析。将剩余的裂解液在IP-裂解缓冲液中稀释20倍或通过用IP-裂解缓冲液平衡的Bio-Spin色谱柱(Bio-Rad)以降低SDS浓度。然后,将裂解液与抗FLAG(M2)珠在4℃下温育4小时或过夜。如描述的清洗珠用于FLAG-标签的蛋白纯化,并在2%SDS样品缓冲液中煮沸。如所指示的,通过与抗FLAG、抗SUMO2/3、抗SUMO1、抗泛素及其它抗体的免疫印迹分析来自珠的蛋白。为了更好地比较泛素化或SUMO化物质的水平,通常将含有类似水平的未修饰蛋白的d-IP产物用于免疫印迹分析。

体外SUMO化测定

用于体外泛素化和SUMO化反应的组分购自Boston Biochem。在30μl含有纯化的HA-Atxn1 82Q-FLAG(600ng/200nM)、FLAG-PML(对于图4D,50和200ng或22和90nM;对于图4E,100ng或45nM)或FLAG-PML M6(100ng或45nM)、SAE1/SAE2(125nM)、Ubc9(1μM)、His-SUMO2(25μM)、Hsp70(420ng/200nM)、Hsp40(240ng/200nM)和BSA(0.1μg/ml)的反应缓冲液(50mM Tris pH 7.5、5.0mM Mg2+-ATP和2.5mMDTT)中,在37℃下进行体外SUMO化测定。通过加入30μl含有2%SDS和50mM DTT的IP-裂解缓冲液并在95℃下加热10分钟使反应混合物变性。保存1等份加热的反应混合物用于免疫印迹分析,并将其余部分在无SDS的IP-裂解缓冲液中稀释20倍。通过抗HA珠(Roche)使HA-Atxn1-FLAG免疫沉淀并使用抗SUMO2/3抗体分析SUMO2/3修饰。

体外泛素化测定

在10μl含有纯化的GST-RNF4蛋白(250ng/530nM)、UBE1(125nM)、UbcH5a(625nM)、泛素(2.5μg/30μΜ)和Mg2+-ATP(2.5mM)的反应缓冲液(50mM Tris pH 7.5和2.5mM DTT)中,在37℃下进行RNF4自泛素化的体外测定。将反应混合物在95℃下加热10分钟并通过免疫印迹分析。

对于SUMO化的Atxn1 82Q的体外泛素化,制备SUMO化且未修饰的Atxn1 82Q蛋白混合物作为泛素化反应底物。将与FLAG-Atxn1 82Q-HA(1.5μg/300nM)结合的M2珠和对照M2珠与总体积50μl的含有5mM ATP的Mg2+-ATP-能量再生溶液(Boston Biochem)中的0.75μM SAE1/SAE2、12.5μM Ubc9、125μM His-SUMO2和2.5mM DTT混合。为了实现足够的Atxn1 82Q SUMO化,在37℃下反应24小时,并在12小时后更换反应缓冲液。然后,用具有另外的0、0.5和1M KCl的IP-裂解缓冲液和泛素化反应缓冲液(50mM Tris pH 7.5和150mM NaCl)依次清洗珠(Tang et al.,2006,Nat Cell Biol,8:855-862)。

然后,将Atxn1 82Q珠和对照珠在37℃下与处于反应缓冲液(50mMTris-HCl pH 7.5,150mM NaCl和2.5mM DTT)中的20μl体积的含有GST-rRNF4(0、40、160和500ng或0、43、170和530nM)、UBE1(100nM)、UbcH5a(500nM)、泛素(5μg/30μΜ)和Mg2+-ATP(2.5mM)的泛素化反应混合物温育1小时。然后,将珠与上清液分离并用IP-裂解缓冲液清洗。通过加入含有2%SDS和50mM DTT的IP-裂解缓冲液并在95℃下加热10分钟使Atxn1 82Q变性并释放。稀释后,用M2珠使Atxn182Q免疫沉淀。通过免疫印迹分析来自反应的IP产物和上清液。

小鼠育种和基因型分型

提供了内含通过浦肯野细胞特异性启动子元件驱动的具有82个CAG重复的人SCA1编码区的杂合B05转基因小鼠(Atxn1tg/-)(Burright et al.,1995,Cell,82:937-948)。提供了PML-/-小鼠(Wang et al.,1998,Science,279:1547-1551)。使Atxn1tg/-小鼠(在FVB背景上)与PML-/-(在129Sv背景上)交配。使来自F1代的PML+/-:Atxn1tg/-小鼠与PML-/-或PML+/+交配以产生用于转棒测试和病理的小鼠。交配方案不影响PML+或PML+/-:Atxn1tg/-小鼠F2代的转棒表现、聚集体形成、分子层厚度或树突分枝。如描述的(Burright et al.,1995,Cell,82:937-948)(对于Atxn1)或者根据NCI Mouse Repository的建议(对于PML)通过PCR确定小鼠基因型。

加速转棒测试

使用加速转棒装置(47600,Ugo Basile,Italy)来测量运动协调性和平衡。仅使用未进行过实验的动物。每只动物每天测试三次,连续进行4天,每次试验之间休息1小时。对于每次试验,将小鼠置于4-80rpm的加速转棒上10分钟。记录它们从转棒上摔下的等待时间(以秒为单位)。

小鼠小脑的免疫染色和病理学分析

用指明的抗体对石蜡包埋的小脑中矢状面切片染色并使用Leica SP5II激光扫描共聚焦显微镜显象,或者使用苏木精染色并使用Olympus BX51显微镜显象。

如前所述(有修改)进行免疫组织化学和免疫荧光(Duda et al.,2000,J Neuropathol Exp Neurol,59:830-841;Emmer et al.,2011,J Biol Chem 286:35104-35118)。将石蜡包埋的小脑切成10-μm切片。对于分子层测量,每只小鼠分析间隔100μm的三个苏木精染色的中矢状面切片。将每个切片在原裂处的20个测量取平均值。

为了定量浦肯野细胞树突分枝,用抗浦肯野细胞-特异性蛋白质钙结合蛋白的抗体对小脑的中矢状面切片染色(小鼠,CB-955,1:250;Sigma)。用Leica SP5II激光扫描共聚焦显微镜将20个0.5μm的光学切片累加。将最亮的连续12个切片(6μm)投影用于最大强度。使用ImageJ绘制来自前顶裂的相同区域的荧光强度谱。

为了定量浦肯野细胞,用抗钙结合蛋白抗体对中矢状面切片染色并将相当的区域用于细胞计数。通过使用ImageJ沿浦肯野细胞体细胞中心绘制分割线来测量浦肯野细胞层的长度。对于每只小鼠,测量沿约30mm的350-900个神经元。通过将细胞数目除以浦肯野细胞层的长度确定浦肯野细胞密度。

为了确定具有聚集体的浦肯野细胞的数目,用抗泛素(小鼠,Ubi-1,MAB1510,1:2,000;Millipore)抗体对中矢状面切片染色。对每只小鼠来自相同脑区域的300个或以上的细胞计数。使用安装有DP71Olympus数码相机的Olympus BX51显微镜采集图像。

将每个基因型的四只小鼠用于1岁小鼠的浦肯野细胞计数,将每个基因型的两只PML+/+小鼠用于树突分枝的定量,并且将每个基因型的三只小鼠用于其它研究。

统计分析

当适用时,通过卡方检验和学生t检验分析具有聚集体的细胞数目。通过具有重复测量的双因素ANOVA和学生t检验分析行为得分和小脑病理。使用Prism5软件或Microsoft Excel 2008分析所有数据。

现将描述实验结果。

PML促进致病Ataxin-1蛋白的蛋白酶体降解

SCA1是以进行性共济失调和神经元丧失,特别是小脑浦肯野细胞丧失为特征的致死性神经障碍。其是由SCA1基因产物Ataxin-1(Atxn1)中polyQ延伸的扩增所造成的(Orr and Zoghbi,2007,Annu Rev Neurosci,30:575-621)。为了研究PML在消除细胞核错误折叠的蛋白质中的作用,产生细胞培养模型,其中在HeLa细胞中表达C端融合至增强的绿色荧光蛋白的具有82个邻接谷氨酰胺的致病性Atxn1蛋白Atxn1 82Q-GFP。与人SCA1患者和小鼠SCA1转基因模型中的致病性Atxn1蛋白类似(Skinner et al.,1997,Nature,389:971-974),Atxn1 82Q-GFP定位至细胞核,并且在显微镜可见的包涵体中显示出具有显著更高浓度的扩散定位模式(图1A和图1B)。Atxn1 82Q-GFP还在细胞裂解液中获得了NP-40可溶(可溶性的或NS)和NP-40不溶的(聚集体)物质两者。后者可以进一步分为SDS可溶(SS)和SDS耐受(SR)物质(图1C)。

与先前报道一致(Skinner et al.,1997,Nature,389:971-974),内源PML与Atxn1 82Q-GFP包涵体共定位,在邻近于它们的体内积累并且还在其中分布(图1A)。PML以几种同种型表达(Nisole et al.,2013,Front Oncol,3:125)。检验了5种主要的PML同种型(I、II、III、IV和VI)并且发现所有5种均与Atxn1-GFP包涵体共定位(图8A)。对于后续分析,选择常用的同种型IV(此后除非另作说明,否则将其称为PML)。

当与Atxn1 82Q共表达时,PML显著降低了Atxn1 82Q-GFP核包涵体的尺寸(图1B)。其还降低了Atxn1 82Q-GFP蛋白的稳态水平,特别是聚集的SS和SR物质(图1C,左图)。为了评价内源PML(所有同种型)的影响,使用两个独立的小干扰RNA(siRNA)将其降低。这显著增加Atxn1 82Q-GFP的水平,特别是聚集的物质(图1C,右图)。沉默的PML还增加FLAG标签的Atxn1 82Q蛋白的稳态水平(图8B)。可以通过PML的siRNA抗型逆转PML siRNA对Atxn1 82Q的影响(图8C),从而排除siRNA的脱靶效应。

为了评价PML是否特异性降低致病性Atxn1蛋白,使用了非致病的Ataxin-1蛋白Atxn1 30Q。PML的强制表达不会降低Atxn1 30Q-GFP的丰度,而PML的降低也不会使其显著增加(图1D),强调了PML对致病性Atxn1蛋白的选择性影响。

PML不会抑制Atxn1 82Q基因的转录(图8D)。为了确定PML是否促进Atxn1 82Q蛋白的降解,进行了脉冲追踪测定。在没有共转染的PML的情况下,总[35S]标记的Atxn1 82Q蛋白相当稳定,并且在18小时中,其水平仅降低~20%。通过对比,在存在PML的情况下,总[35S]Atxn1 82Q蛋白不稳定,并且在相同时间段内,其水平降低~80%(图1E)。

使用环己酰亚胺(CHX)进行实验以阻断蛋白质合成并研究预先存在的Atxn1 82Q蛋白的降解。PML的强制表达加速了聚集的Atxn1 82Q的降解,将其半衰期从8小时降至2小时,同时对可溶性Atxn1 82Q具有极小的影响(图1F)。相反,沉默的PML延长了聚集的Atxn1 82Q的半衰期,并且对可溶性Atxn1 82Q的半衰期影响程度较小(图1G)。蛋白酶体抑制剂MG132显著降低了PML除去聚集的Atxn1 82Q的能力(图1H)。相反,PML不会改变Atxn1 30Q的半衰期(图8E)。总的来说,这些结果表明PML靶向致病性Atxn1蛋白,但不靶向正常Atxn1蛋白来进行蛋白酶体降解。

PML在降解核错误折叠的蛋白质中的一般作用

为了评价PML是否在降解核中错误折叠的蛋白质中起广泛作用,测试了与神经变性有关的两种其它蛋白:(1)HD基因的第一外显子编码的亨廷顿蛋白(Htt)的致病片段Httex1p 97QP(Steffan et al.,2004,Science,304:100-104);和(2)TAR DNA结合蛋白43(TDP-43),其与肌萎缩性侧索硬化(ALS,也称为鲁盖瑞氏症)和具有泛素化包涵体的额颞叶脑退行性病变(FTLD-U)两者有关(Chen-Plotkin et al.,2010,Nat Rev Neurosci,6:211-220)。Httex1p 97QP在细胞核和细胞质两者中形成显微镜可见的包涵体(图1I),而TDP-43主要在细胞核中形成包涵体。PML降低细胞核,但不降低细胞质Httex1p 97QP包涵体(图1I),并且减少聚集的Httex1p97QP的量(图1J,泳道1和2)。PML还降低聚集的,但不降低可溶的TDP-43的量(图1K)。

为了扩展这些分析,使用了模型分子伴侣底物萤火虫荧光素酶(FlucDM)的结构不稳定的突变体,其作为细胞PQC系统能力探针开发(Gupta et al.,2011,Nat Methods,8:879-884)。内源PML与FlucDM的核形式(nFlucDM-GFP)部分共定位,其形成包涵体,但不与野生型对应物(nFlucWT-GFP)共定位,其显示出扩散的定位(图8F)。沉默的PML显著增加聚集的nFlucDM-GFP的水平(图8G)并且延长了总nFlucDM-GFP蛋白的半衰期(图1L)。总的来说,这些结果表明PML有利于除去哺乳动物细胞核中的多个错误折叠的蛋白质。

通过PML上的不同位点识别错误折叠的蛋白质

为了研究PML降解错误折叠的蛋白质的机制,首先检验了PML是否能够直接识别这些蛋白质。对于这些实验,使用了致病(103Q)和非致病(25Q)的Htt片段,各自均融合至谷胱甘肽S-转移酶(GST)。在使用纯化的重组蛋白的体外测定中,固定化FLAG-PML,而非对照蛋白FLAG-GFP拉下GST-Htt 103Q(图2A),表明PML和Htt 103Q之间特异性和直接的相互作用。FLAG-PML也拉下GST-Htt 25Q。然而,这种相互作用显著弱于PML-Htt 103Q相互作用(图2A)。在交换实验中,固定化GST-Htt 103Q蛋白与FLAG-PML的相互作用比固定化GST-Htt 25Q要更强烈(图9A)。识别广泛的错误折叠的蛋白质的Hsp70和Hsp40不增加PML-Htt 103Q的相互作用(图2A)。这些结果表明PML可以直接与polyQ蛋白结合并优先与致病形式结合。

还检验了PML是否选择性结合至变性的荧光素酶。使固定在Ni-NTA珠上的6xHis标签的荧光素酶用脲变性或者保持天然形式。变性的而非天然的荧光素酶与GST-PML特异性相互作用,并且Hsp70/Hsp40系统不会增强这种相互作用(图2B)。因此,PML可以直接识别错误折叠的荧光素酶,但不能识别天然荧光素酶。

为了理解PML与错误折叠的蛋白质之间相互作用的分子基础,设法确定PML的底物识别位点(SRS)以及这些SRS识别的底物上的结构特征。先前已显示以依赖于其长度的方式,polyQ和侧接区域形成了CC结构,其有利于polyQ蛋白组装为寡聚或聚集状态,并且还介导了polyQ蛋白与含有CC的蛋白质的相互作用(Fiumara et al.,2010,Cell,143:1121-1135)。因此,假设PML通过其在TRIM/RBCC基序内的CC区域与致病性polyQ蛋白相互作用。构建了一组PML片段(F1-F5),其中每个片段含有或缺少CC区域(图9B)。含有CC区域的片段(F1)与Htt103Q相互作用,而缺少该区域的两个片段(F2和F3)不会(图9B和图9C)。此外,CC区域单独(F4)结合至Htt 103Q,而从整个PML蛋白删除该区域(F5或ΔCC)大大降低了这种结合。因此,PML几乎只通过CC区域识别Htt 103Q。与全长PML类似,PML CC对致病性Htt 103Q显示出明显优先于不致病Htt 25Q的结合(图2C)。PML CC还与另一种致病性Htt构建体Htt 52Q强相互作用(图2C)。因此,PML CC可能构成SRS(称为SRS1)。

为了测试PML CC是否识别Htt蛋白中的同源CC结构,将预测参与CC形成的Htt 52Q中的残基突变,得到Htt 52Q cc-(图9D)。先前显示类似的突变降低了Htt 72Q中CC结构的形成(Fiumara et al.,2010,Cell,143:1121-1135)。事实上,与Htt 52Q相比,Htt 52Q cc-显示出显著降低的形成聚集体的倾向和与PML CC显著更弱的相互作用(图2C)。因此,PML CC/SRS1可能与致病性Htt蛋白上的CC结构相互作用。

如果PML也促进非polyQ蛋白,如荧光素酶和TDP-43的降解(图1和图8),这是因为PML可能含有可以识别错误折叠的蛋白质上的非CC结构特征的至少另一种SRS。为了测试这种可能性,检验了一组PML片段与变性荧光素酶的相互作用。尽管单独的CC区域可以与变性荧光素酶相互作用,但是在缺少该区域但含有C端(aa 361-633)的两个片段(F2和F5)中也观察到了显著的相互作用水平(图9B和图10A)。使用其它C端内缺失构建体(F6-F18,图9B和图10B),发现63个氨基酸的延伸(aa571-633)对结合至变性荧光素酶是足够的。该延伸的NH2-或COOH-末端缺失消除了结合(图10)。因此,PML的最后63个氨基酸可能构成另一种SRS(称为SRS2)。

为了研究PML SRS2可以识别的荧光素酶中的线性序列,将纯化的PML SRS2用于筛选代表荧光素酶完整序列的纤维素结合的肽文库。该文库由180个肽组成,每个肽含有与相邻的肽重叠10个氨基酸的13个氨基酸残基。与分子伴侣,如Hsp70和ClpB(Rudiger et al.,1997,EMBO J,16:1501-1507;Schlieker et al.,2004,Nat Struct Mol Biol,11:607-615)类似,PML SRS2仅结合至这些肽的亚型(图2D),表明其识别具有不同氨基酸组成的肽的能力。相对于该文库中全部的肽,PML SRS2-相互作用肽中所有20个氨基酸的相对出现分析显示PML SRS2强烈倾向于芳香族(Phe、Trp和Tyr)和带正电荷的(Arg和Lys)残基,并且不倾向于带负电荷的残基(Asp和Glu)(图2E)。除SRS2对Leu和His具有额外的优先性(ClpB不倾向于它们)外,这种氨基酸优先性类似于ClpB(Schlieker et al.,2004,Nat Struct Mol Biol,11:607-615)。

为了比较,测试了PML CC/SRS1与该肽文库的结合。与该区域识别更高级结构而不是线性序列的倾向一致,PML CC/SRS1微弱结合至少量肽(图9E)。基于这些结果,推断PML含有可以识别错误折叠的蛋白质的至少两个区域:TRIM/RBCC基序内的CC区域(SRS1)和其C端的63个氨基酸延伸(SRS2),其可以识别CC结构以及分别在芳香族和碱性氨基酸两者中富含的暴露的肽。

Atxn1 82Q降解中SUMO化的参与

进行实验以研究在识别时,PML如何促进错误折叠的蛋白质的降解。通常通过SUMO修饰与神经变性有关的错误折叠的蛋白质,尽管这种修饰的作用仍不清楚(Martin et al.,2007,Nat Rev Neurosci,8:948-959)。哺乳动物细胞表达三种主要的SUMO蛋白,SUMO1-SUMO3。SUMO2和SUMO3在它们的序列中几乎彼此相同(统称为SUMO2/3)并且与SUMO1约50%相同(Wilkinson and Henley,2010,Biochem J,428:133-145)。研究了这些SUMO蛋白对Atxn1 82Q的修饰以及它们在Atxn1 82Q降解中的作用。

通过外源(Riley et al.,2005,J Biol Chem,280:21942-21948)和内源(图3A,左图)SUMO1两者修饰Atxn1 82Q,并且这种修饰弱于对Atxn1 30Q的修饰(Riley et al.,2005,J Biol Chem,280:21942-21948)。还通过内源SUMO2/3修饰Atxn1 82Q(图3A,右图),并且在细胞核中与GFP-SUMO2/3共定位(图3B)。Atxn1 82Q中与SUMO1和SUMO2/3结合的位点可以是不同的,因为具有受损的SUMO1结合的突变Atxn1 82Q,Atxn1 82Q(5KR)(Riley et al.,2005,J Biol Chem,280:21942-21948),显示在SUMO2/3结合中无缺陷(图11A)。

值得注意的,与Atxn1 30Q相比,内源SUMO2/3对Atxn1 82Q的修饰更强(图3C),将这些Atxn1蛋白的不同响应与PML介导的降解相关联(图1和图8)。类似地,通过内源SUMO2/3修饰TDP-43(图11B)。SUMO 2/3还修饰FlucDM以及另一种结构不稳定的荧光素酶突变体FlucSM(Gupta et al.,2011,Nat Methods,8:879-884)。这些荧光素酶突变体的SUMO2/3修饰比野生型荧光素酶的更强(图11C)。

此外,蛋白酶体抑制增强Atxn1 82Q与GFP-SUMO2/3的共定位(图3B)。其还增加SUMO2/3修饰的Atxn1 82Q以及泛素化Atxn1 82Q,但不会增加SUMO1修饰的Atxn1 82Q(图3A和图3D,泳道2和3)。同样地,蛋白酶体抑制增加SUMO2/3修饰的TDP-43和荧光素酶突变体(图11B和图11C)。

为了评价SUMO蛋白在Atxn1 82Q的泛素化和蛋白酶体降解中的作用,使用siRNA分别使SUMO2/3和SUMO1沉默。沉默的SUMO2/3而非SUMO1有效降低了Atxn1 82Q的泛素化(图3D,泳道4-7)。沉默的SUMO2/3还增加Atxn1 82Q的水平,特别是聚集的形式,但是不能增加对照蛋白GFP的水平(图11D-图11F),并且其减弱了PML除去聚集的Atxn1 82Q的能力(图3E)。

SUMO2和SUMO3含有使得能够形成聚合链的内部SUMO化共有位点。SUMO1不含该位点,并且当结合至SUMO2/3链时,其可以终止链延伸(Wilkinson and Henley,2010,Biochem J,428:133-145)。因此,将两种其它策略用于抑制SUMO2/3对Atxn1 82Q的修饰。首先,使SUMO1过表达。这强烈降低了Atxn1 82Q与SUMO2/3的结合,并且同时降低了Atxn182Q与泛素的受损的结合(图3D,泳道8和9)和其通过PML的降解(图3F,左图)。第二,使用在链形成中有缺陷的SUMO2突变体SUMO2KR。SUMO2KR的过表达有效降低了SUMO2/3修饰的Atxn1 82Q物质的量,特别是具有高分子量的那些。其还降低了泛素修饰的Atxn1 82Q物质(图3D,泳道12和13)并且减弱了PML降解Atxn1 82Q的能力(图3F,右图)。此外,使用了SUMO化-缺陷型Htt突变体Httex1p 97QP(KR)(Steffan et al.,2004,Science,304:100-104),并且发现其耐受PML介导的降解(图1J)。总的来说,这些结果显示Atxn1 82Q以及可能的其它错误折叠的蛋白的泛素化和降解取决于SUMO2/3对它们的修饰。

PML作为Atxn1 82Q的SUMO E3连接酶

先前显示PML具有增强SUMO化效率和特异性的SUMO E3连接酶活性(Chu and Yang,2011,Oncogene,30:1108-1116)。因此,检验了PML是否促进Atxn1 82Q的SUMO化。当在细胞中与Atxn1 82Q共表达时,在不存在和存在蛋白酶体抑制剂MG132的情况下,PML强烈增加Atxn182Q的SUMO2/3修饰(图4A)。相反,在这些条件下,沉默的PML显著降低SUMO2/3修饰的Atxn1 82Q(图4B)。类似地,沉默的PML降低nFlucDM的SUMO2/3修饰(图4C)。

为了确认PML对Atxn1 82Q的SUMO E3活性,使用纯化的重组蛋白进行体外SUMO化测定。在不存在PML的情况下,SUMO2微弱修饰Atxn182Q(图4D和图4E),这与可以在没有SUMO E3连接酶的情况下体外进行SUMO化的先前观察结果一致(Wilkinson and Henley,2010,Biochem J,428:133-145)。值得注意的,PML以剂量依赖性的式增加Atxn1 82Q SUMO化(图4D和图4E)。相反,SUMO E3缺陷型突变体PML M6(Chu and Yang,2011,Oncogene,30:1108-1116)则不能(图4E和图11G);PML M6在减少聚集的Atxn1 82Q方面也是无效的(图4F)。这些结果表明PML是Atxn182Q的SUMO E3连接酶,并且该活性参与了Atxn1 82Q降解。

RNF4在降解错误折叠的蛋白质中的作用

RNF4是具有四个串联的SUMO相互作用基序(SIM)的RING域泛素连接酶,其可以识别并泛素化与聚-SUMO2/3链结合的蛋白(Sun et al.,2007,EMBO J,26:4102-4112)。然而,RNF4在降解错误折叠的蛋白质方面的作用仍不明确。发现强制的RNF4表达强烈降低细胞裂解液中聚集的Atxn1 82Q的稳态水平以及细胞核中Atxn1 82Q包涵体的数目(图12A)。RNF4还缩短了聚集但不可溶的Atxn1 82Q的半衰期(图5B,泳道1-12;图5C和图12B)。相反,单独或组合使用三种siRNA降低内源RNF4增加了细胞裂解液中总的和聚集的Atxn1 82Q蛋白以及细胞核中Atxn1 82Q包涵体(图5E)。RNF4的siRNA耐受形式可以逆转RNF4降低的影响(图12C),表明siRNA的特异性。

此外,内源和外源RNF4蛋白两者通常显示出扩散的细胞核分布模式,并且与Atxn1 82Q包涵体具有极小或中等的共定位。然而,在蛋白酶体阻断时,RNF4在Atxn1 82Q包涵体中高度富集(图5F和图12E),可能反映了RNF4在清除Atxn1 82Q中的降低作用。与其对Atxn1 82Q的作用相反,RNF4不会降低Atxn1 30Q的水平(图5G)。总的来说,这些结果表明了RNF4在消除致病性Atxn1蛋白中的作用。

为了评估RNF4对错误折叠的蛋白质的一般作用,对Httex1p 97QP、TDP-43和nFlucDM进行测试。RNF4的强制表达显著降低了Httex1p97QP,特别是聚集的形式,同时对Httex1p 97QP(KR)的具有弱的多的作用(图5H)。类似地,RNF4的强制表达降低了TDP-43的水平(图12F),而沉默的RNF4增加具有核包涵体的TDP-43表达细胞的百分比(图12G和图12H)。在蛋白酶体抑制后,内源RNF4在TDP-43包涵体中变得高度浓缩(图12I),这类似于在相同条件下其在Atxn1 82Q包涵体中的积累(图5F)。此外,沉默的RNF4延长了nFlucDM的半衰期(图5I)。总的来说,这些观察结果表明RNF4在多种错误折叠的蛋白质的降解中起重要作用。

RNF4介导SUMO2/3修饰的Atxn1 82Q的泛素化和降解

与PML类似,RNF4消除错误折叠的蛋白质的能力取决于SUMO2/3,因为这种能力在缺少SUMO2/3的细胞中受损,但在缺少SUMO1的细胞中不受损(图13A)。值得注意的,相对于未修饰的蛋白,RNF4的强制表达优选地降低SUMO2/3修饰的Atxn1 82Q和nFlucDM(图6A和图6B)。相反地,沉默的RNF4增加SUMO2/3修饰的Atxn1 82Q(图6C)并且增加GFP-SUMO2与Atxn1 82Q包涵体的定位(图13B)。这些结果表明RNF4靶向SUMO2/3修饰的错误折叠的蛋白质用于降解。

为了确认RNF4使SUMO2/3结合的错误折叠的蛋白质泛素化,使用未修饰的和SUMO2修饰的Atxn1 82Q蛋白的混合物进行体外泛素化测定(图6D)。在存在增加剂量的RNF4的情况下,具有相对低分子量的SUMO2修饰的Atxn1 82Q蛋白(图6E,泳道1、6和9)逐渐转化为同样被泛素修饰的高分子量物质(泳道2-4、7-9和12-14)。相反,未修饰的Atxn1 82Q蛋白未泛素化(泳道1-4)。因此,RNF4是SUMO2/3修饰的Atxn1 82Q蛋白的泛素连接酶,但对于未修饰的Atxn1 82Q蛋白不是。

RNF4具有泛素连接酶和SUMO结合活性(Sun et al.,2007,EMBO J,26:4102-4112)。为了确定这些活性在降解Atxn1 82Q中的参与,产生泛素连接酶(CS和CS1)或SUMO结合(SIMm)活性缺陷的RNF4突变体(图13C)。虽然丧失了它们的泛素E3活性(图13D),但是CS和CS1仍能与Atxn1 82Q包涵体共定位(图6F)。另一方面,SIMm保留了显著水平的泛素连接酶活性(图13E),但是不能与Atxn1 82Q包涵体共定位(图6G)。RNF4突变体种类均不能除去聚集的Atxn1 82Q(图6H)。总的来说,这些结果表明RNF4通过其SIM区域结合至SUMO2/3修饰的错误折叠的蛋白质并且通过其连接酶活性使这些蛋白泛素化。

尽管PML的强制表达导致对照细胞内聚集的Atxn1 82Q的有效清除,但是这种能力在RNF4缺失的细胞中大大减弱(图6I)。相互地,RNF4的强制表达尽管在加速对照细胞中Atxn1 82Q的降解中非常有效,但是在PML缺失的细胞中不能(图5B,泳道19-24相对于泳道7-12;和图5C)。此外,在显示出高Atxn1 82Q水平的PML缺失的细胞中,沉默的RNF4不会进一步增加Atxn1 82Q的水平(图13F)。这些结果表明PML和RNF4在Atxn1 82Q降解中的相互依赖性。

SCA1的小鼠模型中PML缺陷加剧了行为和病理表型

上述结果显示了通过顺序的PML介导的SUMO化和RNF4介导的泛素化降解Atxn1 82Q和可能的其它核错误折叠的蛋白质的PQC系统。为了研究该系统的生理学作用,使用了SCA1(B05)小鼠模型,其在小脑浦肯野细胞中表达Atxn1 82Q转基因(Atxn1tg/-)。与人SCA1患者类似,B05小鼠随年龄增长发展出共济失调和神经学异常(Burright et al.,1995,Cell,82:937-948)。小鼠中RNF4的丧失导致了胚胎致死(Hu et al.,2010,Proc Natl Acad Sci,107:15087-15092),从而使得无法分析其对B05小鼠的影响。然而,PML敲除的(PML-/-)小鼠是活的并且看起来发育正常(Wang et al.,1998,Science,279:1547-1551)。将B05小鼠与PML-/-和PML-野生型(PML+/+)小鼠杂交,并且对所有基因型(PML+/+、PML+/-和PML-/-、PML+/+:Atxn1tg/-、PML+/-:Atxn1tg/-和PML-/-Atxn1tg/-)的同窝小鼠比较运动表现和神经病理学。

使用逐渐加速的速度的转棒装置评价运动表现,包括平衡、调协和耐力。为了确定任何潜在的行为缺陷是否是由于逐渐降低的能力所造成的,与发育损害相反,检验不同年龄的小鼠。为了排除长期运动记忆的影响,仅使用未进行过实验的动物,每只动物连续测试4天。

在7周龄时,所有小鼠在转棒上的表现类似(图7A)。尽管在不同基因型的小鼠中观察到了一些差异,但是它们不是统计学显著的(ANOVAp=0.53),表明PML-/-小鼠在它们的运动功能中不具有预先存在的损害。在11周龄时,所有缺少Atxn1 82Q转基因的小鼠(PML+/+、PML+/-,和PML-/-)仍未在它们的表现中显示出统计学差异(ANOVA p=0.33)(图7B)并且PML+/+和PML+/+:Atxn1tg/-表现也类似。这些观察结果表明在该年龄单独的PML缺陷或Atxn1 82Q转基因表达不足以导致运动缺陷。感兴趣地,与PML+/+:Atxn1tg/-或PML-/-小鼠相比,PML-/-:Atxn1tg/-在转棒表现中显示出严重的损害。尽管这三组动物在连续4天测试开始时相当,但是不同于另外两组,PML-/-:Atxn1tg/-小鼠随时间显示出最低的改善。PML-/-:Atxn1tg/-小鼠在转棒上缺乏改善使得联想到晚期的Atxn1tg/-小鼠(Clark et al.,1997,J Neurosci,17:7385-7395)。PML杂合的对应部分(PML+/-:Atxn1tg/-小鼠)在转棒上显示出中等的损害(对于三个Atxn1tg/-组,ANOVA p=0.0004)(图7B)。因此,PML缺陷加剧了Atxn1tg/-小鼠的运动缺陷。

Atxn1tg/-小鼠的主要神经病理学表型是浦肯野细胞,小脑皮层顶层(分子层)的成分的退化。这种退化开始表现为分子层收缩和浦肯野细胞树突萎缩,并且随后表现为浦肯野细胞体丧失(Burright et al.,1995,Cell,82:937-948;Clark et al.,1997,J Neurosci,17:7385-7395)。12周龄时,与PML+/+小鼠相比,PML+/-和PML-/-小鼠仅显示出微小且统计学不显著的分子层收缩,而PML+/+:Atxn1tg/-小鼠显示出明显的收缩(图7C和图7D)。由于PML+/+:Atxn1tg/-小鼠在转棒上的表现与PML+/+小鼠类似(图7B),因此PML+/+:Atxn1tg/-小鼠中的神经变性可能未达到临界阈值。先前已观察到了SCA1转基因模型的行为和病理表型之间的这种非线性相关性(Gehrking et al.,2011,Hum Mol Genet,20:2204-2212)。重要地,与PML+/+:Atxn1tg/-小鼠相比,PML+/-:Atxn1tg/-和PML-/-:Atxn1tg/-小鼠在分子层厚度方面分别显示出中等和强烈的进一步减小(图7C和图7D)。这与这些动物在转棒上表现变差有关(图7B)。因此,PML缺陷加剧了Atxn1tg/-小鼠中分子层的收缩。

还通过用抗浦肯野细胞特异性蛋白钙结合蛋白的抗体的免疫荧光染色检验了浦肯野细胞的树突分枝。12周龄时,含有Atxn1 82Q转基因的所有组中浦肯野细胞树突的荧光强度降低至使得无法准确比较的极低水平(图14A和图14B)。值得注意的,与PML+/+同窝小鼠相比,PML-/-小鼠显示出浦肯野细胞树突分枝的强烈减少,而PML+/-小鼠显示出中等减少(图14A和图14B)。这些结果表明PML本身在保护抵抗神经变性中具有作用。

尽管与PML缺陷有关的分子层变薄和浦肯野细胞树突丧失,但是在不同基因型的12周大动物中未观察到浦肯野细胞群体的显著性差异(图14C)。1岁时,与PML+/+小鼠相比,PML-/-小鼠在浦肯野细胞数目方面仅显示出轻度(11.0%)且统计学不显著(p=0.107)的减少,但是PML+/+:Atxn1tg/-小鼠显示出显著减少(图7E和图7F)。值得注意的,与PML+/+:Atxn1tg/-小鼠相比,PML-/-:Atxn1tg/-小鼠在浦肯野细胞密度方面显示出显著的进一步减少,并且PML+/-:Atxn1tg/-小鼠显示出中等细胞损失(图7E和图7F)。此外,这些结果表明PML缺陷使Atxn1 82Q转基因所引起的神经病理学缺陷恶化。

B05小鼠的神经变性伴随有浦肯野细胞中泛素阳性的Atxn1 82Q包涵体的形成(Clark et al.,1997,J Neurosci,17:7385-7395)。为了确定PML对Atxn1 82Q核包涵体的影响,在12周龄的小鼠中定量具有这些包涵体的浦肯野细胞。单独的PML缺陷不会导致聚集体的形成(图14D),但是其显著增加Atxn1 82Q转基因小鼠中含有聚集体的浦肯野细胞的数目(图7G和图7H)。总的来说,这些结果表明内源PML在防止SCA1动物中错误折叠的蛋白质积累和抑制这种神经退行性疾病发展方面起作用。

降解错误折叠的蛋白质的PQC系统

本文提供了哺乳动物细胞核中降解错误折叠的蛋白质的PQC系统的证据。该系统包括识别分枝PML,其选择性结合至错误折叠的蛋白质并用聚SUMO2/3链标记这些蛋白,和效应因子分枝RNF4,其使SUMO化的错误折叠的蛋白质泛素化并靶向它们用于蛋白酶体降解。这种中继系统可能提供了哺乳动物细胞中错误折叠的蛋白质和蛋白酶体之间的关键连接,并且其可以在保护不受神经退行性及其它蛋白质构象病中起到重要作用(图7I)。

PML对错误折叠的蛋白质的选择性识别

这种系统灵敏的选择性在于含有至少两个SRS的PML。SRS1由TRIM/RBCC基序内的CC区域组成,其有利于致病性polyQ蛋白以及可能的其它错误折叠的蛋白质上的CC结构。SRS2由C端63个氨基酸组成,其识别富含芳香族(Phe、Trp和Tyr)和带正电荷的(Arg和Lys)氨基酸的短肽。SRS2类似于细菌的Hsp100ClpB(Schlieker et al.,2004,Nat Struct Mol Biol,11:607-615),除了SRS2还有利于含Leu(经常在错误折叠的蛋白质中暴露并且通过Hsp70接合的残基)的肽(Rudiger et al.,1997,EMBO J,16:1501-1507)。因此,SRS2显示出ClpB和Hsp70的混合底物特异性。这些观察结果表明PML可以识别通常存在于错误折叠的蛋白质中的结构或区域。

SUMO化在降解错误折叠的蛋白质中的作用

与SUMO结合是主要的翻译后修饰,其发生在多种蛋白质上并且是大部分真核生命所必需的。然而,除了其改变蛋白质-蛋白质相互作用的一般性外,SUMO化的生理功能仍是难以捉摸的(Wilkinson and Henley,2010,Biochem J,428:133-145)。PML-RNF4系统的主要特征是在泛素化之前SUMO2/3修饰的参与(图3、图4和图11)。先前观察到通过蛋白质-变性应激显著增加SUMO2/3与细胞蛋白质的结合(Saitoh and Hinchey,2000,J Biol Chem,275:6252-6258)。当前研究中所显示的证据提供了对这种观察结果的解释,并且表明SUMO2/3修饰的主要生理功能可能是与泛素化一致作用以有利于错误折叠的蛋白质的降解。

SUMO结合增加蛋白溶解度(Panavas et al.,2009,Methods Mol Biol,497:303-317)。由于聚集的蛋白质不能被蛋白酶体有效降解(Verhoef et al.,2002,Hum Mol Genet,11:2689-2700),因此增加蛋白溶解度可以是泛素化之前PML所赋予的有益作用。此外,SUMO化程度可以使得能够对是否选择给定的错误折叠的蛋白质用于再折叠或降解进行“分类决策”。与这种设想一致,与单一的SUMO结合似乎足以增加蛋白质溶解度(Panavas et al.,2009,Methods Mol Biol,497:303-317),并因此可以有利于再折叠。相反,对于用于泛素化和降解的通过RNF4上四个串联的SIM的有效识别,需要与SUMO2/3链结合(Tatham et al.,2008,Nat Cell Biol,10:538-546)。在不成功的再折叠尝试后,可以形成这些链。

已报道错误折叠的蛋白质的SUMO化促进或抑制神经退行性疾病(Martin et al.,2007,Nat Rev Neurosci,8:948-959)。通过SUMO1和SUMO2/3在错误折叠的蛋白质去除中的不同功能(图3)以及通过SUMO化功能之间的二分性:增加异常蛋白质的溶解度(可以增加其毒性)和促进其降解,这些表面上对立的观察结果可以统一。因此,SUMO化增加的结局可能取决于它是否可以通过细胞降解能力匹配。

潜在的主要PQC系统

细胞核中的蛋白质可以内含突变或者承受急性和慢性损害,如其他处的蛋白质一样。细胞核高度拥挤的环境可能使得维持蛋白质量特别有挑战性。预期泛素-蛋白酶体途径是细胞核中主要的降解系统,其中自体吞噬已知不运行。先前的研究显示几种泛素连接酶,如酵母San1和Doa10以及哺乳动物UHRF-2和E6-AP参与细胞核错误折叠的蛋白质的降解(Cummings,1999,Neuron,24:879-892;Deng and Hochstrasser,2006,Nature,443:827-831;Gardner et al.,2005,Cell,120:803-815;Iwata et al.,2009,J Biol Chem,284:9796-9803)。尽管如此,PML的主要细胞核定位以及PML和RNF4对不同错误折叠的核蛋白的有效作用表明PML-RNF4系统可能是哺乳动物细胞核中主要的PQC系统。

TRIM蛋白家族是多细胞动物中共有的,从秀丽隐杆线虫(C.elegans)中的约20个成员至小鼠和人中的超过70个(Ozato et al.,2008,Nat Rev Immunol,8:849-860)。先前表明至少一些其它的TRIM蛋白也具有SUMO E3活性(Chu and Yang,2011,Oncogene,30:1108-116)。考虑到除细胞核之外,它们定位至细胞质(Ozato et al.,2008,Nat Rev Immunol,8:849-860),推测TRIM蛋白也参与细胞质中的PQC。在进化过程中TRIM蛋白的快速扩增可能部分是对长寿命动物细胞中蛋白质量管理复杂性增加的响应。

RNF4在脊椎动物中是保守的(Sun et al.,2007,EMBO J,26:4102-4112)。SUMO依赖性泛素连接酶也存在于低等真核物种中(Sun et al.,2007,EMBO J,26:4102-4112),并且参与至少一种突变酵母转录因子的降解(Wang and Prelich,2009,Mol Cell Biol,29:1694-1706)。因此,还可能的是类似于PML-RNF4系统的系统可能在维持这些生物中的蛋白质量中起作用。

PML-RNF4系统和神经变性

PML-/-小鼠的广泛特征为多种表型,包括致瘤作用(Wang et al.,1998,Science,279:1547-1551)。本研究表明了PML在保护免受神经变性中的作用(图7和图14)。包括SCA和HD在内的神经退行性病症通常是迟发性疾病。积累的证据表明衰老期间PQC中的进行性衰退(Balch et al.,2008,Science,319:916-919)。PML-RNF4对与SCA1、HD和ALS有关的致病性蛋白以及PML缺陷对SCA1小鼠模型进展的强烈影响,以及患有多种神经退行性疾病的患者中神经元包涵体中PML的积累(Skinner et al.,1997,Nature,389,971-974;Takahashi et al.,2003,Neurobiol Dis,13:230-237)表明PML-RNF4系统的缺陷或功能障碍可以在这些疾病中起作用。因此,PML-RNF4系统和类似系统将在它们的治疗中成为有价值的靶标。

实施例2:TRIM蛋白可以识别错误折叠的蛋白质并促进它们的降解

含三结构域(TRIM)的家族由多细胞动物细胞中大量的蛋白质组成,其范围从秀丽隐杆线虫中的约20个至小鼠和人中的超过70个。这些蛋白质在它们的N端共有特征性的TRIM或RBCC基序,其由RING域、一个或两个B盒(与RING域类似,通过锌离子配位)和卷曲螺旋区组成。在这之后是在TRIM蛋白中更加可变的C端区域并且含有不同的基序(Hatakeyama,2011,Nat Rev Cancer,11:792-804;Ozato et al.,2008,Nat Rev Immunol,8:849-860)。TRIM蛋白调节一系列细胞过程,包括保护免受癌症和病毒感染的那些。生物化学上,一些TRIM蛋白显示出泛素E3连接酶活性,这归因于TRIM/RBCC区域内的RING域。另外,至少一些TRIM蛋白具有E3连接酶用于蛋白与SUMO(小类泛素修饰因子)结合。然而,关于SUMO E3的TRIM家族的底物仍存在未解决的问题。

如上所述,据证实PML(早幼粒细胞性白血病蛋白质;也称为TRIM19)在错误折叠的蛋白质的除去中起到重要作用。PML开始鉴别为参与与大部分急性早幼粒细胞白血病有关的t(15;17)染色体翻译的基因产物。其是PML核体的主要结构和同名组分。已表明PML能够特异性结合至并促进一系列错误折叠的蛋白质的降解。通过不同的区域,PML可以识别错误折叠的蛋白质中存在的共同特征,包括富含芳香族氨基酸残基和卷曲螺旋结构的肽。然后,PML用聚-SUMO2/3链通过其SUMO E3活性标记错误折叠的蛋白质。这允许SUMO靶向的泛素连接酶(STUbL)RNF4识别修饰的错误折叠的蛋白,RNF4使错误折叠的蛋白质泛素化并靶向它们以用于蛋白酶体中的后续降解。PML在蛋白质量控制中的作用对于保护抵抗神经退行性疾病是重要的,因为PML缺陷加剧了脊髓小脑性共济失调1型(SCA1)(由ataxin-1中多聚谷氨酰胺(polyQ)延伸的扩增引起的进行性和致死性疾病)小鼠模型的行为和神经病理学表型。考虑到存在大量TRIM蛋白,使用PML获得的结果引起了其它TRIM蛋白,如PML是否能够识别并降解错误折叠的蛋白质的重要问题。在本文所提供的实验中,分析了一组TRIM蛋白并且据观察在TRIM蛋白中识别和降解错误折叠的蛋白质的能力是普遍的,表明在后生动物细胞中该家族在蛋白质量控制中的重要作用。

现将描述在这些实验中使用的材料和方法。

质粒

通过PCR在pRK5中制备FLAG-TRIM27、FLAG-TRIM32和FLAG-TRIM5δ。PCR扩增的模板购自Open Biosystems,并且相应的基因登录号分别为BC013580、BC003154和CV029096。所有三种基因是人源的。先前描述了以下质粒:FLAG-PML(同种型IV)(Chu and Yang,2011,Oncogene,30:1108-116);Atxn1 82Q-GFP、FLAG-Atxn1 82Q和HA-Httex1p97QP(Guo et al.,2014,Mol Cell,55(1):15-30);FLAG-TRIM11(Ishikawa et al.,2006,FEBS Lett,580:4784-4792);FLAG-TRIM22(长形式)(Barr et al.,2008);HA-TRIM39(Lee et al.,2009,Exp Cell Res,315:1313-1325);HA-标签的TRIM1、2、3、4、5、6、8、9、10、11、12、13、14、18、20、21、22、23、24、25、26、27、28、29、30、31和32(Reymond et al.,2001,EMBO J,20:2140-2151;Uchil et al.,2008,2008,PLoS Pathog,4:e16);和其余的TRIM表达质粒,其表达具有N端HA标签或C端V5标签的蛋白(Versteeg et al.,2013,Immunity,38:384-398)。为了分析TRIM蛋白对Atxn1 82Q和Httex1p 97QP水平的影响,将Trim11(NM_145214.2)、Trim15(NM_033229.2)、Trim28(NM_005762.2)、Trim34(NM_021616.5)、Trim39(NM_021253.3)、Trim42(NM_152616.4)、Trim43(NM_138800.1)、Trim65(NM_173547.3)、Trim67(NM_001004342.3)、Trim70(NM_001037330.1)、Trim71(NM_001039111.2)和Trim75(NM_001033429.2)构建到含有5'-HA标签的pcDNA 3.1(-)载体中。获得所有其它Trim质粒(Versteeg et al.,2013,Immunity,38:384-398)。

siRNA

SUMO2/3siRNA(Santa Cruz sc-37167)是三种不同的siRNA双螺旋的混合,具有有义链序列5'-CCCAUUCCUUUAUUGUACA-3'(SEQ ID NO:157)、5'-CAGAGAAUGACCACAUCAA-3'(SEQ ID NO:158)和5'-CAGUUAUGUUGUCGUGUAU-3'(SEQ ID NO:159)。

TRIM27siRNA购自Qiagen,其有义链序列为5'-AACTCTTAGGCCTAACCCAGA-3'(SEQ ID NO:162)。

细胞培养和转染

HeLa细胞得自ATCC。PML+/+和PML-/-MEF细胞来源于具有相应基因型的小鼠胚胎。将细胞保持在标准培养条件下。根据生产商的说明,使用Lipofectamine 2000将DNA质粒转染至细胞,并且使用Lipofectamine 2000或RNAiMAX(Invitrogen)在连续两天在两轮中将siRNA转染到细胞中。当转染DNA和siRNA两者时,在第二轮siRNA转染后1天转染DNA。在上次转染后24小时,以7.5-10μM(最终浓度)加入MG132(Sigma)4-5小时。

免疫荧光

将在盖玻片上培养的细胞用4%多聚甲醛固定15分钟,用0.2%Triton X-100透性化15分钟,依次与第一和第二抗体温育。第一抗体为抗HA、抗FLAG(小鼠mAb M2,1:2,000)(Sigma)和抗TRIM27抗体。第二抗体为FITC结合的抗鼠、抗兔(Zymed)和抗山羊(Invitrogen)IgG;德克萨斯红结合的抗鼠和抗兔IgG(Vector labs);和罗丹明红-X结合的抗山羊抗体(Jackson ImmunoResearch Labs)。之后,用含有DAPI(Vector Labs)的培养基装载细胞,并用Nikon Eclipse E800或者Olympus IX81显微镜采集图像。

细胞裂解液分级、免疫印迹和过滤器阻滞测定

如描述的制备样品(Guo et al.,2014,Mol Cell,55(1):15-30)。简要地,将细胞在冰上在补充有250IU/ml核酸酶(Sigma)、1mM PMSF、1×完全蛋白酶混合物(Roche)和20mM N-乙基马来酰亚胺(NEM;Sigma)的NP-40裂解缓冲液(50mM Tris,pH 8.8、100mM NaCl、5mM MgCl2、0.5%NP-40、2mM DTT)中裂解。将细胞裂解液以13,000rpm在4℃下离心15分钟。通过SDS-PAGE和免疫印迹分析含有NP-40可溶(NS)蛋白的上清液。将颗粒在补充有250IU/ml核酸酶、1mM PMSF、1×完全蛋白酶混合物和20mM NEM的颗粒缓冲液(20mM Tris,pH 8.0、15mM MgCl2、2mM DTT)中再悬浮。将颗粒部分在2%SDS加50mM DTT中煮沸并通过SDS-PAGE溶解。通过免疫印迹检测进入凝胶(SDS可溶的,SS)的蛋白质。对于过滤器阻滞(点印迹)测定,将一部分煮沸的颗粒施加于孔径0.2μm的膜过滤器,并且通过免疫印迹法分析保留在过滤器上的SDS-耐受性(SR)聚集体。第一抗体为:抗HA(兔,Y-11,1:500)(Santa Cruz Biotechnology);抗FLAG(小鼠,M2,1:7,500)和抗肌动蛋白(兔,1:10,000)(Sigma);抗GFP(小鼠,1:4,000)(Clonetech);和SUMO2/3(兔,1:250,Abgent)。第二抗体结合至HRP(Santa Cruz Biotechnology),或用IRD Fluor 800或IRD Fluor 680(LI-COR,Inc.)标记。使用ECL试剂进行免疫印迹并使用ImageJ分析,或者使用Odyssey红外成像系统扫描并使用Image Studio Lite(LI-COR,Inc.)分析。

蛋白质纯化和体外SUMO化测定

如前描述的(Tang et al.,2006,Nat Cell Biol,8:855-862;Tang et al.,2004,J Biol Chem,279:20369-20377)(有修改),在293T细胞中表达FLAG-TRIM27和HA-Atxn1 82Q-FLAG并通过抗-FLAG M2珠(Sigma)纯化。将细胞在补充有1mM PMSF和1×完全蛋白酶混合物的IP-裂解缓冲液(50mM Tris,pH 7.5、150mM NaCl、0.5%Triton X-100、0.5%NP-40和2mM DTT)中裂解。对于TRIM27纯化,IP-裂解缓冲液还补充有20μM ZnCl2。将裂解液与抗FLAG M2珠在4℃下温育4小时至过夜。用含有0、0.5和1M KCl的IP-裂解缓冲液以及用洗脱缓冲液(50mM Tris,pH 7.5、150mM NaCl和2mM DTT)清洗M2珠。在含有0.1-0.3mg/ml 3×FLAG肽(Sigma)的洗脱缓冲液中洗脱结合的蛋白质。

用于体外SUMO化反应的其它组分购自Boston Biochem。在30μl含有纯化的HA-Atxn1 82Q-FLAG(600ng/200nM)、FLAG-TRIM27、SAE1/SAE2(125nM)、Ubc9(1μM)、His-SUMO2(25μM)和BSA(0.1μg/ml)的反应缓冲液(50mM Tris pH 7.5、5.0mM Mg2+-ATP和2.5mM DTT)中,在37℃下进行体外SUMO化测定1.5小时。通过加入30μl含有2%SDS和50mM DTT的IP-裂解缓冲液并在95℃下加热10分钟使反应混合物变性。保存1等份加热的反应混合物用于免疫印迹分析,并将其余部分在无SDS的IP-裂解缓冲液中稀释20倍。通过抗HA珠(Roche)使HA-Atxn1 82Q-FLAG免疫沉淀并使用抗-SUMO2/3抗体分析SUMO2/3修饰。

现将描述实验结果。

TRIM27、TRIM32和TRIM5δ与致病性Ataxin-1和亨廷顿蛋白的共定位

先前显示PML/TRIM19可以识别并促进多种细胞核错误折叠的蛋白质的降解,包括Atxn1 82Q,与脊髓小脑性共济失调1型(SCA1)有关的具有82个谷氨酰胺延伸的致病性ataxin 1蛋白(Guo et al.,2014,Mol Cell,55(1):15-30)。人和小鼠中TRIM蛋白家族由基于保守TRIM/RBCC基序的C端区域的结构特征可以辨别的超过70个成员组成(Ozato et al.,2008,Nat Rev Immunol,8:849-860)。大部分TRIM蛋白在该区域中包含一个或多个保守域,其中最常见的包括PRY-SPRY域(在~40种TRIM蛋白中存在)和多个NHL域(在4种TRIM蛋白中存在)。另外,一些TRIM蛋白在C端区域中不包含可识别的基序。为了检验其它人TRIM蛋白,如PML是否能够识别错误折叠的蛋白质,首先选择了TRIM27(具有PRY-SPRY域)、TRIM32(具有多个NHL域)和TRIM5δ(无已知的域的TRIM5的短拼接变体)。当在HeLa细胞中表达时,这三种TRIM蛋白显示重叠但不同的细胞定位模式:TRIM27和TRIM32定位在细胞质和细胞核中,而TRIM5δ仅定位在细胞质中。尽管如此,所有三种蛋白在它们各自的隔室内在斑点体(speckled body)中浓缩(图15A)。

进行实验以检验TRIM27、TRIM32和TRIM5δ与Atxn1 82Q以及与和HD有关的含有97个谷氨酰胺的延伸的亨廷顿蛋白(Htt)片段(Httex1p97Q)的共定位。作为增强的绿色荧光蛋白(GFP)的融合蛋白表达的Atxn182Q仅在核中形成包涵体,而HA标签的httex1p 97QP在细胞核和细胞质两者中形成包涵体(Guo et al.,2014,Mol Cell,55(1):15-30)(图15A和图15B)。外源TRIM27和TRIM32与Atxn1 82Q-GFP包涵体在细胞核中共定位(图15A),而与HA-Httex1p 97QP包涵体在细胞核和细胞质两者中共定位(图15B和图15C)。此外,内源TRIM27也与Atxn1 82QGFP包涵体共定位(图15D)。外源TRIM5δ仅与细胞质HA-Httex1p 97QP包涵体共定位(图15B和图15C)。因此,即使在它们的C端序列中存在差异,但是TRIM27、TRIM32和TRIM5δ能够识别在它们各自细胞隔室中形成的错误折叠的蛋白质。

TRIM27、TRIM32和TRIM5δ降低了不溶性和可溶性Atxn1 82Q蛋白的水平

为了检验这些TRIM蛋白是否能够降低错误折叠的蛋白质的水平,将它们中的每一种与FLAG标签的Atxn1 82Q在HeLa细胞中表达。分析细胞裂解液中NP40可溶(可溶或NS)和NP-40不溶,但SDS-可溶(聚集体或SS)的FLAG-Atxn1 82Q的水平。值得注意的,每种TRIM蛋白都能够降低可溶性和聚集的Atxn1 82Q蛋白的水平(图16A和图16B)。TRIM27和TRIM32还降低了耐受NB-40和SDS(SR)两者的Atxn1 82Q蛋白的水平,其通过过滤器阻滞测定检测并且可能代表了β-淀粉样蛋白结构(见下文)。

当以类似水平表达时,所有三种TRIM蛋白,特别是TRIM32在降低可溶性Atxn1 82Q水平方面显示出强于PML的活性(图16A)。此外,尽管TRIM27和TRIM5δ的活性多少弱于PML,但是TRIM32在减少聚集的Atxn1 82Q方面的活性与PML相当(图16A和图16B)。选择TRIM27用于进一步分析。使用siRNA降低内源TRIM27的表达,这导致可溶性和聚集的Atnx1 82Q蛋白的水平显著升高(图16C)。总的来说,这些结果表明与PML类似,TRIM27、TRIM32和TRIM5δ能够除去错误折叠的蛋白质。与这些TRIM蛋白相反,作为SUMO E3的PIAS家族成员的PIASy不能降低可溶性或聚集的Atxn1 82Q的水平(图16D)。

TRIM27和TRIM32对Atxn1 82Q的影响与PML无关

TRIM27在PML核体中部分共定位(Cao et al.,1998,J Cell Sci,111(Pt10):1319-1329)(图17A)。为了检验TRIM27和另一种核定位的TRIM蛋白TRIM32是否依赖PML来降解错误折叠的蛋白质,使用了PML野生型(PML+/+)和PML缺陷型(PML-/-)小鼠胚胎成纤维细胞(MEF)。尽管TRIM27与PML核体共定位,但是即使在不存在PML的情况下,TRIM27也存在于Atxn1 82Q聚集体中(图17B)。与PML+/+MEF相比,PML-/-中的SDS不溶性Atxn1 82Q聚集体的水平显著更高,这与PML在除去Atxn182Q中的作用一致(Guo et al.,2014,Mol Cell,55(1):15-30)。重新引入PML降低了聚集体Atxn1 82Q的水平。值得注意的,在PML+/+和PML-/-细胞两者中,对于减少聚集的Atxn1 82Q,TRIM27和TRIM32与PML一样有效(图17C)。总的来说,这些结果表明TRIM27和TRIM32可以不依赖PML而消除Atxn1 82Q。

TRIM27、TRIM32和TRIM5δ靶向Atxn1 82Q用于SUMO2/3依赖性方式的蛋白酶体降解

先前发现PML通过其SUMO E3活性促进错误折叠的蛋白质的蛋白酶体降解(Guo et al.,2014,Mol Cell,55(1):15-30)。当用蛋白酶体抑制剂MG132处理细胞时,TRIM27、TRIM32和TRIM5δ减少可溶性并且特别是不溶性Atxn1 82Q蛋白的能力显著受损,这表明这三种TRIM蛋白也靶向Atxn1 82Q用于蛋白酶体降解(图18A-图18C)。另外,当用siRNA降低内源SUMO2/3时,TRIM27、TRIM32和TRIM5δ不再能有效降低Atxn182Q的水平(图18D-图18F)。因此,这些TRIM蛋白在降解Atxn1 82Q中也依赖SUMO2/3。

使用TRIM27举例,然后检验了PML以外的TRIM蛋白是否可以用作Atxn1 82Q的SUMO E3连接酶。当与SUMO化反应组分,包括SUMOE1(SAE1/SAE2)、E2(Ubc9)、SUMO2和ATP温育时,Atxn1 82Q极少结合至SUMO2。然而,Atxn1 82Q与SUMO2的结合以TRIM27剂量依赖性的方式增加(图18G),表明TRIM27是Atxn1 82Q的SUMO E3。

与错误折叠的蛋白质共定位是TRIM蛋白的普遍性质

这些观察结果提示,对其余TRIM蛋白测试了它们识别Atxn1 82Q的能力。除了存在于小鼠而不是人中的两种(TRIM12和TRIM30)之外,所有其它TRIM蛋白是人来源的。用HA、Flag或V5表位标记每种TRIM,并将其与Atxn1 82Q-EGFP在HeLa细胞中共表达。

先前对TRIM蛋白亚型的调查显示这些蛋白质与细胞质和细胞核两者中的隔室有关(Reymond et al.,2001,EMBO J,20:2140-2151)。在本发明所测试的TRIM蛋白中,11种TRIM蛋白(TRIM42、43、53、59-61、67、70-72和75)通过免疫荧光无法检测。在剩余的63种TRIM蛋白中,它们中的五种(TRIM22、28、33、65和66)仅在细胞核中定位,其定位模式类似于PML。然而,不能排除在本发明中未测试的这些TRIM蛋白的某些同种型可以存在于细胞质中的可能性。例如,对于PML显示了这些胞质同种型。27种TRIM蛋白(TRIM1-5、7、9、10、13、18、20、24、25、29、34、36、37、39、45-47、50、54、63、69和76)主要或仅在细胞质中定位。27种TRIM蛋白(TRIM6、8、11、12、16、21、23、27、30-32、35、38、40、44、48、49、51、52、55、56、58、62、64、68、73和74)以显著的量存在于细胞核和细胞质两者中。一些TRIM蛋白在细胞核或细胞质中形成点状或丝状结构,而其它TRIM蛋白分散定位,并且它们中的一些显示出核周定位。通常,TRIM蛋白在相同或不同细胞中显示出这些定位模式的组合(图19和表2)。

在大多数细胞中,14种TRIM蛋白(TRIM6、8、11、19、21、22、27、28、30、32、33、35、38和51),包括PML/TRIM19与细胞核Atxn182Q包涵体共定位(图19和表2),其占部分存在或仅存在于细胞核中的32种TRIM蛋白中的几乎43%。这些TRIM蛋白来自于具有不同C端序列的不同亚组。如TRIM27,6种蛋白质(TRIM6、11、21、22、35和38)是亚组IV的成员,其通常在PRY基序之前包含SPRY基序。如PML,TRIM8是组V的成员,其不包含已知的域。TRIM28和TRIM33是组VI的成员,其包含PHD-BR基序。TRIM32属于亚组VII,其具有五个NHL重复。TRIM51认为是类TRIM蛋白,其缺少两个B盒,但是含有RING域和卷曲螺旋区;如亚组IV成员一样,它还含有SPRY域。总的来说,这些结果显示即使在它们的C端存在差异,但是大量TRIM蛋白具有识别错误折叠的蛋白质的能力。

通过其它TRIM蛋白介导的错误折叠的蛋白质的降解

为了检验TRIM蛋白在蛋白质量控制中的作用,测试了除TRIM53(假基因)和TRIM57(与TRIM59相同)外所有的人TRIM蛋白以及TRIM12和TRIM30(小鼠来源)降解Atxn1 82Q的能力。35种人TRIM蛋白(TRIM3、4、5、6、7、9、11、13-17、19、20、21、24、25、28、29、34、39、43-46、49、50、52、58、59、65、67、70、74和75)和小鼠TRIM30能够不同程度降低Atxn1 82Q的NS和/或SS部分的水平(图20A和表2)。一些TRIM蛋白显示出强活性,包括TRIM4、5、9、11、16、17、20、30、39、43、65、70、75(图20A和表2),并且活性最强的似乎是TRIM11(图16A,图20A和表2)。值得注意的,在本测定中如上所示能够降低Atxn182Q的TRIM27和TRIM32未显示出效果。这种差异可能是由于表达水平所造成的。在上述实验中,将TRIM27和TRIM32克隆到质粒pRK5中。但是在本部分的实验中,将TRIM27和TRIM32克隆到pcDNA中。与来自pcDNA质粒的表达相比,一致观察到来自pRK5质粒的更强的表达。因此,即使对于未显示出降解Atxn1 82Q的活性的那些,当它们的表达水平升高时,它们可能会显示活性。因此,推断促进Atxn1 82Q降解的能力在TRIM蛋白中是普遍的。

值得注意的,几种TRIM蛋白增加,而不是抑制Atxn1 82Q的表达,包括TRIM26、33、42、47、48、66、69和76。这表明不同TRIM蛋白对Atxn1 82Q不同的作用。

同时,还检验了TRIM蛋白对Httex1p 97QP的作用(图20B)。仅有少数TRIM蛋白能够降低NS和/或SS部分中Httex1p 97QP的水平,包括TRIM3、11、30、68、74和75。大部分TRIM蛋白能够增加Httex1p 97QP的水平,包括TRIM1、4、6-10、12-15、21、23-28、32-39、41-47、49-51、54-56、58、60-67、69-73、76和77。不同于Atxn1 82Q,仅有一小部分表达Httex1p 97QP的细胞含有包涵体。一致地,与相同部分中Atxn1 82Q的份数相比,SS部分中Httex1p 97QP蛋白的份数极少。另外,在上述研究中,PML(在pRK5质粒中克隆)能够降低Httex1p 97QP的水平,而此处PML(在pcDNA中克隆)不能。因此,如它们对Atxn1 82Q的影响一样,TRIM蛋白对Httex1p 97QP的影响还可能受它们表达水平的影响。这些结果表明了TRIM蛋白对不同错误折叠的蛋白质的不同影响,其中它们可以受其表达水平的影响。

TRIM蛋白

先前已显示PML能够消除哺乳动物细胞核中存在的一系列错误折叠的蛋白质。此处所提供的实验系统分析了几乎所有TRIM蛋白并且发现未预期地它们中大量的蛋白可以识别错误折叠的蛋白质。然而,该数字可能是低估的,因为主要使用了单一错误折叠的蛋白质Atxn1 82Q。在PML中已确定了至少两个底物识别位点(SRS),RBCC基序内的卷曲螺旋区和包含最后60个氨基酸的区域。这些SRS分别可以识别存在于一些变性蛋白中的卷曲螺旋结构和富含芳香酸残基(Phe、Trp和Tyr)的肽。TRIM蛋白在它们的C端区域中差异最大(Hatakeyama,2011,Nat Rev Cancer,11:792-804;Ozato et al.,2008,Nat Rev Immunol,8:849-860)。可能的是其它TRIM蛋白可以具有识别具有不同结构特征的错误折叠的蛋白质的特异性能力。另外,可能的是多种TRIM蛋白可以识别细胞质中错误折叠的蛋白质。因此,本发明的观察结果强调了该大家族在蛋白质量控制中的关键重要性。

值得注意的,不论其细胞质定位,TRIM5δ能够促进Atxn1 82Q降解。可能的是在引入细胞核之前,TRIM5δ可以识别细胞质中可溶性的错误折叠的Atxn1 82Q。另外,TRIM27和TRIM32显示出降解可溶性Atxn1 82Q的较强能力,而PML显示较低的活性。这可能与不同于PML,TRIM27和TRIM32部分定位至其中Atxn1 82Q蛋白(以及几乎所有其它蛋白)产生的细胞质的事实有关。因此,通过识别去往核的错误折叠的蛋白质,胞质TRIM蛋白可以在核的蛋白质量控制中起到重要作用。

PML以依赖于其SUMO E3活性的方式降解错误折叠的蛋白质(Guo et al.,2014,Mol Cell,55(1):15-30)。同样地,TRIM5δ、TRIM27和TRIM32依赖SUMO以除去Atxn1 82Q。体外测定还确认了TRIM27对Atxn1 82Q的SUMO E3活性。因此,TRIM蛋白可以使用类似的机制清除具有错误折叠的蛋白质的细胞。用于降解错误折叠的蛋白质的最相关的SUMO蛋白是SUMO2/3。通过SUMO2/3形成的多聚链有利于通过RNF4上的多个SIM的识别(Tatham et al.,2008,Nat Cell Biol,10:538-546)。与在蛋白质量控制中的作用一致,非应激细胞中的SUMO2/3主要是非结合形式,但在蛋白损伤应激后结合至靶蛋白(Golebiowski et al.,2009,Sci Signal,2:ra24;Saitoh and Hinchey,2000,J Biol Chem,275:6252-6258)。尽管如此,考虑到TRIM蛋白的多样性以及与其中一些有关的泛素E3连接酶活性(Meroni and Diez-Roux,2005,Bioessays,27:1147-1157),仍可能的是一些TRIM蛋白可以主要作为泛素E3起作用以用于错误折叠的蛋白质。

目前,不清楚一些TRIM蛋白,如Atxn1 82Q为何可以降解错误折叠的蛋白质,而其它的不可以。另外,在所测试的TRIM蛋白中,TRIM11具有明显更强的降低Atxn1 82Q的活性。仍需确定可能解释其活性的原因。一些TRIM蛋白可以增加Atxn1 82Q并且特别是Httex1p 97QP的表达水平。这进一步强调了这些蛋白的功能多样性。TRIM蛋白可以用作分子伴侣以防止蛋白错误折叠并且作为解聚酶以溶解先前形成的蛋白聚集体。这些活性可以部分解释TRIM蛋白稳定化Atxn1 82Q并且特别是Httex1p97QP的效果。这表明了在与错误折叠的蛋白质有关的疾病疗法中TRIM蛋白的另一种重要用途。一些人类疾病与具有部分功能的突变蛋白的降解紧密相关。显著的实例是囊性纤维化(CF),它是由基因囊性纤维化跨膜传导调节剂(CFTR)中的突变所引起的。这些突变在细胞质中降解。正在发展疗法以稳定突变的CFTR,允许突变的CFTR达到膜以实现其功能。设想使用TRIM蛋白可以在CF及与部分功能蛋白降解有关的其它疾病中实现该目标。

实施例3:将重组TRIM11递送至哺乳动物细胞促进错误折叠的蛋白质降解

如上所述,表明含有三结构域(TRIM)家族的成员在错误折叠的蛋白质的识别和降解中起重要作用。开始,使用PML/TRIM19作为实例,表明PML可以通过识别通常存在于错误折叠的蛋白质中的结构特征的不同区域来特异性结合至错误折叠的蛋白质。然后,PML使用其SUMO E3活性使用聚-SUMO2/3链标记错误折叠的蛋白质。这使得SUMO化-靶标泛素连接酶(STUbL)RNF4能够识别错误折叠的蛋白质,并对错误折叠的蛋白质进行后续泛素化和蛋白酶体降解。还显示这种PML-RNF4介导的顺序SUMO化和泛素化系统在保护抵抗神经退行性疾病中起重要作用。

随后,研究了所有已知的人TRIM蛋白中的绝大部分并且发现它们中的大多数能够定位至错误折叠的蛋白质,如致病性ataxin 1(Atxn1 82Q)和亨廷顿蛋白(Htt 97Q)所形成的包涵体。通过测试代表性TRIM蛋白,显示多种TRIM蛋白还能够以SUMO依赖性方式降解错误折叠的蛋白质。值得注意的,在所测试的TRIM蛋白中,一种TRIM蛋白TRIM11显示出特别强的降低错误折叠的蛋白质的活性。

在本文所提供的实验中,作为降解与神经变性有关的胞内错误折叠的蛋白质的试剂,设法开发了重组TRIM11。对于此,使用HIV Tat来源的肽,其能够将蛋白质递送到哺乳动物细胞中(图22)。将TRIM11与Tat来源的肽融合,在细菌中表达融合蛋白,并使用亲合树脂,然后通过凝胶过滤柱将蛋白纯化至均一。为了比较,同时还纯化了Tat-肽融合的SUMO2蛋白。

用Atxn1 82Q-GFP、Atxn1 30Q或Htt 97Q-GFP转染HeLa细胞,并且随后与重组TRIM11或SUMO2蛋白一起温育。如图21A所示,用TRIM11处理HeLa细胞导致Atxn1 82Q水平强烈降低。其还导致Htt 97Q水平强烈降低(图21B)。相反,SUMO2对Atxn1 82Q水平的影响极小(图21C)。这些数据表明将TRIM11递送到哺乳动物细胞会降低错误折叠的蛋白质的水平,从而提供了将TRIM-RNF4系统靶向神经退行性及其它蛋白质错误折叠疾病的疗法的证明或概念证据。

实施例4:SCA1和HD患者中RNF4向神经元包涵体的定位

小鼠中的RNF4缺陷导致胚胎死亡(Hu et al.,2010,PNAS107:15087-92),使得无法分析其缺陷对小鼠神经变性模型的影响。先前研究显示PML与SUMO和泛素一起与多聚谷氨酰胺疾病患者中的神经元包涵体共定位(Dorval and Fraser;2006,J BIol Chem 281:9919-24;Martin et al.,2007,Nat Rev Neurosci 8:948-59;Skinner et al.,1997,Nature 389:971-4;Takahasi et al.,2003,Neurobiol Dis 13:230-70),表明PML参与这些疾病。本文所提供的数据分析了人SCA1和HD患者死后脑组织中RNF4的定位。

现将描述在这些实验中使用的材料和方法。

由哈佛大学脑组织资源中心(Harvard Brain Tissue Resource Center)提供HD患者组织(苍白球):获得AN06564、AN12127和AN12029(观察到聚集体)和AN09048、AN19685、AN14942、AN13612和AN17467(未观察到聚集体);从国家共济失调基金会(National Ataxia Foundation)获得了SCA1患者组织(基础点)。如前描述的(有修改)进行免疫组织化学和免疫荧光(Duda et al.,2000,J Neuropathol Exp Neurol 59:830-41;Emmer et al.,2011,J Biol Chem 286:35104-18)。将HD和SCA1患者脑在石蜡中包埋并切成6μm切片。将切片对所指示的抗RNF4(#1:兔,11-25,1:300,Sigma;#2:山羊,C15,1:25,Santa Cruz)、抗亨廷顿蛋白(小鼠,MAB5374,1:500;Millipore)、抗泛素(小鼠,Ubi-1,MAB1510,1:2,000;Millipore)和抗polyQ(小鼠MAB1574,1C2,1:1,000;Millipore)染色。对照兔抗体用于确认抗RNF4染色的特异性。

现将描述实验结果。

当在无包涵体的患者神经元中检测时,RNF4倾向于在整个细胞核中扩散分布或形成细胞核灶点(图23)。在检验的SCA1的两种情况下,细胞核包涵体对抗polyQ抗体(1C2)存在反应性。RNF4分别存在于16个1C2-反应性细胞核包涵体中的5个以及17个中的4个中(图24)。

在HD患者中,在所检验的8个患者样品中的3个中,在苍白球区中存在对抗亨廷顿蛋白或抗泛素抗体反应性的细胞质包涵体。这与上述观察结果一致:细胞核包涵体在成年发病的HD病例中是少有的(DiFiglia et al.,1997,Science 277:1990-3)。尽管如此,基于使用两种单独抗RNF4抗体的结果,RNF4与约20%苍白球区中的包涵体共定位(图25和图26)。含有RNF4的包涵体倾向于显示出被亨廷顿蛋白或泛素信号的环围绕的RNF4免疫反应性(图25和图26)。SCA1和HD患者中RNF4与polyQ聚集体的部分共定位使得联想到PML与polyQ聚集体的部分共定位(Skinner et al.,1997,Nature 389:971-4;Takahashi et al.,2003,Neurobiol Dis 13:230-7),并且其可以反映受疾病影响的神经元中PML/TRIM-RNF4系统的阻断。

实施例5:TRIM蛋白的分子伴侣和蛋白解聚酶活性

本文描述的数据表明TRIM11可以用作分子伴侣和解聚酶。在一个方面,TRIM11可以防止聚集体形成。另一方面,TRIM11不仅能使应激诱导的非淀粉样蛋白聚集体(荧光素酶和GFP聚集体)再折叠,而且还能够解聚淀粉样蛋白聚集体(Atxn1 82Q聚集体和α-突触核蛋白原纤维)。此外,本文还显示TRIM11还是可以SUMO化Atxin1 82Q并后续降解的SUMO E3连接酶。因此,TRIM11可以用作蛋白质解聚和降解之间的联系。

现将描述在这些实验中使用的材料和方法。

siRNA转染

小鼠TRIM11siRNA(sc-76735)购自Santa Cruz。在第一天,将海马神经元细胞(15000个细胞/孔)在具有平板培养基的96孔板中平板接种。在第二天(平板接种后约18小时),将平板培养基完全更换为神经元培养基。在第三天,通过Lipofectamine RNAiMAX将siRNA(1pmol)转染至细胞。根据生产商的说明进行转染方法。

细胞培养物

用Mccoy 5A培养基培养HCT16细胞。用RPMI 1640培养基培养A549细胞。用DMEM培养基培养HeLa和HEK293T细胞。

这些培养基均含有10%胎牛血清(FBS)。获得原代神经元细胞。根据先前描述的方法培养神经元细胞。简要地,首先用平板培养基(组合神经基础培养基、B27、GlutaMAX、青霉素/链霉素和10%FBS,通过指定的稀释比)将细胞平板接种。18-24h后,将平板培养基完全更换为神经元培养基(组合神经基础培养基、B27、GlutaMAX和青霉素/链霉素,通过指定的稀释比)。除通知之外,将所有细胞保持在37℃下加湿的5%CO2气氛中。

免疫荧光

将在盖玻片上铺板的细胞在室温下用4%多聚甲醛固定10分钟。将细胞在20℃下用甲醇进一步透性化10分钟。用PBS清洗细胞,然后通过2%牛血清白蛋白(BSA)在室温下封闭30分钟。将细胞与第一抗体在4℃温育过夜,然后与荧光第二抗体在室温下温育1小时。最后,用含有4',6-二脒基-2-苯基吲哚(DAPI)(Vector Laboratories;H-1200)的封固剂将盖玻片封固至载玻片。对于抗体稀释,应用抗HA(1:100)、抗p-a-Syn(1:100)和抗p62(1:200)。

抗体

抗TRIM11(ABC926)抗体购自Millipore。单克隆抗Flag抗体、兔Flag抗体和抗Flag琼脂糖珠购自Sigma。抗HA亲合基质(克隆3F10,11815016001)购自Roche。抗HSF1(sc-9144)购自Santa Cruz。抗a-突触核蛋白磷酸-Ser129(825701)购自Biolegend。抗Hsp70(ADI-SPA-810-D)购自Enzo。抗a-突触核蛋白(2642)和抗Hsp90(4874)购自Cell signaling technology。

蛋白质分级、过滤器阻滞测定和免疫印迹

将细胞用裂解缓冲液(50mM Tris,pH 8.8、100mM NaCl、5mM MgCl2、0.5%IGEPAL CA-630、1mM DTT、250IU/ml核酸酶(Sigma)、1mM PMSF和1×完全蛋白酶混合物(Roche))在冰上裂解30分钟。通过在4℃下以13000rpm离心15-20分钟获得上清液。将颗粒在颗粒缓冲液(20mM Tris,pH 8.0、15mM MgCl2、1mM DTT、250IU/ml核酸酶、1mM PMSF和1×完全蛋白酶混合物)中在冰上进一步再悬浮30分钟,然后与2%SDS缓冲液直接煮沸。通过Bradford测定(Bio-Rad Labs)测量蛋白浓度。通过SDS-PAGE对所有蛋白质样品进行免疫印迹。将上清液视为可溶性部分。将颗粒视为不溶性部分(SDS可溶)。对另一部分进行过滤器阻滞测定。简要地,使颗粒样品通过0.2μM孔径的膜过滤,从而通过免疫印迹分析保留在膜上的SDS耐受性聚集体。

蛋白质纯化

在293T细胞中转染Flag-Atxn1-82Q-HA并通过抗Flag M2珠纯化。为了获得高度纯化的蛋白,用增加浓度的NaCl彻底清洗珠。

在纯化的BL21DE3细胞中表达His荧光素酶。为了产生固定化的天然或变性的荧光素酶,根据生产商的说明,首先从细菌细胞裂解液中纯化天然荧光素酶。然后,通过将天然荧光素酶与8M脲温育5分钟产生变性的荧光素酶。

体内和体外SUMO化测定

对于体内SUMO化测定,用指明的质粒转染细胞。48小时后,在补充有2%SDS和50mM DTT的裂解缓冲液(50mM Tris-Cl,pH 7.4、150mM NaCl、0.5%Trition、1mM DTT、1mM PMSF和1×完全蛋白酶混合物)中裂解细胞。将细胞裂解液在95℃下进一步煮沸10分钟。保存1等份用于输入。将其余的细胞裂解液在裂解缓冲液中稀释10倍,然后与抗HA珠在4℃下温育过夜。彻底清洗珠,并用指示的抗体通过免疫印迹法分析。对于体外SUMO化测定,His-SUMO-2(UL-753)、UbcH9(E2-465)和SUMO E1(E-315)购自Boston Biochem。

质粒

通过将KpnI/XbaI TRIM11cDNA插入到KpnI/XbaI消化的pcDNA3.1-Flag载体中来构建Flag-TRIM11。通过定点突变产生Flag-TRIM11突变(12/13EE至12/13AA)。通过将BglII/SalI TRIM11cDNA插入到BamHI/SalI消化的pGEX-1λT载体中来构建GST-TRIM11。通过将Hsp70和TRIM11cDNA插入到pEGFP-C1载体中来构建GFP-Hsp70和GFP-TRIM11。通过DNA测序验证所有构建体。

荧光素酶再活化测定

为了产生聚集体,将萤火虫荧光素酶(100nM)在荧光素酶再折叠缓冲液(LRB;25mM HEPES-KOH[pH 7.4],150mM KAOc,10mM MgAOc,10mM DTT)中在45℃下加热8-10分钟。将聚集的荧光素酶(10nM)与指定浓度的TRIM11或其它蛋白质在25℃下温育90分钟。对于Hsp104/Hsp70-Hsp40二分子伴侣系统,需要5mM ATP和ATP再生系统(1mM磷酸肌酸、0.5uM肌酸激酶)。对于体内荧光素酶再折叠测定,用野生型荧光素酶瞬时转染96孔板中的细胞。24小时后,将细胞在42℃或45℃下分别加热1小时或30分钟。在热冲击之前,向培养基中加入20μg/ml的环己酰亚胺。热冲击后,将细胞转移到37℃的温育箱中另外保持1.5或3小时。使用Promega荧光素酶系统测量荧光素酶活性。

GFP解聚测定

为了产生聚集体,将GFP(4.5μM)在缓冲液A(20mM Tris-HCl,pH 7.5、100mM KCl、20mM MgCl2、5mM DTT、0.1mM EDTA、10%(v/v)甘油)中在85℃下温育15分钟。将GFP聚集体(0.45μM)与不同量的指定的蛋白在25℃下温育60分钟。通过在395nm激发时,在510nm处测量荧光来检测GFP聚集体的解聚(Infinite M200pro)。

试剂

荧光素酶(l9506)、Mg-ATP(A9187)、磷酸肌酸(P1937)和肌酸激酶(C3755)购自Sigma。KRIBB11(385570)购自Millipore。β-淀粉样蛋白(1-42)(RP10017)购自GenScript。

现将描述实验结果。

PML(TRIM19)使RNF4可以识别的错误折叠的蛋白质SUMO化以通过蛋白酶体降解。然而,TRIM11是否也是SUMO E3连接酶仍是未知的。根据TRIM的结构分析,对于其泛素E3连接酶活性,需要两种高度保守的TRIM25的谷氨酸盐(Glu9和Glu10)。因此,产生了缺少E3连接酶活性的突变的TRIM11(Glu12/Glu13至Ala12/Ala13)(图27A和图27B)。体内SUMO化测定显示通过野生型TRIM11以及PML使Atxn1 82Q有效SUMO化(图27A)。为了进一步确认,体外SUMO化测定显示通过TRIM11可以使纯化的Atxn1 82Q显著SUMO化。因此,TRIM11也是Atxn1 82Q的SUMO E3连接酶。在细胞中,如Hsp70一样,TRIM11主要定位在细胞质中(图28A和图28B)。感兴趣地,可以将TRIM11或Hsp70补充至Atxn1 82Q的聚集体(图28C和图28D),表明TRIM11和Atxn1 82Q之间可能存在相互作用。为了测试这点,拉下分析显示TRIM11选择性结合至Atxn1 82Q(图29A)。重要地,TRIM11优先与致病型Atxn1 82Q相互作用,而不是Atxn1 30Q(图29B)。因此,假设TRIM11具有选择性结合至错误折叠的蛋白质的潜力。为了研究该预测,进一步产生天然或变性荧光素酶珠,然后进行拉下试验。如图29C所示,与对照GST蛋白相反,TIRM11特异性结合至变性荧光素酶,这表示TRIM11能够结合错误折叠的蛋白质。

已表明分子伴侣Hsp70可以抑制SCA1小鼠中病理相关的Atxn1 82Q的表达。另外,Hsp70降低了Atxn1 82Q的去污剂不溶性部分的蛋白水平,这与本文所提供的结果一致(图28E)。然后,研究对于聚集体形成,TRIM11和Hsp70之间是否存在相似性。为了研究这点,通过去污剂分级分析Atxn182Q的溶解特征。如图28E所示,如Hsp70一样,TRIM11降低了Atxn1 82Q的去污剂不溶性部分,表明TRIM11可以控制蛋白聚集。此外,MG132处理适当增加不溶性Atxn1 82Q部分(图28E),这与蛋白酶体是控制Atxn182Q聚集体形成所必需的先前报道一致。感兴趣地,当Atxn1 82Q与野生型TRIM11或突变体TRIM11共表达时,与野生型TRIM11相比,突变体TRIM11降低Atxn1 82Q的去污剂不溶性部分的能力较低(图28F),表明TRIM11的E3连接酶活性是Atxn1 82Q聚集体减少所需的。为了进一步测试TRIM11是否对细胞中淀粉样蛋白样聚集体有影响,将野生型或突变体TRIM11转染到细胞中并且ThT染色显示野生型和突变体TRIM11均可以下调淀粉样蛋白样聚集体的水平(图28G)。Atxn1 82Q的稳定过表达适当增加ThT染色(图30A)。类似地,TRIM11还可以降低稳定细胞中Atxn182Q的去污剂不溶性部分(图30B)。重要地,野生型TRIM11降低细胞聚集体的能力比突变体TRIM11更强(图30C和30D)。

由于Hsp70和TRIM11的相似性,假设TRIM11可以用作控制蛋白质聚集的分子伴侣。通常,分子伴侣具有防止易于聚集的错误折叠的蛋白质的能力。这是控制蛋白聚集的最主要和有效的方法。因此,研究了TRIM11在聚集体形成中的预防功能。在没有分子伴侣的情况下,对热冲击响应,荧光素酶活性快速降低(图31A)。然而,TRIM11和Hsp70的温育可以有效保护荧光素酶免受热失活(图31A)。类似地,TRIM11还保护GFP蛋白抵抗热(图31C)。这些结果表明TRIM11可以用作如同Hsp70的分子伴侣。在细胞中,TRIM11的稳定过表达能够适当保护荧光素酶抵抗热冲击(图31C和图31D)。此外,TRIM11还可以明显恢复荧光素酶的热失活(图31C和图31D),这进一步确认了TRIM11的类分子伴侣功能。重要地,TRIM11过表达不会改变Hsp70的蛋白水平(图31E)。然后,将阿尔茨海默病相关β-淀粉样蛋白(1-42)用作底物来研究TRIM11的预防功能。如图31F所示,通过ThT分析,TRIM11可以抑制淀粉样蛋白纤维形成。另外,纯化Atxn1 82Q以测试聚集体形成过程。对照蛋白GST不能防止Atxn1聚集体形成,而TRIM11有效阻断聚集体形成(图31G)。此外,TRIM11还可以防止Atxn1 82Q的淀粉样蛋白样聚集体(图31G)。已知p53易于体外形成聚集体。如图31H所示,TRIM11显著防止p53变性。有趣地,可以通过TRIM11使p53体外SUMO化(图27C)并且p53的SUMO化可以阻断淀粉样蛋白样聚集体,但促进寡聚物形成。总之,TRIM11可以用作分子伴侣蛋白以防止聚集体形成。

为了确定TRIM11是否可以响应热冲击而上调,使用了HCT116细胞稳定表达载体或TRIM11。在对照稳定细胞中,在热冲击后的恢复期间TRIM11增加(图32A)。然而,在TRIM11表达细胞中,不能上调外源TRIM11蛋白(图32B),表明热诱导的TRIM11上调不是由于蛋白质稳定性变化所造成的。在HeLa细胞中进一步确认了热诱导的TRIM11上调(图32C)。由于多种应激可以诱导蛋白错误折叠,检验了是否可以通过其它应激来诱导TRIM11。如图30D和30E所示,TRIM11的蛋白水平响应于As2O3和H2O2处理而上调。可以响应于热冲击而诱导TRIM11的mRNA水平(图32F),这与Hsp70相似。为了研究热冲击诱导TRIM11的机制,产生了稳定表达HSF1的A549细胞,HSF1是控制热响应性蛋白的关键转录因子。出乎意料地,在有或没有热冲击处理的情况下,HSF1的过表达下调TRIM11的蛋白水平(图32G),表明对于TRIM11转录的调控,可能不需要HSF1。

可以在热冲击响应期间激活另一种关键转录因子p53。p53还能够直接增加TRIM的转录,例如TRIM21和TRIM24。因此,研究了是否可以通过p53对热冲击响应而控制TRIM11。如图33A所示,在p53野生型中TRIM11响应于热冲击而增加,但是在无p53的HCT116细胞中不增加。因此,TRIM11的mRNA水平仅在p53野生型HCT116细胞中增强(图33B),表明p53有助于TRIM11转录。为了进一步确认p53的重要性,在稳定表达p53shRNA的A549细胞中,不能上调TRIM11的蛋白和mRNA水平(图33C)。在p53降低的HCT116细胞中再次确认了类似的现象(图33D)。总的来说,这些结果强烈表示p53可能是响应于热冲击而上调TRIM11的关键因素。HSF1被认为是热应激后对细胞存活的保护措施。化学抑制剂KRIBB11可以用于抑制HSF1活性。如图33E和图33F所示,通过KRIBB11处理,无p53的细胞对热冲击更敏感,这表示p53还有助于对热应激响应的细胞存活。

Hsp70可以促进某些种类的蛋白聚集体溶解。然后,研究TRIM11是否可以解聚蛋白聚集体。如预期的,TRIM11从不溶性聚集体再次溶解了热失活的荧光素酶(图34A)并且以剂量依赖性方式恢复荧光素酶活性(图34B)。另外,使用预形成的GFP聚集体作为底物进一步确认了TRIM11的溶解功能(图34C和图34D)。这些结果表明TRIM11可以解聚失调的聚集体。然后,使用Atxn1 82Q聚集体作为底物进行测定。如图34E所示,TRIM11可以有效地再溶解Atxn1 82Q聚集体。值得注意地,Hsp70/Hsp40二分子伴侣仅将Atxn1 82Q的淀粉样蛋白样结构解聚成颗粒(图34F),表明TRIM11可能以不同方式起作用。如预期的,TRIM11和Hsp104将淀粉样蛋白样结构显著恢复为上清液(图34G)。

为了确定TRIM11的哪个功能域是TRIM11解聚活性所需要的,将加热的荧光素酶聚集体用作底物。如图35B所示,TRIM11可以有效恢复荧光素酶活性的热失活,但是TRIM11的RBC或B30.2片段具有较弱的再激活能力。一致地,在沉降测定中,TRIM11再溶解预形成的荧光素酶聚集体的活性比其两个片段更强(图35C)。此外,与全长TRIM11相比,每个单独的域几乎丢失了再折叠活性(图35D)。此外,全长TRIM11与Atxn1 82Q的相互作用非常强烈,但是RBC或B30.2不结合至Atxn1 82Q(图36A)。类似地,TRIM11的单独的域不结合至Atxn1 82Q(图36B)。这些结果强烈表明TRIM11的解聚功能可能需要与底物结合。

然后,研究了蛋白解聚是否需要SUMO E3连接酶活性。此处将突变体TRIM11用于测试其解聚活性。如图37A所示,如同野生型TRIM11,突变体TRIM11维持了防止荧光素酶热失活的能力。此外,突变体TRIM11还具有与野生型TRIM11类似的恢复变性荧光素酶活性的潜能(图37B)。突变体TRIM11仍能够选择性结合至变性荧光素酶(图37C)。所有这些结果表明TRIM11不依赖其E3活性在体外发挥其解聚功能。值得注意地,突变体TRIM11基本上定位在细胞核中(图37D),这与野生型TRIM11不同。此外,突变体TRIM11还可以与Atxn1 82Q共定位。

先前的研究显示多细胞动物蛋白解聚酶系统Hsp110、Hsp70和Hsp40不能有效解聚淀粉样蛋白。因此,研究了TRIM11是否可以解聚淀粉样蛋白纤维。此处作为应用α-突触核蛋白作为客体。首先,确定TRIM11是否还可以防止α-突触核蛋白纤维形成。如图38A所示,TRIM11以及Hsp70和Hsp104有效抑制α-突触核蛋白淀粉样蛋白纤维的形成(图38A)。另外,抑制是剂量依赖性的方式(图38B)。事实上,电子显微镜(EM)显示通过对照GST蛋白显示了α-突触核蛋白纤维,而通过TRIM11未观察到纤维(图38C)。然后,将α-突触核蛋白纤维用于研究TRIM11的解聚。如图38D所示,通过TRIM11或突变体Hsp104处理,α-突触核蛋白纤维的溶解以剂量依赖性方式增加。因此,使用TRIM11或Hsp104处理,通过ThT荧光,纤维解体是显而易见的(图38E)。

存在超过70个具有共同N端和不同C端的TRIM家族成员。因此,随后研究了其它TRIM蛋白是否如同TRIM11具有类似的功能。因此,选择了TRIM21,因为其与TRIM11共有相同的域。体外拉下试验显示TRIM21也优先结合至变性荧光素酶(图39A)。类似地,TRIM21可以适当溶解加热的荧光素酶聚集体(图39B)并恢复荧光素酶的热失活(图39C)。TRIM21也能够保护荧光素酶抵抗热失活(图39D)。PML可以通过蛋白酶体降解Atxn1 82Q聚集体。因此,为了研究Atxn1聚集体的解聚和降解是否通过PML相联系,从293T细胞纯化PML(图40A)。如图40B所示,PML可以以剂量依赖性方式恢复GFP荧光的热失活。然而,恢复能力弱于TRIM11(图40C)。还从293T细胞纯化PML的一些片段(图40D)。感兴趣地,PML的一个片段(361-633)可能是加热的GFP聚集体再激活所必需的(图40E)。

为了更好地理解TRIM11在控制α-突触核蛋白聚集体形成中的作用,将小鼠原代海马神经元用作模型。与GST或TRIM11一起体外产生α-突触核蛋白原纤维(PEF)(GST-PEF和TRIM11-PEF)。加入PEF后两周,通过PEF聚集体诱导约30%神经元的死亡(图41A)。然而,TRIM11-PEF对神经元具有较低的毒性(图41A)。事实上,免疫荧光显示存在概括了帕金森氏症脑中路易氏小体特征的显著的α-突触核蛋白聚集体,而在加入TRIM11-PEF后没有大的聚集体(图41B),这进一步显示TRIM11具有防止α-突触核蛋白纤维形成的功能。值得注意地,在两类神经元(皮层和海马神经元)中,可以对热冲击响应上调TRIM11(图42A和图42B)。此外,在海马神经元中TRIM11的mRNA水平也得到增加(图42C)。有趣地,TRIM11可能参与控制神经退行性疾病。

本文引用的每一个专利、专利申请和专利公开的公开以其全部内容通过引证并入本文。尽管已参考具体实施方式公开了本发明,但显而易见的是本领域技术人员在不背离本发明的真实精神和范围的情况下可以设计本发明的其它实施方式和变化。所附权利要求旨在视为包含所有这些实施方式和等价变化。

序列表

<110> 宾夕法尼亚大学理事会(The Trustees Of The University Of Pennsylvania)

杨小鲁(Yang, Xiaolu)

郭骊骊(Guo, Lili)

<120> 用于降解错误折叠蛋白的组合物和方法

<130> 046483-6149-00-WO.605285

<150> US 62/168,309

<151> 2015-05-29

<160> 169

<170> PatentIn version 3.5

<210> 1

<211> 2208

<212> DNA

<213> 智人(Homo sapiens)

<400> 1

atgggtgaaa gcccagcctc cgtggttctt aatgcctcag gaggactatt ttcactaaag 60

atggaaacac tggagtctga attgacctgt ccaatctgcc tagagttgtt tgaagacccc 120

cttctgctcc cttgtgctca cagcctctgc ttcagctgtg cccatcgcat tttggtatca 180

agctgcagct ctggtgaatc cattgaaccc attactgctt tccagtgtcc tacctgcagg 240

tatgttatct cgctgaacca ccggggcctg gatggcctca agaggaatgt gactctgcag 300

aacattattg atcgcttcca gaaggcttca gtcagtgggc ccaattcccc tagtgagagc 360

cgccgggaaa ggacttacag gcccaccact gccatgtcta gcgagcgaat tgcttgccaa 420

ttctgtgagc aggacccgcc aagggatgca gtaaaaacat gcatcacctg tgaggtctcc 480

tactgtgacc gttgcctgcg ggccacgcac cccaacaaga aacctttcac cagccaccgc 540

ctggtggaac cagtgccaga cacacatctt cgagggatca cctgcctgga ccatgagaat 600

gagaaagtga acatgtactg tgtatctgat gaccaattga tctgtgcctt atgcaaactg 660

gtgggtcgtc accgagacca tcaggtcgca tccctgaatg atcgatttga gaaactcaag 720

caaactctgg agatgaacct caccaacctg gttaagcgca acagcgaact agaaaatcaa 780

atggccaaac taatacagat ctgccagcag gttgaggtga atactgctat gcatgaggca 840

aaacttatgg aagaatgtga cgagttggta gagatcatcc agcagaggaa gcaaatgatc 900

gctgtcaaaa tcaaagagac aaaggttatg aaactgagaa agttggcaca gcaggttgct 960

aattgccgcc agtgtcttga acggtcaaca gtcctcatca accaagctga gcatatcctg 1020

aaagaaaatg accaggcacg gtttctacag tctgcaaaaa atattgctga gagggtcgct 1080

atggcaactg catcttctca agttctgatt ccagacatca attttaatga tgcctttgaa 1140

aactttgctt tagatttttc cagagaaaag aaactgctag aggggttaga ttatttaaca 1200

gccccaaacc caccatctat ccgagaagaa ctctgtactg cctcccatga caccattaca 1260

gtccactgga tctcggatga tgagttcagc atcagctcct atgagcttca gtacaccata 1320

ttcactggcc aggctaactt catcagtaag tcatggtgta gttggggcct gtggccagag 1380

ataaggaaat gtaaggaagc agtaagctgc tcaagattgg ccggggcgcc acgaggcctg 1440

tataattcag tagacagctg gatgattgtt cccaacatta aacagaacca ttacacagtg 1500

catggactcc agagcgggac tcgctacatc ttcatcgtta aagccataaa ccaagccggc 1560

agccggaaca gtgaacctac ccgactaaaa acaaacagcc aaccctttaa attggatccc 1620

aaaatgactc acaagaagtt gaagatctcc aatgatggat tgcagatgga gaaggatgaa 1680

agctctctaa agaagagcca caccccagag aggtttagtg gcacagggtg ctatggggca 1740

gcaggaaata tattcattga cagtggctgc cactattggg aggtggtcat gggttcctca 1800

acatggtatg caattggcat tgcctacaaa tcagctccaa agaatgaatg gattggcaag 1860

aatgcctcct catgggtctt ctctcgctgc aatagtaact tcgtggtgag acacaacaac 1920

aaggaaatgc tggtggatgt gcccccacac ctgaagcgtc tgggtgtcct cctggattat 1980

gacaacaata tgctgtcttt ctatgaccca gctaactctc tccatcttca tacttttgat 2040

gtgaccttca ttcttccagt ttgtccaaca tttacaatct ggaacaaatc cctaatgatc 2100

ctgtctggct tgcctgcccc agattttatt gattaccctg agcggcagga atgcaactgc 2160

aggcctcaag aatcccctta tgtttctggg atgaaaacct gtcattaa 2208

<210> 2

<211> 735

<212> PRT

<213> 智人(Homo sapiens)

<400> 2

Met Gly Glu Ser Pro Ala Ser Val Val Leu Asn Ala Ser Gly Gly Leu

1 5 10 15

Phe Ser Leu Lys Met Glu Thr Leu Glu Ser Glu Leu Thr Cys Pro Ile

20 25 30

Cys Leu Glu Leu Phe Glu Asp Pro Leu Leu Leu Pro Cys Ala His Ser

35 40 45

Leu Cys Phe Ser Cys Ala His Arg Ile Leu Val Ser Ser Cys Ser Ser

50 55 60

Gly Glu Ser Ile Glu Pro Ile Thr Ala Phe Gln Cys Pro Thr Cys Arg

65 70 75 80

Tyr Val Ile Ser Leu Asn His Arg Gly Leu Asp Gly Leu Lys Arg Asn

85 90 95

Val Thr Leu Gln Asn Ile Ile Asp Arg Phe Gln Lys Ala Ser Val Ser

100 105 110

Gly Pro Asn Ser Pro Ser Glu Ser Arg Arg Glu Arg Thr Tyr Arg Pro

115 120 125

Thr Thr Ala Met Ser Ser Glu Arg Ile Ala Cys Gln Phe Cys Glu Gln

130 135 140

Asp Pro Pro Arg Asp Ala Val Lys Thr Cys Ile Thr Cys Glu Val Ser

145 150 155 160

Tyr Cys Asp Arg Cys Leu Arg Ala Thr His Pro Asn Lys Lys Pro Phe

165 170 175

Thr Ser His Arg Leu Val Glu Pro Val Pro Asp Thr His Leu Arg Gly

180 185 190

Ile Thr Cys Leu Asp His Glu Asn Glu Lys Val Asn Met Tyr Cys Val

195 200 205

Ser Asp Asp Gln Leu Ile Cys Ala Leu Cys Lys Leu Val Gly Arg His

210 215 220

Arg Asp His Gln Val Ala Ser Leu Asn Asp Arg Phe Glu Lys Leu Lys

225 230 235 240

Gln Thr Leu Glu Met Asn Leu Thr Asn Leu Val Lys Arg Asn Ser Glu

245 250 255

Leu Glu Asn Gln Met Ala Lys Leu Ile Gln Ile Cys Gln Gln Val Glu

260 265 270

Val Asn Thr Ala Met His Glu Ala Lys Leu Met Glu Glu Cys Asp Glu

275 280 285

Leu Val Glu Ile Ile Gln Gln Arg Lys Gln Met Ile Ala Val Lys Ile

290 295 300

Lys Glu Thr Lys Val Met Lys Leu Arg Lys Leu Ala Gln Gln Val Ala

305 310 315 320

Asn Cys Arg Gln Cys Leu Glu Arg Ser Thr Val Leu Ile Asn Gln Ala

325 330 335

Glu His Ile Leu Lys Glu Asn Asp Gln Ala Arg Phe Leu Gln Ser Ala

340 345 350

Lys Asn Ile Ala Glu Arg Val Ala Met Ala Thr Ala Ser Ser Gln Val

355 360 365

Leu Ile Pro Asp Ile Asn Phe Asn Asp Ala Phe Glu Asn Phe Ala Leu

370 375 380

Asp Phe Ser Arg Glu Lys Lys Leu Leu Glu Gly Leu Asp Tyr Leu Thr

385 390 395 400

Ala Pro Asn Pro Pro Ser Ile Arg Glu Glu Leu Cys Thr Ala Ser His

405 410 415

Asp Thr Ile Thr Val His Trp Ile Ser Asp Asp Glu Phe Ser Ile Ser

420 425 430

Ser Tyr Glu Leu Gln Tyr Thr Ile Phe Thr Gly Gln Ala Asn Phe Ile

435 440 445

Ser Lys Ser Trp Cys Ser Trp Gly Leu Trp Pro Glu Ile Arg Lys Cys

450 455 460

Lys Glu Ala Val Ser Cys Ser Arg Leu Ala Gly Ala Pro Arg Gly Leu

465 470 475 480

Tyr Asn Ser Val Asp Ser Trp Met Ile Val Pro Asn Ile Lys Gln Asn

485 490 495

His Tyr Thr Val His Gly Leu Gln Ser Gly Thr Arg Tyr Ile Phe Ile

500 505 510

Val Lys Ala Ile Asn Gln Ala Gly Ser Arg Asn Ser Glu Pro Thr Arg

515 520 525

Leu Lys Thr Asn Ser Gln Pro Phe Lys Leu Asp Pro Lys Met Thr His

530 535 540

Lys Lys Leu Lys Ile Ser Asn Asp Gly Leu Gln Met Glu Lys Asp Glu

545 550 555 560

Ser Ser Leu Lys Lys Ser His Thr Pro Glu Arg Phe Ser Gly Thr Gly

565 570 575

Cys Tyr Gly Ala Ala Gly Asn Ile Phe Ile Asp Ser Gly Cys His Tyr

580 585 590

Trp Glu Val Val Met Gly Ser Ser Thr Trp Tyr Ala Ile Gly Ile Ala

595 600 605

Tyr Lys Ser Ala Pro Lys Asn Glu Trp Ile Gly Lys Asn Ala Ser Ser

610 615 620

Trp Val Phe Ser Arg Cys Asn Ser Asn Phe Val Val Arg His Asn Asn

625 630 635 640

Lys Glu Met Leu Val Asp Val Pro Pro His Leu Lys Arg Leu Gly Val

645 650 655

Leu Leu Asp Tyr Asp Asn Asn Met Leu Ser Phe Tyr Asp Pro Ala Asn

660 665 670

Ser Leu His Leu His Thr Phe Asp Val Thr Phe Ile Leu Pro Val Cys

675 680 685

Pro Thr Phe Thr Ile Trp Asn Lys Ser Leu Met Ile Leu Ser Gly Leu

690 695 700

Pro Ala Pro Asp Phe Ile Asp Tyr Pro Glu Arg Gln Glu Cys Asn Cys

705 710 715 720

Arg Pro Gln Glu Ser Pro Tyr Val Ser Gly Met Lys Thr Cys His

725 730 735

<210> 3

<211> 2316

<212> DNA

<213> 智人(Homo sapiens)

<400> 3

atgcacagga gtggccgtta tggaacgcag cagcagcgtg cagggtcaaa gacagccggc 60

cccccatgtc agtggtctag gatggccagt gaaggcacca acatcccaag tcctgtggtg 120

cgccagattg acaagcagtt tctgatttgc agtatatgcc tggaacggta caagaatccc 180

aaggttctcc cctgtctgca cactttctgc gagaggtgcc tgcagaacta cattcctgcc 240

cacagtttaa ccctctcctg cccagtgtgc cgccagacct ccatcctgcc cgagaaaggg 300

gtggccgcgc tccagaacaa tttcttcatc acaaacctga tggacgtgct gcagcgaact 360

ccaggcagca acgctgagga gtcttccatc ctggagacag tcactgctgt ggctgcggga 420

aagcctctct cttgcccaaa ccacgatggg aatgtgatgg aattttactg ccagtcctgt 480

gagactgcca tgtgtcggga gtgcacggag ggggagcacg cagagcaccc cacagttcca 540

ctcaaggatg tggtggaaca gcacaaggcc tcgctccagg tccagctgga tgctgtcaac 600

aaaaggctcc cagaaataga ttctgctctt cagttcatct ctgaaatcat tcatcagtta 660

accaaccaaa aggccagcat cgtggatgac attcattcca cctttgatga gctccagaag 720

actttaaatg tgcgcaagag tgtgctgctt atggaattgg aggtcaacta tggcctcaaa 780

cacaaagtcc tccagtcgca gctggatact ctgctccagg ggcaggagag cattaagagc 840

tgcagcaact tcacagcgca ggccctcaac catggcacgg agaccgaggt cctactggtg 900

aagaagcaga tgagcgagaa gctgaacgag ctggccgacc aggacttccc cttgcacccg 960

cgggagaacg accagctgga tttcatcgtg gaaaccgagg ggctgaagaa gtccatccac 1020

aacctcggga cgatcttaac caccaacgcc gttgcctcag agacagtggc cacgggcgag 1080

gggctgcggc agaccatcat cgggcagccc atgtccgtca ccatcaccac caaggacaaa 1140

gacggtgagc tgtgcaaaac cggcaacgcc tacctcaccg ccgaactgag cacccccgac 1200

gggagcgtgg cagacgggga gatcctggac aacaagaacg gcacctatga gtttttgtac 1260

actgtccaga aggaagggga ctttaccctg tctctgagac tctatgacca gcacatccga 1320

ggcagcccgt ttaagctgaa agtgatccga tccgctgatg tgtctcccac cacagaaggc 1380

gtgaagaggc gcgttaagtc cccggggagc ggccacgtca agcagaaagc tgtgaaaaga 1440

cccgcaagca tgtacagcac tggaaaacga aaagagaatc ccatcgaaga cgatttgatc 1500

tttcgagtgg gtaccaaagg aagaaataaa ggagagttta caaatcttca gggggtagct 1560

gcatctacaa atggaaagat attaattgca gacagtaaca accaatgtgt gcagatattt 1620

tccaatgatg gccagttcaa aagtcgtttt ggcatacggg gacgctctcc ggggcagctg 1680

cagcggccca caggagtggc tgtacatccc agtggggaca taatcattgc cgattatgat 1740

aataaatggg tcagcatttt ctcctccgat gggaaattta agacaaaaat tggatcagga 1800

aagctgatgg gacccaaagg agtttctgtg gaccgcaatg ggcacattat tgttgtggac 1860

aacaaggcgt gctgcgtgtt tatcttccag ccaaacggga aaatagtcac caggtttggt 1920

agccgaggaa atggggacag gcagtttgca ggtccccatt ttgcagctgt aaatagcaat 1980

aatgagatta ttattacaga tttccataat cattctgtca aggtgtttaa tcaggaagga 2040

gaattcatgt tgaagtttgg ctcaaatgga gaaggaaatg ggcagtttaa tgctccaaca 2100

ggtgtagcag tggattcaaa tggaaacatc attgtggccg actggggaaa cagcaggatc 2160

caggtttttg atgggagtgg atcatttttg tcctacatta acacatctgc tgacccactc 2220

tatggccccc aaggcctggc cctaacttca gatggtcatg ttgtggttgc agactctgga 2280

aatcactgtt tcaaagtcta tcgatactta cagtaa 2316

<210> 4

<211> 771

<212> PRT

<213> 智人(Homo sapiens)

<400> 4

Met His Arg Ser Gly Arg Tyr Gly Thr Gln Gln Gln Arg Ala Gly Ser

1 5 10 15

Lys Thr Ala Gly Pro Pro Cys Gln Trp Ser Arg Met Ala Ser Glu Gly

20 25 30

Thr Asn Ile Pro Ser Pro Val Val Arg Gln Ile Asp Lys Gln Phe Leu

35 40 45

Ile Cys Ser Ile Cys Leu Glu Arg Tyr Lys Asn Pro Lys Val Leu Pro

50 55 60

Cys Leu His Thr Phe Cys Glu Arg Cys Leu Gln Asn Tyr Ile Pro Ala

65 70 75 80

His Ser Leu Thr Leu Ser Cys Pro Val Cys Arg Gln Thr Ser Ile Leu

85 90 95

Pro Glu Lys Gly Val Ala Ala Leu Gln Asn Asn Phe Phe Ile Thr Asn

100 105 110

Leu Met Asp Val Leu Gln Arg Thr Pro Gly Ser Asn Ala Glu Glu Ser

115 120 125

Ser Ile Leu Glu Thr Val Thr Ala Val Ala Ala Gly Lys Pro Leu Ser

130 135 140

Cys Pro Asn His Asp Gly Asn Val Met Glu Phe Tyr Cys Gln Ser Cys

145 150 155 160

Glu Thr Ala Met Cys Arg Glu Cys Thr Glu Gly Glu His Ala Glu His

165 170 175

Pro Thr Val Pro Leu Lys Asp Val Val Glu Gln His Lys Ala Ser Leu

180 185 190

Gln Val Gln Leu Asp Ala Val Asn Lys Arg Leu Pro Glu Ile Asp Ser

195 200 205

Ala Leu Gln Phe Ile Ser Glu Ile Ile His Gln Leu Thr Asn Gln Lys

210 215 220

Ala Ser Ile Val Asp Asp Ile His Ser Thr Phe Asp Glu Leu Gln Lys

225 230 235 240

Thr Leu Asn Val Arg Lys Ser Val Leu Leu Met Glu Leu Glu Val Asn

245 250 255

Tyr Gly Leu Lys His Lys Val Leu Gln Ser Gln Leu Asp Thr Leu Leu

260 265 270

Gln Gly Gln Glu Ser Ile Lys Ser Cys Ser Asn Phe Thr Ala Gln Ala

275 280 285

Leu Asn His Gly Thr Glu Thr Glu Val Leu Leu Val Lys Lys Gln Met

290 295 300

Ser Glu Lys Leu Asn Glu Leu Ala Asp Gln Asp Phe Pro Leu His Pro

305 310 315 320

Arg Glu Asn Asp Gln Leu Asp Phe Ile Val Glu Thr Glu Gly Leu Lys

325 330 335

Lys Ser Ile His Asn Leu Gly Thr Ile Leu Thr Thr Asn Ala Val Ala

340 345 350

Ser Glu Thr Val Ala Thr Gly Glu Gly Leu Arg Gln Thr Ile Ile Gly

355 360 365

Gln Pro Met Ser Val Thr Ile Thr Thr Lys Asp Lys Asp Gly Glu Leu

370 375 380

Cys Lys Thr Gly Asn Ala Tyr Leu Thr Ala Glu Leu Ser Thr Pro Asp

385 390 395 400

Gly Ser Val Ala Asp Gly Glu Ile Leu Asp Asn Lys Asn Gly Thr Tyr

405 410 415

Glu Phe Leu Tyr Thr Val Gln Lys Glu Gly Asp Phe Thr Leu Ser Leu

420 425 430

Arg Leu Tyr Asp Gln His Ile Arg Gly Ser Pro Phe Lys Leu Lys Val

435 440 445

Ile Arg Ser Ala Asp Val Ser Pro Thr Thr Glu Gly Val Lys Arg Arg

450 455 460

Val Lys Ser Pro Gly Ser Gly His Val Lys Gln Lys Ala Val Lys Arg

465 470 475 480

Pro Ala Ser Met Tyr Ser Thr Gly Lys Arg Lys Glu Asn Pro Ile Glu

485 490 495

Asp Asp Leu Ile Phe Arg Val Gly Thr Lys Gly Arg Asn Lys Gly Glu

500 505 510

Phe Thr Asn Leu Gln Gly Val Ala Ala Ser Thr Asn Gly Lys Ile Leu

515 520 525

Ile Ala Asp Ser Asn Asn Gln Cys Val Gln Ile Phe Ser Asn Asp Gly

530 535 540

Gln Phe Lys Ser Arg Phe Gly Ile Arg Gly Arg Ser Pro Gly Gln Leu

545 550 555 560

Gln Arg Pro Thr Gly Val Ala Val His Pro Ser Gly Asp Ile Ile Ile

565 570 575

Ala Asp Tyr Asp Asn Lys Trp Val Ser Ile Phe Ser Ser Asp Gly Lys

580 585 590

Phe Lys Thr Lys Ile Gly Ser Gly Lys Leu Met Gly Pro Lys Gly Val

595 600 605

Ser Val Asp Arg Asn Gly His Ile Ile Val Val Asp Asn Lys Ala Cys

610 615 620

Cys Val Phe Ile Phe Gln Pro Asn Gly Lys Ile Val Thr Arg Phe Gly

625 630 635 640

Ser Arg Gly Asn Gly Asp Arg Gln Phe Ala Gly Pro His Phe Ala Ala

645 650 655

Val Asn Ser Asn Asn Glu Ile Ile Ile Thr Asp Phe His Asn His Ser

660 665 670

Val Lys Val Phe Asn Gln Glu Gly Glu Phe Met Leu Lys Phe Gly Ser

675 680 685

Asn Gly Glu Gly Asn Gly Gln Phe Asn Ala Pro Thr Gly Val Ala Val

690 695 700

Asp Ser Asn Gly Asn Ile Ile Val Ala Asp Trp Gly Asn Ser Arg Ile

705 710 715 720

Gln Val Phe Asp Gly Ser Gly Ser Phe Leu Ser Tyr Ile Asn Thr Ser

725 730 735

Ala Asp Pro Leu Tyr Gly Pro Gln Gly Leu Ala Leu Thr Ser Asp Gly

740 745 750

His Val Val Val Ala Asp Ser Gly Asn His Cys Phe Lys Val Tyr Arg

755 760 765

Tyr Leu Gln

770

<210> 5

<211> 2235

<212> DNA

<213> 智人(Homo sapiens)

<400> 5

atggcaaaga gggaggacag ccctggccca gaggtccagc caatggacaa gcagttcctg 60

gtatgcagca tctgcctgga tcggtaccag tgccccaagg ttcttccttg cctgcacacc 120

ttctgtgaga gatgtctcca aaactatatc cctgcccaga gcctgacgct atcctgtcca 180

gtatgccggc agacgtccat cctcccagag cagggcgtct cggcactgca gaacaacttc 240

ttcatcagca gcctcatgga ggcaatgcag caggcacctg atggggccca cgacccggag 300

gacccccacc ccctcagtgt agtggctggc cgccctctct cctgccccaa ccatgaaggc 360

aagacgatgg agttttactg tgaggcctgt gagacggcca tgtgtggtga gtgccgcgcc 420

ggggagcatc gtgagcatgg cacagtgctg ctgagggatg tggtggagca gcacaaggcg 480

gccctgcagc gccagctcga ggctgtgcgt ggccgattgc cacagctgtc cgcagcaatt 540

gccttagtcg ggggcatcag ccagcagctg caggagcgca aggcagaggc cctggcccag 600

atcagtgcag cgttcgagga cctggagcaa gcactgcagc agcgcaagca ggctctggtc 660

agcgacctgg agaccatttg tggggccaaa cagaaggtgt tgcaaagcca gctggacaca 720

ctgcgccagg gtcaggaaca catcggcagt agctgcagct ttgcagagca ggcactgcgc 780

ctgggctcgg ccccggaggt gttgctggtg cgcaagcaca tgcgagagcg gctggctgca 840

ttggcggcac aggccttccc ggagcggcca catgagaatg cacagctgga actggtcctt 900

gaggtggacg gtctgcggcg atcggtgctc aatctgggcg cactgctcac cacgagcgcc 960

actgcacacg aaacggtggc cacgggagag ggcctgcgcc aggcgctagt gggccagcct 1020

gcctcgctca ctgtcactac caaagacaag gacgggcggt tggtgcgcac aggcagcgct 1080

gagctgcgtg cagagatcac cggcccggac ggcacgcgcc ttccggtgcc agtggtggac 1140

cacaagaatg gcacatatga gctagtgtac acagcgcgca cggaaggcga gctgctcctc 1200

tcggtgctgc tctacggaca gccagtgcgc ggcagcccct tccgcgtgcg tgccctgcgt 1260

ccgggggacc tgccaccttc cccggacgat gtgaagcgcc gtgtcaagtc ccctggcggc 1320

cccggcagcc atgtgcgcca gaaggcagtg cgtaggccca gctccatgta cagcacaggc 1380

ggcaaacgaa aggacaaccc aattgaggat gagctcgtct tccgtgttgg cagtcgtgga 1440

agggagaaag gtgaattcac caatttacaa ggtgtgtccg cagccagcag cggccgcatc 1500

gtggtagcag acagcaacaa ccagtgtatt caggttttct ccaatgaggg ccagttcaag 1560

ttccgttttg gggtccgagg acgctcacct gggcagctgc agcgccccac aggtgtggca 1620

gtggacacca atggagacat aattgtggca gactatgaca accgttgggt cagcatcttc 1680

tcccctgagg gcaagttcaa gaccaagatt ggagctggcc gcctcatggg ccccaaggga 1740

gtggccgtag accggaatgg acatatcatt gtggtcgaca acaagtcttg ctgcgtcttt 1800

accttccagc ccaatggcaa actggttggc cgttttgggg gccgtggggc cactgaccgc 1860

cactttgcag ggccccattt tgtggctgtg aacaacaaga atgaaattgt agtaacggac 1920

ttccataacc attcagtgaa ggtgtacagt gccgatggag agttcctctt caagtttggc 1980

tcccatggcg agggcaatgg gcagttcaat gcccccacag gagtagctgt ggactccaat 2040

ggaaacatca ttgtggctga ctggggcaac agccgcatcc aggtattcga cagctctggc 2100

tccttcctgt cctatatcaa cacatctgca gaaccactgt atggtccaca gggcctggca 2160

ctgacctcgg atggccatgt ggtggtggct gatgctggca accactgctt taaagcctat 2220

cgctacctcc agtag 2235

<210> 6

<211> 744

<212> PRT

<213> 智人(Homo sapiens)

<400> 6

Met Ala Lys Arg Glu Asp Ser Pro Gly Pro Glu Val Gln Pro Met Asp

1 5 10 15

Lys Gln Phe Leu Val Cys Ser Ile Cys Leu Asp Arg Tyr Gln Cys Pro

20 25 30

Lys Val Leu Pro Cys Leu His Thr Phe Cys Glu Arg Cys Leu Gln Asn

35 40 45

Tyr Ile Pro Ala Gln Ser Leu Thr Leu Ser Cys Pro Val Cys Arg Gln

50 55 60

Thr Ser Ile Leu Pro Glu Gln Gly Val Ser Ala Leu Gln Asn Asn Phe

65 70 75 80

Phe Ile Ser Ser Leu Met Glu Ala Met Gln Gln Ala Pro Asp Gly Ala

85 90 95

His Asp Pro Glu Asp Pro His Pro Leu Ser Val Val Ala Gly Arg Pro

100 105 110

Leu Ser Cys Pro Asn His Glu Gly Lys Thr Met Glu Phe Tyr Cys Glu

115 120 125

Ala Cys Glu Thr Ala Met Cys Gly Glu Cys Arg Ala Gly Glu His Arg

130 135 140

Glu His Gly Thr Val Leu Leu Arg Asp Val Val Glu Gln His Lys Ala

145 150 155 160

Ala Leu Gln Arg Gln Leu Glu Ala Val Arg Gly Arg Leu Pro Gln Leu

165 170 175

Ser Ala Ala Ile Ala Leu Val Gly Gly Ile Ser Gln Gln Leu Gln Glu

180 185 190

Arg Lys Ala Glu Ala Leu Ala Gln Ile Ser Ala Ala Phe Glu Asp Leu

195 200 205

Glu Gln Ala Leu Gln Gln Arg Lys Gln Ala Leu Val Ser Asp Leu Glu

210 215 220

Thr Ile Cys Gly Ala Lys Gln Lys Val Leu Gln Ser Gln Leu Asp Thr

225 230 235 240

Leu Arg Gln Gly Gln Glu His Ile Gly Ser Ser Cys Ser Phe Ala Glu

245 250 255

Gln Ala Leu Arg Leu Gly Ser Ala Pro Glu Val Leu Leu Val Arg Lys

260 265 270

His Met Arg Glu Arg Leu Ala Ala Leu Ala Ala Gln Ala Phe Pro Glu

275 280 285

Arg Pro His Glu Asn Ala Gln Leu Glu Leu Val Leu Glu Val Asp Gly

290 295 300

Leu Arg Arg Ser Val Leu Asn Leu Gly Ala Leu Leu Thr Thr Ser Ala

305 310 315 320

Thr Ala His Glu Thr Val Ala Thr Gly Glu Gly Leu Arg Gln Ala Leu

325 330 335

Val Gly Gln Pro Ala Ser Leu Thr Val Thr Thr Lys Asp Lys Asp Gly

340 345 350

Arg Leu Val Arg Thr Gly Ser Ala Glu Leu Arg Ala Glu Ile Thr Gly

355 360 365

Pro Asp Gly Thr Arg Leu Pro Val Pro Val Val Asp His Lys Asn Gly

370 375 380

Thr Tyr Glu Leu Val Tyr Thr Ala Arg Thr Glu Gly Glu Leu Leu Leu

385 390 395 400

Ser Val Leu Leu Tyr Gly Gln Pro Val Arg Gly Ser Pro Phe Arg Val

405 410 415

Arg Ala Leu Arg Pro Gly Asp Leu Pro Pro Ser Pro Asp Asp Val Lys

420 425 430

Arg Arg Val Lys Ser Pro Gly Gly Pro Gly Ser His Val Arg Gln Lys

435 440 445

Ala Val Arg Arg Pro Ser Ser Met Tyr Ser Thr Gly Gly Lys Arg Lys

450 455 460

Asp Asn Pro Ile Glu Asp Glu Leu Val Phe Arg Val Gly Ser Arg Gly

465 470 475 480

Arg Glu Lys Gly Glu Phe Thr Asn Leu Gln Gly Val Ser Ala Ala Ser

485 490 495

Ser Gly Arg Ile Val Val Ala Asp Ser Asn Asn Gln Cys Ile Gln Val

500 505 510

Phe Ser Asn Glu Gly Gln Phe Lys Phe Arg Phe Gly Val Arg Gly Arg

515 520 525

Ser Pro Gly Gln Leu Gln Arg Pro Thr Gly Val Ala Val Asp Thr Asn

530 535 540

Gly Asp Ile Ile Val Ala Asp Tyr Asp Asn Arg Trp Val Ser Ile Phe

545 550 555 560

Ser Pro Glu Gly Lys Phe Lys Thr Lys Ile Gly Ala Gly Arg Leu Met

565 570 575

Gly Pro Lys Gly Val Ala Val Asp Arg Asn Gly His Ile Ile Val Val

580 585 590

Asp Asn Lys Ser Cys Cys Val Phe Thr Phe Gln Pro Asn Gly Lys Leu

595 600 605

Val Gly Arg Phe Gly Gly Arg Gly Ala Thr Asp Arg His Phe Ala Gly

610 615 620

Pro His Phe Val Ala Val Asn Asn Lys Asn Glu Ile Val Val Thr Asp

625 630 635 640

Phe His Asn His Ser Val Lys Val Tyr Ser Ala Asp Gly Glu Phe Leu

645 650 655

Phe Lys Phe Gly Ser His Gly Glu Gly Asn Gly Gln Phe Asn Ala Pro

660 665 670

Thr Gly Val Ala Val Asp Ser Asn Gly Asn Ile Ile Val Ala Asp Trp

675 680 685

Gly Asn Ser Arg Ile Gln Val Phe Asp Ser Ser Gly Ser Phe Leu Ser

690 695 700

Tyr Ile Asn Thr Ser Ala Glu Pro Leu Tyr Gly Pro Gln Gly Leu Ala

705 710 715 720

Leu Thr Ser Asp Gly His Val Val Val Ala Asp Ala Gly Asn His Cys

725 730 735

Phe Lys Ala Tyr Arg Tyr Leu Gln

740

<210> 7

<211> 1503

<212> DNA

<213> 智人(Homo sapiens)

<400> 7

atggaagctg aggacatcca ggaggagttg acctgcccca tctgcctgga ctatttccag 60

gacccggtgt ccatcgagtg cggccacaac ttctgccgcg gctgcctgca ccgcaactgg 120

gcgccgggcg gcggcccgtt cccctgcccc gaatgtcggc acccatcggc gcccgccgcg 180

ctgcgaccca actgggccct ggccaggctg actgagaaga cgcagcgccg gcgcctgggc 240

cccgtgcccc cgggcctgtg cggccgccac tgggagccgc tgcggctctt ctgcgaggac 300

gaccagcggc cagtgtgcct ggtgtgcagg gagtcccagg agcaccagac tcacgccatg 360

gcacccatcg acgaggcctt cgagagctac cggacaggta actttgacat ccacgtggat 420

gaatggaaga gaagactaat taggctgctc ttgtaccatt ttaagcagga ggagaaactt 480

cttaagtctc agcgtaatct cgtggccaag atgaagaaag tcatgcattt acaggatgta 540

gaagtgaaga acgccacaca gtggaaggat aagataaaga gtcagcgaat gagaatcagc 600

acggagtttt caaagctgca caacttcctg gttgaagaag aggacctgtt tcttcagaga 660

ttgaacaaag aagaagaaga gacgaagaag aagctgaatg agaacacgtt aaaactcaat 720

caaactatcg cttcattgaa gaagctcatc ttagaggtgg gggagaagag ccaggctccc 780

accctggagc tgcttcagaa tccaaaagaa gtgttgacca ggagtgagat ccaggatgtg 840

aactattctc ttgaagctgt aaaggtgaag acagtgtgcc agataccatt gatgaaggaa 900

atgctaaagc gattccaagt ggctgtaaac ctagctgaag acacagctca tcccaaactc 960

gtcttctccc aggaagggag atacgtgaaa aatacagcat cagccagttc ttggccagtg 1020

ttttcttcag catggaacta ctttgctgga tggaggaatc ctcagaagac tgcttttgta 1080

gagagatttc agcacttacc ctgtgttctg ggaaaaaacg ttttcacctc agggaaacat 1140

tactgggaag ttgagagtag agatagtctg gaggttgctg ttggggtgtg tcgggaggac 1200

gtcatgggaa ttactgatcg ttcaaaaatg tccccagatg tgggcatctg ggcgatttat 1260

tggagtgctg ctggctattg gcccttgata ggcttccctg gaactcccac ccagcaagag 1320

ccagctctcc accgagtggg ggtttacctg gatcgtggga ctgggaatgt ctccttctac 1380

agcgctgtgg acggagtgca cctgcacacc ttttcttgtt cttctgtctc acgcctccgg 1440

ccattttttt ggttgagtcc attagcatct ttagtcattc caccagtgac tgataggaaa 1500

tga 1503

<210> 8

<211> 500

<212> PRT

<213> 智人(Homo sapiens)

<400> 8

Met Glu Ala Glu Asp Ile Gln Glu Glu Leu Thr Cys Pro Ile Cys Leu

1 5 10 15

Asp Tyr Phe Gln Asp Pro Val Ser Ile Glu Cys Gly His Asn Phe Cys

20 25 30

Arg Gly Cys Leu His Arg Asn Trp Ala Pro Gly Gly Gly Pro Phe Pro

35 40 45

Cys Pro Glu Cys Arg His Pro Ser Ala Pro Ala Ala Leu Arg Pro Asn

50 55 60

Trp Ala Leu Ala Arg Leu Thr Glu Lys Thr Gln Arg Arg Arg Leu Gly

65 70 75 80

Pro Val Pro Pro Gly Leu Cys Gly Arg His Trp Glu Pro Leu Arg Leu

85 90 95

Phe Cys Glu Asp Asp Gln Arg Pro Val Cys Leu Val Cys Arg Glu Ser

100 105 110

Gln Glu His Gln Thr His Ala Met Ala Pro Ile Asp Glu Ala Phe Glu

115 120 125

Ser Tyr Arg Thr Gly Asn Phe Asp Ile His Val Asp Glu Trp Lys Arg

130 135 140

Arg Leu Ile Arg Leu Leu Leu Tyr His Phe Lys Gln Glu Glu Lys Leu

145 150 155 160

Leu Lys Ser Gln Arg Asn Leu Val Ala Lys Met Lys Lys Val Met His

165 170 175

Leu Gln Asp Val Glu Val Lys Asn Ala Thr Gln Trp Lys Asp Lys Ile

180 185 190

Lys Ser Gln Arg Met Arg Ile Ser Thr Glu Phe Ser Lys Leu His Asn

195 200 205

Phe Leu Val Glu Glu Glu Asp Leu Phe Leu Gln Arg Leu Asn Lys Glu

210 215 220

Glu Glu Glu Thr Lys Lys Lys Leu Asn Glu Asn Thr Leu Lys Leu Asn

225 230 235 240

Gln Thr Ile Ala Ser Leu Lys Lys Leu Ile Leu Glu Val Gly Glu Lys

245 250 255

Ser Gln Ala Pro Thr Leu Glu Leu Leu Gln Asn Pro Lys Glu Val Leu

260 265 270

Thr Arg Ser Glu Ile Gln Asp Val Asn Tyr Ser Leu Glu Ala Val Lys

275 280 285

Val Lys Thr Val Cys Gln Ile Pro Leu Met Lys Glu Met Leu Lys Arg

290 295 300

Phe Gln Val Ala Val Asn Leu Ala Glu Asp Thr Ala His Pro Lys Leu

305 310 315 320

Val Phe Ser Gln Glu Gly Arg Tyr Val Lys Asn Thr Ala Ser Ala Ser

325 330 335

Ser Trp Pro Val Phe Ser Ser Ala Trp Asn Tyr Phe Ala Gly Trp Arg

340 345 350

Asn Pro Gln Lys Thr Ala Phe Val Glu Arg Phe Gln His Leu Pro Cys

355 360 365

Val Leu Gly Lys Asn Val Phe Thr Ser Gly Lys His Tyr Trp Glu Val

370 375 380

Glu Ser Arg Asp Ser Leu Glu Val Ala Val Gly Val Cys Arg Glu Asp

385 390 395 400

Val Met Gly Ile Thr Asp Arg Ser Lys Met Ser Pro Asp Val Gly Ile

405 410 415

Trp Ala Ile Tyr Trp Ser Ala Ala Gly Tyr Trp Pro Leu Ile Gly Phe

420 425 430

Pro Gly Thr Pro Thr Gln Gln Glu Pro Ala Leu His Arg Val Gly Val

435 440 445

Tyr Leu Asp Arg Gly Thr Gly Asn Val Ser Phe Tyr Ser Ala Val Asp

450 455 460

Gly Val His Leu His Thr Phe Ser Cys Ser Ser Val Ser Arg Leu Arg

465 470 475 480

Pro Phe Phe Trp Leu Ser Pro Leu Ala Ser Leu Val Ile Pro Pro Val

485 490 495

Thr Asp Arg Lys

500

<210> 9

<211> 1482

<212> DNA

<213> 智人(Homo sapiens)

<400> 9

atggcttctg gaatcctggt taatgtaaag gaggaggtga cctgccccat ctgcctggaa 60

ctcctgacac aacccctgag cctggactgc ggccacagct tctgccaagc atgcctcact 120

gcaaaccaca agaagtccat gctagacaaa ggagagagta gctgccctgt gtgccggatc 180

agttaccagc ctgagaacat acggcctaat cggcatgtag ccaacatagt ggagaagctc 240

agggaggtca agttgagccc agaggggcag aaagttgatc attgtgcacg ccatggagag 300

aaacttctac tcttctgtca ggaggacggg aaggtcattt gctggctttg tgagcggtct 360

caggagcacc gtggtcacca cacgttcctc acagaggagg ttgcccggga gtaccaagtg 420

aagctccagg cagctctgga gatgctgagg cagaagcagc aggaagctga agagttagaa 480

gctgacatca gagaagagaa agcttcctgg aagactcaaa tacagtatga caaaaccaac 540

gtcttggcag attttgagca actgagagac atcctggact gggaggagag caatgagctg 600

caaaacctgg agaaggagga ggaagacatt ctgaaaagcc ttacgaactc tgaaactgag 660

atggtgcagc agacccagtc cctgagagag ctcatctcag atctggagca tcggctgcag 720

gggtcagtga tggagctgct tcagggtgtg gatggcgtca taaaaaggac ggagaacgtg 780

accttgaaga agccagaaac ttttccaaaa aatcaaagga gagtgtttcg agctcctgat 840

ctgaaaggaa tgctagaagt gtttagagag ctgacagatg tccgacgcta ctgggttgat 900

gtgacagtgg ctccaaacaa catttcatgt gctgtcattt ctgaagataa gagacaagtg 960

agctctccga aaccacagat aatatatggg gcacgaggga caagatacca gacatttgtg 1020

aatttcaatt attgtactgg catcctgggc tctcaaagta tcacatcagg gaaacattac 1080

tgggaggtag acgtgtccaa gaaaactgct tggatcctgg gggtatgtgc tggcttccaa 1140

cctgatgcaa tgtgtaatat tgaaaaaaat gaaaattatc aacctaaata cggctactgg 1200

gttatagggt tagaggaagg agttaaatgt agtgctttcc aggatagttc cttccatact 1260

ccttctgttc ctttcattgt gcccctctct gtgattattt gtcctgatcg tgttggagtt 1320

ttcctagact atgaggcttg cactgtctca ttcttcaata tcacaaacca tggatttctc 1380

atctataagt tttctcactg ttctttttct cagcctgtat ttccatattt aaatcctaga 1440

aaatgtggag tccccatgac tctgtgctca ccaagctctt ga 1482

<210> 10

<211> 493

<212> PRT

<213> 智人(Homo sapiens)

<400> 10

Met Ala Ser Gly Ile Leu Val Asn Val Lys Glu Glu Val Thr Cys Pro

1 5 10 15

Ile Cys Leu Glu Leu Leu Thr Gln Pro Leu Ser Leu Asp Cys Gly His

20 25 30

Ser Phe Cys Gln Ala Cys Leu Thr Ala Asn His Lys Lys Ser Met Leu

35 40 45

Asp Lys Gly Glu Ser Ser Cys Pro Val Cys Arg Ile Ser Tyr Gln Pro

50 55 60

Glu Asn Ile Arg Pro Asn Arg His Val Ala Asn Ile Val Glu Lys Leu

65 70 75 80

Arg Glu Val Lys Leu Ser Pro Glu Gly Gln Lys Val Asp His Cys Ala

85 90 95

Arg His Gly Glu Lys Leu Leu Leu Phe Cys Gln Glu Asp Gly Lys Val

100 105 110

Ile Cys Trp Leu Cys Glu Arg Ser Gln Glu His Arg Gly His His Thr

115 120 125

Phe Leu Thr Glu Glu Val Ala Arg Glu Tyr Gln Val Lys Leu Gln Ala

130 135 140

Ala Leu Glu Met Leu Arg Gln Lys Gln Gln Glu Ala Glu Glu Leu Glu

145 150 155 160

Ala Asp Ile Arg Glu Glu Lys Ala Ser Trp Lys Thr Gln Ile Gln Tyr

165 170 175

Asp Lys Thr Asn Val Leu Ala Asp Phe Glu Gln Leu Arg Asp Ile Leu

180 185 190

Asp Trp Glu Glu Ser Asn Glu Leu Gln Asn Leu Glu Lys Glu Glu Glu

195 200 205

Asp Ile Leu Lys Ser Leu Thr Asn Ser Glu Thr Glu Met Val Gln Gln

210 215 220

Thr Gln Ser Leu Arg Glu Leu Ile Ser Asp Leu Glu His Arg Leu Gln

225 230 235 240

Gly Ser Val Met Glu Leu Leu Gln Gly Val Asp Gly Val Ile Lys Arg

245 250 255

Thr Glu Asn Val Thr Leu Lys Lys Pro Glu Thr Phe Pro Lys Asn Gln

260 265 270

Arg Arg Val Phe Arg Ala Pro Asp Leu Lys Gly Met Leu Glu Val Phe

275 280 285

Arg Glu Leu Thr Asp Val Arg Arg Tyr Trp Val Asp Val Thr Val Ala

290 295 300

Pro Asn Asn Ile Ser Cys Ala Val Ile Ser Glu Asp Lys Arg Gln Val

305 310 315 320

Ser Ser Pro Lys Pro Gln Ile Ile Tyr Gly Ala Arg Gly Thr Arg Tyr

325 330 335

Gln Thr Phe Val Asn Phe Asn Tyr Cys Thr Gly Ile Leu Gly Ser Gln

340 345 350

Ser Ile Thr Ser Gly Lys His Tyr Trp Glu Val Asp Val Ser Lys Lys

355 360 365

Thr Ala Trp Ile Leu Gly Val Cys Ala Gly Phe Gln Pro Asp Ala Met

370 375 380

Cys Asn Ile Glu Lys Asn Glu Asn Tyr Gln Pro Lys Tyr Gly Tyr Trp

385 390 395 400

Val Ile Gly Leu Glu Glu Gly Val Lys Cys Ser Ala Phe Gln Asp Ser

405 410 415

Ser Phe His Thr Pro Ser Val Pro Phe Ile Val Pro Leu Ser Val Ile

420 425 430

Ile Cys Pro Asp Arg Val Gly Val Phe Leu Asp Tyr Glu Ala Cys Thr

435 440 445

Val Ser Phe Phe Asn Ile Thr Asn His Gly Phe Leu Ile Tyr Lys Phe

450 455 460

Ser His Cys Ser Phe Ser Gln Pro Val Phe Pro Tyr Leu Asn Pro Arg

465 470 475 480

Lys Cys Gly Val Pro Met Thr Leu Cys Ser Pro Ser Ser

485 490

<210> 11

<211> 1551

<212> DNA

<213> 智人(Homo sapiens)

<400> 11

atgtgcgggt cagagaggat tctacaggca ggaaacatct tagaaatcag ggttgggcag 60

gcaggagcca ggagagtagc tacaatgact tcaccagtac tggtggacat acgagaagag 120

gtgacctgcc ctatctgcct ggagctccta acagaacccc tgagcataga ctgtggccac 180

agcttctgcc aagcctgcat cacaccaaat ggcagggaat cagtgattgg tcaagaaggg 240

gaaagaagct gccctgtgtg ccagaccagc taccagccag ggaacctgcg gcctaatcgg 300

catctggcca acatagtgag gcggctcaga gaggtagtgt tgggccctgg gaagcagctg 360

aaagcagttc tttgtgcaga ccatggagaa aaactgcagc tcttctgtca ggaggatggg 420

aaggtcattt gctggctttg tgagcggtct caggagcacc gtggtcacca cacgttcctc 480

gtggaggagg ttgcccagga gtaccaggag aagtttcagg agtctctaaa gaagctgaag 540

aacgaggagc aggaagctga gaagctaaca gcttttatca gagagaagaa aacatcctgg 600

aagaatcaga tggagcctga gagatgcagg atccagacag agtttaatca gctgcgaaat 660

atcctagaca gagtggagca acgggagctg aaaaagctgg aacaggaaga gaagaagggg 720

ctacgaatta tagaagaggc tgagaatgat ctggtccacc agacccagtc gctgcgagag 780

ctcatctcgg atctggagcg tcgatgtcag gggtcaacaa tggagctgct gcaggatgtg 840

agtgatgtca cagaaaggag tgagttctgg accctgagga agccagaagc tctccctaca 900

aagctgagaa gtatgttccg agccccagat ctgaaaagga tgctgcgagt gtgtagagag 960

ctgacagatg tccaaagcta ctgggttgac gtgaccctga atccacacac agctaattta 1020

aatcttgtcc tggctaaaaa ccggagacaa gtgaggtttg tgggagctaa agtatctgga 1080

ccttcctgtc tggaaaagca ttatgactgt agtgtcctgg gctcccagca cttctcctct 1140

ggtaagcatt actgggaggt agatgtggcc aagaagactg cctggatcct gggggtatgc 1200

agcaattcac tgggacctac attctctttc aaccattttg ctcaaaatca cagtgcttac 1260

tccaggtatc agcctcagag tggatactgg gtgattgggt tacagcataa ccatgaatat 1320

agggcctatg aggattcttc cccttccctg cttctctcca tgacagtgcc ccctcgccgt 1380

gttggggttt tcttagatta tgaggctggt actgtctcct tttataatgt cacaaaccat 1440

ggcttcccca tctacacttt ctctaaatat tactttccca ctactctttg tccatatttt 1500

aatccttgca actgtgtaat tcctatgacc ctgcgtcgtc caagctcttg a 1551

<210> 12

<211> 516

<212> PRT

<213> 智人(Homo sapiens)

<400> 12

Met Cys Gly Ser Glu Arg Ile Leu Gln Ala Gly Asn Ile Leu Glu Ile

1 5 10 15

Arg Val Gly Gln Ala Gly Ala Arg Arg Val Ala Thr Met Thr Ser Pro

20 25 30

Val Leu Val Asp Ile Arg Glu Glu Val Thr Cys Pro Ile Cys Leu Glu

35 40 45

Leu Leu Thr Glu Pro Leu Ser Ile Asp Cys Gly His Ser Phe Cys Gln

50 55 60

Ala Cys Ile Thr Pro Asn Gly Arg Glu Ser Val Ile Gly Gln Glu Gly

65 70 75 80

Glu Arg Ser Cys Pro Val Cys Gln Thr Ser Tyr Gln Pro Gly Asn Leu

85 90 95

Arg Pro Asn Arg His Leu Ala Asn Ile Val Arg Arg Leu Arg Glu Val

100 105 110

Val Leu Gly Pro Gly Lys Gln Leu Lys Ala Val Leu Cys Ala Asp His

115 120 125

Gly Glu Lys Leu Gln Leu Phe Cys Gln Glu Asp Gly Lys Val Ile Cys

130 135 140

Trp Leu Cys Glu Arg Ser Gln Glu His Arg Gly His His Thr Phe Leu

145 150 155 160

Val Glu Glu Val Ala Gln Glu Tyr Gln Glu Lys Phe Gln Glu Ser Leu

165 170 175

Lys Lys Leu Lys Asn Glu Glu Gln Glu Ala Glu Lys Leu Thr Ala Phe

180 185 190

Ile Arg Glu Lys Lys Thr Ser Trp Lys Asn Gln Met Glu Pro Glu Arg

195 200 205

Cys Arg Ile Gln Thr Glu Phe Asn Gln Leu Arg Asn Ile Leu Asp Arg

210 215 220

Val Glu Gln Arg Glu Leu Lys Lys Leu Glu Gln Glu Glu Lys Lys Gly

225 230 235 240

Leu Arg Ile Ile Glu Glu Ala Glu Asn Asp Leu Val His Gln Thr Gln

245 250 255

Ser Leu Arg Glu Leu Ile Ser Asp Leu Glu Arg Arg Cys Gln Gly Ser

260 265 270

Thr Met Glu Leu Leu Gln Asp Val Ser Asp Val Thr Glu Arg Ser Glu

275 280 285

Phe Trp Thr Leu Arg Lys Pro Glu Ala Leu Pro Thr Lys Leu Arg Ser

290 295 300

Met Phe Arg Ala Pro Asp Leu Lys Arg Met Leu Arg Val Cys Arg Glu

305 310 315 320

Leu Thr Asp Val Gln Ser Tyr Trp Val Asp Val Thr Leu Asn Pro His

325 330 335

Thr Ala Asn Leu Asn Leu Val Leu Ala Lys Asn Arg Arg Gln Val Arg

340 345 350

Phe Val Gly Ala Lys Val Ser Gly Pro Ser Cys Leu Glu Lys His Tyr

355 360 365

Asp Cys Ser Val Leu Gly Ser Gln His Phe Ser Ser Gly Lys His Tyr

370 375 380

Trp Glu Val Asp Val Ala Lys Lys Thr Ala Trp Ile Leu Gly Val Cys

385 390 395 400

Ser Asn Ser Leu Gly Pro Thr Phe Ser Phe Asn His Phe Ala Gln Asn

405 410 415

His Ser Ala Tyr Ser Arg Tyr Gln Pro Gln Ser Gly Tyr Trp Val Ile

420 425 430

Gly Leu Gln His Asn His Glu Tyr Arg Ala Tyr Glu Asp Ser Ser Pro

435 440 445

Ser Leu Leu Leu Ser Met Thr Val Pro Pro Arg Arg Val Gly Val Phe

450 455 460

Leu Asp Tyr Glu Ala Gly Thr Val Ser Phe Tyr Asn Val Thr Asn His

465 470 475 480

Gly Phe Pro Ile Tyr Thr Phe Ser Lys Tyr Tyr Phe Pro Thr Thr Leu

485 490 495

Cys Pro Tyr Phe Asn Pro Cys Asn Cys Val Ile Pro Met Thr Leu Arg

500 505 510

Arg Pro Ser Ser

515

<210> 13

<211> 666

<212> DNA

<213> 智人(Homo sapiens)

<400> 13

atggcggctg tgggaccgcg gaccggcccc ggaaccggcg ccgaggctct agcgctggcg 60

gcagagctgc agggcgaggc gacgtgctcc atctgcctag agctctttcg tgagccggtg 120

tccgtcgagt gcggccacag cttctgccgc gcctgcatag ggcgctgctg ggagcgcccg 180

ggcgcggggt ctgttggggc cgccacccgc gcgcccccct tcccactgcc ctgtccgcag 240

tgccgcgagc ccgcgcgccc cagtcagctg cggcccaacc ggcagctggc ggcagtggcc 300

acgctcctgc ggcgcttcag cctgcccgcg gctgccccgg gagagcacgg gtctcaggcg 360

gccgcggccc gggcagcggc tgcccgctgc gggcagcatg gcgaaccctt caagctctac 420

tgccaggacg acggacgcgc catctgcgtg gtgtgcgacc gcgcccgcga gcaccgcgag 480

cacgccgtgc tgccgctgga cgaggcggtg caggaggcca aggagctctt ggagtccagg 540

ctgagggtct tgaagaagga actggaggac tgtgaggtgt tccggtccac ggaaaagaag 600

gagagcaagg agctgctggt gagccaggca cccgcaggcc ccccgtggga cattacagag 660

gcctga 666

<210> 14

<211> 221

<212> PRT

<213> 智人(Homo sapiens)

<400> 14

Met Ala Ala Val Gly Pro Arg Thr Gly Pro Gly Thr Gly Ala Glu Ala

1 5 10 15

Leu Ala Leu Ala Ala Glu Leu Gln Gly Glu Ala Thr Cys Ser Ile Cys

20 25 30

Leu Glu Leu Phe Arg Glu Pro Val Ser Val Glu Cys Gly His Ser Phe

35 40 45

Cys Arg Ala Cys Ile Gly Arg Cys Trp Glu Arg Pro Gly Ala Gly Ser

50 55 60

Val Gly Ala Ala Thr Arg Ala Pro Pro Phe Pro Leu Pro Cys Pro Gln

65 70 75 80

Cys Arg Glu Pro Ala Arg Pro Ser Gln Leu Arg Pro Asn Arg Gln Leu

85 90 95

Ala Ala Val Ala Thr Leu Leu Arg Arg Phe Ser Leu Pro Ala Ala Ala

100 105 110

Pro Gly Glu His Gly Ser Gln Ala Ala Ala Ala Arg Ala Ala Ala Ala

115 120 125

Arg Cys Gly Gln His Gly Glu Pro Phe Lys Leu Tyr Cys Gln Asp Asp

130 135 140

Gly Arg Ala Ile Cys Val Val Cys Asp Arg Ala Arg Glu His Arg Glu

145 150 155 160

His Ala Val Leu Pro Leu Asp Glu Ala Val Gln Glu Ala Lys Glu Leu

165 170 175

Leu Glu Ser Arg Leu Arg Val Leu Lys Lys Glu Leu Glu Asp Cys Glu

180 185 190

Val Phe Arg Ser Thr Glu Lys Lys Glu Ser Lys Glu Leu Leu Val Ser

195 200 205

Gln Ala Pro Ala Gly Pro Pro Trp Asp Ile Thr Glu Ala

210 215 220

<210> 15

<211> 1656

<212> DNA

<213> 智人(Homo sapiens)

<400> 15

atggcggaga attggaagaa ctgcttcgag gaggagctca tctgccctat ctgcctgcac 60

gttttcgtgg agccagtgca gctgccgtgc aaacacaact tctgccgggg ctgcatcggc 120

gaggcgtggg ccaaggacag cggcctcgta cgctgcccag agtgcaacca ggcctacaac 180

cagaagccgg gcctggagaa gaacctgaag ctcaccaaca tcgtggagaa gttcaatgcc 240

ctgcacgtgg agaagccgcc ggcggcgctg cactgcgtgt tctgccgccg cggccccccg 300

ctgcccgcgc agaaggtctg cctgcgctgc gaggcgccct gctgccagtc ccacgtgcag 360

acgcacctgc agcagccctc caccgcccgc gggcacctcc tggtggaggc ggacgacgtg 420

cgggcctgga gctgcccgca gcacaacgcc taccgcctct accactgcga ggccgagcag 480

gtggccgtgt gccagtactg ctgctactac agcggcgcgc atcagggaca ctcggtgtgc 540

gacgtggaga tccgaaggaa tgaaatccgg aagatgctca tgaagcagca ggaccggctg 600

gaggagcgag agcaggacat tgaggaccag ctgtacaaac tcgagtcaga caagcgcctg 660

gtggaggaga aagtgaacca actgaaggag gaagttcggc tgcagtacga gaagctgcac 720

cagctgctgg acgaggacct gcggcagaca gtggaggtcc tagacaaggc ccaggccaag 780

ttctgcagcg agaacgcagc gcaggcgctg cacctcgggg agcgcatgca ggaggccaag 840

aagctgctgg gctccctgca gctgctcttt gataagacgg aggatgtcag cttcatgaag 900

aacaccaagt ctgtgaaaat cctgatggac aggacccaga cctgcacgag cagcagcctt 960

tcccccacta agatcggcca cctgaactcc aagctcttcc tgaacgaagt ggccaagaag 1020

gagaagcagc tgcggaaaat gctagaaggc cccttcagca cgccggtgcc cttcctgcag 1080

agtgtccccc tgtacccttg cggcgtgagc agctctgggg cggaaaagcg caagcactca 1140

acggccttcc cagaggccag tttcctagag acgtcgtcgg gccctgtggg cggccagtac 1200

ggggcggcgg gcacagccag cggtgagggc cagtctgggc agcccctggg gccctgcagc 1260

tccacgcagc acttggtggc cctgccgggc ggcgcccaac cagtgcactc aagccccgtg 1320

ttccccccat cgcagtatcc caatggctcc gccgcccagc agcccatgct cccccagtat 1380

ggcggccgca agattctcgt ctgttctgtg gacaactgtt actgttcttc cgtggccaac 1440

catggcggcc accagcccta cccccgctcc ggccactttc cctggacagt gccctcgcag 1500

gagtactcac acccgctccc gcccacaccc tccgtccccc agtcccttcc cagcctggcg 1560

gtcagagact ggcttgacgc ctcccagcag cccggccacc aggatttcta cagggtgtat 1620

gggcagccgt ccaccaaaca ctacgtgacg agctaa 1656

<210> 16

<211> 551

<212> PRT

<213> 智人(Homo sapiens)

<400> 16

Met Ala Glu Asn Trp Lys Asn Cys Phe Glu Glu Glu Leu Ile Cys Pro

1 5 10 15

Ile Cys Leu His Val Phe Val Glu Pro Val Gln Leu Pro Cys Lys His

20 25 30

Asn Phe Cys Arg Gly Cys Ile Gly Glu Ala Trp Ala Lys Asp Ser Gly

35 40 45

Leu Val Arg Cys Pro Glu Cys Asn Gln Ala Tyr Asn Gln Lys Pro Gly

50 55 60

Leu Glu Lys Asn Leu Lys Leu Thr Asn Ile Val Glu Lys Phe Asn Ala

65 70 75 80

Leu His Val Glu Lys Pro Pro Ala Ala Leu His Cys Val Phe Cys Arg

85 90 95

Arg Gly Pro Pro Leu Pro Ala Gln Lys Val Cys Leu Arg Cys Glu Ala

100 105 110

Pro Cys Cys Gln Ser His Val Gln Thr His Leu Gln Gln Pro Ser Thr

115 120 125

Ala Arg Gly His Leu Leu Val Glu Ala Asp Asp Val Arg Ala Trp Ser

130 135 140

Cys Pro Gln His Asn Ala Tyr Arg Leu Tyr His Cys Glu Ala Glu Gln

145 150 155 160

Val Ala Val Cys Gln Tyr Cys Cys Tyr Tyr Ser Gly Ala His Gln Gly

165 170 175

His Ser Val Cys Asp Val Glu Ile Arg Arg Asn Glu Ile Arg Lys Met

180 185 190

Leu Met Lys Gln Gln Asp Arg Leu Glu Glu Arg Glu Gln Asp Ile Glu

195 200 205

Asp Gln Leu Tyr Lys Leu Glu Ser Asp Lys Arg Leu Val Glu Glu Lys

210 215 220

Val Asn Gln Leu Lys Glu Glu Val Arg Leu Gln Tyr Glu Lys Leu His

225 230 235 240

Gln Leu Leu Asp Glu Asp Leu Arg Gln Thr Val Glu Val Leu Asp Lys

245 250 255

Ala Gln Ala Lys Phe Cys Ser Glu Asn Ala Ala Gln Ala Leu His Leu

260 265 270

Gly Glu Arg Met Gln Glu Ala Lys Lys Leu Leu Gly Ser Leu Gln Leu

275 280 285

Leu Phe Asp Lys Thr Glu Asp Val Ser Phe Met Lys Asn Thr Lys Ser

290 295 300

Val Lys Ile Leu Met Asp Arg Thr Gln Thr Cys Thr Ser Ser Ser Leu

305 310 315 320

Ser Pro Thr Lys Ile Gly His Leu Asn Ser Lys Leu Phe Leu Asn Glu

325 330 335

Val Ala Lys Lys Glu Lys Gln Leu Arg Lys Met Leu Glu Gly Pro Phe

340 345 350

Ser Thr Pro Val Pro Phe Leu Gln Ser Val Pro Leu Tyr Pro Cys Gly

355 360 365

Val Ser Ser Ser Gly Ala Glu Lys Arg Lys His Ser Thr Ala Phe Pro

370 375 380

Glu Ala Ser Phe Leu Glu Thr Ser Ser Gly Pro Val Gly Gly Gln Tyr

385 390 395 400

Gly Ala Ala Gly Thr Ala Ser Gly Glu Gly Gln Ser Gly Gln Pro Leu

405 410 415

Gly Pro Cys Ser Ser Thr Gln His Leu Val Ala Leu Pro Gly Gly Ala

420 425 430

Gln Pro Val His Ser Ser Pro Val Phe Pro Pro Ser Gln Tyr Pro Asn

435 440 445

Gly Ser Ala Ala Gln Gln Pro Met Leu Pro Gln Tyr Gly Gly Arg Lys

450 455 460

Ile Leu Val Cys Ser Val Asp Asn Cys Tyr Cys Ser Ser Val Ala Asn

465 470 475 480

His Gly Gly His Gln Pro Tyr Pro Arg Ser Gly His Phe Pro Trp Thr

485 490 495

Val Pro Ser Gln Glu Tyr Ser His Pro Leu Pro Pro Thr Pro Ser Val

500 505 510

Pro Gln Ser Leu Pro Ser Leu Ala Val Arg Asp Trp Leu Asp Ala Ser

515 520 525

Gln Gln Pro Gly His Gln Asp Phe Tyr Arg Val Tyr Gly Gln Pro Ser

530 535 540

Thr Lys His Tyr Val Thr Ser

545 550

<210> 17

<211> 2133

<212> DNA

<213> 智人(Homo sapiens)

<400> 17

atggaggaga tggaagagga gttgaaatgc cccgtgtgcg gctccttcta tcgggagccc 60

atcatcctgc cctgctctca caatttgtgt caggcgtgcg cccgcaacat cctggtgcag 120

accccagagt ctgaatcccc ccagagccat cgggccgcgg gctccggggt ctccgactat 180

gactatctgg acctggacaa gatgagccta tacagcgagg cggacagcgg ctatggctcc 240

tacggggggt tcgccagcgc ccccactacc ccgtgccaga agtcccccaa cggcgtccgc 300

gtgtttcccc cggctatgcc gccaccggcc acccacttgt caccggccct ggccccggtg 360

ccccgcaact cctgtatcac ctgcccccag tgtcaccgca gcctcatcct ggatgaccgg 420

gggctccgcg gcttccccaa gaatcgcgta ctggaagggg taattgaccg ctaccagcag 480

agcaaagccg cggccctcaa gtgccagctc tgcgagaagg cgcccaagga agccaccgtc 540

atgtgcgaac agtgcgatgt cttctactgc gatccgtgcc gcctgcgctg ccacccgccc 600

cgggggcccc tagccaagca ccgcctggtg cccccggccc agggtcgtgt gagccggagg 660

ctgagcccac gcaaggtctc cacctgcaca gaccacgagc tggagaacca cagcatgtac 720

tgcgtgcaat gcaagatgcc cgtgtgctac cagtgcttgg aggagggcaa acactccagc 780

cacgaagtca aggctctggg ggccatgtgg aaactacata agagccagct ctcccaggcg 840

ctgaacggac tgtcagacag ggccaaagaa gccaaggagt ttctggtaca gctgcgcaac 900

atggtccagc agatccagga gaacagtgtg gagtttgaag cctgtctggt ggcccaatgt 960

gatgccctca tcgatgccct caacagaaga aaagcccagc tgctggcccg cgtcaacaag 1020

gagcatgagc acaagctgaa ggtggttcga gatcagatct ctcactgcac agtgaaattg 1080

cgccagacca caggtctcat ggagtactgc ttggaggtga ttaaggaaaa tgatcctagt 1140

ggttttttgc agatttctga cgccctcata agaagagtgc acctgactga ggatcagtgg 1200

ggtaaaggca cactcactcc aaggatgacc acggactttg acttgagtct ggacaacagc 1260

cctctgctgc aatccatcca ccagctggat ttcgtgcaag tgaaagcttc ctctccagtc 1320

ccagcaaccc ctatcctaca gctggaggaa tgttgtaccc acaacaacag cgctacgttg 1380

tcctggaaac agccacctct gtccacggtg cccgccgatg gatacattct ggagctggat 1440

gatggcaacg gtggtcaatt ccgggaggtg tatgtgggga aggagacaat gtgcactgtg 1500

gatggtcttc acttcaacag cacatacaac gctcgggtca aggccttcaa caaaacagga 1560

gtcagcccgt acagcaagac cctggtcctc caaacgtctg aggtggcctg gtttgctttc 1620

gaccctggct cggcgcactc ggacatcatc ctctccaatg acaacctgac agtgacctgt 1680

agtagctatg atgaccgggt ggtgctaggg aagactggct tctccaaggg catccactac 1740

tgggagctca cggtagatcg ctatgacaac caccctgatc ctgcctttgg tgtggctcgc 1800

atggacgtga tgaaggatgt gatgttagga aaagacgaca aagcttgggc aatgtatgtg 1860

gacaataacc ggagctggtt catgcacaac aactcgcaca ccaacagaac tgagggaggg 1920

atcacaaaag gggccacaat tggggtcctc ctcgacttaa atagaaaaaa cttgacattt 1980

tttatcaacg atgaacaaca aggtcccata gcatttgata acgtggaggg cctcttcttc 2040

cctgcggtca gcctgaacag gaacgtgcag gtcacgctgc acaccgggct cccagtcccc 2100

gacttctact ccagcagagc atcaatagcc taa 2133

<210> 18

<211> 710

<212> PRT

<213> 智人(Homo sapiens)

<400> 18

Met Glu Glu Met Glu Glu Glu Leu Lys Cys Pro Val Cys Gly Ser Phe

1 5 10 15

Tyr Arg Glu Pro Ile Ile Leu Pro Cys Ser His Asn Leu Cys Gln Ala

20 25 30

Cys Ala Arg Asn Ile Leu Val Gln Thr Pro Glu Ser Glu Ser Pro Gln

35 40 45

Ser His Arg Ala Ala Gly Ser Gly Val Ser Asp Tyr Asp Tyr Leu Asp

50 55 60

Leu Asp Lys Met Ser Leu Tyr Ser Glu Ala Asp Ser Gly Tyr Gly Ser

65 70 75 80

Tyr Gly Gly Phe Ala Ser Ala Pro Thr Thr Pro Cys Gln Lys Ser Pro

85 90 95

Asn Gly Val Arg Val Phe Pro Pro Ala Met Pro Pro Pro Ala Thr His

100 105 110

Leu Ser Pro Ala Leu Ala Pro Val Pro Arg Asn Ser Cys Ile Thr Cys

115 120 125

Pro Gln Cys His Arg Ser Leu Ile Leu Asp Asp Arg Gly Leu Arg Gly

130 135 140

Phe Pro Lys Asn Arg Val Leu Glu Gly Val Ile Asp Arg Tyr Gln Gln

145 150 155 160

Ser Lys Ala Ala Ala Leu Lys Cys Gln Leu Cys Glu Lys Ala Pro Lys

165 170 175

Glu Ala Thr Val Met Cys Glu Gln Cys Asp Val Phe Tyr Cys Asp Pro

180 185 190

Cys Arg Leu Arg Cys His Pro Pro Arg Gly Pro Leu Ala Lys His Arg

195 200 205

Leu Val Pro Pro Ala Gln Gly Arg Val Ser Arg Arg Leu Ser Pro Arg

210 215 220

Lys Val Ser Thr Cys Thr Asp His Glu Leu Glu Asn His Ser Met Tyr

225 230 235 240

Cys Val Gln Cys Lys Met Pro Val Cys Tyr Gln Cys Leu Glu Glu Gly

245 250 255

Lys His Ser Ser His Glu Val Lys Ala Leu Gly Ala Met Trp Lys Leu

260 265 270

His Lys Ser Gln Leu Ser Gln Ala Leu Asn Gly Leu Ser Asp Arg Ala

275 280 285

Lys Glu Ala Lys Glu Phe Leu Val Gln Leu Arg Asn Met Val Gln Gln

290 295 300

Ile Gln Glu Asn Ser Val Glu Phe Glu Ala Cys Leu Val Ala Gln Cys

305 310 315 320

Asp Ala Leu Ile Asp Ala Leu Asn Arg Arg Lys Ala Gln Leu Leu Ala

325 330 335

Arg Val Asn Lys Glu His Glu His Lys Leu Lys Val Val Arg Asp Gln

340 345 350

Ile Ser His Cys Thr Val Lys Leu Arg Gln Thr Thr Gly Leu Met Glu

355 360 365

Tyr Cys Leu Glu Val Ile Lys Glu Asn Asp Pro Ser Gly Phe Leu Gln

370 375 380

Ile Ser Asp Ala Leu Ile Arg Arg Val His Leu Thr Glu Asp Gln Trp

385 390 395 400

Gly Lys Gly Thr Leu Thr Pro Arg Met Thr Thr Asp Phe Asp Leu Ser

405 410 415

Leu Asp Asn Ser Pro Leu Leu Gln Ser Ile His Gln Leu Asp Phe Val

420 425 430

Gln Val Lys Ala Ser Ser Pro Val Pro Ala Thr Pro Ile Leu Gln Leu

435 440 445

Glu Glu Cys Cys Thr His Asn Asn Ser Ala Thr Leu Ser Trp Lys Gln

450 455 460

Pro Pro Leu Ser Thr Val Pro Ala Asp Gly Tyr Ile Leu Glu Leu Asp

465 470 475 480

Asp Gly Asn Gly Gly Gln Phe Arg Glu Val Tyr Val Gly Lys Glu Thr

485 490 495

Met Cys Thr Val Asp Gly Leu His Phe Asn Ser Thr Tyr Asn Ala Arg

500 505 510

Val Lys Ala Phe Asn Lys Thr Gly Val Ser Pro Tyr Ser Lys Thr Leu

515 520 525

Val Leu Gln Thr Ser Glu Val Ala Trp Phe Ala Phe Asp Pro Gly Ser

530 535 540

Ala His Ser Asp Ile Ile Leu Ser Asn Asp Asn Leu Thr Val Thr Cys

545 550 555 560

Ser Ser Tyr Asp Asp Arg Val Val Leu Gly Lys Thr Gly Phe Ser Lys

565 570 575

Gly Ile His Tyr Trp Glu Leu Thr Val Asp Arg Tyr Asp Asn His Pro

580 585 590

Asp Pro Ala Phe Gly Val Ala Arg Met Asp Val Met Lys Asp Val Met

595 600 605

Leu Gly Lys Asp Asp Lys Ala Trp Ala Met Tyr Val Asp Asn Asn Arg

610 615 620

Ser Trp Phe Met His Asn Asn Ser His Thr Asn Arg Thr Glu Gly Gly

625 630 635 640

Ile Thr Lys Gly Ala Thr Ile Gly Val Leu Leu Asp Leu Asn Arg Lys

645 650 655

Asn Leu Thr Phe Phe Ile Asn Asp Glu Gln Gln Gly Pro Ile Ala Phe

660 665 670

Asp Asn Val Glu Gly Leu Phe Phe Pro Ala Val Ser Leu Asn Arg Asn

675 680 685

Val Gln Val Thr Leu His Thr Gly Leu Pro Val Pro Asp Phe Tyr Ser

690 695 700

Ser Arg Ala Ser Ile Ala

705 710

<210> 19

<211> 1446

<212> DNA

<213> 智人(Homo sapiens)

<400> 19

atggcctctg ctgcctctgt gaccagcctg gcagatgaag tcaactgccc catctgtcag 60

ggtaccctga gggagccggt cactatcgac tgcggccaca acttctgccg ggcctgcctt 120

acccgctact gtgagatacc aggcccagac ctggaggagt cccctacttg cccactctgc 180

aaagaaccct tccgtcctgg gagcttccgg cccaactggc agctggctaa cgtggtggag 240

aacattgagc gcctccagct ggtgtccaca ctgggtttgg gagaggagga tgtctgccaa 300

gagcacggag agaagatcta cttcttctgt gaggatgatg agatgcagtt gtgcgtggtg 360

tgccgggagg ctggggagca cgctacccac accatgcgct tcctggagga tgcagcggct 420

ccctataggg aacaaatcca taagtgtctt aaatgtctaa gaaaagagag agaggagatt 480

caagaaatcc agtcaagaga aaataaaagg atgcaagtcc tcctgactca ggtgtccacc 540

aagagacaac aggtgatttc tgagttcgca cacctgagga agtttctaga ggaacagcag 600

agcatcctct tagcacaatt ggagagccag gatggggaca tcttgaggca acgggatgaa 660

tttgatttgc tggttgctgg ggagatctgc cggtttagtg ctcttattga agaactggag 720

gagaagaatg agaggccagc aagggagctc ctgacggaca tcagaagcac tctaataaga 780

tgtgaaacca gaaagtgccg gaaaccggtg gctgtgtcgc cagagctggg ccagaggatt 840

cgggactttc cccagcaggc cctcccgctg cagagggaga tgaagatgtt tctggaaaaa 900

ctatgctttg agttggacta tgagccagct cacatttctc tagaccctca gacttcccac 960

cccaagctcc tcttgtccga ggaccaccag cgagctcagt tctcctacaa atggcagaac 1020

tcaccagaca acccccaacg ttttgaccgg gccacctgtg ttctggccca cactggcatc 1080

acagggggga gacacacgtg ggtggtgagt atagacctgg cccatggggg cagctgcacc 1140

gtgggcgtgg tgagcgagga tgtgcagcgg aagggggagc ttcggctgcg gccagaggag 1200

ggggtgtggg ctgtgaggct ggcttggggc ttcgtctcgg ctctgggctc cttccccaca 1260

cggctgaccc tgaaggagca gccccggcag gtgagggtgt ctcttgacta tgaggtgggc 1320

tgggtgacct tcaccaacgc tgtcacccga gagcccatct acaccttcac tgcctccttc 1380

actaggaagg tcattccctt ctttgggctc tggggccgag ggtccagttt ctccctgagc 1440

tcctga 1446

<210> 20

<211> 481

<212> PRT

<213> 智人(Homo sapiens)

<400> 20

Met Ala Ser Ala Ala Ser Val Thr Ser Leu Ala Asp Glu Val Asn Cys

1 5 10 15

Pro Ile Cys Gln Gly Thr Leu Arg Glu Pro Val Thr Ile Asp Cys Gly

20 25 30

His Asn Phe Cys Arg Ala Cys Leu Thr Arg Tyr Cys Glu Ile Pro Gly

35 40 45

Pro Asp Leu Glu Glu Ser Pro Thr Cys Pro Leu Cys Lys Glu Pro Phe

50 55 60

Arg Pro Gly Ser Phe Arg Pro Asn Trp Gln Leu Ala Asn Val Val Glu

65 70 75 80

Asn Ile Glu Arg Leu Gln Leu Val Ser Thr Leu Gly Leu Gly Glu Glu

85 90 95

Asp Val Cys Gln Glu His Gly Glu Lys Ile Tyr Phe Phe Cys Glu Asp

100 105 110

Asp Glu Met Gln Leu Cys Val Val Cys Arg Glu Ala Gly Glu His Ala

115 120 125

Thr His Thr Met Arg Phe Leu Glu Asp Ala Ala Ala Pro Tyr Arg Glu

130 135 140

Gln Ile His Lys Cys Leu Lys Cys Leu Arg Lys Glu Arg Glu Glu Ile

145 150 155 160

Gln Glu Ile Gln Ser Arg Glu Asn Lys Arg Met Gln Val Leu Leu Thr

165 170 175

Gln Val Ser Thr Lys Arg Gln Gln Val Ile Ser Glu Phe Ala His Leu

180 185 190

Arg Lys Phe Leu Glu Glu Gln Gln Ser Ile Leu Leu Ala Gln Leu Glu

195 200 205

Ser Gln Asp Gly Asp Ile Leu Arg Gln Arg Asp Glu Phe Asp Leu Leu

210 215 220

Val Ala Gly Glu Ile Cys Arg Phe Ser Ala Leu Ile Glu Glu Leu Glu

225 230 235 240

Glu Lys Asn Glu Arg Pro Ala Arg Glu Leu Leu Thr Asp Ile Arg Ser

245 250 255

Thr Leu Ile Arg Cys Glu Thr Arg Lys Cys Arg Lys Pro Val Ala Val

260 265 270

Ser Pro Glu Leu Gly Gln Arg Ile Arg Asp Phe Pro Gln Gln Ala Leu

275 280 285

Pro Leu Gln Arg Glu Met Lys Met Phe Leu Glu Lys Leu Cys Phe Glu

290 295 300

Leu Asp Tyr Glu Pro Ala His Ile Ser Leu Asp Pro Gln Thr Ser His

305 310 315 320

Pro Lys Leu Leu Leu Ser Glu Asp His Gln Arg Ala Gln Phe Ser Tyr

325 330 335

Lys Trp Gln Asn Ser Pro Asp Asn Pro Gln Arg Phe Asp Arg Ala Thr

340 345 350

Cys Val Leu Ala His Thr Gly Ile Thr Gly Gly Arg His Thr Trp Val

355 360 365

Val Ser Ile Asp Leu Ala His Gly Gly Ser Cys Thr Val Gly Val Val

370 375 380

Ser Glu Asp Val Gln Arg Lys Gly Glu Leu Arg Leu Arg Pro Glu Glu

385 390 395 400

Gly Val Trp Ala Val Arg Leu Ala Trp Gly Phe Val Ser Ala Leu Gly

405 410 415

Ser Phe Pro Thr Arg Leu Thr Leu Lys Glu Gln Pro Arg Gln Val Arg

420 425 430

Val Ser Leu Asp Tyr Glu Val Gly Trp Val Thr Phe Thr Asn Ala Val

435 440 445

Thr Arg Glu Pro Ile Tyr Thr Phe Thr Ala Ser Phe Thr Arg Lys Val

450 455 460

Ile Pro Phe Phe Gly Leu Trp Gly Arg Gly Ser Ser Phe Ser Leu Ser

465 470 475 480

Ser

<210> 21

<211> 1407

<212> DNA

<213> 智人(Homo sapiens)

<400> 21

atggccgccc ccgacctgtc caccaacctc caggaggagg ccacctgcgc catctgcctc 60

gactacttca cggatccggt gatgaccgac tgcggccaca acttctgccg cgagtgcatc 120

cggcgctgct ggggccagcc cgagggcccg tacgcgtgcc ccgagtgccg cgagctgtcc 180

ccgcagagga acctgcggcc caaccgcccg cttgctaaga tggccgagat ggcgcggcgc 240

ctgcacccgc cgtcgccggt cccgcagggc gtgtgccccg cgcaccgcga gccactggcc 300

gccttctgtg gcgacgagct gcgcctcctg tgtgcggcct gcgagcgctc tggggagcac 360

tgggcgcacc gcgtgcggcc gctgcaggac gcggccgaag acctcaaggc gaagctggag 420

aagtcactgg agcatctccg gaagcagatg caggatgcgt tgctgttcca agcccaggcg 480

gatgagacct gcgtcttgtg gcagaagatg gtggagagcc agcggcagaa cgtgctgggt 540

gagttcgagc gtcttcgccg tttgctggca gaggaggagc agcagctgct gcagaggctg 600

gaggaggagg agctggaggt gctgccccgg ctgcgggagg gcgcagccca cctaggccag 660

cagagcgccc acctagctga gctcatcgcc gagctcgagg gccgctgcca gctgcctgct 720

ctggggctgc tgcaggacat caaggacgcc ctgcgcaggg tccaggatgt gaagctgcag 780

cccccagaag ttgtgcctat ggagctgagg accgtgtgca gggtcccggg actggtagag 840

acactgcgga ggtttcgagg ggacgtgacc ttggacccgg acaccgccaa ccctgagctg 900

atcctgtctg aagacaggcg gagcgtgcag cggggggacc tacggcaggc cctgccggac 960

agcccagagc gctttgaccc cggcccctgc gtgctgggcc aggagcgctt cacctcaggc 1020

cgccactact gggaggtgga ggttggggac cgcaccagct gggccctggg ggtgtgcagg 1080

gagaacgtga acaggaagga gaagggcgag ctgtccgcgg gcaacggctt ctggatcctg 1140

gtcttcctgg ggagctatta caattcctcg gaacgggcct tggctccact ccgggaccca 1200

cccaggcgcg tggggatctt tctggactac gaggctggac atctctcttt ctacagtgcc 1260

accgatgggt cactgctatt catctttccc gagatcccct tctcggggac gctgcggccc 1320

ctcttctcac ccctgtccag cagcccgacc ccgatgacta tctgccggcc gaaaggtggg 1380

tccggggaca ccctggctcc ccagtga 1407

<210> 22

<211> 468

<212> PRT

<213> 智人(Homo sapiens)

<400> 22

Met Ala Ala Pro Asp Leu Ser Thr Asn Leu Gln Glu Glu Ala Thr Cys

1 5 10 15

Ala Ile Cys Leu Asp Tyr Phe Thr Asp Pro Val Met Thr Asp Cys Gly

20 25 30

His Asn Phe Cys Arg Glu Cys Ile Arg Arg Cys Trp Gly Gln Pro Glu

35 40 45

Gly Pro Tyr Ala Cys Pro Glu Cys Arg Glu Leu Ser Pro Gln Arg Asn

50 55 60

Leu Arg Pro Asn Arg Pro Leu Ala Lys Met Ala Glu Met Ala Arg Arg

65 70 75 80

Leu His Pro Pro Ser Pro Val Pro Gln Gly Val Cys Pro Ala His Arg

85 90 95

Glu Pro Leu Ala Ala Phe Cys Gly Asp Glu Leu Arg Leu Leu Cys Ala

100 105 110

Ala Cys Glu Arg Ser Gly Glu His Trp Ala His Arg Val Arg Pro Leu

115 120 125

Gln Asp Ala Ala Glu Asp Leu Lys Ala Lys Leu Glu Lys Ser Leu Glu

130 135 140

His Leu Arg Lys Gln Met Gln Asp Ala Leu Leu Phe Gln Ala Gln Ala

145 150 155 160

Asp Glu Thr Cys Val Leu Trp Gln Lys Met Val Glu Ser Gln Arg Gln

165 170 175

Asn Val Leu Gly Glu Phe Glu Arg Leu Arg Arg Leu Leu Ala Glu Glu

180 185 190

Glu Gln Gln Leu Leu Gln Arg Leu Glu Glu Glu Glu Leu Glu Val Leu

195 200 205

Pro Arg Leu Arg Glu Gly Ala Ala His Leu Gly Gln Gln Ser Ala His

210 215 220

Leu Ala Glu Leu Ile Ala Glu Leu Glu Gly Arg Cys Gln Leu Pro Ala

225 230 235 240

Leu Gly Leu Leu Gln Asp Ile Lys Asp Ala Leu Arg Arg Val Gln Asp

245 250 255

Val Lys Leu Gln Pro Pro Glu Val Val Pro Met Glu Leu Arg Thr Val

260 265 270

Cys Arg Val Pro Gly Leu Val Glu Thr Leu Arg Arg Phe Arg Gly Asp

275 280 285

Val Thr Leu Asp Pro Asp Thr Ala Asn Pro Glu Leu Ile Leu Ser Glu

290 295 300

Asp Arg Arg Ser Val Gln Arg Gly Asp Leu Arg Gln Ala Leu Pro Asp

305 310 315 320

Ser Pro Glu Arg Phe Asp Pro Gly Pro Cys Val Leu Gly Gln Glu Arg

325 330 335

Phe Thr Ser Gly Arg His Tyr Trp Glu Val Glu Val Gly Asp Arg Thr

340 345 350

Ser Trp Ala Leu Gly Val Cys Arg Glu Asn Val Asn Arg Lys Glu Lys

355 360 365

Gly Glu Leu Ser Ala Gly Asn Gly Phe Trp Ile Leu Val Phe Leu Gly

370 375 380

Ser Tyr Tyr Asn Ser Ser Glu Arg Ala Leu Ala Pro Leu Arg Asp Pro

385 390 395 400

Pro Arg Arg Val Gly Ile Phe Leu Asp Tyr Glu Ala Gly His Leu Ser

405 410 415

Phe Tyr Ser Ala Thr Asp Gly Ser Leu Leu Phe Ile Phe Pro Glu Ile

420 425 430

Pro Phe Ser Gly Thr Leu Arg Pro Leu Phe Ser Pro Leu Ser Ser Ser

435 440 445

Pro Thr Pro Met Thr Ile Cys Arg Pro Lys Gly Gly Ser Gly Asp Thr

450 455 460

Leu Ala Pro Gln

465

<210> 23

<211> 894

<212> DNA

<213> 小鼠(Mus musculus)

<400> 23

atggcttcac aattcatgaa gaatttaaag gaggaggtga cctgtcctgt ctgtctgaac 60

ctgatggtga aacctgtgag tgcagattgt ggtcacacct tctgccaagg ctgcatcacg 120

ttgtactttg aatccatcaa atgcgataag aaagtgttca tttgccctgt gtgccgaatt 180

agttaccagt ttagcaatct gaggcctaat cgaaatgtgg ccaacatagt agagaggctc 240

aaaatgttca agcccagccc agaagaggag cagaaagtgt ttaactgtgc aagacatgga 300

aagaaactcc agctcttctg taggaaggac atgatggcca tctgctggct ttgtgagcga 360

tctcaggagc accgtggtca caaaacagct ctcattgaag aggtggccca ggagtacaag 420

gagcagctgc aggtagttct gcaaaggctg atggcagata agaaaaaatt tgaaaactgg 480

aaagatgacc ttcagaagga tagaacttac tgggagaatc aaatacagaa agatgtgcag 540

aatgttcggt cagagtttaa acgaatgagg gatatcatgg actctgagga gaagaaggaa 600

ttgcagaagc tgaggcaaga gaaggaagac attctcaaca acctggcaga gtctgaaagt 660

gagcatgctc agcagagcaa gttgctagaa gacttcatct cagatgtgga acatcagtta 720

cagtgctcag acatagaaat actgcagggt gtggagaaca tcatagaacg gagtcatact 780

ttttcgatga agaagcccaa agccatcgcc agggaacaaa gaaagttccg agcccctgac 840

ctgcaaggca tgctgcaagt gctgcaagag gtcacagagg ctcatcgcta ctag 894

<210> 24

<211> 297

<212> PRT

<213> 小鼠(Mus musculus)

<400> 24

Met Ala Ser Gln Phe Met Lys Asn Leu Lys Glu Glu Val Thr Cys Pro

1 5 10 15

Val Cys Leu Asn Leu Met Val Lys Pro Val Ser Ala Asp Cys Gly His

20 25 30

Thr Phe Cys Gln Gly Cys Ile Thr Leu Tyr Phe Glu Ser Ile Lys Cys

35 40 45

Asp Lys Lys Val Phe Ile Cys Pro Val Cys Arg Ile Ser Tyr Gln Phe

50 55 60

Ser Asn Leu Arg Pro Asn Arg Asn Val Ala Asn Ile Val Glu Arg Leu

65 70 75 80

Lys Met Phe Lys Pro Ser Pro Glu Glu Glu Gln Lys Val Phe Asn Cys

85 90 95

Ala Arg His Gly Lys Lys Leu Gln Leu Phe Cys Arg Lys Asp Met Met

100 105 110

Ala Ile Cys Trp Leu Cys Glu Arg Ser Gln Glu His Arg Gly His Lys

115 120 125

Thr Ala Leu Ile Glu Glu Val Ala Gln Glu Tyr Lys Glu Gln Leu Gln

130 135 140

Val Val Leu Gln Arg Leu Met Ala Asp Lys Lys Lys Phe Glu Asn Trp

145 150 155 160

Lys Asp Asp Leu Gln Lys Asp Arg Thr Tyr Trp Glu Asn Gln Ile Gln

165 170 175

Lys Asp Val Gln Asn Val Arg Ser Glu Phe Lys Arg Met Arg Asp Ile

180 185 190

Met Asp Ser Glu Glu Lys Lys Glu Leu Gln Lys Leu Arg Gln Glu Lys

195 200 205

Glu Asp Ile Leu Asn Asn Leu Ala Glu Ser Glu Ser Glu His Ala Gln

210 215 220

Gln Ser Lys Leu Leu Glu Asp Phe Ile Ser Asp Val Glu His Gln Leu

225 230 235 240

Gln Cys Ser Asp Ile Glu Ile Leu Gln Gly Val Glu Asn Ile Ile Glu

245 250 255

Arg Ser His Thr Phe Ser Met Lys Lys Pro Lys Ala Ile Ala Arg Glu

260 265 270

Gln Arg Lys Phe Arg Ala Pro Asp Leu Gln Gly Met Leu Gln Val Leu

275 280 285

Gln Glu Val Thr Glu Ala His Arg Tyr

290 295

<210> 25

<211> 1224

<212> DNA

<213> 智人(Homo sapiens)

<400> 25

atggagctgc ttgaagaaga tctcacatgc cctatttgtt gtagtctgtt tgatgatcca 60

cgggttttgc cttgctccca caacttctgc aaaaaatgct tagaaggtat cttagaaggg 120

agtgtgcgga attccttgtg gagaccagct ccattcaagt gtcctacatg ccgtaaggaa 180

acttcagcta ctggaattaa tagcctgcag gttaattact ccctgaaggg tattgtggaa 240

aagtataaca agatcaagat ctctcccaaa atgccagtat gcaaaggaca cttggggcag 300

cctctcaaca ttttctgcct gactgatatg cagctgattt gtgggatctg tgctactcgt 360

ggggagcaca ccaaacatgt cttctgttct attgaagatg cctatgctca ggaaagggat 420

gcctttgagt ccctcttcca gagctttgag acctggcgtc ggggagatgc tctttctcgc 480

ttggatacct tggaaactag taagaggaaa tccctacagt tactgactaa agattcagat 540

aaagtgaagg aattttttga gaagttacaa cacacactgg atcaaaagaa gaatgaaatt 600

ctgtctgact ttgagaccat gaaacttgct gttatgcaag catatgaccc agagatcaac 660

aaactcaaca ccatcttgca ggagcaacgg atggccttta acattgctga ggctttcaaa 720

gatgtgtcag aacccattgt atttctgcaa cagatgcagg agtttagaga gaaaatcaaa 780

gtaatcaagg aaactccttt acctccctct aatttgcctg caagcccttt aatgaagaac 840

tttgatacca gtcagtggga agacataaaa ctagtcgatg tggataaact ttctttgcct 900

caagacactg gcacattcat tagcaagatt ccctggagct tttataagtt atttttgcta 960

atccttctgc ttggccttgt cattgtcttt ggtcctacca tgttcctaga atggtcatta 1020

tttgatgacc tggcaacttg gaaaggctgt ctttcaaact tcagttccta tctgactaaa 1080

acagccgatt tcatagaaca atcagttttt tactgggaac aggtgacaga tgggtttttc 1140

attttcaatg aaagattcaa gaattttact ttggtggtac tgaacaatgt ggcagaattt 1200

gtgtgcaaat ataaactatt ataa 1224

<210> 26

<211> 407

<212> PRT

<213> 智人(Homo sapiens)

<400> 26

Met Glu Leu Leu Glu Glu Asp Leu Thr Cys Pro Ile Cys Cys Ser Leu

1 5 10 15

Phe Asp Asp Pro Arg Val Leu Pro Cys Ser His Asn Phe Cys Lys Lys

20 25 30

Cys Leu Glu Gly Ile Leu Glu Gly Ser Val Arg Asn Ser Leu Trp Arg

35 40 45

Pro Ala Pro Phe Lys Cys Pro Thr Cys Arg Lys Glu Thr Ser Ala Thr

50 55 60

Gly Ile Asn Ser Leu Gln Val Asn Tyr Ser Leu Lys Gly Ile Val Glu

65 70 75 80

Lys Tyr Asn Lys Ile Lys Ile Ser Pro Lys Met Pro Val Cys Lys Gly

85 90 95

His Leu Gly Gln Pro Leu Asn Ile Phe Cys Leu Thr Asp Met Gln Leu

100 105 110

Ile Cys Gly Ile Cys Ala Thr Arg Gly Glu His Thr Lys His Val Phe

115 120 125

Cys Ser Ile Glu Asp Ala Tyr Ala Gln Glu Arg Asp Ala Phe Glu Ser

130 135 140

Leu Phe Gln Ser Phe Glu Thr Trp Arg Arg Gly Asp Ala Leu Ser Arg

145 150 155 160

Leu Asp Thr Leu Glu Thr Ser Lys Arg Lys Ser Leu Gln Leu Leu Thr

165 170 175

Lys Asp Ser Asp Lys Val Lys Glu Phe Phe Glu Lys Leu Gln His Thr

180 185 190

Leu Asp Gln Lys Lys Asn Glu Ile Leu Ser Asp Phe Glu Thr Met Lys

195 200 205

Leu Ala Val Met Gln Ala Tyr Asp Pro Glu Ile Asn Lys Leu Asn Thr

210 215 220

Ile Leu Gln Glu Gln Arg Met Ala Phe Asn Ile Ala Glu Ala Phe Lys

225 230 235 240

Asp Val Ser Glu Pro Ile Val Phe Leu Gln Gln Met Gln Glu Phe Arg

245 250 255

Glu Lys Ile Lys Val Ile Lys Glu Thr Pro Leu Pro Pro Ser Asn Leu

260 265 270

Pro Ala Ser Pro Leu Met Lys Asn Phe Asp Thr Ser Gln Trp Glu Asp

275 280 285

Ile Lys Leu Val Asp Val Asp Lys Leu Ser Leu Pro Gln Asp Thr Gly

290 295 300

Thr Phe Ile Ser Lys Ile Pro Trp Ser Phe Tyr Lys Leu Phe Leu Leu

305 310 315 320

Ile Leu Leu Leu Gly Leu Val Ile Val Phe Gly Pro Thr Met Phe Leu

325 330 335

Glu Trp Ser Leu Phe Asp Asp Leu Ala Thr Trp Lys Gly Cys Leu Ser

340 345 350

Asn Phe Ser Ser Tyr Leu Thr Lys Thr Ala Asp Phe Ile Glu Gln Ser

355 360 365

Val Phe Tyr Trp Glu Gln Val Thr Asp Gly Phe Phe Ile Phe Asn Glu

370 375 380

Arg Phe Lys Asn Phe Thr Leu Val Val Leu Asn Asn Val Ala Glu Phe

385 390 395 400

Val Cys Lys Tyr Lys Leu Leu

405

<210> 27

<211> 1329

<212> DNA

<213> 智人(Homo sapiens)

<400> 27

atggcgggcg cggcgaccgg gagccggacc cctgggaggt cggagcttgt cgagggatgc 60

ggctggcgct gcccggagca tggcgaccgc gtggctgagc tcttctgtcg ccgctgccgc 120

cgctgcgtgt gcgcgctttg cccggtgctg ggcgcgcacc gtggccaccc tgtgggcctg 180

gcgctggagg cagcggtgca cgtgcagaaa ctcagccaag aatgtttaaa gcagctggca 240

atcaagaagc agcagcacat tgacaacata acccagatag aagatgccac cgagaagctc 300

aaggctaatg cagagtcaag taaaacctgg ctgaagggga aattcactga actcagatta 360

ctacttgacg aagaggaagc gctggccaag aaattcattg ataaaaacac gcagcttacc 420

ctccaggtgt acagggaaca agctgactct tgcagagagc aacttgacat catgaatgat 480

ctctccaaca gggtctggag tatcagccag gagcccgatc ctgtccagag gcttcaggca 540

tacacggcca ccgagcagga gatgcagcag cagatgagcc tcggggagct gtgccatccc 600

gtgcccctct cctttgagcc cgtcaagagc ttctttaagg gcctcgtgga agccgtggag 660

agtacattac agacgccatt ggacattcgc cttaaggaaa gcataaactg ccagctctca 720

gacccttcca gcaccaagcc aggtaccttg ttgaaaacca gcccctcacc agagcgatcg 780

ctattgctga aatacgcgcg cacgcccacg ctggatcctg acacgatgca cgcgcgcctg 840

cgcctgtccg ccgatcgcct gacggtgcgc tgcggcctgc tgggcagcct ggggcccgtg 900

cccgtgctgc ggttcgacgc gctctggcaa gtgctggctc gtgactgctt cgccaccggc 960

cgccactact gggaggttga cgtgcaggag gcgggcgccg gctggtgggt gggcgcggcc 1020

tacgcctccc ttcggcgccg cggggcctcg gccgccgccc gcctgggctg caaccgccag 1080

tcctggtgcc tcaagcgcta cgaccttgag tactgggcct tccacgacgg ccagcgcagc 1140

cgcctgcggc cccgcgacga cctcgaccgg ctcggcgtct tcctggacta cgaggccggc 1200

gtcctcgcct tctacgacgt gacgggcggc atgagccacc tgcatacctt ccgcgccacg 1260

ttccaggagc cgctctaccc ggccctgcgg ctctgggagg gggccatcag catcccccgg 1320

ctgccctag 1329

<210> 28

<211> 442

<212> PRT

<213> 智人(Homo sapiens)

<400> 28

Met Ala Gly Ala Ala Thr Gly Ser Arg Thr Pro Gly Arg Ser Glu Leu

1 5 10 15

Val Glu Gly Cys Gly Trp Arg Cys Pro Glu His Gly Asp Arg Val Ala

20 25 30

Glu Leu Phe Cys Arg Arg Cys Arg Arg Cys Val Cys Ala Leu Cys Pro

35 40 45

Val Leu Gly Ala His Arg Gly His Pro Val Gly Leu Ala Leu Glu Ala

50 55 60

Ala Val His Val Gln Lys Leu Ser Gln Glu Cys Leu Lys Gln Leu Ala

65 70 75 80

Ile Lys Lys Gln Gln His Ile Asp Asn Ile Thr Gln Ile Glu Asp Ala

85 90 95

Thr Glu Lys Leu Lys Ala Asn Ala Glu Ser Ser Lys Thr Trp Leu Lys

100 105 110

Gly Lys Phe Thr Glu Leu Arg Leu Leu Leu Asp Glu Glu Glu Ala Leu

115 120 125

Ala Lys Lys Phe Ile Asp Lys Asn Thr Gln Leu Thr Leu Gln Val Tyr

130 135 140

Arg Glu Gln Ala Asp Ser Cys Arg Glu Gln Leu Asp Ile Met Asn Asp

145 150 155 160

Leu Ser Asn Arg Val Trp Ser Ile Ser Gln Glu Pro Asp Pro Val Gln

165 170 175

Arg Leu Gln Ala Tyr Thr Ala Thr Glu Gln Glu Met Gln Gln Gln Met

180 185 190

Ser Leu Gly Glu Leu Cys His Pro Val Pro Leu Ser Phe Glu Pro Val

195 200 205

Lys Ser Phe Phe Lys Gly Leu Val Glu Ala Val Glu Ser Thr Leu Gln

210 215 220

Thr Pro Leu Asp Ile Arg Leu Lys Glu Ser Ile Asn Cys Gln Leu Ser

225 230 235 240

Asp Pro Ser Ser Thr Lys Pro Gly Thr Leu Leu Lys Thr Ser Pro Ser

245 250 255

Pro Glu Arg Ser Leu Leu Leu Lys Tyr Ala Arg Thr Pro Thr Leu Asp

260 265 270

Pro Asp Thr Met His Ala Arg Leu Arg Leu Ser Ala Asp Arg Leu Thr

275 280 285

Val Arg Cys Gly Leu Leu Gly Ser Leu Gly Pro Val Pro Val Leu Arg

290 295 300

Phe Asp Ala Leu Trp Gln Val Leu Ala Arg Asp Cys Phe Ala Thr Gly

305 310 315 320

Arg His Tyr Trp Glu Val Asp Val Gln Glu Ala Gly Ala Gly Trp Trp

325 330 335

Val Gly Ala Ala Tyr Ala Ser Leu Arg Arg Arg Gly Ala Ser Ala Ala

340 345 350

Ala Arg Leu Gly Cys Asn Arg Gln Ser Trp Cys Leu Lys Arg Tyr Asp

355 360 365

Leu Glu Tyr Trp Ala Phe His Asp Gly Gln Arg Ser Arg Leu Arg Pro

370 375 380

Arg Asp Asp Leu Asp Arg Leu Gly Val Phe Leu Asp Tyr Glu Ala Gly

385 390 395 400

Val Leu Ala Phe Tyr Asp Val Thr Gly Gly Met Ser His Leu His Thr

405 410 415

Phe Arg Ala Thr Phe Gln Glu Pro Leu Tyr Pro Ala Leu Arg Leu Trp

420 425 430

Glu Gly Ala Ile Ser Ile Pro Arg Leu Pro

435 440

<210> 29

<211> 1398

<212> DNA

<213> 智人(Homo sapiens)

<400> 29

atgcccgcaa ccccgtccct gaaggtggtc catgagctgc ctgcctgtac cctctgtgcg 60

gggccgctgg aggatgcggt gaccattccc tgtggacaca ccttctgccg gctctgcctc 120

cccgcgctct cccagatggg ggcccaatcc tcgggcaaga tcctgctctg cccgctctgc 180

caagaggagg agcaggcaga gactcccatg gcccctgtgc ccctgggccc gctgggagaa 240

acttactgcg aggagcacgg cgagaagatc tacttcttct gcgagaacga tgccgagttc 300

ctctgtgtgt tctgcaggga gggtcccacg caccaggcgc acaccgtggg gttcctggac 360

gaggccattc agccctaccg ggatcgtctc aggagtcgac tggaagctct gagcacggag 420

agagatgaga ttgaggatgt aaagtgtcaa gaagaccaga agcttcaagt gctgctgact 480

cagatcgaaa gcaagaagca tcaggtggaa acagcttttg agaggctgca gcaggagctg 540

gagcagcagc gatgtctcct gctggccagg ctgagggagc tggagcagca gatttggaag 600

gagagggatg aatatatcac aaaggtctct gaggaagtca cccggcttgg agcccaggtc 660

aaggagctgg aggagaagtg tcagcagcca gcaagtgagc ttctacaaga tgtcagagtc 720

aaccagagca ggtgtgagat gaagactttt gtgagtcctg aggccatttc tcctgacctt 780

gtcaagaaga tccgtgattt ccacaggaaa atactcaccc tcccagagat gatgaggatg 840

ttctcagaaa acttggcgca tcatctggaa atagattcag gggtcatcac tctggaccct 900

cagaccgcca gccggagcct ggttctctcg gaagacagga agtcagtgag gtacacccgg 960

cagaagaaga gcctgccaga cagccccctg cgcttcgacg gcctcccggc ggttctgggc 1020

ttcccgggct tctcctccgg gcgccaccgc tggcaggttg acctgcagct gggcgacggc 1080

ggcggctgca cggtgggggt ggccggggag ggggtgagga ggaagggaga gatgggactc 1140

agcgccgagg acggcgtctg ggccgtgatc atctcgcacc agcagtgctg ggccagcacc 1200

tccccgggca ccgacctgcc gctgagcgag atcccgcgcg gcgtgagagt cgccctggac 1260

tacgaggcgg ggcaggtgac cctccacaac gcccagaccc aggagcccat cttcaccttc 1320

actgcctctt tctccggcaa agtcttccct ttctttgccg tctggaaaaa aggttcctgc 1380

cttacgctga aaggctga 1398

<210> 30

<211> 465

<212> PRT

<213> 智人(Homo sapiens)

<400> 30

Met Pro Ala Thr Pro Ser Leu Lys Val Val His Glu Leu Pro Ala Cys

1 5 10 15

Thr Leu Cys Ala Gly Pro Leu Glu Asp Ala Val Thr Ile Pro Cys Gly

20 25 30

His Thr Phe Cys Arg Leu Cys Leu Pro Ala Leu Ser Gln Met Gly Ala

35 40 45

Gln Ser Ser Gly Lys Ile Leu Leu Cys Pro Leu Cys Gln Glu Glu Glu

50 55 60

Gln Ala Glu Thr Pro Met Ala Pro Val Pro Leu Gly Pro Leu Gly Glu

65 70 75 80

Thr Tyr Cys Glu Glu His Gly Glu Lys Ile Tyr Phe Phe Cys Glu Asn

85 90 95

Asp Ala Glu Phe Leu Cys Val Phe Cys Arg Glu Gly Pro Thr His Gln

100 105 110

Ala His Thr Val Gly Phe Leu Asp Glu Ala Ile Gln Pro Tyr Arg Asp

115 120 125

Arg Leu Arg Ser Arg Leu Glu Ala Leu Ser Thr Glu Arg Asp Glu Ile

130 135 140

Glu Asp Val Lys Cys Gln Glu Asp Gln Lys Leu Gln Val Leu Leu Thr

145 150 155 160

Gln Ile Glu Ser Lys Lys His Gln Val Glu Thr Ala Phe Glu Arg Leu

165 170 175

Gln Gln Glu Leu Glu Gln Gln Arg Cys Leu Leu Leu Ala Arg Leu Arg

180 185 190

Glu Leu Glu Gln Gln Ile Trp Lys Glu Arg Asp Glu Tyr Ile Thr Lys

195 200 205

Val Ser Glu Glu Val Thr Arg Leu Gly Ala Gln Val Lys Glu Leu Glu

210 215 220

Glu Lys Cys Gln Gln Pro Ala Ser Glu Leu Leu Gln Asp Val Arg Val

225 230 235 240

Asn Gln Ser Arg Cys Glu Met Lys Thr Phe Val Ser Pro Glu Ala Ile

245 250 255

Ser Pro Asp Leu Val Lys Lys Ile Arg Asp Phe His Arg Lys Ile Leu

260 265 270

Thr Leu Pro Glu Met Met Arg Met Phe Ser Glu Asn Leu Ala His His

275 280 285

Leu Glu Ile Asp Ser Gly Val Ile Thr Leu Asp Pro Gln Thr Ala Ser

290 295 300

Arg Ser Leu Val Leu Ser Glu Asp Arg Lys Ser Val Arg Tyr Thr Arg

305 310 315 320

Gln Lys Lys Ser Leu Pro Asp Ser Pro Leu Arg Phe Asp Gly Leu Pro

325 330 335

Ala Val Leu Gly Phe Pro Gly Phe Ser Ser Gly Arg His Arg Trp Gln

340 345 350

Val Asp Leu Gln Leu Gly Asp Gly Gly Gly Cys Thr Val Gly Val Ala

355 360 365

Gly Glu Gly Val Arg Arg Lys Gly Glu Met Gly Leu Ser Ala Glu Asp

370 375 380

Gly Val Trp Ala Val Ile Ile Ser His Gln Gln Cys Trp Ala Ser Thr

385 390 395 400

Ser Pro Gly Thr Asp Leu Pro Leu Ser Glu Ile Pro Arg Gly Val Arg

405 410 415

Val Ala Leu Asp Tyr Glu Ala Gly Gln Val Thr Leu His Asn Ala Gln

420 425 430

Thr Gln Glu Pro Ile Phe Thr Phe Thr Ala Ser Phe Ser Gly Lys Val

435 440 445

Phe Pro Phe Phe Ala Val Trp Lys Lys Gly Ser Cys Leu Thr Leu Lys

450 455 460

Gly

465

<210> 31

<211> 1695

<212> DNA

<213> 智人(Homo sapiens)

<400> 31

atggctgagt tggatctaat ggctccaggg ccactgccca gggccactgc tcagccccca 60

gcccctctca gcccagactc tgggtcaccc agcccagatt ctgggtcagc cagcccagtg 120

gaagaagagg acgtgggctc ctcggagaag cttggcaggg agacggagga acaggacagc 180

gactctgcag agcaggggga tcctgctggt gaggggaaag aggtcctgtg tgacttctgc 240

cttgatgaca ccagaagagt gaaggcagtg aagtcctgtc taacctgcat ggtgaattac 300

tgtgaagagc acttgcagcc gcatcaggtg aacatcaaac tgcaaagcca cctgctgacc 360

gagccagtga aggaccacaa ctggcgatac tgccctgccc accacagccc actgtctgcc 420

ttctgctgcc ctgatcagca gtgcatctgc caggactgtt gccaggagca cagtggccac 480

accatagtct ccctggatgc agcccgcagg gacaaggagg ctgaactcca gtgcacccag 540

ttagacttgg agcggaaact caagttgaat gaaaatgcca tctccaggct ccaggctaac 600

caaaagtctg ttctggtgtc ggtgtcagag gtcaaagcgg tggctgaaat gcagtttggg 660

gaactccttg ctgctgtgag gaaggcccag gccaatgtga tgctcttctt agaggagaag 720

gagcaagctg cgctgagcca ggccaacggt atcaaggccc acctggagta caggagtgcc 780

gagatggaga agagcaagca ggagctggag aggatggcgg ccatcagcaa cactgtccag 840

ttcttggagg agtactgcaa gtttaagaac actgaagaca tcaccttccc tagtgtttac 900

gtagggctga aggataaact ctcgggcatc cgcaaagtta tcacggaatc cactgtacac 960

ttaatccagt tgctggagaa ctataagaaa aagctccagg agttttccaa ggaagaggag 1020

tatgacatca gaactcaagt gtctgccgtt gttcagcgca aatattggac ttccaaacct 1080

gagcccagca ccagggaaca gttcctccaa tatgcgtatg acatcacgtt tgacccggac 1140

acagcacaca agtatctccg gctgcaggag gagaaccgca aggtcaccaa caccacgccc 1200

tgggagcatc cctacccgga cctccccagc aggttcctgc actggcggca ggtgctgtcc 1260

cagcagagtc tgtacctgca caggtactat tttgaggtgg agatcttcgg ggcaggcacc 1320

tatgttggcc tgacctgcaa aggcatcgac cggaaagggg aggagcgcaa cagttgcatt 1380

tccggaaaca acttctcctg gagcctccaa tggaacggga aggagttcac ggcctggtac 1440

agtgacatgg agaccccact caaagctggc cctttccgga ggctcggggt ctatatcgac 1500

ttcccgggag ggatcctttc cttctatggc gtagagtatg ataccatgac tctggttcac 1560

aagtttgcct gcaaattttc agaaccagtc tatgctgcct tctggctttc caagaaggaa 1620

aacgccatcc ggattgtaga tctgggagag gaacccgaga agccagcacc gtccttggtg 1680

gggactgctc cctag 1695

<210> 32

<211> 564

<212> PRT

<213> 智人(Homo sapiens)

<400> 32

Met Ala Glu Leu Asp Leu Met Ala Pro Gly Pro Leu Pro Arg Ala Thr

1 5 10 15

Ala Gln Pro Pro Ala Pro Leu Ser Pro Asp Ser Gly Ser Pro Ser Pro

20 25 30

Asp Ser Gly Ser Ala Ser Pro Val Glu Glu Glu Asp Val Gly Ser Ser

35 40 45

Glu Lys Leu Gly Arg Glu Thr Glu Glu Gln Asp Ser Asp Ser Ala Glu

50 55 60

Gln Gly Asp Pro Ala Gly Glu Gly Lys Glu Val Leu Cys Asp Phe Cys

65 70 75 80

Leu Asp Asp Thr Arg Arg Val Lys Ala Val Lys Ser Cys Leu Thr Cys

85 90 95

Met Val Asn Tyr Cys Glu Glu His Leu Gln Pro His Gln Val Asn Ile

100 105 110

Lys Leu Gln Ser His Leu Leu Thr Glu Pro Val Lys Asp His Asn Trp

115 120 125

Arg Tyr Cys Pro Ala His His Ser Pro Leu Ser Ala Phe Cys Cys Pro

130 135 140

Asp Gln Gln Cys Ile Cys Gln Asp Cys Cys Gln Glu His Ser Gly His

145 150 155 160

Thr Ile Val Ser Leu Asp Ala Ala Arg Arg Asp Lys Glu Ala Glu Leu

165 170 175

Gln Cys Thr Gln Leu Asp Leu Glu Arg Lys Leu Lys Leu Asn Glu Asn

180 185 190

Ala Ile Ser Arg Leu Gln Ala Asn Gln Lys Ser Val Leu Val Ser Val

195 200 205

Ser Glu Val Lys Ala Val Ala Glu Met Gln Phe Gly Glu Leu Leu Ala

210 215 220

Ala Val Arg Lys Ala Gln Ala Asn Val Met Leu Phe Leu Glu Glu Lys

225 230 235 240

Glu Gln Ala Ala Leu Ser Gln Ala Asn Gly Ile Lys Ala His Leu Glu

245 250 255

Tyr Arg Ser Ala Glu Met Glu Lys Ser Lys Gln Glu Leu Glu Arg Met

260 265 270

Ala Ala Ile Ser Asn Thr Val Gln Phe Leu Glu Glu Tyr Cys Lys Phe

275 280 285

Lys Asn Thr Glu Asp Ile Thr Phe Pro Ser Val Tyr Val Gly Leu Lys

290 295 300

Asp Lys Leu Ser Gly Ile Arg Lys Val Ile Thr Glu Ser Thr Val His

305 310 315 320

Leu Ile Gln Leu Leu Glu Asn Tyr Lys Lys Lys Leu Gln Glu Phe Ser

325 330 335

Lys Glu Glu Glu Tyr Asp Ile Arg Thr Gln Val Ser Ala Val Val Gln

340 345 350

Arg Lys Tyr Trp Thr Ser Lys Pro Glu Pro Ser Thr Arg Glu Gln Phe

355 360 365

Leu Gln Tyr Ala Tyr Asp Ile Thr Phe Asp Pro Asp Thr Ala His Lys

370 375 380

Tyr Leu Arg Leu Gln Glu Glu Asn Arg Lys Val Thr Asn Thr Thr Pro

385 390 395 400

Trp Glu His Pro Tyr Pro Asp Leu Pro Ser Arg Phe Leu His Trp Arg

405 410 415

Gln Val Leu Ser Gln Gln Ser Leu Tyr Leu His Arg Tyr Tyr Phe Glu

420 425 430

Val Glu Ile Phe Gly Ala Gly Thr Tyr Val Gly Leu Thr Cys Lys Gly

435 440 445

Ile Asp Arg Lys Gly Glu Glu Arg Asn Ser Cys Ile Ser Gly Asn Asn

450 455 460

Phe Ser Trp Ser Leu Gln Trp Asn Gly Lys Glu Phe Thr Ala Trp Tyr

465 470 475 480

Ser Asp Met Glu Thr Pro Leu Lys Ala Gly Pro Phe Arg Arg Leu Gly

485 490 495

Val Tyr Ile Asp Phe Pro Gly Gly Ile Leu Ser Phe Tyr Gly Val Glu

500 505 510

Tyr Asp Thr Met Thr Leu Val His Lys Phe Ala Cys Lys Phe Ser Glu

515 520 525

Pro Val Tyr Ala Ala Phe Trp Leu Ser Lys Lys Glu Asn Ala Ile Arg

530 535 540

Ile Val Asp Leu Gly Glu Glu Pro Glu Lys Pro Ala Pro Ser Leu Val

545 550 555 560

Gly Thr Ala Pro

<210> 33

<211> 1434

<212> DNA

<213> 智人(Homo sapiens)

<400> 33

atggaggctg tggaactcgc cagaaaactg caggaggaag ctacgtgctc catctgtctg 60

gattacttca cagaccctgt gatgaccacc tgtggccaca acttctgccg agcctgcatc 120

cagctgagct gggaaaaggc gaggggcaag aaggggaggc ggaagcggaa gggctccttc 180

ccctgccccg agtgcagaga gatgtccccg cagaggaacc tgctgcccaa ccggctgctg 240

accaaggtgg ccgagatggc gcagcagcat cctggtctgc agaagcaaga cctgtgccag 300

gagcaccacg agcccctcaa gcttttctgc cagaaggacc agagccccat ctgtgtggtg 360

tgcagggagt cccgggagca ccggctgcac agggtgctgc ccgccgagga ggcagtgcag 420

gggtacaagt tgaagctgga ggaggacatg gagtaccttc gggagcagat caccaggaca 480

gggaatctgc aggccaggga ggagcagagc ttagccgagt ggcagggcaa ggtgaaggag 540

cggagagaac gcattgtgct ggagtttgag aagatgaacc tctacctggt ggaagaagag 600

cagaggctcc tccaggctct ggagacggaa gaagaggaga ctgccagcag gctccgggag 660

agcgtggcct gcctggaccg gcagggtcac tctctggagc tgctgctgct gcagctggag 720

gagcggagca cacaggggcc cctccagatg ctgcaggaca tgaaggaacc cctgagcagg 780

aagaacaacg tgagtgtgca gtgcccagag gttgcccccc caaccagacc caggactgtg 840

tgcagagttc ccggacagat tgaagtgcta agaggctttc tagaggatgt ggtgcctgat 900

gccacctccg cgtaccccta cctcctcctg tatgagagcc gccagaggcg ctacctcggc 960

tcttcgccgg agggcagtgg gttctgcagc aaggaccgat ttgtggctta cccctgtgct 1020

gtgggccaga cggccttctc ctctgggagg cactactggg aggtgggcat gaacatcacc 1080

ggggacgcgt tgtgggccct gggtgtgtgc agggacaacg tgagccggaa agacagggtc 1140

cccaagtgcc ccgaaaacgg cttctgggtg gtgcagctgt ccaaggggac caagtactta 1200

tccaccttct ctgccctaac cccggtcatg ctgatggagc ctcccagcca catgggcatc 1260

ttcctggact tcgaagccgg ggaagtgtcc ttctacagtg taagcgatgg gtcccacctg 1320

cacacctact cccaggccac cttcccaggc cccctgcagc ctttcttctg cctgggggct 1380

ccgaagtctg gtcagatggt catctccaca gtgaccatgt gggtgaaagg atag 1434

<210> 34

<211> 477

<212> PRT

<213> 智人(Homo sapiens)

<400> 34

Met Glu Ala Val Glu Leu Ala Arg Lys Leu Gln Glu Glu Ala Thr Cys

1 5 10 15

Ser Ile Cys Leu Asp Tyr Phe Thr Asp Pro Val Met Thr Thr Cys Gly

20 25 30

His Asn Phe Cys Arg Ala Cys Ile Gln Leu Ser Trp Glu Lys Ala Arg

35 40 45

Gly Lys Lys Gly Arg Arg Lys Arg Lys Gly Ser Phe Pro Cys Pro Glu

50 55 60

Cys Arg Glu Met Ser Pro Gln Arg Asn Leu Leu Pro Asn Arg Leu Leu

65 70 75 80

Thr Lys Val Ala Glu Met Ala Gln Gln His Pro Gly Leu Gln Lys Gln

85 90 95

Asp Leu Cys Gln Glu His His Glu Pro Leu Lys Leu Phe Cys Gln Lys

100 105 110

Asp Gln Ser Pro Ile Cys Val Val Cys Arg Glu Ser Arg Glu His Arg

115 120 125

Leu His Arg Val Leu Pro Ala Glu Glu Ala Val Gln Gly Tyr Lys Leu

130 135 140

Lys Leu Glu Glu Asp Met Glu Tyr Leu Arg Glu Gln Ile Thr Arg Thr

145 150 155 160

Gly Asn Leu Gln Ala Arg Glu Glu Gln Ser Leu Ala Glu Trp Gln Gly

165 170 175

Lys Val Lys Glu Arg Arg Glu Arg Ile Val Leu Glu Phe Glu Lys Met

180 185 190

Asn Leu Tyr Leu Val Glu Glu Glu Gln Arg Leu Leu Gln Ala Leu Glu

195 200 205

Thr Glu Glu Glu Glu Thr Ala Ser Arg Leu Arg Glu Ser Val Ala Cys

210 215 220

Leu Asp Arg Gln Gly His Ser Leu Glu Leu Leu Leu Leu Gln Leu Glu

225 230 235 240

Glu Arg Ser Thr Gln Gly Pro Leu Gln Met Leu Gln Asp Met Lys Glu

245 250 255

Pro Leu Ser Arg Lys Asn Asn Val Ser Val Gln Cys Pro Glu Val Ala

260 265 270

Pro Pro Thr Arg Pro Arg Thr Val Cys Arg Val Pro Gly Gln Ile Glu

275 280 285

Val Leu Arg Gly Phe Leu Glu Asp Val Val Pro Asp Ala Thr Ser Ala

290 295 300

Tyr Pro Tyr Leu Leu Leu Tyr Glu Ser Arg Gln Arg Arg Tyr Leu Gly

305 310 315 320

Ser Ser Pro Glu Gly Ser Gly Phe Cys Ser Lys Asp Arg Phe Val Ala

325 330 335

Tyr Pro Cys Ala Val Gly Gln Thr Ala Phe Ser Ser Gly Arg His Tyr

340 345 350

Trp Glu Val Gly Met Asn Ile Thr Gly Asp Ala Leu Trp Ala Leu Gly

355 360 365

Val Cys Arg Asp Asn Val Ser Arg Lys Asp Arg Val Pro Lys Cys Pro

370 375 380

Glu Asn Gly Phe Trp Val Val Gln Leu Ser Lys Gly Thr Lys Tyr Leu

385 390 395 400

Ser Thr Phe Ser Ala Leu Thr Pro Val Met Leu Met Glu Pro Pro Ser

405 410 415

His Met Gly Ile Phe Leu Asp Phe Glu Ala Gly Glu Val Ser Phe Tyr

420 425 430

Ser Val Ser Asp Gly Ser His Leu His Thr Tyr Ser Gln Ala Thr Phe

435 440 445

Pro Gly Pro Leu Gln Pro Phe Phe Cys Leu Gly Ala Pro Lys Ser Gly

450 455 460

Gln Met Val Ile Ser Thr Val Thr Met Trp Val Lys Gly

465 470 475

<210> 35

<211> 1945

<212> DNA

<213> 智人(Homo sapiens)

<400> 35

atggaaacac tggagtcaga actgacctgc cctatttgtc tggagctctt tgaggaccct 60

cttctactgc cctgcgcaca cagcctctgc ttcaactgcg cccaccgcat cctagtatca 120

cactgtgcca ccaacgagtc tgtggagtcc atcaccgcct tccagtgccc cacctgccgg 180

catgtcatca ccctcagcca gcgaggtcta gacgggctca agcgcaacgt caccctacag 240

aacatcatcg acaggttcca gaaagcatca gtgagcgggc ccaactctcc cagcgagacc 300

cgtcgggagc gggcctttga cgccaacacc atgacctccg ccgagaaggt cctctgccag 360

ttttgtgacc aggatcctgc ccaggacgct gtgaagacct gtgtcacttg tgaagtatcc 420

tactgtgacg agtgcctgaa agccactcac ccgaataaga agccctttac aggccatcgt 480

ctgattgagc caattccgga ctctcacatc cgggggctga tgtgcttgga gcatgaggat 540

gagaaggtga atatgtactg tgtgaccgat gaccagttaa tctgtgcctt gtgtaaactg 600

gttgggcggc accgcgatca tcaggtggca gctttgagtg agcgctatga caaattgaag 660

caaaacttag agagtaacct caccaacctt attaagagga acacagaact ggagaccctt 720

ttggctaaac tcatccaaac ctgtcaacat gttgaagtca atgcatcacg tcaagaagcc 780

aaattgacag aggagtgtga tcttctcatt gagatcattc agcaaagacg acagattatt 840

ggaaccaaga tcaaagaagg gaaggtgatg aggcttcgca aactggctca gcagattgca 900

aactgcaaac agtgcattga gcggtcagca tcactcatct cccaagcgga acactctctg 960

aaggagaatg atcatgcgcg tttcctacag actgctaaga atatcaccga gagagtctcc 1020

atggcaactg catcctccca ggttctaatt cctgaaatca acctcaatga cacatttgac 1080

acctttgcct tagatttttc ccgagagaag aaactgctag aatgtctgga ttaccttaca 1140

gctcccaacc ctcccacaat tagagaagag ctctgcacag cttcatatga caccatcact 1200

gtgcattgga cctccgatga tgagttcagc gtggtctcct acgagctcca gtacaccata 1260

ttcaccggac aagccaacgt cgttagtctg tgtaattcgg ctgatagctg gatgatagta 1320

cccaacatca agcagaacca ctacacggtg cacggtctgc agagcggcac caagtacatc 1380

ttcatggtca aggccatcaa ccaggcgggc agccgcagca gtgagcctgg gaagttgaag 1440

acaaacagcc aaccatttaa actggatccc aaatctgctc atcgaaaact gaaggtgtcc 1500

catgataact tgacagtaga acgtgatgag tcatcatcca agaagagtca cacacctgaa 1560

cgcttcacca gccaggggag ctatggagta gctggaaatg tgtttattga tagtggccgg 1620

cattattggg aagtggtcat aagtggaagc acatggtatg ccattggtct tgcttacaaa 1680

tcagccccga agcatgaatg gattgggaag aactctgctt cctgggcgct ctgccgctgc 1740

aacaataact gggtggtgag acacaatagc aaggaaatcc ccattgagcc tgccccccac 1800

ctccggcgcg tgggcatcct gctggactat gataacggct ctatcgcctt ttatgatgct 1860

ttgaactcca tccacctcta caccttcgac gtcgcatttg cgcagcctgt ttgccccacc 1920

ttcaccgtgt ggaacaagtg tctga 1945

<210> 36

<211> 667

<212> PRT

<213> 智人(Homo sapiens)

<400> 36

Met Glu Thr Leu Glu Ser Glu Leu Thr Cys Pro Ile Cys Leu Glu Leu

1 5 10 15

Phe Glu Asp Pro Leu Leu Leu Pro Cys Ala His Ser Leu Cys Phe Asn

20 25 30

Cys Ala His Arg Ile Leu Val Ser His Cys Ala Thr Asn Glu Ser Val

35 40 45

Glu Ser Ile Thr Ala Phe Gln Cys Pro Thr Cys Arg His Val Ile Thr

50 55 60

Leu Ser Gln Arg Gly Leu Asp Gly Leu Lys Arg Asn Val Thr Leu Gln

65 70 75 80

Asn Ile Ile Asp Arg Phe Gln Lys Ala Ser Val Ser Gly Pro Asn Ser

85 90 95

Pro Ser Glu Thr Arg Arg Glu Arg Ala Phe Asp Ala Asn Thr Met Thr

100 105 110

Ser Ala Glu Lys Val Leu Cys Gln Phe Cys Asp Gln Asp Pro Ala Gln

115 120 125

Asp Ala Val Lys Thr Cys Val Thr Cys Glu Val Ser Tyr Cys Asp Glu

130 135 140

Cys Leu Lys Ala Thr His Pro Asn Lys Lys Pro Phe Thr Gly His Arg

145 150 155 160

Leu Ile Glu Pro Ile Pro Asp Ser His Ile Arg Gly Leu Met Cys Leu

165 170 175

Glu His Glu Asp Glu Lys Val Asn Met Tyr Cys Val Thr Asp Asp Gln

180 185 190

Leu Ile Cys Ala Leu Cys Lys Leu Val Gly Arg His Arg Asp His Gln

195 200 205

Val Ala Ala Leu Ser Glu Arg Tyr Asp Lys Leu Lys Gln Asn Leu Glu

210 215 220

Ser Asn Leu Thr Asn Leu Ile Lys Arg Asn Thr Glu Leu Glu Thr Leu

225 230 235 240

Leu Ala Lys Leu Ile Gln Thr Cys Gln His Val Glu Val Asn Ala Ser

245 250 255

Arg Gln Glu Ala Lys Leu Thr Glu Glu Cys Asp Leu Leu Ile Glu Ile

260 265 270

Ile Gln Gln Arg Arg Gln Ile Ile Gly Thr Lys Ile Lys Glu Gly Lys

275 280 285

Val Met Arg Leu Arg Lys Leu Ala Gln Gln Ile Ala Asn Cys Lys Gln

290 295 300

Cys Ile Glu Arg Ser Ala Ser Leu Ile Ser Gln Ala Glu His Ser Leu

305 310 315 320

Lys Glu Asn Asp His Ala Arg Phe Leu Gln Thr Ala Lys Asn Ile Thr

325 330 335

Glu Arg Val Ser Met Ala Thr Ala Ser Ser Gln Val Leu Ile Pro Glu

340 345 350

Ile Asn Leu Asn Asp Thr Phe Asp Thr Phe Ala Leu Asp Phe Ser Arg

355 360 365

Glu Lys Lys Leu Leu Glu Cys Leu Asp Tyr Leu Thr Ala Pro Asn Pro

370 375 380

Pro Thr Ile Arg Glu Glu Leu Cys Thr Ala Ser Tyr Asp Thr Ile Thr

385 390 395 400

Val His Trp Thr Ser Asp Asp Glu Phe Ser Val Val Ser Tyr Glu Leu

405 410 415

Gln Tyr Thr Ile Phe Thr Gly Gln Ala Asn Val Val Ser Leu Cys Asn

420 425 430

Ser Ala Asp Ser Trp Met Ile Val Pro Asn Ile Lys Gln Asn His Tyr

435 440 445

Thr Val His Gly Leu Gln Ser Gly Thr Lys Tyr Ile Phe Met Val Lys

450 455 460

Ala Ile Asn Gln Ala Gly Ser Arg Ser Ser Glu Pro Gly Lys Leu Lys

465 470 475 480

Thr Asn Ser Gln Pro Phe Lys Leu Asp Pro Lys Ser Ala His Arg Lys

485 490 495

Leu Lys Val Ser His Asp Asn Leu Thr Val Glu Arg Asp Glu Ser Ser

500 505 510

Ser Lys Lys Ser His Thr Pro Glu Arg Phe Thr Ser Gln Gly Ser Tyr

515 520 525

Gly Val Ala Gly Asn Val Phe Ile Asp Ser Gly Arg His Tyr Trp Glu

530 535 540

Val Val Ile Ser Gly Ser Thr Trp Tyr Ala Ile Gly Leu Ala Tyr Lys

545 550 555 560

Ser Ala Pro Lys His Glu Trp Ile Gly Lys Asn Ser Ala Ser Trp Ala

565 570 575

Leu Cys Arg Cys Asn Asn Asn Trp Val Val Arg His Asn Ser Lys Glu

580 585 590

Ile Pro Ile Glu Pro Ala Pro His Leu Arg Arg Val Gly Ile Leu Leu

595 600 605

Asp Tyr Asp Asn Gly Ser Ile Ala Phe Tyr Asp Ala Leu Asn Ser Ile

610 615 620

His Leu Tyr Thr Phe Asp Val Ala Phe Ala Gln Pro Val Cys Pro Thr

625 630 635 640

Phe Thr Val Trp Asn Lys Cys Leu Thr Ile Ile Thr Gly Leu Pro Ile

645 650 655

Pro Asp His Leu Asp Cys Thr Glu Gln Leu Pro

660 665

<210> 37

<211> 2649

<212> DNA

<213> 智人(Homo sapiens)

<400> 37

atggagcctg cacccgcccg atctccgagg ccccagcagg accccgcccg gccccaggag 60

cccaccatgc ctccccccga gaccccctct gaaggccgcc agcccagccc cagccccagc 120

cctacagagc gagcccccgc ttcggaggag gagttccagt ttctgcgctg ccagcaatgc 180

caggcggaag ccaagtgccc gaagctgctg ccttgtctgc acacgctgtg ctcaggatgc 240

ctggaggcgt cgggcatgca gtgccccatc tgccaggcgc cctggcccct aggtgcagac 300

acacccgccc tggataacgt ctttttcgag agtctgcagc ggcgcctgtc ggtgtaccgg 360

cagattgtgg atgcgcaggc tgtgtgcacc cgctgcaaag agtcggccga cttctggtgc 420

tttgagtgcg agcagctcct ctgcgccaag tgcttcgagg cacaccagtg gttcctcaag 480

cacgaggccc ggcccctagc agagctgcgc aaccagtcgg tgcgtgagtt cctggacggc 540

acccgcaaga ccaacaacat cttctgctcc aaccccaacc accgcacccc tacgctgacc 600

agcatctact gccgaggatg ttccaagccg ctgtgctgct cgtgcgcgct ccttgacagc 660

agccacagtg agctcaagtg cgacatcagc gcagagatcc agcagcgaca ggaggagctg 720

gacgccatga cgcaggcgct gcaggagcag gatagtgcct ttggcgcggt tcacgcgcag 780

atgcacgcgg ccgtcggcca gctgggccgc gcgcgtgccg agaccgagga gctgatccgc 840

gagcgcgtgc gccaggtggt agctcacgtg cgggctcagg agcgcgagct gctggaggct 900

gtggacgcgc ggtaccagcg cgactacgag gagatggcca gtcggctggg ccgcctggat 960

gctgtgctgc agcgcatccg cacgggcagc gcgctggtgc agaggatgaa gtgctacgcc 1020

tcggaccagg aggtgctgga catgcacggt ttcctgcgcc aggcgctctg ccgcctgcgc 1080

caggaggagc cccagagcct gcaagctgcc gtgcgcaccg atggcttcga cgagttcaag 1140

gtgcgcctgc aggacctcag ctcttgcatc acccagggga aagatgcagc tgtatccaag 1200

aaagccagcc cagaggctgc cagcactccc agggacccta ttgacgttga cctgcccgag 1260

gaggcagaga gagtgaaggc ccaggttcag gccctggggc tggctgaagc ccagcctatg 1320

gctgtggtac agtcagtgcc cggggcacac cccgtgccag tgtacgcctt ctccatcaaa 1380

ggcccttcct atggagagga tgtctccaat acaacgacag cccagaagag gaagtgcagc 1440

cagacccagt gccccaggaa ggtcatcaag atggagtctg aggaggggaa ggaggcaagg 1500

ttggctcgga gctccccgga gcagcccagg cccagcacct ccaaggcagt ctcaccaccc 1560

cacctggatg gaccgcctag ccccaggagc cccgtcatag gaagtgaggt cttcctgccc 1620

aacagcaacc acgtggccag tggcgccggg gaggcagagg aacgcgttgt ggtgatcagc 1680

agctcggaag actcagatgc cgaaaactcg tcctcccgag agctggatga cagcagcagt 1740

gagtccagtg acctccagct ggaaggcccc agcaccctca gggtcctgga cgagaacctt 1800

gctgaccccc aagcagaaga cagacctctg gttttctttg acctcaagat tgacaatgaa 1860

acccagaaga ttagccagct ggctgcggtg aaccgggaaa gcaagttccg cgtggtcatc 1920

cagcctgaag ccttcttcag catctactcc aaggccgtgt ccctggaggt ggggctgcag 1980

cacttcctca gctttctgag ctccatgcgc cgccctatct tggcctgcta caagctgtgg 2040

gggcctggcc tcccaaactt cttccgggcc ctggaggaca ttaacaggct gtgggaattc 2100

caggaggcca tctcgggctt cctggctgcc ctgcctctca tccgggagcg tgtgcccggg 2160

gccagcagct tcaaactcaa gaacctggcc cagacctacc tggcgagaaa catgagcgag 2220

cgcagcgcca tggctgccgt gctggccatg cgtgacctgt gccgcctcct cgaggtctcc 2280

ccgggccccc agctggccca gcatgtctac cccttcagta gcctgcagtg ctttgcctcc 2340

ctgcagcccc tggtgcaggc agctgtgctg ccccgggctg aggcccgcct cctggcccta 2400

cacaacgtga gcttcatgga gctgctgagt gcacaccgcc gtgaccggca ggggggcctg 2460

aagaagtaca gccgctatct aagcctgcag accaccacgt tgccccctgc ccagcctgct 2520

ttcaacctgc aggctctggg cacctacttt gaaggcctgt tggagggtcc ggcgctggca 2580

cgggcagaag gagtctccac cccacttgct ggccgtggct tggcagagag ggcctcccag 2640

cagagctga 2649

<210> 38

<211> 882

<212> PRT

<213> 智人(Homo sapiens)

<400> 38

Met Glu Pro Ala Pro Ala Arg Ser Pro Arg Pro Gln Gln Asp Pro Ala

1 5 10 15

Arg Pro Gln Glu Pro Thr Met Pro Pro Pro Glu Thr Pro Ser Glu Gly

20 25 30

Arg Gln Pro Ser Pro Ser Pro Ser Pro Thr Glu Arg Ala Pro Ala Ser

35 40 45

Glu Glu Glu Phe Gln Phe Leu Arg Cys Gln Gln Cys Gln Ala Glu Ala

50 55 60

Lys Cys Pro Lys Leu Leu Pro Cys Leu His Thr Leu Cys Ser Gly Cys

65 70 75 80

Leu Glu Ala Ser Gly Met Gln Cys Pro Ile Cys Gln Ala Pro Trp Pro

85 90 95

Leu Gly Ala Asp Thr Pro Ala Leu Asp Asn Val Phe Phe Glu Ser Leu

100 105 110

Gln Arg Arg Leu Ser Val Tyr Arg Gln Ile Val Asp Ala Gln Ala Val

115 120 125

Cys Thr Arg Cys Lys Glu Ser Ala Asp Phe Trp Cys Phe Glu Cys Glu

130 135 140

Gln Leu Leu Cys Ala Lys Cys Phe Glu Ala His Gln Trp Phe Leu Lys

145 150 155 160

His Glu Ala Arg Pro Leu Ala Glu Leu Arg Asn Gln Ser Val Arg Glu

165 170 175

Phe Leu Asp Gly Thr Arg Lys Thr Asn Asn Ile Phe Cys Ser Asn Pro

180 185 190

Asn His Arg Thr Pro Thr Leu Thr Ser Ile Tyr Cys Arg Gly Cys Ser

195 200 205

Lys Pro Leu Cys Cys Ser Cys Ala Leu Leu Asp Ser Ser His Ser Glu

210 215 220

Leu Lys Cys Asp Ile Ser Ala Glu Ile Gln Gln Arg Gln Glu Glu Leu

225 230 235 240

Asp Ala Met Thr Gln Ala Leu Gln Glu Gln Asp Ser Ala Phe Gly Ala

245 250 255

Val His Ala Gln Met His Ala Ala Val Gly Gln Leu Gly Arg Ala Arg

260 265 270

Ala Glu Thr Glu Glu Leu Ile Arg Glu Arg Val Arg Gln Val Val Ala

275 280 285

His Val Arg Ala Gln Glu Arg Glu Leu Leu Glu Ala Val Asp Ala Arg

290 295 300

Tyr Gln Arg Asp Tyr Glu Glu Met Ala Ser Arg Leu Gly Arg Leu Asp

305 310 315 320

Ala Val Leu Gln Arg Ile Arg Thr Gly Ser Ala Leu Val Gln Arg Met

325 330 335

Lys Cys Tyr Ala Ser Asp Gln Glu Val Leu Asp Met His Gly Phe Leu

340 345 350

Arg Gln Ala Leu Cys Arg Leu Arg Gln Glu Glu Pro Gln Ser Leu Gln

355 360 365

Ala Ala Val Arg Thr Asp Gly Phe Asp Glu Phe Lys Val Arg Leu Gln

370 375 380

Asp Leu Ser Ser Cys Ile Thr Gln Gly Lys Asp Ala Ala Val Ser Lys

385 390 395 400

Lys Ala Ser Pro Glu Ala Ala Ser Thr Pro Arg Asp Pro Ile Asp Val

405 410 415

Asp Leu Pro Glu Glu Ala Glu Arg Val Lys Ala Gln Val Gln Ala Leu

420 425 430

Gly Leu Ala Glu Ala Gln Pro Met Ala Val Val Gln Ser Val Pro Gly

435 440 445

Ala His Pro Val Pro Val Tyr Ala Phe Ser Ile Lys Gly Pro Ser Tyr

450 455 460

Gly Glu Asp Val Ser Asn Thr Thr Thr Ala Gln Lys Arg Lys Cys Ser

465 470 475 480

Gln Thr Gln Cys Pro Arg Lys Val Ile Lys Met Glu Ser Glu Glu Gly

485 490 495

Lys Glu Ala Arg Leu Ala Arg Ser Ser Pro Glu Gln Pro Arg Pro Ser

500 505 510

Thr Ser Lys Ala Val Ser Pro Pro His Leu Asp Gly Pro Pro Ser Pro

515 520 525

Arg Ser Pro Val Ile Gly Ser Glu Val Phe Leu Pro Asn Ser Asn His

530 535 540

Val Ala Ser Gly Ala Gly Glu Ala Glu Glu Arg Val Val Val Ile Ser

545 550 555 560

Ser Ser Glu Asp Ser Asp Ala Glu Asn Ser Ser Ser Arg Glu Leu Asp

565 570 575

Asp Ser Ser Ser Glu Ser Ser Asp Leu Gln Leu Glu Gly Pro Ser Thr

580 585 590

Leu Arg Val Leu Asp Glu Asn Leu Ala Asp Pro Gln Ala Glu Asp Arg

595 600 605

Pro Leu Val Phe Phe Asp Leu Lys Ile Asp Asn Glu Thr Gln Lys Ile

610 615 620

Ser Gln Leu Ala Ala Val Asn Arg Glu Ser Lys Phe Arg Val Val Ile

625 630 635 640

Gln Pro Glu Ala Phe Phe Ser Ile Tyr Ser Lys Ala Val Ser Leu Glu

645 650 655

Val Gly Leu Gln His Phe Leu Ser Phe Leu Ser Ser Met Arg Arg Pro

660 665 670

Ile Leu Ala Cys Tyr Lys Leu Trp Gly Pro Gly Leu Pro Asn Phe Phe

675 680 685

Arg Ala Leu Glu Asp Ile Asn Arg Leu Trp Glu Phe Gln Glu Ala Ile

690 695 700

Ser Gly Phe Leu Ala Ala Leu Pro Leu Ile Arg Glu Arg Val Pro Gly

705 710 715 720

Ala Ser Ser Phe Lys Leu Lys Asn Leu Ala Gln Thr Tyr Leu Ala Arg

725 730 735

Asn Met Ser Glu Arg Ser Ala Met Ala Ala Val Leu Ala Met Arg Asp

740 745 750

Leu Cys Arg Leu Leu Glu Val Ser Pro Gly Pro Gln Leu Ala Gln His

755 760 765

Val Tyr Pro Phe Ser Ser Leu Gln Cys Phe Ala Ser Leu Gln Pro Leu

770 775 780

Val Gln Ala Ala Val Leu Pro Arg Ala Glu Ala Arg Leu Leu Ala Leu

785 790 795 800

His Asn Val Ser Phe Met Glu Leu Leu Ser Ala His Arg Arg Asp Arg

805 810 815

Gln Gly Gly Leu Lys Lys Tyr Ser Arg Tyr Leu Ser Leu Gln Thr Thr

820 825 830

Thr Leu Pro Pro Ala Gln Pro Ala Phe Asn Leu Gln Ala Leu Gly Thr

835 840 845

Tyr Phe Glu Gly Leu Leu Glu Gly Pro Ala Leu Ala Arg Ala Glu Gly

850 855 860

Val Ser Thr Pro Leu Ala Gly Arg Gly Leu Ala Glu Arg Ala Ser Gln

865 870 875 880

Gln Ser

<210> 39

<211> 2346

<212> DNA

<213> 智人(Homo sapiens)

<400> 39

atggctaaga cccctagtga ccatctgctg tccaccctgg aggagctggt gccctatgac 60

ttcgagaagt tcaagttcaa gctgcagaac accagtgtgc agaaggagca ctccaggatc 120

ccccggagcc agatccagag agccaggccg gtgaagatgg ccactctgct ggtcacctac 180

tatggggaag agtacgccgt gcagctcacc ctgcaggtcc tgcgggccat caaccagcgc 240

ctgctggccg aggagctcca cagggcagcc attcaggaat attccacaca agaaaacggc 300

acagatgatt ccgcagcgtc cagctccctg ggggagaaca agcccaggag cctgaagact 360

ccagaccacc ccgaggggaa cgaggggaac ggccctcggc cgtacggggg cggagctgcc 420

agcctgcggt gcagccagcc cgaggccggg agggggctgt cgaggaagcc cctgagcaaa 480

cgcagagaga aggcctcgga gggcctggac gcgcagggca agcctcggac ccggagcccg 540

gccctgccgg gcgggagaag ccccggcccc tgcagggcgc tagagggggg ccaggccgag 600

gtccggctgc gcagaaacgc cagctccgcg gggaggctgc aggggctggc ggggggcgcc 660

ccggggcaga aggagtgcag gcccttcgaa gtgtacctgc cctcgggaaa gatgcgacct 720

agaagccttg aggtcaccat ttctacaggg gagaaggcgc ccgcaaatcc agaaattctc 780

ctgactctag aggaaaagac agctgcgaat ctggactcgg caacagaacc ccgggcaagg 840

cccactccgg atggaggggc atctgcggac ctgaaggaag gccctggaaa tccagaacat 900

tcggtcaccg gaaggccacc agacacggct gcgagtcccc gctgccacgc ccaggaagga 960

gacccagttg acggtacctg tgtgcgtgat tcctgcagct tccccgaggc agtttctggg 1020

cacccccagg cctcaggcag ccgctcacct ggctgccccc ggtgccagga ctcccatgaa 1080

aggaagagcc cgggaagcct aagcccccag cccctgccac agtgtaagcg ccacctgaag 1140

caggtccagc tgctcttctg tgaggatcac gatgagccca tctgcctcat ctgcagtctg 1200

agtcaggagc accaaggcca ccgggtgcgc cccattgagg aggtcgccct ggaacacaag 1260

aagaaaattc agaagcagct ggagcatctg aagaagctga gaaaatcagg ggaggagcag 1320

cgatcctatg gggaggagaa ggcagtgagc tttctgaaac aaactgaagc gctgaagcag 1380

cgggtgcaga ggaagctgga gcaggtgtac tacttcctgg agcagcaaga gcatttcttt 1440

gtggcctcac tggaggacgt gggccagatg gttgggcaga tcaggaaggc atatgacacc 1500

cgcgtatccc aggacatcgc cctgctcgat gcgctgattg gggaactgga ggccaaggag 1560

tgccagtcag aatgggaact tctgcaggac attggagaca tcttgcacag ggctaagaca 1620

gtgcctgtcc ctgaaaagtg gaccactcct caagagataa aacaaaagat ccaactcctc 1680

caccagaagt cagagtttgt ggagaagagc acaaagtact tctcagaaac cctgcgttca 1740

gaaatggaaa tgttcaatgt tccagagctg attggcgctc aggcacatgc tgttaatgtg 1800

attctggatg cagaaaccgc ttaccccaac ctcatcttct ctgatgatct gaagagtgtt 1860

agacttggaa acaagtggga gaggctgcct gatggcccgc aaagatttga cagctgtatc 1920

attgttctgg gctctccgag tttcctctct ggccgccgtt actgggaggt ggaggttgga 1980

gacaagacag catggatcct gggagcctgc aagacatcca taagcaggaa agggaacatg 2040

actctgtcgc cagagaatgg ctactgggtg gtgataatga tgaaggaaaa tgagtaccag 2100

gcgtccagcg ttcccccgac ccgcctgcta ataaaggagc ctcccaagcg tgtgggcatc 2160

ttcgtggact acagagttgg aagcatctcc ttttacaatg tgacagccag atcccacatc 2220

tatacattcg ccagctgctc tttctctggg ccccttcaac ctatcttcag ccctgggaca 2280

cgtgatggag ggaagaacac agctcctctg actatctgtc cagtgggtgg tcaggggcct 2340

gactga 2346

<210> 40

<211> 781

<212> PRT

<213> 智人(Homo sapiens)

<400> 40

Met Ala Lys Thr Pro Ser Asp His Leu Leu Ser Thr Leu Glu Glu Leu

1 5 10 15

Val Pro Tyr Asp Phe Glu Lys Phe Lys Phe Lys Leu Gln Asn Thr Ser

20 25 30

Val Gln Lys Glu His Ser Arg Ile Pro Arg Ser Gln Ile Gln Arg Ala

35 40 45

Arg Pro Val Lys Met Ala Thr Leu Leu Val Thr Tyr Tyr Gly Glu Glu

50 55 60

Tyr Ala Val Gln Leu Thr Leu Gln Val Leu Arg Ala Ile Asn Gln Arg

65 70 75 80

Leu Leu Ala Glu Glu Leu His Arg Ala Ala Ile Gln Glu Tyr Ser Thr

85 90 95

Gln Glu Asn Gly Thr Asp Asp Ser Ala Ala Ser Ser Ser Leu Gly Glu

100 105 110

Asn Lys Pro Arg Ser Leu Lys Thr Pro Asp His Pro Glu Gly Asn Glu

115 120 125

Gly Asn Gly Pro Arg Pro Tyr Gly Gly Gly Ala Ala Ser Leu Arg Cys

130 135 140

Ser Gln Pro Glu Ala Gly Arg Gly Leu Ser Arg Lys Pro Leu Ser Lys

145 150 155 160

Arg Arg Glu Lys Ala Ser Glu Gly Leu Asp Ala Gln Gly Lys Pro Arg

165 170 175

Thr Arg Ser Pro Ala Leu Pro Gly Gly Arg Ser Pro Gly Pro Cys Arg

180 185 190

Ala Leu Glu Gly Gly Gln Ala Glu Val Arg Leu Arg Arg Asn Ala Ser

195 200 205

Ser Ala Gly Arg Leu Gln Gly Leu Ala Gly Gly Ala Pro Gly Gln Lys

210 215 220

Glu Cys Arg Pro Phe Glu Val Tyr Leu Pro Ser Gly Lys Met Arg Pro

225 230 235 240

Arg Ser Leu Glu Val Thr Ile Ser Thr Gly Glu Lys Ala Pro Ala Asn

245 250 255

Pro Glu Ile Leu Leu Thr Leu Glu Glu Lys Thr Ala Ala Asn Leu Asp

260 265 270

Ser Ala Thr Glu Pro Arg Ala Arg Pro Thr Pro Asp Gly Gly Ala Ser

275 280 285

Ala Asp Leu Lys Glu Gly Pro Gly Asn Pro Glu His Ser Val Thr Gly

290 295 300

Arg Pro Pro Asp Thr Ala Ala Ser Pro Arg Cys His Ala Gln Glu Gly

305 310 315 320

Asp Pro Val Asp Gly Thr Cys Val Arg Asp Ser Cys Ser Phe Pro Glu

325 330 335

Ala Val Ser Gly His Pro Gln Ala Ser Gly Ser Arg Ser Pro Gly Cys

340 345 350

Pro Arg Cys Gln Asp Ser His Glu Arg Lys Ser Pro Gly Ser Leu Ser

355 360 365

Pro Gln Pro Leu Pro Gln Cys Lys Arg His Leu Lys Gln Val Gln Leu

370 375 380

Leu Phe Cys Glu Asp His Asp Glu Pro Ile Cys Leu Ile Cys Ser Leu

385 390 395 400

Ser Gln Glu His Gln Gly His Arg Val Arg Pro Ile Glu Glu Val Ala

405 410 415

Leu Glu His Lys Lys Lys Ile Gln Lys Gln Leu Glu His Leu Lys Lys

420 425 430

Leu Arg Lys Ser Gly Glu Glu Gln Arg Ser Tyr Gly Glu Glu Lys Ala

435 440 445

Val Ser Phe Leu Lys Gln Thr Glu Ala Leu Lys Gln Arg Val Gln Arg

450 455 460

Lys Leu Glu Gln Val Tyr Tyr Phe Leu Glu Gln Gln Glu His Phe Phe

465 470 475 480

Val Ala Ser Leu Glu Asp Val Gly Gln Met Val Gly Gln Ile Arg Lys

485 490 495

Ala Tyr Asp Thr Arg Val Ser Gln Asp Ile Ala Leu Leu Asp Ala Leu

500 505 510

Ile Gly Glu Leu Glu Ala Lys Glu Cys Gln Ser Glu Trp Glu Leu Leu

515 520 525

Gln Asp Ile Gly Asp Ile Leu His Arg Ala Lys Thr Val Pro Val Pro

530 535 540

Glu Lys Trp Thr Thr Pro Gln Glu Ile Lys Gln Lys Ile Gln Leu Leu

545 550 555 560

His Gln Lys Ser Glu Phe Val Glu Lys Ser Thr Lys Tyr Phe Ser Glu

565 570 575

Thr Leu Arg Ser Glu Met Glu Met Phe Asn Val Pro Glu Leu Ile Gly

580 585 590

Ala Gln Ala His Ala Val Asn Val Ile Leu Asp Ala Glu Thr Ala Tyr

595 600 605

Pro Asn Leu Ile Phe Ser Asp Asp Leu Lys Ser Val Arg Leu Gly Asn

610 615 620

Lys Trp Glu Arg Leu Pro Asp Gly Pro Gln Arg Phe Asp Ser Cys Ile

625 630 635 640

Ile Val Leu Gly Ser Pro Ser Phe Leu Ser Gly Arg Arg Tyr Trp Glu

645 650 655

Val Glu Val Gly Asp Lys Thr Ala Trp Ile Leu Gly Ala Cys Lys Thr

660 665 670

Ser Ile Ser Arg Lys Gly Asn Met Thr Leu Ser Pro Glu Asn Gly Tyr

675 680 685

Trp Val Val Ile Met Met Lys Glu Asn Glu Tyr Gln Ala Ser Ser Val

690 695 700

Pro Pro Thr Arg Leu Leu Ile Lys Glu Pro Pro Lys Arg Val Gly Ile

705 710 715 720

Phe Val Asp Tyr Arg Val Gly Ser Ile Ser Phe Tyr Asn Val Thr Ala

725 730 735

Arg Ser His Ile Tyr Thr Phe Ala Ser Cys Ser Phe Ser Gly Pro Leu

740 745 750

Gln Pro Ile Phe Ser Pro Gly Thr Arg Asp Gly Gly Lys Asn Thr Ala

755 760 765

Pro Leu Thr Ile Cys Pro Val Gly Gly Gln Gly Pro Asp

770 775 780

<210> 41

<211> 1428

<212> DNA

<213> 智人(Homo sapiens)

<400> 41

atggcttcag cagcacgctt gacaatgatg tgggaggagg tcacatgccc tatctgcctg 60

gaccccttcg tggagcctgt gagcatcgag tgtggccaca gcttctgcca ggaatgcatc 120

tctcaggttg ggaaaggtgg gggcagcgtc tgtcctgtgt gccggcagcg ctttctgctc 180

aagaatctcc ggcccaatcg acagctagcc aacatggtga acaaccttaa agaaatcagc 240

caggaggcca gagagggcac acagggggaa cggtgtgcag tgcatggaga gagacttcac 300

ctgttctgtg agaaagatgg gaaggccctt tgctgggtat gtgcccagtc tcggaaacac 360

cgtgaccacg ccatggtccc tcttgaggag gctgcacagg agtaccagga gaagctccag 420

gtggcattag gggaactgag aagaaagcag gagttggctg agaagttgga agtggaaatt 480

gcaataaaga gagcagactg gaagaaaaca gtggaaacac agaaatctag gattcacgca 540

gagtttgtgc agcaaaaaaa cttcctggtt gaagaagaac agaggcagct gcaggagctg 600

gagaaggatg agagggagca gctgagaatc ctgggggaga aagaggccaa gctggcccag 660

cagagccagg ccctacagga gctcatctca gagctagatc gaaggtgcca cagctcagca 720

ctggaactgc tgcaggaggt gataattgtc ctggaaagga gtgagtcctg gaacctgaag 780

gacctggata ttacctctcc agaactcagg agtgtgtgcc atgtgccagg gctgaagaag 840

atgctgagga catgtgcagt ccacatcact ctggatccag acacagccaa tccgtggctg 900

atactttcag aagatcggag acaagtgagg cttggagaca cccagcagag catacctgga 960

aatgaagaga gatttgatag ttatcctatg gtcctgggtg cccagcactt tcactctgga 1020

aaacattact gggaggtaga tgtgacagga aaggaggcct gggacctggg tgtctgcaga 1080

gactctgtgc gcaggaaggg gcactttttg cttagttcca agagtggctt ctggacaatt 1140

tggttgtgga acaaacaaaa atatgaggct ggcacctacc cccagactcc cctccacctt 1200

caggtgcctc catgccaagt tgggattttc ctggactatg aggctggcat ggtctccttc 1260

tacaacatca ctgaccatgg ctccctcatc tactccttct ctgaatgtgc ctttacagga 1320

cctctgcggc ccttcttcag tcctggtttc aatgatggag gaaaaaacac agcccctcta 1380

accctctgtc cactgaatat tggatcacaa ggatccactg actattga 1428

<210> 42

<211> 475

<212> PRT

<213> 智人(Homo sapiens)

<400> 42

Met Ala Ser Ala Ala Arg Leu Thr Met Met Trp Glu Glu Val Thr Cys

1 5 10 15

Pro Ile Cys Leu Asp Pro Phe Val Glu Pro Val Ser Ile Glu Cys Gly

20 25 30

His Ser Phe Cys Gln Glu Cys Ile Ser Gln Val Gly Lys Gly Gly Gly

35 40 45

Ser Val Cys Pro Val Cys Arg Gln Arg Phe Leu Leu Lys Asn Leu Arg

50 55 60

Pro Asn Arg Gln Leu Ala Asn Met Val Asn Asn Leu Lys Glu Ile Ser

65 70 75 80

Gln Glu Ala Arg Glu Gly Thr Gln Gly Glu Arg Cys Ala Val His Gly

85 90 95

Glu Arg Leu His Leu Phe Cys Glu Lys Asp Gly Lys Ala Leu Cys Trp

100 105 110

Val Cys Ala Gln Ser Arg Lys His Arg Asp His Ala Met Val Pro Leu

115 120 125

Glu Glu Ala Ala Gln Glu Tyr Gln Glu Lys Leu Gln Val Ala Leu Gly

130 135 140

Glu Leu Arg Arg Lys Gln Glu Leu Ala Glu Lys Leu Glu Val Glu Ile

145 150 155 160

Ala Ile Lys Arg Ala Asp Trp Lys Lys Thr Val Glu Thr Gln Lys Ser

165 170 175

Arg Ile His Ala Glu Phe Val Gln Gln Lys Asn Phe Leu Val Glu Glu

180 185 190

Glu Gln Arg Gln Leu Gln Glu Leu Glu Lys Asp Glu Arg Glu Gln Leu

195 200 205

Arg Ile Leu Gly Glu Lys Glu Ala Lys Leu Ala Gln Gln Ser Gln Ala

210 215 220

Leu Gln Glu Leu Ile Ser Glu Leu Asp Arg Arg Cys His Ser Ser Ala

225 230 235 240

Leu Glu Leu Leu Gln Glu Val Ile Ile Val Leu Glu Arg Ser Glu Ser

245 250 255

Trp Asn Leu Lys Asp Leu Asp Ile Thr Ser Pro Glu Leu Arg Ser Val

260 265 270

Cys His Val Pro Gly Leu Lys Lys Met Leu Arg Thr Cys Ala Val His

275 280 285

Ile Thr Leu Asp Pro Asp Thr Ala Asn Pro Trp Leu Ile Leu Ser Glu

290 295 300

Asp Arg Arg Gln Val Arg Leu Gly Asp Thr Gln Gln Ser Ile Pro Gly

305 310 315 320

Asn Glu Glu Arg Phe Asp Ser Tyr Pro Met Val Leu Gly Ala Gln His

325 330 335

Phe His Ser Gly Lys His Tyr Trp Glu Val Asp Val Thr Gly Lys Glu

340 345 350

Ala Trp Asp Leu Gly Val Cys Arg Asp Ser Val Arg Arg Lys Gly His

355 360 365

Phe Leu Leu Ser Ser Lys Ser Gly Phe Trp Thr Ile Trp Leu Trp Asn

370 375 380

Lys Gln Lys Tyr Glu Ala Gly Thr Tyr Pro Gln Thr Pro Leu His Leu

385 390 395 400

Gln Val Pro Pro Cys Gln Val Gly Ile Phe Leu Asp Tyr Glu Ala Gly

405 410 415

Met Val Ser Phe Tyr Asn Ile Thr Asp His Gly Ser Leu Ile Tyr Ser

420 425 430

Phe Ser Glu Cys Ala Phe Thr Gly Pro Leu Arg Pro Phe Phe Ser Pro

435 440 445

Gly Phe Asn Asp Gly Gly Lys Asn Thr Ala Pro Leu Thr Leu Cys Pro

450 455 460

Leu Asn Ile Gly Ser Gln Gly Ser Thr Asp Tyr

465 470 475

<210> 43

<211> 1497

<212> DNA

<213> 智人(Homo sapiens)

<400> 43

atggatttct cagtaaaggt agacatagag aaggaggtga cctgccccat ctgcctggag 60

ctcctgacag aacctctgag cctagattgt ggccacagct tctgccaagc ctgcatcact 120

gcaaagatca aggagtcagt gatcatctca agaggggaaa gcagctgtcc tgtgtgtcag 180

accagattcc agcctgggaa cctccgacct aatcggcatc tggccaacat agttgagaga 240

gtcaaagagg tcaagatgag cccacaggag gggcagaaga gagatgtctg tgagcaccat 300

ggaaaaaaac tccagatctt ctgtaaggag gatggaaaag tcatttgctg ggtttgtgaa 360

ctgtctcagg aacaccaagg tcaccaaaca ttccgcataa acgaggtggt caaggaatgt 420

caggaaaagc tgcaggtagc cctgcagagg ctgataaagg aggatcaaga ggctgagaag 480

ctggaagatg acatcagaca agagagaacc gcctggaaga attatatcca gatcgagaga 540

cagaagattc tgaaagggtt caatgaaatg agagtcatct tggacaatga ggagcagaga 600

gagctgcaaa agctggagga aggtgaggtg aatgtgctgg ataacctggc agcagctaca 660

gaccagctgg tccagcagag gcaggatgcc agcacgctca tctcagatct ccagcggagg 720

ttgaggggat cgtcagtaga gatgctgcag gatgtgattg acgtcatgaa aaggagtgaa 780

agctggacat tgaagaagcc aaaatctgtt tccaagaaac taaagagtgt attccgagta 840

ccagatctga gtgggatgct gcaagttctt aaagagctga cagatgtcca gtactactgg 900

gtggacgtga tgctgaatcc aggcagtgcc acttcgaatg ttgctatttc tgtggatcag 960

agacaagtga aaactgtacg cacctgcaca tttaagaatt caaatccatg tgatttttct 1020

gcttttggtg tcttcggctg ccaatatttc tcttcgggga aatattactg ggaagtagat 1080

gtgtctggaa agattgcctg gatcctgggc gtacacagta aaataagtag tctgaataaa 1140

aggaagagct ctgggtttgc ttttgatcca agtgtaaatt attcaaaagt ttactccaga 1200

tatagacctc aatatggcta ctgggttata ggattacaga atacatgtga atataatgct 1260

tttgaggact cctcctcttc tgatcccaag gttttgactc tctttatggc tgtgcctccc 1320

tgtcgtattg gggttttcct agactatgag gcaggcattg tctcattttt caatgtcaca 1380

aaccacggag cactcatcta caagttctct ggatgtcgct tttctcgacc tgcttatccg 1440

tatttcaatc cttggaactg cctagtcccc atgactgtgt gcccaccgag ctcctga 1497

<210> 44

<211> 498

<212> PRT

<213> 智人(Homo sapiens)

<400> 44

Met Asp Phe Ser Val Lys Val Asp Ile Glu Lys Glu Val Thr Cys Pro

1 5 10 15

Ile Cys Leu Glu Leu Leu Thr Glu Pro Leu Ser Leu Asp Cys Gly His

20 25 30

Ser Phe Cys Gln Ala Cys Ile Thr Ala Lys Ile Lys Glu Ser Val Ile

35 40 45

Ile Ser Arg Gly Glu Ser Ser Cys Pro Val Cys Gln Thr Arg Phe Gln

50 55 60

Pro Gly Asn Leu Arg Pro Asn Arg His Leu Ala Asn Ile Val Glu Arg

65 70 75 80

Val Lys Glu Val Lys Met Ser Pro Gln Glu Gly Gln Lys Arg Asp Val

85 90 95

Cys Glu His His Gly Lys Lys Leu Gln Ile Phe Cys Lys Glu Asp Gly

100 105 110

Lys Val Ile Cys Trp Val Cys Glu Leu Ser Gln Glu His Gln Gly His

115 120 125

Gln Thr Phe Arg Ile Asn Glu Val Val Lys Glu Cys Gln Glu Lys Leu

130 135 140

Gln Val Ala Leu Gln Arg Leu Ile Lys Glu Asp Gln Glu Ala Glu Lys

145 150 155 160

Leu Glu Asp Asp Ile Arg Gln Glu Arg Thr Ala Trp Lys Asn Tyr Ile

165 170 175

Gln Ile Glu Arg Gln Lys Ile Leu Lys Gly Phe Asn Glu Met Arg Val

180 185 190

Ile Leu Asp Asn Glu Glu Gln Arg Glu Leu Gln Lys Leu Glu Glu Gly

195 200 205

Glu Val Asn Val Leu Asp Asn Leu Ala Ala Ala Thr Asp Gln Leu Val

210 215 220

Gln Gln Arg Gln Asp Ala Ser Thr Leu Ile Ser Asp Leu Gln Arg Arg

225 230 235 240

Leu Arg Gly Ser Ser Val Glu Met Leu Gln Asp Val Ile Asp Val Met

245 250 255

Lys Arg Ser Glu Ser Trp Thr Leu Lys Lys Pro Lys Ser Val Ser Lys

260 265 270

Lys Leu Lys Ser Val Phe Arg Val Pro Asp Leu Ser Gly Met Leu Gln

275 280 285

Val Leu Lys Glu Leu Thr Asp Val Gln Tyr Tyr Trp Val Asp Val Met

290 295 300

Leu Asn Pro Gly Ser Ala Thr Ser Asn Val Ala Ile Ser Val Asp Gln

305 310 315 320

Arg Gln Val Lys Thr Val Arg Thr Cys Thr Phe Lys Asn Ser Asn Pro

325 330 335

Cys Asp Phe Ser Ala Phe Gly Val Phe Gly Cys Gln Tyr Phe Ser Ser

340 345 350

Gly Lys Tyr Tyr Trp Glu Val Asp Val Ser Gly Lys Ile Ala Trp Ile

355 360 365

Leu Gly Val His Ser Lys Ile Ser Ser Leu Asn Lys Arg Lys Ser Ser

370 375 380

Gly Phe Ala Phe Asp Pro Ser Val Asn Tyr Ser Lys Val Tyr Ser Arg

385 390 395 400

Tyr Arg Pro Gln Tyr Gly Tyr Trp Val Ile Gly Leu Gln Asn Thr Cys

405 410 415

Glu Tyr Asn Ala Phe Glu Asp Ser Ser Ser Ser Asp Pro Lys Val Leu

420 425 430

Thr Leu Phe Met Ala Val Pro Pro Cys Arg Ile Gly Val Phe Leu Asp

435 440 445

Tyr Glu Ala Gly Ile Val Ser Phe Phe Asn Val Thr Asn His Gly Ala

450 455 460

Leu Ile Tyr Lys Phe Ser Gly Cys Arg Phe Ser Arg Pro Ala Tyr Pro

465 470 475 480

Tyr Phe Asn Pro Trp Asn Cys Leu Val Pro Met Thr Val Cys Pro Pro

485 490 495

Ser Ser

<210> 45

<211> 1725

<212> DNA

<213> 智人(Homo sapiens)

<400> 45

atggctaccc tggttgtaaa caagctcgga gcgggagtag acagtggccg gcagggcagc 60

cgggggacag ctgtagtgaa ggtgctagag tgtggagttt gtgaagatgt cttttctttg 120

caaggagaca aagttccccg tcttttgctt tgtggccata ccgtctgtca tgactgtctc 180

actcgcctac ctcttcatgg aagagcaatc cgttgcccat ttgatcgaca agtaacagac 240

ctaggtgatt caggtgtctg gggattgaaa aaaaattttg ctttattgga gcttttggaa 300

cgactgcaga atgggcctat tggtcagtat ggagctgcag aagaatccat tgggatatct 360

ggagagagca tcattcgttg tgatgaagat gaagctcacc ttgcctctgt atattgcact 420

gtgtgtgcaa ctcatttgtg ctctgagtgt tctcaagtta ctcattctac aaagacatta 480

gcaaagcaca ggcgagttcc tctagctgat aaacctcatg agaaaactat gtgctctcag 540

caccaggtgc atgccattga gtttgtttgc ttggaagaag gttgtcaaac tagcccactc 600

atgtgctgtg tctgcaaaga atatggaaaa caccagggtc acaagcattc agtattggaa 660

ccagaagcta atcagatccg agcatcaatt ttagatatgg ctcactgcat acggaccttc 720

acagaggaaa tctcagatta ttccagaaaa ttagttggaa ttgtgcagca cattgaagga 780

ggagaacaaa tcgtggaaga tggaattgga atggctcaca cagaacatgt accagggact 840

gcagagaatg cccggtcatg tattcgagct tatttttatg atctacatga aactctgtgt 900

cgtcaagaag aaatggctct aagtgttgtt gatgctcatg ttcgtgaaaa attgatttgg 960

ctcaggcagc aacaagaaga tatgactatt ttgttgtcag aggtttctgc agcctgcctc 1020

cactgtgaaa agactttgca gcaggatgat tgtagagttg tcttggcaaa acaggaaatt 1080

acaaggttac tggaaacatt gcagaaacag cagcagcagt ttacagaagt tgcagatcac 1140

attcagttgg atgccagcat ccctgtcact tttacaaagg ataatcgagt tcacattgga 1200

ccaaaaatgg aaattcgggt cgttacgtta ggattggatg gtgctggaaa aactactatc 1260

ttgtttaagt taaaacagga tgaattcatg cagcccattc caacaattgg ttttaacgtg 1320

gaaactgtag aatataaaaa tctaaaattc actatttggg atgtaggtgg aaaacacaaa 1380

ttaagaccat tgtggaaaca ttattacctc aatactcaag ctgttgtgtt tgttgtagat 1440

agcagtcata gagacagaat tagtgaagca cacagcgaac ttgcaaagtt gttaacggaa 1500

aaagaactcc gagatgctct gctcctgatt tttgctaaca aacaggatgt tgctggagca 1560

ctgtcagtag aagaaatcac tgaactactc agtctccata aattatgctg tggccgtagc 1620

tggtatattc agggctgtga tgctcgaagt ggtatgggac tgtatgaagg gttggactgg 1680

ctctcacggc aacttgtagc tgctggagta ttggatgttg cttga 1725

<210> 46

<211> 574

<212> PRT

<213> 智人(Homo sapiens)

<400> 46

Met Ala Thr Leu Val Val Asn Lys Leu Gly Ala Gly Val Asp Ser Gly

1 5 10 15

Arg Gln Gly Ser Arg Gly Thr Ala Val Val Lys Val Leu Glu Cys Gly

20 25 30

Val Cys Glu Asp Val Phe Ser Leu Gln Gly Asp Lys Val Pro Arg Leu

35 40 45

Leu Leu Cys Gly His Thr Val Cys His Asp Cys Leu Thr Arg Leu Pro

50 55 60

Leu His Gly Arg Ala Ile Arg Cys Pro Phe Asp Arg Gln Val Thr Asp

65 70 75 80

Leu Gly Asp Ser Gly Val Trp Gly Leu Lys Lys Asn Phe Ala Leu Leu

85 90 95

Glu Leu Leu Glu Arg Leu Gln Asn Gly Pro Ile Gly Gln Tyr Gly Ala

100 105 110

Ala Glu Glu Ser Ile Gly Ile Ser Gly Glu Ser Ile Ile Arg Cys Asp

115 120 125

Glu Asp Glu Ala His Leu Ala Ser Val Tyr Cys Thr Val Cys Ala Thr

130 135 140

His Leu Cys Ser Glu Cys Ser Gln Val Thr His Ser Thr Lys Thr Leu

145 150 155 160

Ala Lys His Arg Arg Val Pro Leu Ala Asp Lys Pro His Glu Lys Thr

165 170 175

Met Cys Ser Gln His Gln Val His Ala Ile Glu Phe Val Cys Leu Glu

180 185 190

Glu Gly Cys Gln Thr Ser Pro Leu Met Cys Cys Val Cys Lys Glu Tyr

195 200 205

Gly Lys His Gln Gly His Lys His Ser Val Leu Glu Pro Glu Ala Asn

210 215 220

Gln Ile Arg Ala Ser Ile Leu Asp Met Ala His Cys Ile Arg Thr Phe

225 230 235 240

Thr Glu Glu Ile Ser Asp Tyr Ser Arg Lys Leu Val Gly Ile Val Gln

245 250 255

His Ile Glu Gly Gly Glu Gln Ile Val Glu Asp Gly Ile Gly Met Ala

260 265 270

His Thr Glu His Val Pro Gly Thr Ala Glu Asn Ala Arg Ser Cys Ile

275 280 285

Arg Ala Tyr Phe Tyr Asp Leu His Glu Thr Leu Cys Arg Gln Glu Glu

290 295 300

Met Ala Leu Ser Val Val Asp Ala His Val Arg Glu Lys Leu Ile Trp

305 310 315 320

Leu Arg Gln Gln Gln Glu Asp Met Thr Ile Leu Leu Ser Glu Val Ser

325 330 335

Ala Ala Cys Leu His Cys Glu Lys Thr Leu Gln Gln Asp Asp Cys Arg

340 345 350

Val Val Leu Ala Lys Gln Glu Ile Thr Arg Leu Leu Glu Thr Leu Gln

355 360 365

Lys Gln Gln Gln Gln Phe Thr Glu Val Ala Asp His Ile Gln Leu Asp

370 375 380

Ala Ser Ile Pro Val Thr Phe Thr Lys Asp Asn Arg Val His Ile Gly

385 390 395 400

Pro Lys Met Glu Ile Arg Val Val Thr Leu Gly Leu Asp Gly Ala Gly

405 410 415

Lys Thr Thr Ile Leu Phe Lys Leu Lys Gln Asp Glu Phe Met Gln Pro

420 425 430

Ile Pro Thr Ile Gly Phe Asn Val Glu Thr Val Glu Tyr Lys Asn Leu

435 440 445

Lys Phe Thr Ile Trp Asp Val Gly Gly Lys His Lys Leu Arg Pro Leu

450 455 460

Trp Lys His Tyr Tyr Leu Asn Thr Gln Ala Val Val Phe Val Val Asp

465 470 475 480

Ser Ser His Arg Asp Arg Ile Ser Glu Ala His Ser Glu Leu Ala Lys

485 490 495

Leu Leu Thr Glu Lys Glu Leu Arg Asp Ala Leu Leu Leu Ile Phe Ala

500 505 510

Asn Lys Gln Asp Val Ala Gly Ala Leu Ser Val Glu Glu Ile Thr Glu

515 520 525

Leu Leu Ser Leu His Lys Leu Cys Cys Gly Arg Ser Trp Tyr Ile Gln

530 535 540

Gly Cys Asp Ala Arg Ser Gly Met Gly Leu Tyr Glu Gly Leu Asp Trp

545 550 555 560

Leu Ser Arg Gln Leu Val Ala Ala Gly Val Leu Asp Val Ala

565 570

<210> 47

<211> 3153

<212> DNA

<213> 智人(Homo sapiens)

<400> 47

atggaggtgg cggtggagaa ggcggtggcg gcggcggcag cggcctcggc tgcggcctcc 60

ggggggccct cggcggcgcc gagcggggag aacgaggccg agagtcggca gggcccggac 120

tcggagcgcg gcggcgaggc ggcccggctc aacctgttgg acacttgcgc cgtgtgccac 180

cagaacatcc agagccgggc gcccaagctg ctgccctgcc tgcactcttt ctgccagcgc 240

tgcctgcccg cgccccagcg ctacctcatg ctgcccgcgc ccatgctggg ctcggccgag 300

accccgccac ccgtccctgc ccccggctcg ccggtcagcg gctcgtcgcc gttcgccacc 360

caagttggag tcattcgttg cccagtttgc agccaagaat gtgcagagag acacatcata 420

gataactttt ttgtgaagga cactactgag gttcccagca gtacagtaga aaagtcaaat 480

caggtatgta caagctgtga ggacaacgca gaagccaatg ggttttgtgt agagtgtgtt 540

gaatggctct gcaagacgtg tatcagagct catcagaggg taaagttcac aaaagaccac 600

actgtcagac agaaagagga agtatctcca gaggcagttg gtgtcaccag ccagcgacca 660

gtgttttgtc cttttcataa aaaggagcag ctgaagctgt actgtgagac atgtgacaaa 720

ctgacatgtc gagactgtca gttgttagaa cataaagagc atagatacca atttatagaa 780

gaagcttttc agaatcagaa agtgatcata gatacactaa tcaccaaact gatggaaaaa 840

acaaaataca taaaattcac aggaaatcag atccaaaaca gaattattga agtaaatcaa 900

aatcaaaagc aggtggaaca ggatattaaa gttgctatat ttacactgat ggtagaaata 960

aataaaaaag gaaaagctct actgcatcag ttagagagcc ttgcaaagga ccatcgcatg 1020

aaacttatgc aacaacaaca ggaagtggct ggactctcta aacaattgga gcatgtcatg 1080

catttttcta aatgggcagt ttccagtggc agcagtacag cattacttta tagcaaacga 1140

ctgattacat accggttacg gcacctcctt cgtgcaaggt gtgatgcatc cccagtgacc 1200

aacaacacca tccaatttca ctgtgatcct agtttctggg ctcaaaatat catcaactta 1260

ggttctttag taatcgagga taaagagagc cagccacaaa tgcctaagca gaatcctgtc 1320

gtggaacaga attcacagcc accaagtggt ttatcatcaa accagttatc caagttccca 1380

acacagatca gcctagctca attacggctc cagcatatgc agcaacaggt aatggctcag 1440

aggcaacagg tgcaacggag gccagcacct gtgggtttac caaaccctag aatgcagggg 1500

cccatccagc aaccttccat ctctcatcag caaccgcctc cacgtttgat aaactttcag 1560

aatcacagcc ccaaacccaa tggaccagtt cttcctcctc atcctcaaca actgagatat 1620

ccaccaaacc agaacatacc acgacaagca ataaagccaa accccctaca gatggctttc 1680

ttggctcaac aagccataaa acagtggcag atcagcagtg gacagggaac cccatcaact 1740

accaacagca catcctctac tccttccagc cccacgatta ctagtgcagc aggatatgat 1800

ggaaaggctt ttggttcacc tatgatcgat ttgagctcac cagtgggagg gtcttataat 1860

cttccctctc ttccggatat tgactgttca agtactatta tgctggacaa tattgtgagg 1920

aaagatacta atatagatca tggccagcca agaccaccct caaacagaac ggtccagtca 1980

ccaaattcat cagtgccatc tccaggcctt gcaggacctg ttactatgac tagtgtacac 2040

cccccaatac gttcacctag tgcctccagc gttggaagcc gaggaagctc tggctcttcc 2100

agcaaaccag caggagctga ctctacacac aaagtcccag tggtcatgct ggagccaatt 2160

cgaataaaac aagaaaacag tggaccaccg gaaaattatg atttccctgt tgttatagtg 2220

aagcaagaat cagatgaaga atctaggcct caaaatgcca attatccaag aagcatactc 2280

acctccctgc tcttaaatag cagtcagagc tctacttctg aggagactgt gctaagatca 2340

gatgcccctg atagtacagg agatcaacct ggacttcacc aggacaattc ctcaaatgga 2400

aagtctgaat ggttggatcc ttcccagaag tcacctcttc atgttggaga gacaaggaaa 2460

gaggatgacc ccaatgagga ctggtgtgca gtttgtcaaa acggagggga actcctctgc 2520

tgtgaaaagt gccccaaagt attccatctt tcttgtcatg tgcccacatt gacaaatttt 2580

ccaagtggag agtggatttg cactttctgc cgagacttat ctaaaccaga agttgaatat 2640

gattgtgatg ctcccagtca caactcagaa aaaaagaaaa ctgaaggcct tgttaagtta 2700

acacctatag ataaaaggaa gtgtgagcgc ctacttttat ttctttactg ccatgaaatg 2760

agcctggctt ttcaagaccc tgttcctcta actgtgcctg attattacaa aataattaaa 2820

aatccaatgg atttgtcaac catcaagaaa agactacaag aagattattc catgtactca 2880

aaacctgaag attttgtagc tgattttaga ttgatctttc aaaactgtgc tgaattcaat 2940

gagcctgatt cagaagtagc caatgctggt ataaaacttg aaaattattt tgaagaactt 3000

ctaaagaacc tctatccaga aaaaaggttt cccaaaccag aattcaggaa tgaatcagaa 3060

gataataaat ttagtgatga ttcagatgat gactttgtac agccccggaa gaaacgcctc 3120

aaaagcattg aagaacgcca gttgcttaaa taa 3153

<210> 48

<211> 1050

<212> PRT

<213> 智人(Homo sapiens)

<400> 48

Met Glu Val Ala Val Glu Lys Ala Val Ala Ala Ala Ala Ala Ala Ser

1 5 10 15

Ala Ala Ala Ser Gly Gly Pro Ser Ala Ala Pro Ser Gly Glu Asn Glu

20 25 30

Ala Glu Ser Arg Gln Gly Pro Asp Ser Glu Arg Gly Gly Glu Ala Ala

35 40 45

Arg Leu Asn Leu Leu Asp Thr Cys Ala Val Cys His Gln Asn Ile Gln

50 55 60

Ser Arg Ala Pro Lys Leu Leu Pro Cys Leu His Ser Phe Cys Gln Arg

65 70 75 80

Cys Leu Pro Ala Pro Gln Arg Tyr Leu Met Leu Pro Ala Pro Met Leu

85 90 95

Gly Ser Ala Glu Thr Pro Pro Pro Val Pro Ala Pro Gly Ser Pro Val

100 105 110

Ser Gly Ser Ser Pro Phe Ala Thr Gln Val Gly Val Ile Arg Cys Pro

115 120 125

Val Cys Ser Gln Glu Cys Ala Glu Arg His Ile Ile Asp Asn Phe Phe

130 135 140

Val Lys Asp Thr Thr Glu Val Pro Ser Ser Thr Val Glu Lys Ser Asn

145 150 155 160

Gln Val Cys Thr Ser Cys Glu Asp Asn Ala Glu Ala Asn Gly Phe Cys

165 170 175

Val Glu Cys Val Glu Trp Leu Cys Lys Thr Cys Ile Arg Ala His Gln

180 185 190

Arg Val Lys Phe Thr Lys Asp His Thr Val Arg Gln Lys Glu Glu Val

195 200 205

Ser Pro Glu Ala Val Gly Val Thr Ser Gln Arg Pro Val Phe Cys Pro

210 215 220

Phe His Lys Lys Glu Gln Leu Lys Leu Tyr Cys Glu Thr Cys Asp Lys

225 230 235 240

Leu Thr Cys Arg Asp Cys Gln Leu Leu Glu His Lys Glu His Arg Tyr

245 250 255

Gln Phe Ile Glu Glu Ala Phe Gln Asn Gln Lys Val Ile Ile Asp Thr

260 265 270

Leu Ile Thr Lys Leu Met Glu Lys Thr Lys Tyr Ile Lys Phe Thr Gly

275 280 285

Asn Gln Ile Gln Asn Arg Ile Ile Glu Val Asn Gln Asn Gln Lys Gln

290 295 300

Val Glu Gln Asp Ile Lys Val Ala Ile Phe Thr Leu Met Val Glu Ile

305 310 315 320

Asn Lys Lys Gly Lys Ala Leu Leu His Gln Leu Glu Ser Leu Ala Lys

325 330 335

Asp His Arg Met Lys Leu Met Gln Gln Gln Gln Glu Val Ala Gly Leu

340 345 350

Ser Lys Gln Leu Glu His Val Met His Phe Ser Lys Trp Ala Val Ser

355 360 365

Ser Gly Ser Ser Thr Ala Leu Leu Tyr Ser Lys Arg Leu Ile Thr Tyr

370 375 380

Arg Leu Arg His Leu Leu Arg Ala Arg Cys Asp Ala Ser Pro Val Thr

385 390 395 400

Asn Asn Thr Ile Gln Phe His Cys Asp Pro Ser Phe Trp Ala Gln Asn

405 410 415

Ile Ile Asn Leu Gly Ser Leu Val Ile Glu Asp Lys Glu Ser Gln Pro

420 425 430

Gln Met Pro Lys Gln Asn Pro Val Val Glu Gln Asn Ser Gln Pro Pro

435 440 445

Ser Gly Leu Ser Ser Asn Gln Leu Ser Lys Phe Pro Thr Gln Ile Ser

450 455 460

Leu Ala Gln Leu Arg Leu Gln His Met Gln Gln Gln Val Met Ala Gln

465 470 475 480

Arg Gln Gln Val Gln Arg Arg Pro Ala Pro Val Gly Leu Pro Asn Pro

485 490 495

Arg Met Gln Gly Pro Ile Gln Gln Pro Ser Ile Ser His Gln Gln Pro

500 505 510

Pro Pro Arg Leu Ile Asn Phe Gln Asn His Ser Pro Lys Pro Asn Gly

515 520 525

Pro Val Leu Pro Pro His Pro Gln Gln Leu Arg Tyr Pro Pro Asn Gln

530 535 540

Asn Ile Pro Arg Gln Ala Ile Lys Pro Asn Pro Leu Gln Met Ala Phe

545 550 555 560

Leu Ala Gln Gln Ala Ile Lys Gln Trp Gln Ile Ser Ser Gly Gln Gly

565 570 575

Thr Pro Ser Thr Thr Asn Ser Thr Ser Ser Thr Pro Ser Ser Pro Thr

580 585 590

Ile Thr Ser Ala Ala Gly Tyr Asp Gly Lys Ala Phe Gly Ser Pro Met

595 600 605

Ile Asp Leu Ser Ser Pro Val Gly Gly Ser Tyr Asn Leu Pro Ser Leu

610 615 620

Pro Asp Ile Asp Cys Ser Ser Thr Ile Met Leu Asp Asn Ile Val Arg

625 630 635 640

Lys Asp Thr Asn Ile Asp His Gly Gln Pro Arg Pro Pro Ser Asn Arg

645 650 655

Thr Val Gln Ser Pro Asn Ser Ser Val Pro Ser Pro Gly Leu Ala Gly

660 665 670

Pro Val Thr Met Thr Ser Val His Pro Pro Ile Arg Ser Pro Ser Ala

675 680 685

Ser Ser Val Gly Ser Arg Gly Ser Ser Gly Ser Ser Ser Lys Pro Ala

690 695 700

Gly Ala Asp Ser Thr His Lys Val Pro Val Val Met Leu Glu Pro Ile

705 710 715 720

Arg Ile Lys Gln Glu Asn Ser Gly Pro Pro Glu Asn Tyr Asp Phe Pro

725 730 735

Val Val Ile Val Lys Gln Glu Ser Asp Glu Glu Ser Arg Pro Gln Asn

740 745 750

Ala Asn Tyr Pro Arg Ser Ile Leu Thr Ser Leu Leu Leu Asn Ser Ser

755 760 765

Gln Ser Ser Thr Ser Glu Glu Thr Val Leu Arg Ser Asp Ala Pro Asp

770 775 780

Ser Thr Gly Asp Gln Pro Gly Leu His Gln Asp Asn Ser Ser Asn Gly

785 790 795 800

Lys Ser Glu Trp Leu Asp Pro Ser Gln Lys Ser Pro Leu His Val Gly

805 810 815

Glu Thr Arg Lys Glu Asp Asp Pro Asn Glu Asp Trp Cys Ala Val Cys

820 825 830

Gln Asn Gly Gly Glu Leu Leu Cys Cys Glu Lys Cys Pro Lys Val Phe

835 840 845

His Leu Ser Cys His Val Pro Thr Leu Thr Asn Phe Pro Ser Gly Glu

850 855 860

Trp Ile Cys Thr Phe Cys Arg Asp Leu Ser Lys Pro Glu Val Glu Tyr

865 870 875 880

Asp Cys Asp Ala Pro Ser His Asn Ser Glu Lys Lys Lys Thr Glu Gly

885 890 895

Leu Val Lys Leu Thr Pro Ile Asp Lys Arg Lys Cys Glu Arg Leu Leu

900 905 910

Leu Phe Leu Tyr Cys His Glu Met Ser Leu Ala Phe Gln Asp Pro Val

915 920 925

Pro Leu Thr Val Pro Asp Tyr Tyr Lys Ile Ile Lys Asn Pro Met Asp

930 935 940

Leu Ser Thr Ile Lys Lys Arg Leu Gln Glu Asp Tyr Ser Met Tyr Ser

945 950 955 960

Lys Pro Glu Asp Phe Val Ala Asp Phe Arg Leu Ile Phe Gln Asn Cys

965 970 975

Ala Glu Phe Asn Glu Pro Asp Ser Glu Val Ala Asn Ala Gly Ile Lys

980 985 990

Leu Glu Asn Tyr Phe Glu Glu Leu Leu Lys Asn Leu Tyr Pro Glu Lys

995 1000 1005

Arg Phe Pro Lys Pro Glu Phe Arg Asn Glu Ser Glu Asp Asn Lys

1010 1015 1020

Phe Ser Asp Asp Ser Asp Asp Asp Phe Val Gln Pro Arg Lys Lys

1025 1030 1035

Arg Leu Lys Ser Ile Glu Glu Arg Gln Leu Leu Lys

1040 1045 1050

<210> 49

<211> 1893

<212> DNA

<213> 智人(Homo sapiens)

<400> 49

atggcagagc tgtgccccct ggccgaggag ctgtcgtgct ccatctgcct ggagcccttc 60

aaggagccgg tcaccactcc gtgcggccac aacttctgcg ggtcgtgcct gaatgagacg 120

tgggcagtcc agggctcgcc atacctgtgc ccgcagtgcc gcgccgtcta ccaggcgcga 180

ccgcagctgc acaagaacac ggtgctgtgc aacgtggtgg agcagttcct gcaggccgac 240

ctggcccggg agccacccgc cgacgtctgg acgccgcccg cccgcgcctc tgcacccagc 300

ccgaatgccc aggtggcctg cgaccactgc ctgaaggagg ccgccgtgaa gacgtgcttg 360

gtgtgcatgg cctccttctg tcaggagcac ctgcagccgc acttcgacag ccccgccttc 420

caggaccacc cgctgcagcc gcccgttcgc gacctgttgc gccgcaaatg ttcccagcac 480

aatcggctgc gggaattttt ctgccccgag cacagcgagt gcatctgcca catctgcctg 540

gtggagcata agacctgctc tcccgcgtcc ctgagccagg ccagcgccga cctggaggcc 600

accctgaggc acaaactaac tgtcatgtac agtcagatca acggggcgtc gagagcactg 660

gatgatgtga gaaacaggca gcaggatgtg cggatgactg caaacagaaa ggtggagcag 720

ctacaacaag aatacacgga aatgaaggct ctcttggacg cctcagagac cacctcgaca 780

aggaagataa aggaagagga gaagagggtc aacagcaagt ttgacaccat ttatcagatt 840

ctcctcaaga agaagagtga gatccagacc ttgaaggagg agattgaaca gagcctgacc 900

aagagggatg agttcgagtt tctggagaaa gcatcaaaac tgcgaggaat ctcaacaaag 960

ccagtctaca tccccgaggt ggaactgaac cacaagctga taaaaggcat ccaccagagc 1020

accatagacc tcaaaaacga gctgaagcag tgcatcgggc ggctccagga gcccaccccc 1080

agttcaggtg accctggaga gcatgaccca gcgtccacac acaaatccac acgccctgtg 1140

aagaaggtct ccaaagagga aaagaaatcc aagaaacctc cccctgtccc tgccttaccc 1200

agcaagcttc ccacgtttgg agccccggaa cagttagtgg atttaaaaca agctggcttg 1260

gaggctgcag ccaaagccac cagctcacat ccgaactcaa catctctcaa ggccaaggtg 1320

ctggagacct tcctggccaa gtccagacct gagctcctgg agtattacat taaagtcatc 1380

ctggactaca acaccgccca caacaaagtg gctctgtcag agtgctatac agtagcttct 1440

gtggctgaga tgcctcagaa ctaccggccg catccccaga ggttcacata ctgctctcag 1500

gtgctgggcc tgcactgcta caagaagggg atccactact gggaggtgga gctgcagaag 1560

aacaacttct gtggggtagg catctgctac ggaagcatga accggcaggg cccagaaagc 1620

aggctcggcc gcaacagcgc ctcctggtgc gtggagtggt tcaacaccaa gatctctgcc 1680

tggcacaata acgtggagaa aaccctgccc tccaccaagg ccacgcgggt gggcgtgctt 1740

ctcaactgtg accacggctt tgtcatcttc ttcgctgttg ccgacaaggt ccacctgatg 1800

tataagttca gggtggactt tactgaggct ttgtacccgg ctttctgggt attttctgct 1860

ggtgccacac tctccatctg ctcccccaag tag 1893

<210> 50

<211> 630

<212> PRT

<213> 智人(Homo sapiens)

<400> 50

Met Ala Glu Leu Cys Pro Leu Ala Glu Glu Leu Ser Cys Ser Ile Cys

1 5 10 15

Leu Glu Pro Phe Lys Glu Pro Val Thr Thr Pro Cys Gly His Asn Phe

20 25 30

Cys Gly Ser Cys Leu Asn Glu Thr Trp Ala Val Gln Gly Ser Pro Tyr

35 40 45

Leu Cys Pro Gln Cys Arg Ala Val Tyr Gln Ala Arg Pro Gln Leu His

50 55 60

Lys Asn Thr Val Leu Cys Asn Val Val Glu Gln Phe Leu Gln Ala Asp

65 70 75 80

Leu Ala Arg Glu Pro Pro Ala Asp Val Trp Thr Pro Pro Ala Arg Ala

85 90 95

Ser Ala Pro Ser Pro Asn Ala Gln Val Ala Cys Asp His Cys Leu Lys

100 105 110

Glu Ala Ala Val Lys Thr Cys Leu Val Cys Met Ala Ser Phe Cys Gln

115 120 125

Glu His Leu Gln Pro His Phe Asp Ser Pro Ala Phe Gln Asp His Pro

130 135 140

Leu Gln Pro Pro Val Arg Asp Leu Leu Arg Arg Lys Cys Ser Gln His

145 150 155 160

Asn Arg Leu Arg Glu Phe Phe Cys Pro Glu His Ser Glu Cys Ile Cys

165 170 175

His Ile Cys Leu Val Glu His Lys Thr Cys Ser Pro Ala Ser Leu Ser

180 185 190

Gln Ala Ser Ala Asp Leu Glu Ala Thr Leu Arg His Lys Leu Thr Val

195 200 205

Met Tyr Ser Gln Ile Asn Gly Ala Ser Arg Ala Leu Asp Asp Val Arg

210 215 220

Asn Arg Gln Gln Asp Val Arg Met Thr Ala Asn Arg Lys Val Glu Gln

225 230 235 240

Leu Gln Gln Glu Tyr Thr Glu Met Lys Ala Leu Leu Asp Ala Ser Glu

245 250 255

Thr Thr Ser Thr Arg Lys Ile Lys Glu Glu Glu Lys Arg Val Asn Ser

260 265 270

Lys Phe Asp Thr Ile Tyr Gln Ile Leu Leu Lys Lys Lys Ser Glu Ile

275 280 285

Gln Thr Leu Lys Glu Glu Ile Glu Gln Ser Leu Thr Lys Arg Asp Glu

290 295 300

Phe Glu Phe Leu Glu Lys Ala Ser Lys Leu Arg Gly Ile Ser Thr Lys

305 310 315 320

Pro Val Tyr Ile Pro Glu Val Glu Leu Asn His Lys Leu Ile Lys Gly

325 330 335

Ile His Gln Ser Thr Ile Asp Leu Lys Asn Glu Leu Lys Gln Cys Ile

340 345 350

Gly Arg Leu Gln Glu Pro Thr Pro Ser Ser Gly Asp Pro Gly Glu His

355 360 365

Asp Pro Ala Ser Thr His Lys Ser Thr Arg Pro Val Lys Lys Val Ser

370 375 380

Lys Glu Glu Lys Lys Ser Lys Lys Pro Pro Pro Val Pro Ala Leu Pro

385 390 395 400

Ser Lys Leu Pro Thr Phe Gly Ala Pro Glu Gln Leu Val Asp Leu Lys

405 410 415

Gln Ala Gly Leu Glu Ala Ala Ala Lys Ala Thr Ser Ser His Pro Asn

420 425 430

Ser Thr Ser Leu Lys Ala Lys Val Leu Glu Thr Phe Leu Ala Lys Ser

435 440 445

Arg Pro Glu Leu Leu Glu Tyr Tyr Ile Lys Val Ile Leu Asp Tyr Asn

450 455 460

Thr Ala His Asn Lys Val Ala Leu Ser Glu Cys Tyr Thr Val Ala Ser

465 470 475 480

Val Ala Glu Met Pro Gln Asn Tyr Arg Pro His Pro Gln Arg Phe Thr

485 490 495

Tyr Cys Ser Gln Val Leu Gly Leu His Cys Tyr Lys Lys Gly Ile His

500 505 510

Tyr Trp Glu Val Glu Leu Gln Lys Asn Asn Phe Cys Gly Val Gly Ile

515 520 525

Cys Tyr Gly Ser Met Asn Arg Gln Gly Pro Glu Ser Arg Leu Gly Arg

530 535 540

Asn Ser Ala Ser Trp Cys Val Glu Trp Phe Asn Thr Lys Ile Ser Ala

545 550 555 560

Trp His Asn Asn Val Glu Lys Thr Leu Pro Ser Thr Lys Ala Thr Arg

565 570 575

Val Gly Val Leu Leu Asn Cys Asp His Gly Phe Val Ile Phe Phe Ala

580 585 590

Val Ala Asp Lys Val His Leu Met Tyr Lys Phe Arg Val Asp Phe Thr

595 600 605

Glu Ala Leu Tyr Pro Ala Phe Trp Val Phe Ser Ala Gly Ala Thr Leu

610 615 620

Ser Ile Cys Ser Pro Lys

625 630

<210> 51

<211> 1620

<212> DNA

<213> 智人(Homo sapiens)

<400> 51

atggccacgt cagccccact acggagcctg gaagaggagg tgacctgctc catctgtctt 60

gattacctgc gggaccctgt gaccattgac tgtggccacg tcttctgccg cagctgcacc 120

acagacgtcc gccccatctc agggagccgc cccgtctgcc cactctgcaa gaagcctttt 180

aagaaggaga acatccgacc cgtgtggcaa ctggccagcc tggtggagaa cattgagcgg 240

ctgaaggtgg acaagggcag gcagccggga gaggtgaccc gggagcagca ggatgcaaag 300

ttgtgcgagc gacaccgaga gaagctgcac tactactgtg aggacgacgg gaagctgctg 360

tgcgtgatgt gccgggagtc ccgggagcac aggccccaca cggccgtcct catggagaag 420

gccgcccagc cccacaggga aaaaatcctg aaccacctga gtaccctaag gagggacaga 480

gacaaaattc agggcttcca ggcaaaggga gaagctgata tcctggccgc gctgaagaag 540

ctccaggacc agaggcagta cattgtggct gagtttgagc agggtcatca gttcctgagg 600

gagcgggagg aacacctgct ggaacagctg gcgaagctgg agcaggagct cacggagggc 660

agggagaagt tcaagagccg gggcgtcggg gagcttgccc ggctggccct ggtcatctcc 720

gaactggagg gcaaggcgca gcagccagct gcagagctca tgcaggacac gagagacttc 780

ctaaacaggt atccacggaa gaagttctgg gttgggaaac ccattgctcg agtggttaaa 840

aaaaagaccg gagaattctc agataaactc ctctctctgc aacgaggcct gagggaattc 900

caggggaagc tgctgagaga cttggaatat aagacagtga gcgtcaccct ggacccacag 960

tcggccagtg ggtacctgca gctgtcagag gactggaagt gcgtgaccta caccagcctg 1020

tacaagagtg cctacctgca cccccagcag tttgactgtg agcctggggt gctaggcagc 1080

aagggcttca cctggggcaa ggtctactgg gaagtggaag tggagaggga gggctggtct 1140

gaggatgaag aagaggggga tgaggaggaa gagggagaag aggaggagga ggaagaggag 1200

gccggctatg gggatggata tgacgactgg gaaacggacg aagatgagga atcgttgggc 1260

gatgaagagg aagaagagga ggaggaagag gaggaagttc tggaaagctg catggtgggg 1320

gtggctagag actctgtgaa gaggaaggga gacctctccc tgcggccaga ggatggcgtg 1380

tgggcgctgc gcctctcctc ctccggcatc tgggccaaca ccagccccga ggctgagctt 1440

ttcccagcac tgcggccccg gagagtgggc atcgccctgg attatgaagg gggcaccgtg 1500

actttcacca acgcagagtc acaggaactc atctacacct tcactgccac cttcacccgg 1560

cgcctggtcc ccttcctgtg gctcaagtgg ccaggaacac gcctcctgct aagaccctga 1620

<210> 52

<211> 539

<212> PRT

<213> 智人(Homo sapiens)

<400> 52

Met Ala Thr Ser Ala Pro Leu Arg Ser Leu Glu Glu Glu Val Thr Cys

1 5 10 15

Ser Ile Cys Leu Asp Tyr Leu Arg Asp Pro Val Thr Ile Asp Cys Gly

20 25 30

His Val Phe Cys Arg Ser Cys Thr Thr Asp Val Arg Pro Ile Ser Gly

35 40 45

Ser Arg Pro Val Cys Pro Leu Cys Lys Lys Pro Phe Lys Lys Glu Asn

50 55 60

Ile Arg Pro Val Trp Gln Leu Ala Ser Leu Val Glu Asn Ile Glu Arg

65 70 75 80

Leu Lys Val Asp Lys Gly Arg Gln Pro Gly Glu Val Thr Arg Glu Gln

85 90 95

Gln Asp Ala Lys Leu Cys Glu Arg His Arg Glu Lys Leu His Tyr Tyr

100 105 110

Cys Glu Asp Asp Gly Lys Leu Leu Cys Val Met Cys Arg Glu Ser Arg

115 120 125

Glu His Arg Pro His Thr Ala Val Leu Met Glu Lys Ala Ala Gln Pro

130 135 140

His Arg Glu Lys Ile Leu Asn His Leu Ser Thr Leu Arg Arg Asp Arg

145 150 155 160

Asp Lys Ile Gln Gly Phe Gln Ala Lys Gly Glu Ala Asp Ile Leu Ala

165 170 175

Ala Leu Lys Lys Leu Gln Asp Gln Arg Gln Tyr Ile Val Ala Glu Phe

180 185 190

Glu Gln Gly His Gln Phe Leu Arg Glu Arg Glu Glu His Leu Leu Glu

195 200 205

Gln Leu Ala Lys Leu Glu Gln Glu Leu Thr Glu Gly Arg Glu Lys Phe

210 215 220

Lys Ser Arg Gly Val Gly Glu Leu Ala Arg Leu Ala Leu Val Ile Ser

225 230 235 240

Glu Leu Glu Gly Lys Ala Gln Gln Pro Ala Ala Glu Leu Met Gln Asp

245 250 255

Thr Arg Asp Phe Leu Asn Arg Tyr Pro Arg Lys Lys Phe Trp Val Gly

260 265 270

Lys Pro Ile Ala Arg Val Val Lys Lys Lys Thr Gly Glu Phe Ser Asp

275 280 285

Lys Leu Leu Ser Leu Gln Arg Gly Leu Arg Glu Phe Gln Gly Lys Leu

290 295 300

Leu Arg Asp Leu Glu Tyr Lys Thr Val Ser Val Thr Leu Asp Pro Gln

305 310 315 320

Ser Ala Ser Gly Tyr Leu Gln Leu Ser Glu Asp Trp Lys Cys Val Thr

325 330 335

Tyr Thr Ser Leu Tyr Lys Ser Ala Tyr Leu His Pro Gln Gln Phe Asp

340 345 350

Cys Glu Pro Gly Val Leu Gly Ser Lys Gly Phe Thr Trp Gly Lys Val

355 360 365

Tyr Trp Glu Val Glu Val Glu Arg Glu Gly Trp Ser Glu Asp Glu Glu

370 375 380

Glu Gly Asp Glu Glu Glu Glu Gly Glu Glu Glu Glu Glu Glu Glu Glu

385 390 395 400

Ala Gly Tyr Gly Asp Gly Tyr Asp Asp Trp Glu Thr Asp Glu Asp Glu

405 410 415

Glu Ser Leu Gly Asp Glu Glu Glu Glu Glu Glu Glu Glu Glu Glu Glu

420 425 430

Val Leu Glu Ser Cys Met Val Gly Val Ala Arg Asp Ser Val Lys Arg

435 440 445

Lys Gly Asp Leu Ser Leu Arg Pro Glu Asp Gly Val Trp Ala Leu Arg

450 455 460

Leu Ser Ser Ser Gly Ile Trp Ala Asn Thr Ser Pro Glu Ala Glu Leu

465 470 475 480

Phe Pro Ala Leu Arg Pro Arg Arg Val Gly Ile Ala Leu Asp Tyr Glu

485 490 495

Gly Gly Thr Val Thr Phe Thr Asn Ala Glu Ser Gln Glu Leu Ile Tyr

500 505 510

Thr Phe Thr Ala Thr Phe Thr Arg Arg Leu Val Pro Phe Leu Trp Leu

515 520 525

Lys Trp Pro Gly Thr Arg Leu Leu Leu Arg Pro

530 535

<210> 53

<211> 1542

<212> DNA

<213> 智人(Homo sapiens)

<400> 53

atggcctccg ggagtgtggc cgagtgcctg cagcaggaga ccacctgccc cgtgtgcctg 60

cagtacttcg cagagcccat gatgctcgac tgcggccata acatctgttg cgcgtgcctc 120

gcccgctgct ggggcacggc agagactaac gtgtcgtgcc cgcagtgccg ggagaccttc 180

ccgcagaggc acatgcggcc caaccggcac ctggccaacg tgacccaact ggtaaagcag 240

ctgcgcaccg agcggccgtc ggggcccggc ggcgagatgg gcgtgtgcga gaagcaccgc 300

gagcccctga agctgtactg cgaggaggac cagatgccca tctgcgtggt gtgcgaccgc 360

tcccgcgagc accgcggcca cagcgtgctg ccgctcgagg aggcggtgga gggcttcaag 420

gagcaaatcc agaaccagct cgaccattta aaaagagtga aagatttaaa gaagagacgt 480

cgggcccagg gggaacaggc acgagctgaa ctcttgagcc taacccagat ggagagggag 540

aagattgttt gggagtttga gcagctgtat cactccttaa aggagcatga gtatcgcctc 600

ctggcccgcc ttgaggagct agacttggcc atctacaata gcatcaatgg tgccatcacc 660

cagttctctt gcaacatctc ccacctcagc agcctgatcg ctcagctaga agagaagcag 720

cagcagccca ccagggagct cctgcaggac attggggaca cattgagcag ggctgaaaga 780

atcaggattc ctgaaccttg gatcacacct ccagatttgc aagagaaaat ccacattttt 840

gcccaaaaat gtctattctt gacggagagt ctaaagcagt tcacagaaaa aatgcagtca 900

gatatggaga aaatccaaga attaagagag gctcagttat actcagtgga cgtgactctg 960

gacccagaca cggcctaccc cagcctgatc ctctctgata atctgcggca agtgcggtac 1020

agttacctcc aacaggacct gcctgacaac cccgagaggt tcaatctgtt tccctgtgtc 1080

ttgggctctc catgcttcat cgccgggaga cattattggg aggtagaggt gggagataaa 1140

gccaagtgga ccataggtgt ctgtgaagac tcagtgtgca gaaaaggtgg agtaacctca 1200

gccccccaga atggattctg ggcagtgtct ttgtggtatg ggaaagaata ttgggctctt 1260

acctccccaa tgactgccct acccctgcgg accccgctcc agcgggtggg gattttcttg 1320

gactatgatg ctggtgaggt ctccttctac aacgtgacag agaggtgtca caccttcact 1380

ttctctcatg ctaccttttg tgggcctgtc cggccctact tcagtctgag ttactcggga 1440

gggaaaagtg cagctcctct gatcatctgc cccatgagtg ggatagatgg gttttctggc 1500

catgttggga atcatggtca ttccatggag acctcccctt ga 1542

<210> 54

<211> 513

<212> PRT

<213> 智人(Homo sapiens)

<400> 54

Met Ala Ser Gly Ser Val Ala Glu Cys Leu Gln Gln Glu Thr Thr Cys

1 5 10 15

Pro Val Cys Leu Gln Tyr Phe Ala Glu Pro Met Met Leu Asp Cys Gly

20 25 30

His Asn Ile Cys Cys Ala Cys Leu Ala Arg Cys Trp Gly Thr Ala Glu

35 40 45

Thr Asn Val Ser Cys Pro Gln Cys Arg Glu Thr Phe Pro Gln Arg His

50 55 60

Met Arg Pro Asn Arg His Leu Ala Asn Val Thr Gln Leu Val Lys Gln

65 70 75 80

Leu Arg Thr Glu Arg Pro Ser Gly Pro Gly Gly Glu Met Gly Val Cys

85 90 95

Glu Lys His Arg Glu Pro Leu Lys Leu Tyr Cys Glu Glu Asp Gln Met

100 105 110

Pro Ile Cys Val Val Cys Asp Arg Ser Arg Glu His Arg Gly His Ser

115 120 125

Val Leu Pro Leu Glu Glu Ala Val Glu Gly Phe Lys Glu Gln Ile Gln

130 135 140

Asn Gln Leu Asp His Leu Lys Arg Val Lys Asp Leu Lys Lys Arg Arg

145 150 155 160

Arg Ala Gln Gly Glu Gln Ala Arg Ala Glu Leu Leu Ser Leu Thr Gln

165 170 175

Met Glu Arg Glu Lys Ile Val Trp Glu Phe Glu Gln Leu Tyr His Ser

180 185 190

Leu Lys Glu His Glu Tyr Arg Leu Leu Ala Arg Leu Glu Glu Leu Asp

195 200 205

Leu Ala Ile Tyr Asn Ser Ile Asn Gly Ala Ile Thr Gln Phe Ser Cys

210 215 220

Asn Ile Ser His Leu Ser Ser Leu Ile Ala Gln Leu Glu Glu Lys Gln

225 230 235 240

Gln Gln Pro Thr Arg Glu Leu Leu Gln Asp Ile Gly Asp Thr Leu Ser

245 250 255

Arg Ala Glu Arg Ile Arg Ile Pro Glu Pro Trp Ile Thr Pro Pro Asp

260 265 270

Leu Gln Glu Lys Ile His Ile Phe Ala Gln Lys Cys Leu Phe Leu Thr

275 280 285

Glu Ser Leu Lys Gln Phe Thr Glu Lys Met Gln Ser Asp Met Glu Lys

290 295 300

Ile Gln Glu Leu Arg Glu Ala Gln Leu Tyr Ser Val Asp Val Thr Leu

305 310 315 320

Asp Pro Asp Thr Ala Tyr Pro Ser Leu Ile Leu Ser Asp Asn Leu Arg

325 330 335

Gln Val Arg Tyr Ser Tyr Leu Gln Gln Asp Leu Pro Asp Asn Pro Glu

340 345 350

Arg Phe Asn Leu Phe Pro Cys Val Leu Gly Ser Pro Cys Phe Ile Ala

355 360 365

Gly Arg His Tyr Trp Glu Val Glu Val Gly Asp Lys Ala Lys Trp Thr

370 375 380

Ile Gly Val Cys Glu Asp Ser Val Cys Arg Lys Gly Gly Val Thr Ser

385 390 395 400

Ala Pro Gln Asn Gly Phe Trp Ala Val Ser Leu Trp Tyr Gly Lys Glu

405 410 415

Tyr Trp Ala Leu Thr Ser Pro Met Thr Ala Leu Pro Leu Arg Thr Pro

420 425 430

Leu Gln Arg Val Gly Ile Phe Leu Asp Tyr Asp Ala Gly Glu Val Ser

435 440 445

Phe Tyr Asn Val Thr Glu Arg Cys His Thr Phe Thr Phe Ser His Ala

450 455 460

Thr Phe Cys Gly Pro Val Arg Pro Tyr Phe Ser Leu Ser Tyr Ser Gly

465 470 475 480

Gly Lys Ser Ala Ala Pro Leu Ile Ile Cys Pro Met Ser Gly Ile Asp

485 490 495

Gly Phe Ser Gly His Val Gly Asn His Gly His Ser Met Glu Thr Ser

500 505 510

Pro

<210> 55

<211> 2508

<212> DNA

<213> 智人(Homo sapiens)

<400> 55

atggcggcct ccgcggcggc agcctcggca gcagcggcct cggccgcctc tggcagcccg 60

ggcccgggcg agggctccgc tggcggcgaa aagcgctcca ccgccccttc ggccgcagcc 120

tcggcctctg cctcagccgc ggcgtcgtcg cccgcggggg gcggcgccga ggcgctggag 180

ctgctggagc actgcggcgt gtgcagagag cgcctgcgac ccgagaggga gccccgcctg 240

ctgccctgtt tgcactcggc ctgtagtgcc tgcttagggc ccgcggcccc cgccgccgcc 300

aacagctcgg gggacggcgg ggcggcgggc gacggcaccg tggtggactg tcccgtgtgc 360

aagcaacagt gcttctccaa agacatcgtg gagaattatt tcatgcgtga tagtggcagc 420

aaggctgcca ccgacgccca ggatgcgaac cagtgctgca ctagctgtga ggataatgcc 480

ccagccacca gctactgtgt ggagtgctcg gagcctctgt gtgagacctg tgtagaggcg 540

caccagcggg tgaagtacac caaggaccat actgtgcgct ctactgggcc agccaagtct 600

cgggatggtg aacgtactgt ctattgcaac gtacacaagc atgaacccct tgtgctgttt 660

tgtgagagct gtgatactct cacctgccga gactgccagc tcaatgccca caaggaccac 720

cagtaccagt tcttagagga tgcagtgagg aaccagcgca agctcctggc ctcactggtg 780

aagcgccttg gggacaaaca tgcaacattg cagaagagca ccaaggaggt tcgcagctca 840

atccgccagg tgtctgacgt acagaagcgt gtgcaagtgg atgtcaagat ggccatcctg 900

cagatcatga aggagctgaa taagcggggc cgtgtgctgg tcaatgatgc ccagaaggtg 960

actgaggggc agcaggagcg cctggagcgg cagcactgga ccatgaccaa gatccagaag 1020

caccaggagc acattctgcg ctttgcctct tgggctctgg agagtgacaa caacacagcc 1080

cttttgcttt ctaagaagtt gatctacttc cagctgcacc gggccctcaa gatgattgtg 1140

gatcccgtgg agccacatgg cgagatgaag tttcagtggg acctcaatgc ctggaccaag 1200

agtgccgagg cctttggcaa gattgtggca gagcgtcctg gcactaactc aacaggccct 1260

gcacccatgg cccctccaag agccccaggg cccctgagca agcagggctc tggcagcagc 1320

cagcccatgg aggtgcagga aggctatggc tttgggtcag gagatgatcc ctactcaagt 1380

gcagagcccc atgtgtcagg tgtgaaacgg tcccgctcag gtgagggcga ggtgagcggc 1440

cttatgcgca aggtgccacg agtgagcctt gaacgcctgg acctggacct cacagctgac 1500

agccagccac ccgtcttcaa ggtcttccca ggcagtacca ctgaggacta caaccttatt 1560

gttattgaac gtggcgctgc cgctgcagct accggccagc cagggactgc gcctgcagga 1620

acccctggtg ccccacccct ggctggcatg gccattgtca aggaggagga gacggaggct 1680

gccattggag cccctcctac tgccactgag ggccctgaga ccaaacctgt gcttatggct 1740

cttgcggagg gtcctggtgc tgagggtccc cgcctggcct cacctagtgg cagcaccagc 1800

tcagggctgg aggtggtggc tcctgagggt acctcagccc caggtggtgg cccgggaacc 1860

ctggatgaca gtgccaccat ttgccgtgtc tgccagaagc caggcgatct ggttatgtgc 1920

aaccagtgtg agttttgttt ccacctggac tgtcacctgc cggccctgca ggatgtacca 1980

ggggaggagt ggagctgctc actctgccat gtgctccctg acctgaagga ggaggatggc 2040

agcctcagcc tggatggtgc agacagcact ggcgtggtgg ccaagctctc accagccaac 2100

cagcggaaat gtgagcgtgt actgctggcc ctattctgtc acgaaccctg ccgccccctg 2160

catcagctgg ctaccgactc caccttctcc ctggaccagc ccggtggcac cctggatctg 2220

accctgatcc gtgcccgcct ccaggagaag ttgtcacctc cctacagctc cccacaggag 2280

tttgcccagg atgtgggccg catgttcaag caattcaaca agttaactga ggacaaggca 2340

gacgtgcagt ccatcatcgg cctgcagcgc ttcttcgaga cgcgcatgaa cgaggccttc 2400

ggtgacacca agttctctgc tgtgctggtg gagcccccgc cgatgagcct gcctggtgct 2460

ggcctgagtt cccaggagct gtctggtggc cctggtgatg gcccctga 2508

<210> 56

<211> 835

<212> PRT

<213> 智人(Homo sapiens)

<400> 56

Met Ala Ala Ser Ala Ala Ala Ala Ser Ala Ala Ala Ala Ser Ala Ala

1 5 10 15

Ser Gly Ser Pro Gly Pro Gly Glu Gly Ser Ala Gly Gly Glu Lys Arg

20 25 30

Ser Thr Ala Pro Ser Ala Ala Ala Ser Ala Ser Ala Ser Ala Ala Ala

35 40 45

Ser Ser Pro Ala Gly Gly Gly Ala Glu Ala Leu Glu Leu Leu Glu His

50 55 60

Cys Gly Val Cys Arg Glu Arg Leu Arg Pro Glu Arg Glu Pro Arg Leu

65 70 75 80

Leu Pro Cys Leu His Ser Ala Cys Ser Ala Cys Leu Gly Pro Ala Ala

85 90 95

Pro Ala Ala Ala Asn Ser Ser Gly Asp Gly Gly Ala Ala Gly Asp Gly

100 105 110

Thr Val Val Asp Cys Pro Val Cys Lys Gln Gln Cys Phe Ser Lys Asp

115 120 125

Ile Val Glu Asn Tyr Phe Met Arg Asp Ser Gly Ser Lys Ala Ala Thr

130 135 140

Asp Ala Gln Asp Ala Asn Gln Cys Cys Thr Ser Cys Glu Asp Asn Ala

145 150 155 160

Pro Ala Thr Ser Tyr Cys Val Glu Cys Ser Glu Pro Leu Cys Glu Thr

165 170 175

Cys Val Glu Ala His Gln Arg Val Lys Tyr Thr Lys Asp His Thr Val

180 185 190

Arg Ser Thr Gly Pro Ala Lys Ser Arg Asp Gly Glu Arg Thr Val Tyr

195 200 205

Cys Asn Val His Lys His Glu Pro Leu Val Leu Phe Cys Glu Ser Cys

210 215 220

Asp Thr Leu Thr Cys Arg Asp Cys Gln Leu Asn Ala His Lys Asp His

225 230 235 240

Gln Tyr Gln Phe Leu Glu Asp Ala Val Arg Asn Gln Arg Lys Leu Leu

245 250 255

Ala Ser Leu Val Lys Arg Leu Gly Asp Lys His Ala Thr Leu Gln Lys

260 265 270

Ser Thr Lys Glu Val Arg Ser Ser Ile Arg Gln Val Ser Asp Val Gln

275 280 285

Lys Arg Val Gln Val Asp Val Lys Met Ala Ile Leu Gln Ile Met Lys

290 295 300

Glu Leu Asn Lys Arg Gly Arg Val Leu Val Asn Asp Ala Gln Lys Val

305 310 315 320

Thr Glu Gly Gln Gln Glu Arg Leu Glu Arg Gln His Trp Thr Met Thr

325 330 335

Lys Ile Gln Lys His Gln Glu His Ile Leu Arg Phe Ala Ser Trp Ala

340 345 350

Leu Glu Ser Asp Asn Asn Thr Ala Leu Leu Leu Ser Lys Lys Leu Ile

355 360 365

Tyr Phe Gln Leu His Arg Ala Leu Lys Met Ile Val Asp Pro Val Glu

370 375 380

Pro His Gly Glu Met Lys Phe Gln Trp Asp Leu Asn Ala Trp Thr Lys

385 390 395 400

Ser Ala Glu Ala Phe Gly Lys Ile Val Ala Glu Arg Pro Gly Thr Asn

405 410 415

Ser Thr Gly Pro Ala Pro Met Ala Pro Pro Arg Ala Pro Gly Pro Leu

420 425 430

Ser Lys Gln Gly Ser Gly Ser Ser Gln Pro Met Glu Val Gln Glu Gly

435 440 445

Tyr Gly Phe Gly Ser Gly Asp Asp Pro Tyr Ser Ser Ala Glu Pro His

450 455 460

Val Ser Gly Val Lys Arg Ser Arg Ser Gly Glu Gly Glu Val Ser Gly

465 470 475 480

Leu Met Arg Lys Val Pro Arg Val Ser Leu Glu Arg Leu Asp Leu Asp

485 490 495

Leu Thr Ala Asp Ser Gln Pro Pro Val Phe Lys Val Phe Pro Gly Ser

500 505 510

Thr Thr Glu Asp Tyr Asn Leu Ile Val Ile Glu Arg Gly Ala Ala Ala

515 520 525

Ala Ala Thr Gly Gln Pro Gly Thr Ala Pro Ala Gly Thr Pro Gly Ala

530 535 540

Pro Pro Leu Ala Gly Met Ala Ile Val Lys Glu Glu Glu Thr Glu Ala

545 550 555 560

Ala Ile Gly Ala Pro Pro Thr Ala Thr Glu Gly Pro Glu Thr Lys Pro

565 570 575

Val Leu Met Ala Leu Ala Glu Gly Pro Gly Ala Glu Gly Pro Arg Leu

580 585 590

Ala Ser Pro Ser Gly Ser Thr Ser Ser Gly Leu Glu Val Val Ala Pro

595 600 605

Glu Gly Thr Ser Ala Pro Gly Gly Gly Pro Gly Thr Leu Asp Asp Ser

610 615 620

Ala Thr Ile Cys Arg Val Cys Gln Lys Pro Gly Asp Leu Val Met Cys

625 630 635 640

Asn Gln Cys Glu Phe Cys Phe His Leu Asp Cys His Leu Pro Ala Leu

645 650 655

Gln Asp Val Pro Gly Glu Glu Trp Ser Cys Ser Leu Cys His Val Leu

660 665 670

Pro Asp Leu Lys Glu Glu Asp Gly Ser Leu Ser Leu Asp Gly Ala Asp

675 680 685

Ser Thr Gly Val Val Ala Lys Leu Ser Pro Ala Asn Gln Arg Lys Cys

690 695 700

Glu Arg Val Leu Leu Ala Leu Phe Cys His Glu Pro Cys Arg Pro Leu

705 710 715 720

His Gln Leu Ala Thr Asp Ser Thr Phe Ser Leu Asp Gln Pro Gly Gly

725 730 735

Thr Leu Asp Leu Thr Leu Ile Arg Ala Arg Leu Gln Glu Lys Leu Ser

740 745 750

Pro Pro Tyr Ser Ser Pro Gln Glu Phe Ala Gln Asp Val Gly Arg Met

755 760 765

Phe Lys Gln Phe Asn Lys Leu Thr Glu Asp Lys Ala Asp Val Gln Ser

770 775 780

Ile Ile Gly Leu Gln Arg Phe Phe Glu Thr Arg Met Asn Glu Ala Phe

785 790 795 800

Gly Asp Thr Lys Phe Ser Ala Val Leu Val Glu Pro Pro Pro Met Ser

805 810 815

Leu Pro Gly Ala Gly Leu Ser Ser Gln Glu Leu Ser Gly Gly Pro Gly

820 825 830

Asp Gly Pro

835

<210> 57

<211> 1767

<212> DNA

<213> 智人(Homo sapiens)

<400> 57

atggaagctg cagatgcctc caggagcaac gggtcgagcc cagaagccag ggatgcccgg 60

agcccgtcgg gccccagtgg cagcctggag aatggcacca aggctgacgg caaggatgcc 120

aagaccacca acgggcacgg cggggaggca gctgagggca agagcctggg cagcgccctg 180

aagccagggg aaggtaggag cgccctgttc gcgggcaatg agtggcggcg acccatcatc 240

cagtttgtcg agtccgggga cgacaagaac tccaactact tcagcatgga ctctatggaa 300

ggcaagaggt cgccgtacgc agggctccag ctgggggctg ccaagaagcc acccgttacc 360

tttgccgaaa agggcgagct gcgcaagtcc attttctcgg agtcccggaa gcccacggtg 420

tccatcatgg agcccgggga gacccggcgg aacagctacc cccgggccga cacgggcctt 480

ttttcacggt ccaagtccgg ctccgaggag gtgctgtgcg actcctgcat cggcaacaag 540

cagaaggcgg tcaagtcctg cctggtgtgc caggcctcct tctgcgagct gcatctcaag 600

ccccacctgg agggcgccgc cttccgagac caccagctgc tcgagcccat ccgggacttt 660

gaggcccgca agtgtcccgt gcatggcaag acgatggagc tcttctgcca gaccgaccag 720

acctgcatct gctacctttg catgttccag gagcacaaga atcatagcac cgtgacagtg 780

gaggaggcca aggccgagaa ggagacggag ctgtcattgc aaaaggagca gctgcagctc 840

aagatcattg agattgagga tgaagctgag aagtggcaga aggagaagga ccgcatcaag 900

agcttcacca ccaatgagaa ggccatcctg gagcagaact tccgggacct ggtgcgggac 960

ctggagaagc aaaaggagga agtgagggct gcgctggagc agcgggagca ggatgctgtg 1020

gaccaagtga aggtgatcat ggatgctctg gatgagagag ccaaggtgct gcatgaggac 1080

aagcagaccc gggagcagct gcatagcatc agcgactctg tgttgtttct gcaggaattt 1140

ggtgcattga tgagcaatta ctctctcccc ccacccctgc ccacctatca tgtcctgctg 1200

gagggggagg gcctgggaca gtcactaggc aacttcaagg acgacctgct caatgtatgc 1260

atgcgccacg ttgagaagat gtgcaaggcg gacctgagcc gtaacttcat tgagaggaac 1320

cacatggaga acggtggtga ccatcgctat gtgaacaact acacgaacag cttcgggggt 1380

gagtggagtg caccggacac catgaagaga tactccatgt acctgacacc caaaggtggg 1440

gtccggacat cataccagcc ctcgtctcct ggccgcttca ccaaggagac cacccagaag 1500

aatttcaaca atctctatgg caccaaaggt aactacacct cccgggtctg ggagtactcc 1560

tccagcattc agaactctga caatgacctg cccgtcgtcc aaggcagctc ctccttctcc 1620

ctgaaaggct atccctccct catgcggagc caaagcccca aggcccagcc ccagacttgg 1680

aaatctggca agcagactat gctgtctcac taccggccat tctacgtcaa caaaggcaac 1740

gggattgggt ccaacgaagc cccatga 1767

<210> 58

<211> 588

<212> PRT

<213> 智人(Homo sapiens)

<400> 58

Met Glu Ala Ala Asp Ala Ser Arg Ser Asn Gly Ser Ser Pro Glu Ala

1 5 10 15

Arg Asp Ala Arg Ser Pro Ser Gly Pro Ser Gly Ser Leu Glu Asn Gly

20 25 30

Thr Lys Ala Asp Gly Lys Asp Ala Lys Thr Thr Asn Gly His Gly Gly

35 40 45

Glu Ala Ala Glu Gly Lys Ser Leu Gly Ser Ala Leu Lys Pro Gly Glu

50 55 60

Gly Arg Ser Ala Leu Phe Ala Gly Asn Glu Trp Arg Arg Pro Ile Ile

65 70 75 80

Gln Phe Val Glu Ser Gly Asp Asp Lys Asn Ser Asn Tyr Phe Ser Met

85 90 95

Asp Ser Met Glu Gly Lys Arg Ser Pro Tyr Ala Gly Leu Gln Leu Gly

100 105 110

Ala Ala Lys Lys Pro Pro Val Thr Phe Ala Glu Lys Gly Glu Leu Arg

115 120 125

Lys Ser Ile Phe Ser Glu Ser Arg Lys Pro Thr Val Ser Ile Met Glu

130 135 140

Pro Gly Glu Thr Arg Arg Asn Ser Tyr Pro Arg Ala Asp Thr Gly Leu

145 150 155 160

Phe Ser Arg Ser Lys Ser Gly Ser Glu Glu Val Leu Cys Asp Ser Cys

165 170 175

Ile Gly Asn Lys Gln Lys Ala Val Lys Ser Cys Leu Val Cys Gln Ala

180 185 190

Ser Phe Cys Glu Leu His Leu Lys Pro His Leu Glu Gly Ala Ala Phe

195 200 205

Arg Asp His Gln Leu Leu Glu Pro Ile Arg Asp Phe Glu Ala Arg Lys

210 215 220

Cys Pro Val His Gly Lys Thr Met Glu Leu Phe Cys Gln Thr Asp Gln

225 230 235 240

Thr Cys Ile Cys Tyr Leu Cys Met Phe Gln Glu His Lys Asn His Ser

245 250 255

Thr Val Thr Val Glu Glu Ala Lys Ala Glu Lys Glu Thr Glu Leu Ser

260 265 270

Leu Gln Lys Glu Gln Leu Gln Leu Lys Ile Ile Glu Ile Glu Asp Glu

275 280 285

Ala Glu Lys Trp Gln Lys Glu Lys Asp Arg Ile Lys Ser Phe Thr Thr

290 295 300

Asn Glu Lys Ala Ile Leu Glu Gln Asn Phe Arg Asp Leu Val Arg Asp

305 310 315 320

Leu Glu Lys Gln Lys Glu Glu Val Arg Ala Ala Leu Glu Gln Arg Glu

325 330 335

Gln Asp Ala Val Asp Gln Val Lys Val Ile Met Asp Ala Leu Asp Glu

340 345 350

Arg Ala Lys Val Leu His Glu Asp Lys Gln Thr Arg Glu Gln Leu His

355 360 365

Ser Ile Ser Asp Ser Val Leu Phe Leu Gln Glu Phe Gly Ala Leu Met

370 375 380

Ser Asn Tyr Ser Leu Pro Pro Pro Leu Pro Thr Tyr His Val Leu Leu

385 390 395 400

Glu Gly Glu Gly Leu Gly Gln Ser Leu Gly Asn Phe Lys Asp Asp Leu

405 410 415

Leu Asn Val Cys Met Arg His Val Glu Lys Met Cys Lys Ala Asp Leu

420 425 430

Ser Arg Asn Phe Ile Glu Arg Asn His Met Glu Asn Gly Gly Asp His

435 440 445

Arg Tyr Val Asn Asn Tyr Thr Asn Ser Phe Gly Gly Glu Trp Ser Ala

450 455 460

Pro Asp Thr Met Lys Arg Tyr Ser Met Tyr Leu Thr Pro Lys Gly Gly

465 470 475 480

Val Arg Thr Ser Tyr Gln Pro Ser Ser Pro Gly Arg Phe Thr Lys Glu

485 490 495

Thr Thr Gln Lys Asn Phe Asn Asn Leu Tyr Gly Thr Lys Gly Asn Tyr

500 505 510

Thr Ser Arg Val Trp Glu Tyr Ser Ser Ser Ile Gln Asn Ser Asp Asn

515 520 525

Asp Leu Pro Val Val Gln Gly Ser Ser Ser Phe Ser Leu Lys Gly Tyr

530 535 540

Pro Ser Leu Met Arg Ser Gln Ser Pro Lys Ala Gln Pro Gln Thr Trp

545 550 555 560

Lys Ser Gly Lys Gln Thr Met Leu Ser His Tyr Arg Pro Phe Tyr Val

565 570 575

Asn Lys Gly Asn Gly Ile Gly Ser Asn Glu Ala Pro

580 585

<210> 59

<211> 1491

<212> DNA

<213> 小鼠(Mus musculus)

<400> 59

atggcctcat cagtcctgga gatgataaag gaggaagtaa cctgtcctat ctgtttggag 60

ctcctgaagg aacctgtgag tgctgattgt aaccacagct tctgcagagc ctgcatcaca 120

ctgaattatg agtccaacag aaacacagac gggaagggca actgccctgt atgccgagtt 180

ccttacccat ttgggaatct gaggcctaat ctacatgtgg ccaacatagt agagaggctc 240

aagggattca agtccattcc agaggaggag cagaaggtga atatctgtgc acaacatgga 300

gagaaactcc ggctcttctg taggaaggac atgatggtca tctgctggct ttgtgagcga 360

tctcaggagc accgtggtca ccaaacagct ctcattgaag aggttgacca agaatacaag 420

gagaagctgc agggagctct gtggaagctg atgaaaaagg caaaaatatg tgatgaatgg 480

caggatgacc ttcaactgca gagagttgac tgggagaacc aaatacagat caatgtagaa 540

aatgttcaga gacagtttaa aggactaaga gacctcctgg actccaagga gaatgaggag 600

ctgcagaagc tgaagaaaga gaagaaagag gttatggaaa agctggaaga gtctgaaaat 660

gagctggagg atcagacaga gttggtgaga gacctcatct cagatgtgga acatcatttg 720

gagctctcaa ccttagaaat gctgcagggt gcaaattgtg tcctgagaag gagtcagtcc 780

ttaagcctgc aacagcccca aactgtcccc caaaagagaa aaagaacatt ccaagctcca 840

gatctgaaag gcatgctgca agtgtatcaa ggactcatgg atatccagca atactgggtt 900

catatgactc tacatgcaag gaacaatgca gtcattgcca ttaacaaaga aaaaagacaa 960

atacagtata gaagttacaa tacggttcca gtttctgaga tctaccattt gggtgtcctg 1020

ggatatccag ctctttcctc agggaagcat tactgggaag tagacatatc tagaagtgat 1080

gcctggctcc tcggattaaa tgacggaaag tgtgctcaac cccaacttca ctcaaaggaa 1140

gaaatgggca tcaaaaaaaa ccttcattct cagatcaaac aaaatgtatt gtttcagcct 1200

aaatgtggct actgggttat agggatgaag aatccgtctg tatacaaggc ctttgatgag 1260

tgttctatca cccacaattc cagtatcctg gtcatctctc tgcctgatcg tcccagtcgt 1320

gtcggagttt tcctggatcg gaaagctggc actctctcat tttatgatgt ttctaactgc 1380

ggtgctctca tctataggtt ctatgaccct gccttccctg ttgaagtcta tccatatttt 1440

aatcctatga aatgttcaga gccaatgact atatgcgggc caccctccta a 1491

<210> 60

<211> 496

<212> PRT

<213> 小鼠(Mus musculus)

<400> 60

Met Ala Ser Ser Val Leu Glu Met Ile Lys Glu Glu Val Thr Cys Pro

1 5 10 15

Ile Cys Leu Glu Leu Leu Lys Glu Pro Val Ser Ala Asp Cys Asn His

20 25 30

Ser Phe Cys Arg Ala Cys Ile Thr Leu Asn Tyr Glu Ser Asn Arg Asn

35 40 45

Thr Asp Gly Lys Gly Asn Cys Pro Val Cys Arg Val Pro Tyr Pro Phe

50 55 60

Gly Asn Leu Arg Pro Asn Leu His Val Ala Asn Ile Val Glu Arg Leu

65 70 75 80

Lys Gly Phe Lys Ser Ile Pro Glu Glu Glu Gln Lys Val Asn Ile Cys

85 90 95

Ala Gln His Gly Glu Lys Leu Arg Leu Phe Cys Arg Lys Asp Met Met

100 105 110

Val Ile Cys Trp Leu Cys Glu Arg Ser Gln Glu His Arg Gly His Gln

115 120 125

Thr Ala Leu Ile Glu Glu Val Asp Gln Glu Tyr Lys Glu Lys Leu Gln

130 135 140

Gly Ala Leu Trp Lys Leu Met Lys Lys Ala Lys Ile Cys Asp Glu Trp

145 150 155 160

Gln Asp Asp Leu Gln Leu Gln Arg Val Asp Trp Glu Asn Gln Ile Gln

165 170 175

Ile Asn Val Glu Asn Val Gln Arg Gln Phe Lys Gly Leu Arg Asp Leu

180 185 190

Leu Asp Ser Lys Glu Asn Glu Glu Leu Gln Lys Leu Lys Lys Glu Lys

195 200 205

Lys Glu Val Met Glu Lys Leu Glu Glu Ser Glu Asn Glu Leu Glu Asp

210 215 220

Gln Thr Glu Leu Val Arg Asp Leu Ile Ser Asp Val Glu His His Leu

225 230 235 240

Glu Leu Ser Thr Leu Glu Met Leu Gln Gly Ala Asn Cys Val Leu Arg

245 250 255

Arg Ser Gln Ser Leu Ser Leu Gln Gln Pro Gln Thr Val Pro Gln Lys

260 265 270

Arg Lys Arg Thr Phe Gln Ala Pro Asp Leu Lys Gly Met Leu Gln Val

275 280 285

Tyr Gln Gly Leu Met Asp Ile Gln Gln Tyr Trp Val His Met Thr Leu

290 295 300

His Ala Arg Asn Asn Ala Val Ile Ala Ile Asn Lys Glu Lys Arg Gln

305 310 315 320

Ile Gln Tyr Arg Ser Tyr Asn Thr Val Pro Val Ser Glu Ile Tyr His

325 330 335

Leu Gly Val Leu Gly Tyr Pro Ala Leu Ser Ser Gly Lys His Tyr Trp

340 345 350

Glu Val Asp Ile Ser Arg Ser Asp Ala Trp Leu Leu Gly Leu Asn Asp

355 360 365

Gly Lys Cys Ala Gln Pro Gln Leu His Ser Lys Glu Glu Met Gly Ile

370 375 380

Lys Lys Asn Leu His Ser Gln Ile Lys Gln Asn Val Leu Phe Gln Pro

385 390 395 400

Lys Cys Gly Tyr Trp Val Ile Gly Met Lys Asn Pro Ser Val Tyr Lys

405 410 415

Ala Phe Asp Glu Cys Ser Ile Thr His Asn Ser Ser Ile Leu Val Ile

420 425 430

Ser Leu Pro Asp Arg Pro Ser Arg Val Gly Val Phe Leu Asp Arg Lys

435 440 445

Ala Gly Thr Leu Ser Phe Tyr Asp Val Ser Asn Cys Gly Ala Leu Ile

450 455 460

Tyr Arg Phe Tyr Asp Pro Ala Phe Pro Val Glu Val Tyr Pro Tyr Phe

465 470 475 480

Asn Pro Met Lys Cys Ser Glu Pro Met Thr Ile Cys Gly Pro Pro Ser

485 490 495

<210> 61

<211> 1278

<212> DNA

<213> 智人(Homo sapiens)

<400> 61

atggccagtg ggcagtttgt gaacaaactg caagaggaag tgatctgccc catctgcctg 60

gacattctgc agaaacctgt caccatcgac tgtgggcaca atttctgcct caaatgcatc 120

actcagattg gggaaacatc atgtggattt ttcaaatgtc ccctctgcaa aacttccgta 180

aggaagaacg caatcaggtt caactcgctg ttgcggaatc tggtggagaa aatccaagct 240

ctacaagcct ctgaggtgca gtccaaaagg aaagaggcta catgcccgag gcaccaggag 300

atgttccact atttctgcga ggatgatggg aagttcctct gttttgtgtg tcgtgaatcc 360

aaggaccaca aatcccataa tgtcagcttg atcgaagaag ctgcccagaa ttatcagggg 420

cagattcaag agcagatcca agtcttgcag caaaaggaga aggagacagt acaagtgaag 480

gcacaaggtg tacacagggt cgatgtcttc acggaccagg tagaacatga gaagcaaagg 540

atcctcacag aatttgaact cctgcatcaa gtcctagagg aggagaagaa tttcctgcta 600

tcacggattt actggctggg tcatgaggga acggaagcgg ggaaacacta tgttgcctcc 660

actgagccac agttgaacga tctcaagaag ctcgttgatt ccctgaagac caagcagaac 720

atgccaccca ggcagctgct ggaggatatc aaagtcgtct tgtgcagaag tgaagagttt 780

cagtttctca acccaacccc tgttcctctg gaactggaga aaaaactcag tgaagcaaaa 840

tcaagacatg actccatcac agggagccta aaaaaattca aagaccaact ccaggctgat 900

aggaaaaaag atgaaaacag attcttcaaa agcatgaata aaaatgacat gaagagctgg 960

ggcttgttac agaaaaataa tcataaaatg aacaaaacct cagagcccgg gtcatcttct 1020

gcaggcggca gaactacatc ggggccacca aatcaccact cttcagcccc atcccactcc 1080

ctgtttcggg cctcgtctgc tgggaaagtc acttttccag tatgtctcct ggcctcttat 1140

gatgagattt ctggtcaagg agcgagctct caggatacga agacatttga cgttgcgctg 1200

tccgaggagc tccatgcggc actgagtgag tggctgacag cgatccgggc ttggttttgt 1260

gaggttcctt caagctaa 1278

<210> 62

<211> 425

<212> PRT

<213> 智人(Homo sapiens)

<400> 62

Met Ala Ser Gly Gln Phe Val Asn Lys Leu Gln Glu Glu Val Ile Cys

1 5 10 15

Pro Ile Cys Leu Asp Ile Leu Gln Lys Pro Val Thr Ile Asp Cys Gly

20 25 30

His Asn Phe Cys Leu Lys Cys Ile Thr Gln Ile Gly Glu Thr Ser Cys

35 40 45

Gly Phe Phe Lys Cys Pro Leu Cys Lys Thr Ser Val Arg Lys Asn Ala

50 55 60

Ile Arg Phe Asn Ser Leu Leu Arg Asn Leu Val Glu Lys Ile Gln Ala

65 70 75 80

Leu Gln Ala Ser Glu Val Gln Ser Lys Arg Lys Glu Ala Thr Cys Pro

85 90 95

Arg His Gln Glu Met Phe His Tyr Phe Cys Glu Asp Asp Gly Lys Phe

100 105 110

Leu Cys Phe Val Cys Arg Glu Ser Lys Asp His Lys Ser His Asn Val

115 120 125

Ser Leu Ile Glu Glu Ala Ala Gln Asn Tyr Gln Gly Gln Ile Gln Glu

130 135 140

Gln Ile Gln Val Leu Gln Gln Lys Glu Lys Glu Thr Val Gln Val Lys

145 150 155 160

Ala Gln Gly Val His Arg Val Asp Val Phe Thr Asp Gln Val Glu His

165 170 175

Glu Lys Gln Arg Ile Leu Thr Glu Phe Glu Leu Leu His Gln Val Leu

180 185 190

Glu Glu Glu Lys Asn Phe Leu Leu Ser Arg Ile Tyr Trp Leu Gly His

195 200 205

Glu Gly Thr Glu Ala Gly Lys His Tyr Val Ala Ser Thr Glu Pro Gln

210 215 220

Leu Asn Asp Leu Lys Lys Leu Val Asp Ser Leu Lys Thr Lys Gln Asn

225 230 235 240

Met Pro Pro Arg Gln Leu Leu Glu Asp Ile Lys Val Val Leu Cys Arg

245 250 255

Ser Glu Glu Phe Gln Phe Leu Asn Pro Thr Pro Val Pro Leu Glu Leu

260 265 270

Glu Lys Lys Leu Ser Glu Ala Lys Ser Arg His Asp Ser Ile Thr Gly

275 280 285

Ser Leu Lys Lys Phe Lys Asp Gln Leu Gln Ala Asp Arg Lys Lys Asp

290 295 300

Glu Asn Arg Phe Phe Lys Ser Met Asn Lys Asn Asp Met Lys Ser Trp

305 310 315 320

Gly Leu Leu Gln Lys Asn Asn His Lys Met Asn Lys Thr Ser Glu Pro

325 330 335

Gly Ser Ser Ser Ala Gly Gly Arg Thr Thr Ser Gly Pro Pro Asn His

340 345 350

His Ser Ser Ala Pro Ser His Ser Leu Phe Arg Ala Ser Ser Ala Gly

355 360 365

Lys Val Thr Phe Pro Val Cys Leu Leu Ala Ser Tyr Asp Glu Ile Ser

370 375 380

Gly Gln Gly Ala Ser Ser Gln Asp Thr Lys Thr Phe Asp Val Ala Leu

385 390 395 400

Ser Glu Glu Leu His Ala Ala Leu Ser Glu Trp Leu Thr Ala Ile Arg

405 410 415

Ala Trp Phe Cys Glu Val Pro Ser Ser

420 425

<210> 63

<211> 1962

<212> DNA

<213> 智人(Homo sapiens)

<400> 63

atggctgcag cagcagcttc tcacctgaac ctggatgccc tccgggaagt gctagaatgc 60

cccatctgca tggagtcctt cacagaagag cagctgcgtc ccaagcttct gcactgtggc 120

cataccatct gccgccagtg cctggagaag ctattggcca gtagcatcaa tggtgtccgc 180

tgtccctttt gcagcaagat tacccgcata accagcttga cccagctgac agacaatctg 240

acagtgctaa agatcattga tacagctggg ctcagcgagg ctgtggggct gctcatgtgt 300

cggtcctgtg ggcggcgtct gccccggcaa ttctgccgga gctgtggttt ggtgttatgt 360

gagccctgcc gggaggcaga ccatcagcct cctggccact gtacactccc tgtcaaagaa 420

gcagctgagg agcggcgtcg ggactttgga gagaagttaa ctcgtctgcg ggaacttatg 480

ggggagctgc agcggcggaa ggcagccttg gaaggtgtct ccaaggacct tcaggcaagg 540

tataaagcag ttctccagga gtatgggcat gaggagcgca gggtccagga tgagctggct 600

cgctctcgga agttcttcac aggctctttg gctgaagttg agaagtccaa tagtcaagtg 660

gtagaggagc agagttacct gcttaacatt gcagaggtgc aggctgtgtc tcgctgtgac 720

tacttcctgg ccaagatcaa gcaggcagat gtagcactac tggaggagac agctgatgag 780

gaggagccag agctcactgc cagcttgcct cgggagctca ccctgcaaga tgtggagctc 840

cttaaggtag gtcatgttgg ccccctccaa attggacaag ctgttaagaa gccccggaca 900

gttaacgtgg aagattcctg ggccatggag gccacagcgt ctgctgcctc tacctctgtt 960

acttttagag agatggacat gagcccggag gaagtggttg ccagccctag ggcctcacct 1020

gctaaacagc ggggtcctga ggcagcctcc aatatccagc agtgcctctt tctcaagaag 1080

atgggggcca aaggcagcac tccaggaatg ttcaatcttc cagtcagtct ctacgtgacc 1140

agtcaaggtg aagtactagt cgctgaccgt ggtaactatc gtatacaagt ctttacccgc 1200

aaaggctttt tgaaggaaat ccgccgcagc cccagtggca ttgatagctt tgtgctaagc 1260

ttccttgggg cagatctacc caacctcact cctctctcag tggcaatgaa ctgccagggg 1320

ctgattggtg tgactgacag ctatgataac tccctcaagg tatatacctt ggatggccac 1380

tgcgtggcct gtcacaggag ccagctgagc aaaccatggg gtatcacagc cttgccatct 1440

ggccagtttg tagtaaccga tgtggaaggt ggaaagcttt ggtgtttcac agttgatcga 1500

ggatcagggg tggtcaaata cagctgccta tgtagtgctg tgcggcccaa atttgtcacc 1560

tgtgatgctg agggcaccgt ctacttcacc cagggcttag gcctcaatct ggagaatcgg 1620

cagaatgagc accacctgga gggtggcttt tccattggct ctgtaggccc tgatgggcag 1680

ctgggtcgcc agattagcca cttcttctcg gagaatgagg atttccgctg cattgctggc 1740

atgtgtgtgg atgctcgtgg tgatctcatc gtggctgaca gtagtcgcaa ggaaattctc 1800

cattttccta agggtggggg ctatagtgtc cttattcgag agggacttac ctgtccggtg 1860

ggcatagccc taactcctaa ggggcagctg ctggtcttgg actgttggga tcattgcatc 1920

aagatctaca gctaccatct gagaagatat tccaccccat ag 1962

<210> 64

<211> 653

<212> PRT

<213> 智人(Homo sapiens)

<400> 64

Met Ala Ala Ala Ala Ala Ser His Leu Asn Leu Asp Ala Leu Arg Glu

1 5 10 15

Val Leu Glu Cys Pro Ile Cys Met Glu Ser Phe Thr Glu Glu Gln Leu

20 25 30

Arg Pro Lys Leu Leu His Cys Gly His Thr Ile Cys Arg Gln Cys Leu

35 40 45

Glu Lys Leu Leu Ala Ser Ser Ile Asn Gly Val Arg Cys Pro Phe Cys

50 55 60

Ser Lys Ile Thr Arg Ile Thr Ser Leu Thr Gln Leu Thr Asp Asn Leu

65 70 75 80

Thr Val Leu Lys Ile Ile Asp Thr Ala Gly Leu Ser Glu Ala Val Gly

85 90 95

Leu Leu Met Cys Arg Ser Cys Gly Arg Arg Leu Pro Arg Gln Phe Cys

100 105 110

Arg Ser Cys Gly Leu Val Leu Cys Glu Pro Cys Arg Glu Ala Asp His

115 120 125

Gln Pro Pro Gly His Cys Thr Leu Pro Val Lys Glu Ala Ala Glu Glu

130 135 140

Arg Arg Arg Asp Phe Gly Glu Lys Leu Thr Arg Leu Arg Glu Leu Met

145 150 155 160

Gly Glu Leu Gln Arg Arg Lys Ala Ala Leu Glu Gly Val Ser Lys Asp

165 170 175

Leu Gln Ala Arg Tyr Lys Ala Val Leu Gln Glu Tyr Gly His Glu Glu

180 185 190

Arg Arg Val Gln Asp Glu Leu Ala Arg Ser Arg Lys Phe Phe Thr Gly

195 200 205

Ser Leu Ala Glu Val Glu Lys Ser Asn Ser Gln Val Val Glu Glu Gln

210 215 220

Ser Tyr Leu Leu Asn Ile Ala Glu Val Gln Ala Val Ser Arg Cys Asp

225 230 235 240

Tyr Phe Leu Ala Lys Ile Lys Gln Ala Asp Val Ala Leu Leu Glu Glu

245 250 255

Thr Ala Asp Glu Glu Glu Pro Glu Leu Thr Ala Ser Leu Pro Arg Glu

260 265 270

Leu Thr Leu Gln Asp Val Glu Leu Leu Lys Val Gly His Val Gly Pro

275 280 285

Leu Gln Ile Gly Gln Ala Val Lys Lys Pro Arg Thr Val Asn Val Glu

290 295 300

Asp Ser Trp Ala Met Glu Ala Thr Ala Ser Ala Ala Ser Thr Ser Val

305 310 315 320

Thr Phe Arg Glu Met Asp Met Ser Pro Glu Glu Val Val Ala Ser Pro

325 330 335

Arg Ala Ser Pro Ala Lys Gln Arg Gly Pro Glu Ala Ala Ser Asn Ile

340 345 350

Gln Gln Cys Leu Phe Leu Lys Lys Met Gly Ala Lys Gly Ser Thr Pro

355 360 365

Gly Met Phe Asn Leu Pro Val Ser Leu Tyr Val Thr Ser Gln Gly Glu

370 375 380

Val Leu Val Ala Asp Arg Gly Asn Tyr Arg Ile Gln Val Phe Thr Arg

385 390 395 400

Lys Gly Phe Leu Lys Glu Ile Arg Arg Ser Pro Ser Gly Ile Asp Ser

405 410 415

Phe Val Leu Ser Phe Leu Gly Ala Asp Leu Pro Asn Leu Thr Pro Leu

420 425 430

Ser Val Ala Met Asn Cys Gln Gly Leu Ile Gly Val Thr Asp Ser Tyr

435 440 445

Asp Asn Ser Leu Lys Val Tyr Thr Leu Asp Gly His Cys Val Ala Cys

450 455 460

His Arg Ser Gln Leu Ser Lys Pro Trp Gly Ile Thr Ala Leu Pro Ser

465 470 475 480

Gly Gln Phe Val Val Thr Asp Val Glu Gly Gly Lys Leu Trp Cys Phe

485 490 495

Thr Val Asp Arg Gly Ser Gly Val Val Lys Tyr Ser Cys Leu Cys Ser

500 505 510

Ala Val Arg Pro Lys Phe Val Thr Cys Asp Ala Glu Gly Thr Val Tyr

515 520 525

Phe Thr Gln Gly Leu Gly Leu Asn Leu Glu Asn Arg Gln Asn Glu His

530 535 540

His Leu Glu Gly Gly Phe Ser Ile Gly Ser Val Gly Pro Asp Gly Gln

545 550 555 560

Leu Gly Arg Gln Ile Ser His Phe Phe Ser Glu Asn Glu Asp Phe Arg

565 570 575

Cys Ile Ala Gly Met Cys Val Asp Ala Arg Gly Asp Leu Ile Val Ala

580 585 590

Asp Ser Ser Arg Lys Glu Ile Leu His Phe Pro Lys Gly Gly Gly Tyr

595 600 605

Ser Val Leu Ile Arg Glu Gly Leu Thr Cys Pro Val Gly Ile Ala Leu

610 615 620

Thr Pro Lys Gly Gln Leu Leu Val Leu Asp Cys Trp Asp His Cys Ile

625 630 635 640

Lys Ile Tyr Ser Tyr His Leu Arg Arg Tyr Ser Thr Pro

645 650

<210> 65

<211> 3384

<212> DNA

<213> 智人(Homo sapiens)

<400> 65

atggcggaaa acaaaggcgg cggcgaggct gagagcggcg gcgggggcag cggcagcgcg 60

ccggtaactg ccggggccgc cgggcccgcc gcgcaggagg cggagccgcc tctcaccgcg 120

gtgctggtgg aggaggagga ggaggaaggc ggcagggccg gcgctgaggg cggcgcggcc 180

gggcccgacg acgggggggt ggccgcggcc tcctcgggct cggcccaggc tgcttcatct 240

cctgcggcct cagtgggcac tggagttgcc gggggcgcag tatcgacgcc ggctccagct 300

ccagcctcgg ctcccgctcc gggtccctcg gcagggccgc ctcctggacc gccagcctcg 360

ctcctggaca cctgcgccgt gtgtcagcag agcttgcaga gccggcgtga ggcggagccc 420

aagctgctgc cctgtcttca ctccttctgc ctgcgctgcc tgcccgagcc ggagcgccag 480

ctcagcgtgc ccatcccggg gggcagcaac ggcgacatcc agcaagttgg tgtaatacgg 540

tgcccagtat gccgccaaga atgcagacag atagaccttg tggataatta ttttgtgaaa 600

gacacatctg aagctcctag cagttctgat gaaaaatcag aacaggtatg tactagttgt 660

gaagacaatg caagtgcagt tggcttttgt gtagaatgtg gagagtggct atgtaagaca 720

tgtatcgaag cacatcaaag agtaaaattt actaaagatc acttgatcag gaagaaagaa 780

gatgtctcag agtctgttgg agcatctggt caacgccctg ttttctgccc tgtacacaaa 840

caagaacagt tgaaactttt ctgtgaaaca tgtgatagat tgacatgtag agactgtcag 900

ctattggaac acaaagaaca taggtatcag tttttggaag aagcttttca aaatcagaag 960

ggtgcaattg agaatctact ggcgaaactt cttgagaaga agaattatgt tcattttgca 1020

gctactcagg tgcagaatag gataaaagaa gtaaatgaga ctaacaaacg agtagaacag 1080

gaaattaaag tggccatttt cacccttatc aatgaaatta ataagaaagg aaaatctctc 1140

ttacaacagc tagagaatgt tacaaaggaa agacagatga agttactaca gcagcagaat 1200

gacatcacag gcctttcccg gcaggtgaag catgttatga acttcacaaa ttgggcaatt 1260

gcaagtggca gcagcacagc actactatac agcaagcgac tgattacttt ccagttgcgt 1320

catattttga aagcacggtg tgatcctgtc cctgctgcta atggagcaat acgtttccat 1380

tgtgatccca ccttctgggc aaagaatgta gtcaatttag gtaatctagt aatagagagt 1440

aaaccagctc ctggttatac tcctaatgtt gtagttgggc aagttcctcc agggacaaac 1500

cacattagta aaacccctgg acagattaac ttagcacagc ttcgactcca gcacatgcaa 1560

caacaagtat atgcacagaa acatcagcag ttgcaacaga tgaggatgca gcaaccacca 1620

gcacctgtac caactacaac aacaacaaca caacagcatc ctagacaagc agcccctcag 1680

atgttacaac aacagcctcc tcgattgatc agtgtgcaaa caatgcaaag aggcaacatg 1740

aactgtggag cttttcaagc ccatcagatg agactggctc agaatgctgc cagaatacca 1800

gggataccca ggcacagcgg ccctcaatat tccatgatgc agccacacct ccaaagacaa 1860

cactcaaacc cagggcatgc tggacccttt cccgtagtat cggtacacaa caccacaatc 1920

aacccaacga gccctactac agcaactatg gcaaatgcaa accgaggtcc caccagccca 1980

tctgttacag caatagagct aatcccctca gttaccaatc cagaaaacct tccatcgctg 2040

ccagatattc cacccataca gttggaagat gctggctcaa gtagtttaga taatctacta 2100

agtagataca tctcaggcag tcacctaccc ccacagccta caagcaccat gaatccttct 2160

ccaggtccct ctgccctttc tccgggatca tcaggtttat ccaattctca cacacctgtg 2220

agacccccaa gtacttctag tactggcagt cgaggcagct gtgggtcatc aggaagaact 2280

gctgagaaga caagtcttag tttcaaatct gatcaggtga aggtcaagca agaacctggg 2340

actgaagatg aaatatgtag cttttcagga ggtgtaaaac aagaaaaaac agaggatggc 2400

aggaggagtg cttgcatgtt gagcagtcct gagagtagct tgacaccacc tctctcaacc 2460

aacctgcatc tagaaagtga attggatgca ttggcaagcc tggaaaacca tgtgaaaatt 2520

gaacctgcag atatgaatga aagctgcaaa cagtcagggc tcagcagcct tgttaatgga 2580

aagtccccaa ttcgaagcct catgcacagg tcggcaagga ttggaggaga tggcaacaat 2640

aaagatgatg acccaaatga agactggtgt gctgtctgcc aaaacggagg agatctcttg 2700

tgctgcgaaa aatgtccaaa ggtctttcat ctaacttgtc atgttccaac actacttagc 2760

tttccaagtg gggactggat atgcacattt tgtagagata ttggaaagcc agaagttgaa 2820

tatgattgtg ataatttgca acatagtaag aaggggaaaa ctgcgcaggg gttaagcccc 2880

gtggaccaaa ggaaatgtga acgtcttctg ctttacctct attgccatga attaagtatt 2940

gaattccagg agcctgttcc tgcttcgata ccaaactact ataaaattat aaagaaacca 3000

atggatttat ccaccgtgaa aaagaagctt cagaaaaaac attcccaaca ctaccaaatc 3060

ccggatgact ttgtggccga tgtccgtttg atcttcaaga actgtgaaag gtttaatgaa 3120

atgatgaaag ttgttcaagt ttatgcagac acacaagaga ttaatttgaa ggctgattca 3180

gaagtagctc aggcagggaa agcagttgca ttgtactttg aagataaact cacagagatc 3240

tactcagaca ggaccttcgc acctttgcca gagtttgagc aggaagagga tgatggtgag 3300

gtaactgagg actctgatga agactttata cagccccgca gaaaacgcct aaagtcagat 3360

gagagaccag tacatataaa gtaa 3384

<210> 66

<211> 1127

<212> PRT

<213> 智人(Homo sapiens)

<400> 66

Met Ala Glu Asn Lys Gly Gly Gly Glu Ala Glu Ser Gly Gly Gly Gly

1 5 10 15

Ser Gly Ser Ala Pro Val Thr Ala Gly Ala Ala Gly Pro Ala Ala Gln

20 25 30

Glu Ala Glu Pro Pro Leu Thr Ala Val Leu Val Glu Glu Glu Glu Glu

35 40 45

Glu Gly Gly Arg Ala Gly Ala Glu Gly Gly Ala Ala Gly Pro Asp Asp

50 55 60

Gly Gly Val Ala Ala Ala Ser Ser Gly Ser Ala Gln Ala Ala Ser Ser

65 70 75 80

Pro Ala Ala Ser Val Gly Thr Gly Val Ala Gly Gly Ala Val Ser Thr

85 90 95

Pro Ala Pro Ala Pro Ala Ser Ala Pro Ala Pro Gly Pro Ser Ala Gly

100 105 110

Pro Pro Pro Gly Pro Pro Ala Ser Leu Leu Asp Thr Cys Ala Val Cys

115 120 125

Gln Gln Ser Leu Gln Ser Arg Arg Glu Ala Glu Pro Lys Leu Leu Pro

130 135 140

Cys Leu His Ser Phe Cys Leu Arg Cys Leu Pro Glu Pro Glu Arg Gln

145 150 155 160

Leu Ser Val Pro Ile Pro Gly Gly Ser Asn Gly Asp Ile Gln Gln Val

165 170 175

Gly Val Ile Arg Cys Pro Val Cys Arg Gln Glu Cys Arg Gln Ile Asp

180 185 190

Leu Val Asp Asn Tyr Phe Val Lys Asp Thr Ser Glu Ala Pro Ser Ser

195 200 205

Ser Asp Glu Lys Ser Glu Gln Val Cys Thr Ser Cys Glu Asp Asn Ala

210 215 220

Ser Ala Val Gly Phe Cys Val Glu Cys Gly Glu Trp Leu Cys Lys Thr

225 230 235 240

Cys Ile Glu Ala His Gln Arg Val Lys Phe Thr Lys Asp His Leu Ile

245 250 255

Arg Lys Lys Glu Asp Val Ser Glu Ser Val Gly Ala Ser Gly Gln Arg

260 265 270

Pro Val Phe Cys Pro Val His Lys Gln Glu Gln Leu Lys Leu Phe Cys

275 280 285

Glu Thr Cys Asp Arg Leu Thr Cys Arg Asp Cys Gln Leu Leu Glu His

290 295 300

Lys Glu His Arg Tyr Gln Phe Leu Glu Glu Ala Phe Gln Asn Gln Lys

305 310 315 320

Gly Ala Ile Glu Asn Leu Leu Ala Lys Leu Leu Glu Lys Lys Asn Tyr

325 330 335

Val His Phe Ala Ala Thr Gln Val Gln Asn Arg Ile Lys Glu Val Asn

340 345 350

Glu Thr Asn Lys Arg Val Glu Gln Glu Ile Lys Val Ala Ile Phe Thr

355 360 365

Leu Ile Asn Glu Ile Asn Lys Lys Gly Lys Ser Leu Leu Gln Gln Leu

370 375 380

Glu Asn Val Thr Lys Glu Arg Gln Met Lys Leu Leu Gln Gln Gln Asn

385 390 395 400

Asp Ile Thr Gly Leu Ser Arg Gln Val Lys His Val Met Asn Phe Thr

405 410 415

Asn Trp Ala Ile Ala Ser Gly Ser Ser Thr Ala Leu Leu Tyr Ser Lys

420 425 430

Arg Leu Ile Thr Phe Gln Leu Arg His Ile Leu Lys Ala Arg Cys Asp

435 440 445

Pro Val Pro Ala Ala Asn Gly Ala Ile Arg Phe His Cys Asp Pro Thr

450 455 460

Phe Trp Ala Lys Asn Val Val Asn Leu Gly Asn Leu Val Ile Glu Ser

465 470 475 480

Lys Pro Ala Pro Gly Tyr Thr Pro Asn Val Val Val Gly Gln Val Pro

485 490 495

Pro Gly Thr Asn His Ile Ser Lys Thr Pro Gly Gln Ile Asn Leu Ala

500 505 510

Gln Leu Arg Leu Gln His Met Gln Gln Gln Val Tyr Ala Gln Lys His

515 520 525

Gln Gln Leu Gln Gln Met Arg Met Gln Gln Pro Pro Ala Pro Val Pro

530 535 540

Thr Thr Thr Thr Thr Thr Gln Gln His Pro Arg Gln Ala Ala Pro Gln

545 550 555 560

Met Leu Gln Gln Gln Pro Pro Arg Leu Ile Ser Val Gln Thr Met Gln

565 570 575

Arg Gly Asn Met Asn Cys Gly Ala Phe Gln Ala His Gln Met Arg Leu

580 585 590

Ala Gln Asn Ala Ala Arg Ile Pro Gly Ile Pro Arg His Ser Gly Pro

595 600 605

Gln Tyr Ser Met Met Gln Pro His Leu Gln Arg Gln His Ser Asn Pro

610 615 620

Gly His Ala Gly Pro Phe Pro Val Val Ser Val His Asn Thr Thr Ile

625 630 635 640

Asn Pro Thr Ser Pro Thr Thr Ala Thr Met Ala Asn Ala Asn Arg Gly

645 650 655

Pro Thr Ser Pro Ser Val Thr Ala Ile Glu Leu Ile Pro Ser Val Thr

660 665 670

Asn Pro Glu Asn Leu Pro Ser Leu Pro Asp Ile Pro Pro Ile Gln Leu

675 680 685

Glu Asp Ala Gly Ser Ser Ser Leu Asp Asn Leu Leu Ser Arg Tyr Ile

690 695 700

Ser Gly Ser His Leu Pro Pro Gln Pro Thr Ser Thr Met Asn Pro Ser

705 710 715 720

Pro Gly Pro Ser Ala Leu Ser Pro Gly Ser Ser Gly Leu Ser Asn Ser

725 730 735

His Thr Pro Val Arg Pro Pro Ser Thr Ser Ser Thr Gly Ser Arg Gly

740 745 750

Ser Cys Gly Ser Ser Gly Arg Thr Ala Glu Lys Thr Ser Leu Ser Phe

755 760 765

Lys Ser Asp Gln Val Lys Val Lys Gln Glu Pro Gly Thr Glu Asp Glu

770 775 780

Ile Cys Ser Phe Ser Gly Gly Val Lys Gln Glu Lys Thr Glu Asp Gly

785 790 795 800

Arg Arg Ser Ala Cys Met Leu Ser Ser Pro Glu Ser Ser Leu Thr Pro

805 810 815

Pro Leu Ser Thr Asn Leu His Leu Glu Ser Glu Leu Asp Ala Leu Ala

820 825 830

Ser Leu Glu Asn His Val Lys Ile Glu Pro Ala Asp Met Asn Glu Ser

835 840 845

Cys Lys Gln Ser Gly Leu Ser Ser Leu Val Asn Gly Lys Ser Pro Ile

850 855 860

Arg Ser Leu Met His Arg Ser Ala Arg Ile Gly Gly Asp Gly Asn Asn

865 870 875 880

Lys Asp Asp Asp Pro Asn Glu Asp Trp Cys Ala Val Cys Gln Asn Gly

885 890 895

Gly Asp Leu Leu Cys Cys Glu Lys Cys Pro Lys Val Phe His Leu Thr

900 905 910

Cys His Val Pro Thr Leu Leu Ser Phe Pro Ser Gly Asp Trp Ile Cys

915 920 925

Thr Phe Cys Arg Asp Ile Gly Lys Pro Glu Val Glu Tyr Asp Cys Asp

930 935 940

Asn Leu Gln His Ser Lys Lys Gly Lys Thr Ala Gln Gly Leu Ser Pro

945 950 955 960

Val Asp Gln Arg Lys Cys Glu Arg Leu Leu Leu Tyr Leu Tyr Cys His

965 970 975

Glu Leu Ser Ile Glu Phe Gln Glu Pro Val Pro Ala Ser Ile Pro Asn

980 985 990

Tyr Tyr Lys Ile Ile Lys Lys Pro Met Asp Leu Ser Thr Val Lys Lys

995 1000 1005

Lys Leu Gln Lys Lys His Ser Gln His Tyr Gln Ile Pro Asp Asp

1010 1015 1020

Phe Val Ala Asp Val Arg Leu Ile Phe Lys Asn Cys Glu Arg Phe

1025 1030 1035

Asn Glu Met Met Lys Val Val Gln Val Tyr Ala Asp Thr Gln Glu

1040 1045 1050

Ile Asn Leu Lys Ala Asp Ser Glu Val Ala Gln Ala Gly Lys Ala

1055 1060 1065

Val Ala Leu Tyr Phe Glu Asp Lys Leu Thr Glu Ile Tyr Ser Asp

1070 1075 1080

Arg Thr Phe Ala Pro Leu Pro Glu Phe Glu Gln Glu Glu Asp Asp

1085 1090 1095

Gly Glu Val Thr Glu Asp Ser Asp Glu Asp Phe Ile Gln Pro Arg

1100 1105 1110

Arg Lys Arg Leu Lys Ser Asp Glu Arg Pro Val His Ile Lys

1115 1120 1125

<210> 67

<211> 1467

<212> DNA

<213> 智人(Homo sapiens)

<400> 67

atggcttcaa aaatcttgct taacgtacaa gaggaggtga cctgtcccat ctgcctggag 60

ctgttgacag aacccttgag tctagactgt ggccacagcc tctgccgagc ctgcatcact 120

gtgagcaaca aggaggcagt gaccagcatg ggaggaaaaa gcagctgtcc tgtgtgtggt 180

atcagttact catttgaaca tctacaggct aatcagcatc tggccaacat agtggagaga 240

ctcaaggagg tcaagttgag cccagacaat gggaagaaga gagatctctg tgatcatcat 300

ggagagaaac tcctactctt ctgtaaggag gataggaaag tcatttgctg gctttgtgag 360

cggtctcagg agcaccgtgg tcaccacaca gtcctcacgg aggaagtatt caaggaatgt 420

caggagaaac tccaggcagt cctcaagagg ctgaagaagg aagaggagga agctgagaag 480

ctggaagctg acatcagaga agagaaaact tcctggaagt atcaggtaca aactgagaga 540

caaaggatac aaacagaatt tgatcagctt agaagcatcc taaataatga ggagcagaga 600

gagctgcaaa gattggaaga agaagaaaag aagacgctgg ataagtttgc agaggctgag 660

gatgagctag ttcagcagaa gcagttggtg agagagctca tctcagatgt ggagtgtcgg 720

agtcagtggt caacaatgga gctgctgcag gacatgagtg gaatcatgaa atggagtgag 780

atctggaggc tgaaaaagcc aaaaatggtt tccaagaaac tgaagactgt attccatgct 840

ccagatctga gtaggatgct gcaaatgttt agagaactga cagctgtccg gtgctactgg 900

gtggatgtca cactgaattc agtcaaccta aatttgaatc ttgtcctttc agaagatcag 960

agacaagtga tatctgtgcc aatttggcct tttcagtgtt ataattatgg tgtcttggga 1020

tcccaatatt tctcctctgg gaaacattac tgggaagtgg acgtgtccaa gaaaactgcc 1080

tggatcctgg gggtatactg tagaacatat tcccgccata tgaagtatgt tgttagaaga 1140

tgtgcaaatc gtcaaaatct ttacaccaaa tacagacctc tatttggcta ctgggttata 1200

gggttacaga ataaatgtaa gtatggtgtc tttgaagagt ctttgtcctc tgatcccgag 1260

gttttgactc tctccatggc tgtgcctccc tgccgtgttg gggttttcct cgactatgaa 1320

gcaggcattg tctcattttt caatgtcaca agccatggct ccctcattta caagttctct 1380

aaatgttgct tttctcagcc tgtttatcca tatttcaatc cttggaactg tccagctccc 1440

atgactctat gcccaccaag ctcttga 1467

<210> 68

<211> 488

<212> PRT

<213> 智人(Homo sapiens)

<400> 68

Met Ala Ser Lys Ile Leu Leu Asn Val Gln Glu Glu Val Thr Cys Pro

1 5 10 15

Ile Cys Leu Glu Leu Leu Thr Glu Pro Leu Ser Leu Asp Cys Gly His

20 25 30

Ser Leu Cys Arg Ala Cys Ile Thr Val Ser Asn Lys Glu Ala Val Thr

35 40 45

Ser Met Gly Gly Lys Ser Ser Cys Pro Val Cys Gly Ile Ser Tyr Ser

50 55 60

Phe Glu His Leu Gln Ala Asn Gln His Leu Ala Asn Ile Val Glu Arg

65 70 75 80

Leu Lys Glu Val Lys Leu Ser Pro Asp Asn Gly Lys Lys Arg Asp Leu

85 90 95

Cys Asp His His Gly Glu Lys Leu Leu Leu Phe Cys Lys Glu Asp Arg

100 105 110

Lys Val Ile Cys Trp Leu Cys Glu Arg Ser Gln Glu His Arg Gly His

115 120 125

His Thr Val Leu Thr Glu Glu Val Phe Lys Glu Cys Gln Glu Lys Leu

130 135 140

Gln Ala Val Leu Lys Arg Leu Lys Lys Glu Glu Glu Glu Ala Glu Lys

145 150 155 160

Leu Glu Ala Asp Ile Arg Glu Glu Lys Thr Ser Trp Lys Tyr Gln Val

165 170 175

Gln Thr Glu Arg Gln Arg Ile Gln Thr Glu Phe Asp Gln Leu Arg Ser

180 185 190

Ile Leu Asn Asn Glu Glu Gln Arg Glu Leu Gln Arg Leu Glu Glu Glu

195 200 205

Glu Lys Lys Thr Leu Asp Lys Phe Ala Glu Ala Glu Asp Glu Leu Val

210 215 220

Gln Gln Lys Gln Leu Val Arg Glu Leu Ile Ser Asp Val Glu Cys Arg

225 230 235 240

Ser Gln Trp Ser Thr Met Glu Leu Leu Gln Asp Met Ser Gly Ile Met

245 250 255

Lys Trp Ser Glu Ile Trp Arg Leu Lys Lys Pro Lys Met Val Ser Lys

260 265 270

Lys Leu Lys Thr Val Phe His Ala Pro Asp Leu Ser Arg Met Leu Gln

275 280 285

Met Phe Arg Glu Leu Thr Ala Val Arg Cys Tyr Trp Val Asp Val Thr

290 295 300

Leu Asn Ser Val Asn Leu Asn Leu Asn Leu Val Leu Ser Glu Asp Gln

305 310 315 320

Arg Gln Val Ile Ser Val Pro Ile Trp Pro Phe Gln Cys Tyr Asn Tyr

325 330 335

Gly Val Leu Gly Ser Gln Tyr Phe Ser Ser Gly Lys His Tyr Trp Glu

340 345 350

Val Asp Val Ser Lys Lys Thr Ala Trp Ile Leu Gly Val Tyr Cys Arg

355 360 365

Thr Tyr Ser Arg His Met Lys Tyr Val Val Arg Arg Cys Ala Asn Arg

370 375 380

Gln Asn Leu Tyr Thr Lys Tyr Arg Pro Leu Phe Gly Tyr Trp Val Ile

385 390 395 400

Gly Leu Gln Asn Lys Cys Lys Tyr Gly Val Phe Glu Glu Ser Leu Ser

405 410 415

Ser Asp Pro Glu Val Leu Thr Leu Ser Met Ala Val Pro Pro Cys Arg

420 425 430

Val Gly Val Phe Leu Asp Tyr Glu Ala Gly Ile Val Ser Phe Phe Asn

435 440 445

Val Thr Ser His Gly Ser Leu Ile Tyr Lys Phe Ser Lys Cys Cys Phe

450 455 460

Ser Gln Pro Val Tyr Pro Tyr Phe Asn Pro Trp Asn Cys Pro Ala Pro

465 470 475 480

Met Thr Leu Cys Pro Pro Ser Ser

485

<210> 69

<211> 1482

<212> DNA

<213> 智人(Homo sapiens)

<400> 69

atggagcgga gtcccgacgt gtcccccggg ccttcccgct ccttcaagga ggagttgctc 60

tgcgccgtct gctacgaccc cttccgcgac gcagtcactc tgcgctgcgg ccacaacttc 120

tgccgcgggt gcgtgagccg ctgctgggag gtgcaggtgt cgcccacctg cccagtgtgc 180

aaagaccgcg cgtcacccgc cgacctgcgc accaaccaca ccctcaacaa cctggtggag 240

aagctgctgc gcgaggaggc cgagggcgcg cgctggacca gctaccgctt ctcgcgtgtc 300

tgccgcctgc accgcggaca gctcagcctc ttctgcctcg aggacaagga gctgctgtgc 360

tgctcctgcc aggccgaccc ccgacaccag gggcaccgcg tgcagccggt gaaggacact 420

gcccacgact ttcgggccaa gtgcaggaac atggagcatg cactgcggga gaaggccaag 480

gccttctggg ccatgcggcg ctcctatgag gccatcgcca agcacaatca ggtggaggct 540

gcatggctgg aaggccggat ccggcaggag tttgataagc ttcgcgagtt cttgagagtg 600

gaggagcagg ccattctgga tgccatggcc gaggagacaa ggcagaagca acttctggcc 660

gacgagaaga tgaagcagct cacagaggag acggaggtgc tggcacatga gatcgagcgg 720

ctgcagatgg agatgaagga ggacgacgtt tcttttctca tgaaacacaa gagccgaaaa 780

cgccgactct tctgcaccat ggagccagag ccagtccagc ccggcatgct tatcgatgtc 840

tgcaagtacc tgggctccct gcagtaccgc gtctggaaga agatgcttgc atctgtggaa 900

tctgtaccct tcagctttga ccccaacacc gcagctggct ggctctccgt gtctgacgac 960

ctcaccagcg tcaccaacca tggctaccgc gtgcaggtgg agaacccgga acgcttctcc 1020

tcggcgccct gcctgctggg ctcccgtgtc ttctcacagg gctcgcacgc ctgggaggtg 1080

gcccttgggg ggctgcagag ctggagggtg ggcgtggtac gtgtgcgcca ggactcgggc 1140

gctgagggcc actcacacag ctgctaccac gacacacgct cgggcttctg gtatgtctgc 1200

cgcacgcagg gcgtggaggg ggaccactgc gtgacctcgg acccagccac gtcgcccctg 1260

gtcctggcca tcccacgccg cctgcgtgtg gagctggagt gtgaggaggg cgagctgtct 1320

ttctatgacg cggagcgcca ctgccacctg tacaccttcc acgcccgctt tggggaggtt 1380

cgcccctact tctacctggg gggtgcacgg ggcgccgggc ctccagagcc tttgcgcatc 1440

tgccccttgc acatcagtgt caaggaagaa ctggatggct ga 1482

<210> 70

<211> 493

<212> PRT

<213> 智人(Homo sapiens)

<400> 70

Met Glu Arg Ser Pro Asp Val Ser Pro Gly Pro Ser Arg Ser Phe Lys

1 5 10 15

Glu Glu Leu Leu Cys Ala Val Cys Tyr Asp Pro Phe Arg Asp Ala Val

20 25 30

Thr Leu Arg Cys Gly His Asn Phe Cys Arg Gly Cys Val Ser Arg Cys

35 40 45

Trp Glu Val Gln Val Ser Pro Thr Cys Pro Val Cys Lys Asp Arg Ala

50 55 60

Ser Pro Ala Asp Leu Arg Thr Asn His Thr Leu Asn Asn Leu Val Glu

65 70 75 80

Lys Leu Leu Arg Glu Glu Ala Glu Gly Ala Arg Trp Thr Ser Tyr Arg

85 90 95

Phe Ser Arg Val Cys Arg Leu His Arg Gly Gln Leu Ser Leu Phe Cys

100 105 110

Leu Glu Asp Lys Glu Leu Leu Cys Cys Ser Cys Gln Ala Asp Pro Arg

115 120 125

His Gln Gly His Arg Val Gln Pro Val Lys Asp Thr Ala His Asp Phe

130 135 140

Arg Ala Lys Cys Arg Asn Met Glu His Ala Leu Arg Glu Lys Ala Lys

145 150 155 160

Ala Phe Trp Ala Met Arg Arg Ser Tyr Glu Ala Ile Ala Lys His Asn

165 170 175

Gln Val Glu Ala Ala Trp Leu Glu Gly Arg Ile Arg Gln Glu Phe Asp

180 185 190

Lys Leu Arg Glu Phe Leu Arg Val Glu Glu Gln Ala Ile Leu Asp Ala

195 200 205

Met Ala Glu Glu Thr Arg Gln Lys Gln Leu Leu Ala Asp Glu Lys Met

210 215 220

Lys Gln Leu Thr Glu Glu Thr Glu Val Leu Ala His Glu Ile Glu Arg

225 230 235 240

Leu Gln Met Glu Met Lys Glu Asp Asp Val Ser Phe Leu Met Lys His

245 250 255

Lys Ser Arg Lys Arg Arg Leu Phe Cys Thr Met Glu Pro Glu Pro Val

260 265 270

Gln Pro Gly Met Leu Ile Asp Val Cys Lys Tyr Leu Gly Ser Leu Gln

275 280 285

Tyr Arg Val Trp Lys Lys Met Leu Ala Ser Val Glu Ser Val Pro Phe

290 295 300

Ser Phe Asp Pro Asn Thr Ala Ala Gly Trp Leu Ser Val Ser Asp Asp

305 310 315 320

Leu Thr Ser Val Thr Asn His Gly Tyr Arg Val Gln Val Glu Asn Pro

325 330 335

Glu Arg Phe Ser Ser Ala Pro Cys Leu Leu Gly Ser Arg Val Phe Ser

340 345 350

Gln Gly Ser His Ala Trp Glu Val Ala Leu Gly Gly Leu Gln Ser Trp

355 360 365

Arg Val Gly Val Val Arg Val Arg Gln Asp Ser Gly Ala Glu Gly His

370 375 380

Ser His Ser Cys Tyr His Asp Thr Arg Ser Gly Phe Trp Tyr Val Cys

385 390 395 400

Arg Thr Gln Gly Val Glu Gly Asp His Cys Val Thr Ser Asp Pro Ala

405 410 415

Thr Ser Pro Leu Val Leu Ala Ile Pro Arg Arg Leu Arg Val Glu Leu

420 425 430

Glu Cys Glu Glu Gly Glu Leu Ser Phe Tyr Asp Ala Glu Arg His Cys

435 440 445

His Leu Tyr Thr Phe His Ala Arg Phe Gly Glu Val Arg Pro Tyr Phe

450 455 460

Tyr Leu Gly Gly Ala Arg Gly Ala Gly Pro Pro Glu Pro Leu Arg Ile

465 470 475 480

Cys Pro Leu His Ile Ser Val Lys Glu Glu Leu Asp Gly

485 490

<210> 71

<211> 2187

<212> DNA

<213> 智人(Homo sapiens)

<400> 71

atgtcggagt ctggggagat gagtgaattt ggctacatca tggaattgat agctaaaggc 60

aaggttacca ttaagaatat cgaaagggag ctcatttgcc cagcatgcaa ggagctgttt 120

acccacccat tgattctccc ttgccaacat agtatctgtc ataaatgtgt aaaagaactc 180

ctgctgactc tcgatgattc attcaacgat gtgggatcag acaactccaa tcaaagcagt 240

cctcgacttc ggctcccctc ccctagtatg gataaaattg accgaattaa cagaccaggc 300

tggaagcgca attcattgac cccgaggaca actgttttcc cttgccctgg ctgtgagcat 360

gatgtggatc ttggagaacg aggaatcaat ggtctgtttc gaaacttcac tttggaaact 420

attgtggaaa gatatcgtca agcagctagg gcagccacag ccattatgtg tgacctttgt 480

aaaccaccac ctcaagaatc cacaaaaagc tgcatggact gtagtgcaag ttactgcaat 540

gaatgcttca aaattcatca cccttggggt actataaaag ctcaacatga gtatgttggt 600

ccaactacta acttcagacc caagatttta atgtgcccag aacatgaaac agagagaata 660

aacatgtact gtgaattatg taggaggcca gtttgccatc tgtgtaagtt gggtggtaat 720

catgccaacc accgtgtaac cactatgagc agtgcctaca aaaccttaaa ggaaaagctt 780

tcaaaggata ttgattacct tattggtaag gaaagccagg tgaagagtca aatatctgaa 840

ctaaacttgt taatgaaaga aacagagtgt aatggagaga gggctaaaga agaagcaatt 900

acacattttg aaaagctctt tgaagttctg gaagagagga aatcatctgt tttgaaagca 960

attgactcct ctaagaaact aagattagac aaatttcaga ctcaaatgga agagtaccag 1020

ggacttctag agaacaatgg acttgtggga tatgctcaag aagtgctaaa ggagacagat 1080

cagtcttgct ttgtgcagac agcaaagcag ctccacctca gaatacagaa agccacagaa 1140

tctttgaaga gctttagacc tgcagctcag acttcttttg aagactatgt tgttaatacc 1200

tctaaacaaa cagaacttct tggagaatta tcctttttct ctagtggcat agacgtgcca 1260

gagatcaatg aggaacagag caaagtttat aacaatgcct tgataaattg gcaccatcca 1320

gaaaaggata aagctgatag ctatgttctt gaatatcgga aaatcaatag agatgatgaa 1380

atgtcatgga atgagataga agtgtgtgga acaagtaaaa taattcaaga cttggaaaac 1440

agtagtacct atgctttcag agtaagagct tacaagggtt caatctgtag tccttgcagc 1500

agagaattga ttcttcatac tcctccagct ccagttttca gcttcctctt tgatgaaaaa 1560

tgtggctata ataatgaaca cctcctgctg aacttgaaga gagaccgtgt agagagtaga 1620

gctggattta atcttctgct tgctgcagaa cgcatccaag tgggttatta cacaagctta 1680

gactacatca ttggagatac tggcattaca aaaggaaaac acttctgggc cttccgtgtg 1740

gaaccatatt catacctggt aaaagtggga gttgcttcta gcgataaact acaagaatgg 1800

ctccgttctc cccgggatgc agttagtcca agatatgagc aagacagtgg gcatgacagt 1860

ggaagtgagg atgcctgttt tgattcttca caaccattta ccttagttac tataggcatg 1920

cagaaatttt ttatacccaa gtcacctact tcttctaatg aacctgaaaa tagagttctc 1980

cctatgccaa caagtattgg gattttcctt gactgtgata aaggcaaagt agatttctat 2040

gatatggatc agatgaaatg cctttatgaa cgccaagtgg actgttcaca tacactgtat 2100

ccagcatttg cattaatggg cagtggagga attcagcttg aagaacccat cacagcaaaa 2160

tatctggaat accaagagga catgtag 2187

<210> 72

<211> 728

<212> PRT

<213> 智人(Homo sapiens)

<400> 72

Met Ser Glu Ser Gly Glu Met Ser Glu Phe Gly Tyr Ile Met Glu Leu

1 5 10 15

Ile Ala Lys Gly Lys Val Thr Ile Lys Asn Ile Glu Arg Glu Leu Ile

20 25 30

Cys Pro Ala Cys Lys Glu Leu Phe Thr His Pro Leu Ile Leu Pro Cys

35 40 45

Gln His Ser Ile Cys His Lys Cys Val Lys Glu Leu Leu Leu Thr Leu

50 55 60

Asp Asp Ser Phe Asn Asp Val Gly Ser Asp Asn Ser Asn Gln Ser Ser

65 70 75 80

Pro Arg Leu Arg Leu Pro Ser Pro Ser Met Asp Lys Ile Asp Arg Ile

85 90 95

Asn Arg Pro Gly Trp Lys Arg Asn Ser Leu Thr Pro Arg Thr Thr Val

100 105 110

Phe Pro Cys Pro Gly Cys Glu His Asp Val Asp Leu Gly Glu Arg Gly

115 120 125

Ile Asn Gly Leu Phe Arg Asn Phe Thr Leu Glu Thr Ile Val Glu Arg

130 135 140

Tyr Arg Gln Ala Ala Arg Ala Ala Thr Ala Ile Met Cys Asp Leu Cys

145 150 155 160

Lys Pro Pro Pro Gln Glu Ser Thr Lys Ser Cys Met Asp Cys Ser Ala

165 170 175

Ser Tyr Cys Asn Glu Cys Phe Lys Ile His His Pro Trp Gly Thr Ile

180 185 190

Lys Ala Gln His Glu Tyr Val Gly Pro Thr Thr Asn Phe Arg Pro Lys

195 200 205

Ile Leu Met Cys Pro Glu His Glu Thr Glu Arg Ile Asn Met Tyr Cys

210 215 220

Glu Leu Cys Arg Arg Pro Val Cys His Leu Cys Lys Leu Gly Gly Asn

225 230 235 240

His Ala Asn His Arg Val Thr Thr Met Ser Ser Ala Tyr Lys Thr Leu

245 250 255

Lys Glu Lys Leu Ser Lys Asp Ile Asp Tyr Leu Ile Gly Lys Glu Ser

260 265 270

Gln Val Lys Ser Gln Ile Ser Glu Leu Asn Leu Leu Met Lys Glu Thr

275 280 285

Glu Cys Asn Gly Glu Arg Ala Lys Glu Glu Ala Ile Thr His Phe Glu

290 295 300

Lys Leu Phe Glu Val Leu Glu Glu Arg Lys Ser Ser Val Leu Lys Ala

305 310 315 320

Ile Asp Ser Ser Lys Lys Leu Arg Leu Asp Lys Phe Gln Thr Gln Met

325 330 335

Glu Glu Tyr Gln Gly Leu Leu Glu Asn Asn Gly Leu Val Gly Tyr Ala

340 345 350

Gln Glu Val Leu Lys Glu Thr Asp Gln Ser Cys Phe Val Gln Thr Ala

355 360 365

Lys Gln Leu His Leu Arg Ile Gln Lys Ala Thr Glu Ser Leu Lys Ser

370 375 380

Phe Arg Pro Ala Ala Gln Thr Ser Phe Glu Asp Tyr Val Val Asn Thr

385 390 395 400

Ser Lys Gln Thr Glu Leu Leu Gly Glu Leu Ser Phe Phe Ser Ser Gly

405 410 415

Ile Asp Val Pro Glu Ile Asn Glu Glu Gln Ser Lys Val Tyr Asn Asn

420 425 430

Ala Leu Ile Asn Trp His His Pro Glu Lys Asp Lys Ala Asp Ser Tyr

435 440 445

Val Leu Glu Tyr Arg Lys Ile Asn Arg Asp Asp Glu Met Ser Trp Asn

450 455 460

Glu Ile Glu Val Cys Gly Thr Ser Lys Ile Ile Gln Asp Leu Glu Asn

465 470 475 480

Ser Ser Thr Tyr Ala Phe Arg Val Arg Ala Tyr Lys Gly Ser Ile Cys

485 490 495

Ser Pro Cys Ser Arg Glu Leu Ile Leu His Thr Pro Pro Ala Pro Val

500 505 510

Phe Ser Phe Leu Phe Asp Glu Lys Cys Gly Tyr Asn Asn Glu His Leu

515 520 525

Leu Leu Asn Leu Lys Arg Asp Arg Val Glu Ser Arg Ala Gly Phe Asn

530 535 540

Leu Leu Leu Ala Ala Glu Arg Ile Gln Val Gly Tyr Tyr Thr Ser Leu

545 550 555 560

Asp Tyr Ile Ile Gly Asp Thr Gly Ile Thr Lys Gly Lys His Phe Trp

565 570 575

Ala Phe Arg Val Glu Pro Tyr Ser Tyr Leu Val Lys Val Gly Val Ala

580 585 590

Ser Ser Asp Lys Leu Gln Glu Trp Leu Arg Ser Pro Arg Asp Ala Val

595 600 605

Ser Pro Arg Tyr Glu Gln Asp Ser Gly His Asp Ser Gly Ser Glu Asp

610 615 620

Ala Cys Phe Asp Ser Ser Gln Pro Phe Thr Leu Val Thr Ile Gly Met

625 630 635 640

Gln Lys Phe Phe Ile Pro Lys Ser Pro Thr Ser Ser Asn Glu Pro Glu

645 650 655

Asn Arg Val Leu Pro Met Pro Thr Ser Ile Gly Ile Phe Leu Asp Cys

660 665 670

Asp Lys Gly Lys Val Asp Phe Tyr Asp Met Asp Gln Met Lys Cys Leu

675 680 685

Tyr Glu Arg Gln Val Asp Cys Ser His Thr Leu Tyr Pro Ala Phe Ala

690 695 700

Leu Met Gly Ser Gly Gly Ile Gln Leu Glu Glu Pro Ile Thr Ala Lys

705 710 715 720

Tyr Leu Glu Tyr Gln Glu Asp Met

725

<210> 73

<211> 2895

<212> DNA

<213> 智人(Homo sapiens)

<400> 73

atggatgaac agagcgtgga gagcattgct gaggttttcc gatgtttcat ttgtatggag 60

aaattgcggg atgcacgcct gtgtcctcat tgctccaaac tgtgttgttt cagctgtatt 120

aggcgctggc tgacagagca gagagctcaa tgtcctcatt gccgtgctcc actccagcta 180

cgagaactag taaattgtcg ttgggcagaa gaagtaacac aacagcttga tactcttcaa 240

ctctgcagtc tcaccaaaca tgaagaaaat gaaaaggaca aatgtgaaaa tcaccatgaa 300

aaacttagtg tattttgctg gacttgtaag aagtgtatct gccatcagtg tgcactttgg 360

ggaggaatgc atggcggaca tacctttaaa cctttggcag aaatttatga gcaacacgtc 420

actaaagtga atgaagaggt agccaaactt cgtcggcgtc tcatggaact gatcagctta 480

gttcaagaag tggaaaggaa tgtagaagct gtaagaaatg caaaagatga gcgtgttcgg 540

gaaattagga atgcagtgga gatgatgatt gcacggttag acacacagct gaagaataag 600

cttataacac tgatgggtca gaagacatct ctaacccaag aaacagagct tttggaatcc 660

ttacttcagg aggtggagca ccagttgcgg tcttgtagta agagtgagtt gatatctaag 720

agctcagaga tccttatgat gtttcagcaa gttcatcgga agcccatggc atcttttgtt 780

accactcctg ttccaccaga ctttaccagt gaattagtgc catcttacga ttcagctact 840

tttgttttag agaatttcag cactttgcgt cagagagcag atcctgttta cagtccacct 900

cttcaagttt caggactttg ctggaggtta aaagtttacc cagatggaaa tggagttgtg 960

cgaggttact acttatctgt gtttctggag ctctcagctg gcttgcctga aacttctaaa 1020

tatgaatatc gtgtagagat ggttcaccag tcctgtaatg atcctacaaa aaatatcatt 1080

cgagaatttg catctgactt tgaagttgga gaatgctggg gctataatag atttttccgt 1140

ttggacttac tcgcaaatga aggatacttg aatccacaaa atgatacagt gattttaagg 1200

tttcaggtac gttcaccaac tttctttcaa aaatcccggg accagcattg gtacattact 1260

cagttggaag ctgcacagac tagttatatc caacaaataa acaaccttaa agagagactt 1320

actattgagc tgtctcgaac tcagaagtca agagatttgt caccaccaga taaccatctt 1380

agcccccaaa atgatgatgc tctggagaca cgagctaaga agtctgcatg ctctgacatg 1440

cttctcgaag gtggtcctac tacagcttct gtaagagagg ccaaagagga tgaagaagat 1500

gaggagaaga ttcagaatga agattatcat cacgagcttt cagatggaga tctggatctg 1560

gatcttgttt atgaggatga agtaaatcag ctcgatggca gcagttcctc tgctagttcc 1620

acagcaacaa gtaatacaga agaaaatgat attgatgaag aaactatgtc tggagaaaat 1680

gatgtggaat ataacaacat ggaattagaa gagggagaac tcatggaaga tgcagctgct 1740

gcaggacccg caggtagtag ccatggttat gtgggttcca gtagtagaat atcaagaaga 1800

acacatttat gctccgctgc taccagtagt ttactagaca ttgatccatt aattttaata 1860

catttgttgg accttaagga ccggagcagt atagaaaatt tgtggggctt acagcctcgc 1920

ccacctgctt cacttctgca gcccacagca tcatattctc gaaaagataa agaccaaagg 1980

aagcaacagg caatgtggcg agtgccctct gatttaaaga tgctaaaaag actcaaaact 2040

caaatggccg aagttcgatg tatgaaaact gatgtaaaga atacactttc agaaataaaa 2100

agcagcagtg ctgcttctgg agacatgcag acaagccttt tttctgctga ccaggcagct 2160

ctggctgcat gtggaactga aaactctggc agattgcagg atttgggaat ggaactcctg 2220

gcaaagtcat cagttgccaa ttgttacata cgaaactcca caaataagaa gagtaattcg 2280

cccaagccag ctcgatccag tgtagcaggt agtctatcac ttcgaagagc agtggaccct 2340

ggagaaaata gtcgttcaaa gggagactgt cagactctgt ctgaaggctc cccaggaagc 2400

tctcagtctg ggagcaggca cagttctccc cgagccttga tacatggcag tatcggtgat 2460

attctgccaa aaactgaaga ccggcagtgt aaagctttgg attcagatgc tgttgtggtt 2520

gcagttttca gtggcttgcc tgcggttgag aaaaggagga aaatggtcac cttgggggct 2580

aatgctaaag gaggtcatct ggaaggactg cagatgactg atttggaaaa taattctgaa 2640

actggagagt tacagcctgt actacctgaa ggagcttcag ctgcccctga agaaggaatg 2700

agtagcgaca gtgacattga atgtgacact gagaatgagg agcaggaaga gcataccagt 2760

gtgggcgggt ttcacgactc cttcatggtc atgacacagc ccccggatga agatacacat 2820

tccagttttc ctgatggtga acaaataggc cctgaagatc tcagcttcaa tacagatgaa 2880

aatagtggaa ggtaa 2895

<210> 74

<211> 964

<212> PRT

<213> 智人(Homo sapiens)

<400> 74

Met Asp Glu Gln Ser Val Glu Ser Ile Ala Glu Val Phe Arg Cys Phe

1 5 10 15

Ile Cys Met Glu Lys Leu Arg Asp Ala Arg Leu Cys Pro His Cys Ser

20 25 30

Lys Leu Cys Cys Phe Ser Cys Ile Arg Arg Trp Leu Thr Glu Gln Arg

35 40 45

Ala Gln Cys Pro His Cys Arg Ala Pro Leu Gln Leu Arg Glu Leu Val

50 55 60

Asn Cys Arg Trp Ala Glu Glu Val Thr Gln Gln Leu Asp Thr Leu Gln

65 70 75 80

Leu Cys Ser Leu Thr Lys His Glu Glu Asn Glu Lys Asp Lys Cys Glu

85 90 95

Asn His His Glu Lys Leu Ser Val Phe Cys Trp Thr Cys Lys Lys Cys

100 105 110

Ile Cys His Gln Cys Ala Leu Trp Gly Gly Met His Gly Gly His Thr

115 120 125

Phe Lys Pro Leu Ala Glu Ile Tyr Glu Gln His Val Thr Lys Val Asn

130 135 140

Glu Glu Val Ala Lys Leu Arg Arg Arg Leu Met Glu Leu Ile Ser Leu

145 150 155 160

Val Gln Glu Val Glu Arg Asn Val Glu Ala Val Arg Asn Ala Lys Asp

165 170 175

Glu Arg Val Arg Glu Ile Arg Asn Ala Val Glu Met Met Ile Ala Arg

180 185 190

Leu Asp Thr Gln Leu Lys Asn Lys Leu Ile Thr Leu Met Gly Gln Lys

195 200 205

Thr Ser Leu Thr Gln Glu Thr Glu Leu Leu Glu Ser Leu Leu Gln Glu

210 215 220

Val Glu His Gln Leu Arg Ser Cys Ser Lys Ser Glu Leu Ile Ser Lys

225 230 235 240

Ser Ser Glu Ile Leu Met Met Phe Gln Gln Val His Arg Lys Pro Met

245 250 255

Ala Ser Phe Val Thr Thr Pro Val Pro Pro Asp Phe Thr Ser Glu Leu

260 265 270

Val Pro Ser Tyr Asp Ser Ala Thr Phe Val Leu Glu Asn Phe Ser Thr

275 280 285

Leu Arg Gln Arg Ala Asp Pro Val Tyr Ser Pro Pro Leu Gln Val Ser

290 295 300

Gly Leu Cys Trp Arg Leu Lys Val Tyr Pro Asp Gly Asn Gly Val Val

305 310 315 320

Arg Gly Tyr Tyr Leu Ser Val Phe Leu Glu Leu Ser Ala Gly Leu Pro

325 330 335

Glu Thr Ser Lys Tyr Glu Tyr Arg Val Glu Met Val His Gln Ser Cys

340 345 350

Asn Asp Pro Thr Lys Asn Ile Ile Arg Glu Phe Ala Ser Asp Phe Glu

355 360 365

Val Gly Glu Cys Trp Gly Tyr Asn Arg Phe Phe Arg Leu Asp Leu Leu

370 375 380

Ala Asn Glu Gly Tyr Leu Asn Pro Gln Asn Asp Thr Val Ile Leu Arg

385 390 395 400

Phe Gln Val Arg Ser Pro Thr Phe Phe Gln Lys Ser Arg Asp Gln His

405 410 415

Trp Tyr Ile Thr Gln Leu Glu Ala Ala Gln Thr Ser Tyr Ile Gln Gln

420 425 430

Ile Asn Asn Leu Lys Glu Arg Leu Thr Ile Glu Leu Ser Arg Thr Gln

435 440 445

Lys Ser Arg Asp Leu Ser Pro Pro Asp Asn His Leu Ser Pro Gln Asn

450 455 460

Asp Asp Ala Leu Glu Thr Arg Ala Lys Lys Ser Ala Cys Ser Asp Met

465 470 475 480

Leu Leu Glu Gly Gly Pro Thr Thr Ala Ser Val Arg Glu Ala Lys Glu

485 490 495

Asp Glu Glu Asp Glu Glu Lys Ile Gln Asn Glu Asp Tyr His His Glu

500 505 510

Leu Ser Asp Gly Asp Leu Asp Leu Asp Leu Val Tyr Glu Asp Glu Val

515 520 525

Asn Gln Leu Asp Gly Ser Ser Ser Ser Ala Ser Ser Thr Ala Thr Ser

530 535 540

Asn Thr Glu Glu Asn Asp Ile Asp Glu Glu Thr Met Ser Gly Glu Asn

545 550 555 560

Asp Val Glu Tyr Asn Asn Met Glu Leu Glu Glu Gly Glu Leu Met Glu

565 570 575

Asp Ala Ala Ala Ala Gly Pro Ala Gly Ser Ser His Gly Tyr Val Gly

580 585 590

Ser Ser Ser Arg Ile Ser Arg Arg Thr His Leu Cys Ser Ala Ala Thr

595 600 605

Ser Ser Leu Leu Asp Ile Asp Pro Leu Ile Leu Ile His Leu Leu Asp

610 615 620

Leu Lys Asp Arg Ser Ser Ile Glu Asn Leu Trp Gly Leu Gln Pro Arg

625 630 635 640

Pro Pro Ala Ser Leu Leu Gln Pro Thr Ala Ser Tyr Ser Arg Lys Asp

645 650 655

Lys Asp Gln Arg Lys Gln Gln Ala Met Trp Arg Val Pro Ser Asp Leu

660 665 670

Lys Met Leu Lys Arg Leu Lys Thr Gln Met Ala Glu Val Arg Cys Met

675 680 685

Lys Thr Asp Val Lys Asn Thr Leu Ser Glu Ile Lys Ser Ser Ser Ala

690 695 700

Ala Ser Gly Asp Met Gln Thr Ser Leu Phe Ser Ala Asp Gln Ala Ala

705 710 715 720

Leu Ala Ala Cys Gly Thr Glu Asn Ser Gly Arg Leu Gln Asp Leu Gly

725 730 735

Met Glu Leu Leu Ala Lys Ser Ser Val Ala Asn Cys Tyr Ile Arg Asn

740 745 750

Ser Thr Asn Lys Lys Ser Asn Ser Pro Lys Pro Ala Arg Ser Ser Val

755 760 765

Ala Gly Ser Leu Ser Leu Arg Arg Ala Val Asp Pro Gly Glu Asn Ser

770 775 780

Arg Ser Lys Gly Asp Cys Gln Thr Leu Ser Glu Gly Ser Pro Gly Ser

785 790 795 800

Ser Gln Ser Gly Ser Arg His Ser Ser Pro Arg Ala Leu Ile His Gly

805 810 815

Ser Ile Gly Asp Ile Leu Pro Lys Thr Glu Asp Arg Gln Cys Lys Ala

820 825 830

Leu Asp Ser Asp Ala Val Val Val Ala Val Phe Ser Gly Leu Pro Ala

835 840 845

Val Glu Lys Arg Arg Lys Met Val Thr Leu Gly Ala Asn Ala Lys Gly

850 855 860

Gly His Leu Glu Gly Leu Gln Met Thr Asp Leu Glu Asn Asn Ser Glu

865 870 875 880

Thr Gly Glu Leu Gln Pro Val Leu Pro Glu Gly Ala Ser Ala Ala Pro

885 890 895

Glu Glu Gly Met Ser Ser Asp Ser Asp Ile Glu Cys Asp Thr Glu Asn

900 905 910

Glu Glu Gln Glu Glu His Thr Ser Val Gly Gly Phe His Asp Ser Phe

915 920 925

Met Val Met Thr Gln Pro Pro Asp Glu Asp Thr His Ser Ser Phe Pro

930 935 940

Asp Gly Glu Gln Ile Gly Pro Glu Asp Leu Ser Phe Asn Thr Asp Glu

945 950 955 960

Asn Ser Gly Arg

<210> 75

<211> 1398

<212> DNA

<213> 智人(Homo sapiens)

<400> 75

atggcctcaa ccaccagcac caagaagatg atggaggaag ccacctgctc catctgcctg 60

agcctgatga cgaacccagt aagcatcaac tgtggacaca gctactgcca cttgtgtata 120

acagacttct ttaaaaaccc aagccaaaag caactgaggc aggagacatt ctgctgtccc 180

cagtgtcggg ctccatttca tatggatagc ctccgaccca acaagcagct gggaagcctc 240

attgaagccc tcaaagagac ggatcaagaa atgtcatgtg aggaacacgg agagcagttc 300

cacctgttct gcgaagacga ggggcagctc atctgctggc gctgtgagcg ggcaccacag 360

cacaaagggc acaccacagc tcttgttgaa gacgtatgcc agggctacaa ggaaaagctc 420

cagaaagctg tgacaaaact gaagcaactt gaagacagat gtacggagca gaagctgtcc 480

acagcaatgc gaataactaa atggaaagag aaggtacaga ttcagagaca aaaaatccgg 540

tctgacttta agaatctcca gtgtttccta catgaggaag agaagtctta tctctggagg 600

ctggagaaag aagaacaaca gactctgagt agactgaggg actatgaggc tggtctgggg 660

ctgaagagca atgaactcaa gagccacatc ctggaactgg aggaaaaatg tcagggctca 720

gcccagaaat tgctgcagaa tgtgaatgac actttgagca ggagttgggc tgtgaagctg 780

gaaacatcag aggctgtctc cttggaactt catactatgt gcaatgtttc caagctttac 840

ttcgatgtga agaaaatgtt aaggagtcat caagttagtg tgactctgga tccagataca 900

gctcatcacg aactaattct ctctgaggat cggagacaag tgactcgtgg atacacccag 960

gagaatcagg acacatcttc caggagattt actgccttcc cctgtgtctt gggttgtgaa 1020

ggcttcacct caggaagacg ttactttgaa gtggatgttg gcgaaggaac cggatgggat 1080

ttaggagttt gtatggaaaa tgtgcagagg ggcactggca tgaagcaaga gcctcagtct 1140

ggattctgga ccctcaggct gtgcaaaaag aaaggctatg tagcacttac ttctccccca 1200

acttcccttc atctgcatga gcagcccctg cttgtgggaa tttttctgga ctatgaggcc 1260

ggagttgtat ccttttataa cgggaatact ggctgccaca tctttacttt cccgaaggct 1320

tccttctctg atactctccg gccctatttc caggtttatc aatattctcc tttgtttctg 1380

cctcccccag gtgactaa 1398

<210> 76

<211> 465

<212> PRT

<213> 智人(Homo sapiens)

<400> 76

Met Ala Ser Thr Thr Ser Thr Lys Lys Met Met Glu Glu Ala Thr Cys

1 5 10 15

Ser Ile Cys Leu Ser Leu Met Thr Asn Pro Val Ser Ile Asn Cys Gly

20 25 30

His Ser Tyr Cys His Leu Cys Ile Thr Asp Phe Phe Lys Asn Pro Ser

35 40 45

Gln Lys Gln Leu Arg Gln Glu Thr Phe Cys Cys Pro Gln Cys Arg Ala

50 55 60

Pro Phe His Met Asp Ser Leu Arg Pro Asn Lys Gln Leu Gly Ser Leu

65 70 75 80

Ile Glu Ala Leu Lys Glu Thr Asp Gln Glu Met Ser Cys Glu Glu His

85 90 95

Gly Glu Gln Phe His Leu Phe Cys Glu Asp Glu Gly Gln Leu Ile Cys

100 105 110

Trp Arg Cys Glu Arg Ala Pro Gln His Lys Gly His Thr Thr Ala Leu

115 120 125

Val Glu Asp Val Cys Gln Gly Tyr Lys Glu Lys Leu Gln Lys Ala Val

130 135 140

Thr Lys Leu Lys Gln Leu Glu Asp Arg Cys Thr Glu Gln Lys Leu Ser

145 150 155 160

Thr Ala Met Arg Ile Thr Lys Trp Lys Glu Lys Val Gln Ile Gln Arg

165 170 175

Gln Lys Ile Arg Ser Asp Phe Lys Asn Leu Gln Cys Phe Leu His Glu

180 185 190

Glu Glu Lys Ser Tyr Leu Trp Arg Leu Glu Lys Glu Glu Gln Gln Thr

195 200 205

Leu Ser Arg Leu Arg Asp Tyr Glu Ala Gly Leu Gly Leu Lys Ser Asn

210 215 220

Glu Leu Lys Ser His Ile Leu Glu Leu Glu Glu Lys Cys Gln Gly Ser

225 230 235 240

Ala Gln Lys Leu Leu Gln Asn Val Asn Asp Thr Leu Ser Arg Ser Trp

245 250 255

Ala Val Lys Leu Glu Thr Ser Glu Ala Val Ser Leu Glu Leu His Thr

260 265 270

Met Cys Asn Val Ser Lys Leu Tyr Phe Asp Val Lys Lys Met Leu Arg

275 280 285

Ser His Gln Val Ser Val Thr Leu Asp Pro Asp Thr Ala His His Glu

290 295 300

Leu Ile Leu Ser Glu Asp Arg Arg Gln Val Thr Arg Gly Tyr Thr Gln

305 310 315 320

Glu Asn Gln Asp Thr Ser Ser Arg Arg Phe Thr Ala Phe Pro Cys Val

325 330 335

Leu Gly Cys Glu Gly Phe Thr Ser Gly Arg Arg Tyr Phe Glu Val Asp

340 345 350

Val Gly Glu Gly Thr Gly Trp Asp Leu Gly Val Cys Met Glu Asn Val

355 360 365

Gln Arg Gly Thr Gly Met Lys Gln Glu Pro Gln Ser Gly Phe Trp Thr

370 375 380

Leu Arg Leu Cys Lys Lys Lys Gly Tyr Val Ala Leu Thr Ser Pro Pro

385 390 395 400

Thr Ser Leu His Leu His Glu Gln Pro Leu Leu Val Gly Ile Phe Leu

405 410 415

Asp Tyr Glu Ala Gly Val Val Ser Phe Tyr Asn Gly Asn Thr Gly Cys

420 425 430

His Ile Phe Thr Phe Pro Lys Ala Ser Phe Ser Asp Thr Leu Arg Pro

435 440 445

Tyr Phe Gln Val Tyr Gln Tyr Ser Pro Leu Phe Leu Pro Pro Pro Gly

450 455 460

Asp

465

<210> 77

<211> 1557

<212> DNA

<213> 智人(Homo sapiens)

<400> 77

atggcagaga caagtctgtt agaggctggg gcctctgcag cctctacagc tgcggctttg 60

gagaacttac aggtggaggc gagctgctct gtgtgcctgg agtatctgaa ggaacctgtc 120

atcattgagt gtgggcacaa cttctgcaaa gcttgcatca cccgctggtg ggaggaccta 180

gagagggact tcccttgtcc tgtctgtcga aagacatccc gctaccgcag tctccgacct 240

aatcggcaac taggcagtat ggtggaaatt gccaagcagc tccaggccgt caagcggaag 300

atccgggatg agagcctctg cccccaacac catgaggccc tcagcctttt ctgttatgag 360

gaccaggagg ctgtatgctt gatatgtgca atttcccaca cccaccgggc ccacaccgtt 420

gtgccactgg acgatgctac acaggagtac aaggaaaaac tgcagaagtg tctggagccc 480

ctggaacaga agctgcagga gatcactcgc tgcaagtcct ctgaggagaa gaagcctggt 540

gagctcaaga gactagtgga aagtcgccga cagcagatct tgagggagtt tgaagagctt 600

cataggcggc tggatgaaga gcagcaggtg ttgctttcac gactggaaga agaggaacag 660

gacattctgc agcgactccg agaaaatgct gctcaccttg gggacaagcg ccgggacctg 720

gcccacttgg ctgccgaggt ggagggcaag tgcttacagt caggcttcga gatgcttaag 780

gatgtcaaaa gtaccctgga aaagaatatt cctagaaagt tcggaggctc actctcaacg 840

atctgtccac gggatcataa ggctctcctt ggattagtaa aagaaatcaa cagatgtgaa 900

aaggtgaaga ccatggaggt gacttcagta tccatagagc tggaaaagaa cttcagcaat 960

tttccccgac agtactttgc cctaaggaaa atccttaaac agctaattgc ggatgtgacc 1020

ctggaccctg agacagctca tcctaaccta gtcctgtcag aggatcgtaa gagcgtcaag 1080

ttcgtggaga caagactccg ggatctccct gacacaccaa ggcgtttcac cttctaccct 1140

tgcgtcctgg ctactgaggg tttcacctca ggtcgacact actgggaggt ggaggtgggc 1200

gacaagaccc actgggcagt gggtgtatgc cgggactccg tgagccgaaa gggcgagttg 1260

actccactcc ctgagactgg ctactggcgg gtgcggctat ggaatgggga caaatatgca 1320

gccaccacca caccttttac ccctttgcac atcaaggtga aacccaagcg ggtaggcata 1380

ttcctagact atgaggccgg cacactgtct ttctacaatg tcacagaccg ctctcatatc 1440

tacaccttca ctgatacttt tactgagaaa ctttggcccc tcttctaccc aggcatccgg 1500

gctggacgga agaatgctgc accacttacc atcaggcccc caacagattg ggagtga 1557

<210> 78

<211> 518

<212> PRT

<213> 智人(Homo sapiens)

<400> 78

Met Ala Glu Thr Ser Leu Leu Glu Ala Gly Ala Ser Ala Ala Ser Thr

1 5 10 15

Ala Ala Ala Leu Glu Asn Leu Gln Val Glu Ala Ser Cys Ser Val Cys

20 25 30

Leu Glu Tyr Leu Lys Glu Pro Val Ile Ile Glu Cys Gly His Asn Phe

35 40 45

Cys Lys Ala Cys Ile Thr Arg Trp Trp Glu Asp Leu Glu Arg Asp Phe

50 55 60

Pro Cys Pro Val Cys Arg Lys Thr Ser Arg Tyr Arg Ser Leu Arg Pro

65 70 75 80

Asn Arg Gln Leu Gly Ser Met Val Glu Ile Ala Lys Gln Leu Gln Ala

85 90 95

Val Lys Arg Lys Ile Arg Asp Glu Ser Leu Cys Pro Gln His His Glu

100 105 110

Ala Leu Ser Leu Phe Cys Tyr Glu Asp Gln Glu Ala Val Cys Leu Ile

115 120 125

Cys Ala Ile Ser His Thr His Arg Ala His Thr Val Val Pro Leu Asp

130 135 140

Asp Ala Thr Gln Glu Tyr Lys Glu Lys Leu Gln Lys Cys Leu Glu Pro

145 150 155 160

Leu Glu Gln Lys Leu Gln Glu Ile Thr Arg Cys Lys Ser Ser Glu Glu

165 170 175

Lys Lys Pro Gly Glu Leu Lys Arg Leu Val Glu Ser Arg Arg Gln Gln

180 185 190

Ile Leu Arg Glu Phe Glu Glu Leu His Arg Arg Leu Asp Glu Glu Gln

195 200 205

Gln Val Leu Leu Ser Arg Leu Glu Glu Glu Glu Gln Asp Ile Leu Gln

210 215 220

Arg Leu Arg Glu Asn Ala Ala His Leu Gly Asp Lys Arg Arg Asp Leu

225 230 235 240

Ala His Leu Ala Ala Glu Val Glu Gly Lys Cys Leu Gln Ser Gly Phe

245 250 255

Glu Met Leu Lys Asp Val Lys Ser Thr Leu Glu Lys Asn Ile Pro Arg

260 265 270

Lys Phe Gly Gly Ser Leu Ser Thr Ile Cys Pro Arg Asp His Lys Ala

275 280 285

Leu Leu Gly Leu Val Lys Glu Ile Asn Arg Cys Glu Lys Val Lys Thr

290 295 300

Met Glu Val Thr Ser Val Ser Ile Glu Leu Glu Lys Asn Phe Ser Asn

305 310 315 320

Phe Pro Arg Gln Tyr Phe Ala Leu Arg Lys Ile Leu Lys Gln Leu Ile

325 330 335

Ala Asp Val Thr Leu Asp Pro Glu Thr Ala His Pro Asn Leu Val Leu

340 345 350

Ser Glu Asp Arg Lys Ser Val Lys Phe Val Glu Thr Arg Leu Arg Asp

355 360 365

Leu Pro Asp Thr Pro Arg Arg Phe Thr Phe Tyr Pro Cys Val Leu Ala

370 375 380

Thr Glu Gly Phe Thr Ser Gly Arg His Tyr Trp Glu Val Glu Val Gly

385 390 395 400

Asp Lys Thr His Trp Ala Val Gly Val Cys Arg Asp Ser Val Ser Arg

405 410 415

Lys Gly Glu Leu Thr Pro Leu Pro Glu Thr Gly Tyr Trp Arg Val Arg

420 425 430

Leu Trp Asn Gly Asp Lys Tyr Ala Ala Thr Thr Thr Pro Phe Thr Pro

435 440 445

Leu His Ile Lys Val Lys Pro Lys Arg Val Gly Ile Phe Leu Asp Tyr

450 455 460

Glu Ala Gly Thr Leu Ser Phe Tyr Asn Val Thr Asp Arg Ser His Ile

465 470 475 480

Tyr Thr Phe Thr Asp Thr Phe Thr Glu Lys Leu Trp Pro Leu Phe Tyr

485 490 495

Pro Gly Ile Arg Ala Gly Arg Lys Asn Ala Ala Pro Leu Thr Ile Arg

500 505 510

Pro Pro Thr Asp Trp Glu

515

<210> 79

<211> 777

<212> DNA

<213> 智人(Homo sapiens)

<400> 79

atgatccctt tgcagaagga caaccaggag gagggtgtct gccccatctg ccaggagagc 60

ctgaaggagg ccgtgagcac caactgcgga catctcttct gtcgagtgtg cctgacacag 120

catgtggaga aggcctcagc ctctggggtc ttctgctgcc ccctctgccg gaagccctgt 180

tctgaggagg tgctagggac aggctatatc tgccccaacc accagaagag ggtgtgcagg 240

ttctgtgagg agagcagact tcttctatgt gtggaatgcc tggtgtcccc tgaacacatg 300

tctcatcatg aactgaccat tgaaaatgcc ctcagccact acaaggaacg actcaatcgc 360

cggagcagga agctcagaaa ggacattgca gaacttcagc ggctcaaggc tcagcaggag 420

aagaaactgc aggctctgca gtttcaggta gaccacggga accacaggct ggaggctggg 480

ccggagagcc agcaccaaac cagggaacag ctgggtgccc tccctcagca gtggctgggc 540

cagctggagc acatgccagc agaagcggcc agaatccttg acatctccag ggcagtaaca 600

cagctcagaa gcctggtcat tgatctggaa aggacggcca aggaattaga caccaacaca 660

ctgaagaatg ctggtgactt actgaacagg agtgctccac agaaattaga ggttatttat 720

ccccagttgg agaaaggagt cagtgaattg cttcttcagc cccctcagaa gctctga 777

<210> 80

<211> 258

<212> PRT

<213> 智人(Homo sapiens)

<400> 80

Met Ile Pro Leu Gln Lys Asp Asn Gln Glu Glu Gly Val Cys Pro Ile

1 5 10 15

Cys Gln Glu Ser Leu Lys Glu Ala Val Ser Thr Asn Cys Gly His Leu

20 25 30

Phe Cys Arg Val Cys Leu Thr Gln His Val Glu Lys Ala Ser Ala Ser

35 40 45

Gly Val Phe Cys Cys Pro Leu Cys Arg Lys Pro Cys Ser Glu Glu Val

50 55 60

Leu Gly Thr Gly Tyr Ile Cys Pro Asn His Gln Lys Arg Val Cys Arg

65 70 75 80

Phe Cys Glu Glu Ser Arg Leu Leu Leu Cys Val Glu Cys Leu Val Ser

85 90 95

Pro Glu His Met Ser His His Glu Leu Thr Ile Glu Asn Ala Leu Ser

100 105 110

His Tyr Lys Glu Arg Leu Asn Arg Arg Ser Arg Lys Leu Arg Lys Asp

115 120 125

Ile Ala Glu Leu Gln Arg Leu Lys Ala Gln Gln Glu Lys Lys Leu Gln

130 135 140

Ala Leu Gln Phe Gln Val Asp His Gly Asn His Arg Leu Glu Ala Gly

145 150 155 160

Pro Glu Ser Gln His Gln Thr Arg Glu Gln Leu Gly Ala Leu Pro Gln

165 170 175

Gln Trp Leu Gly Gln Leu Glu His Met Pro Ala Glu Ala Ala Arg Ile

180 185 190

Leu Asp Ile Ser Arg Ala Val Thr Gln Leu Arg Ser Leu Val Ile Asp

195 200 205

Leu Glu Arg Thr Ala Lys Glu Leu Asp Thr Asn Thr Leu Lys Asn Ala

210 215 220

Gly Asp Leu Leu Asn Arg Ser Ala Pro Gln Lys Leu Glu Val Ile Tyr

225 230 235 240

Pro Gln Leu Glu Lys Gly Val Ser Glu Leu Leu Leu Gln Pro Pro Gln

245 250 255

Lys Leu

<210> 81

<211> 1893

<212> DNA

<213> 智人(Homo sapiens)

<400> 81

atggctgccg ttgccatgac acccaaccct gtgcagaccc ttcaggagga ggcggtgtgc 60

gccatctgcc tcgattactt cacggacccc gtgtccatcg gctgcgggca caacttctgc 120

cgagtttgtg taacccagtt gtggggtggg gaggatgagg aggacagaga tgagttagat 180

cgggaggagg aggaggagga cggagaggag gaggaagtgg aggctgtggg ggctggcgcg 240

gggtgggaca cccccatgcg ggatgaagac tacgagggtg acatggagga ggaggtcgag 300

gaggaagaag agggtgtgtt ctggaccagt ggcatgagca ggtccagctg ggacaacatg 360

gactatgtgt gggaggagga ggacgaggag gaagacctgg actactactt gggggacatg 420

gaggaggagg acctgagggg ggaggatgag gaggacgagg aggaagtgct ggaggaggtt 480

gaggaagagg atctagaccc cgtcacccca ctgcccccgc ctccagcccc tcggaggtgc 540

ttcacatgcc ctcagtgccg aaagagcttt cctcggcgga gcttccgccc caacctgcag 600

ctggccaata tggtccaggt gattcggcag atgcacccaa cccctggtcg agggagccgc 660

gtgaccgatc agggcatctg tcccaaacac caagaagccc tgaagctctt ctgcgaggta 720

gacgaagagg ccatctgtgt ggtgtgccga gaatccagga gccacaaaca gcacagcgtg 780

gtgccattgg aggaggtggt gcaggagtac aaggccaaac tgcaggggca cgtggaacca 840

ctgaggaagc acctggaggc agtgcagaag atgaaagcca aggaggagag gcgagtgaca 900

gaactgaaga gccagatgaa gtcagagctg gcagcggtgg cctcggagtt tgggcgactg 960

acacggtttc tggctgaaga gcaggcaggg ctggaacggc gtctcagaga gatgcatgaa 1020

gcccagctgg ggcgtgcggg agccgcggct agtcgccttg cagaacaggc cgcccagctc 1080

agccgcctgc tggcagaggc ccaggagcgg agccagcagg ggggtctccg gctgctccag 1140

gacatcaagg agactttcaa taggtgtgaa gaggtacagc tgcagccccc agaggtctgg 1200

tcccctgacc cgtgccaacc ccatagccat gacttcctga cagatgccat cgtgaggaaa 1260

atgagccgga tgttctgtca ggctgcgaga gtggacctga cgctggaccc tgacacggct 1320

cacccggccc tgatgctgtc ccctgaccgc cggggggtcc gcctggcaga gcggcggcag 1380

gaggttgctg accatcccaa gcgcttctcg gccgactgct gcgtactggg ggcccagggc 1440

ttccgctccg gccggcacta ctgggaggta gaggtgggcg ggcggcgggg ctgggcggtg 1500

ggtgctgccc gtgaatcaac ccatcataag gaaaaggtgg gccctggggg ttcctccgtg 1560

ggcagcgggg atgccagctc ctcgcgccat caccatcgcc gccgccggct ccacctgccc 1620

cagcagcccc tgctccagcg ggaagtgtgg tgcgtgggca ccaacggcaa acgctatcag 1680

gcccagagct ccacagaaca gacgctgctg agccccagtg agaaaccaag gcgctttggt 1740

gtgtacctgg actatgaagc tgggcgcctg ggcttctaca acgcagagac tctagcccac 1800

gtgcacacct tctcggctgc cttcctgggc gagcgtgtct ttcctttctt ccgggtgctc 1860

tccaagggca cccgcatcaa gctctgccct tga 1893

<210> 82

<211> 630

<212> PRT

<213> 智人(Homo sapiens)

<400> 82

Met Ala Ala Val Ala Met Thr Pro Asn Pro Val Gln Thr Leu Gln Glu

1 5 10 15

Glu Ala Val Cys Ala Ile Cys Leu Asp Tyr Phe Thr Asp Pro Val Ser

20 25 30

Ile Gly Cys Gly His Asn Phe Cys Arg Val Cys Val Thr Gln Leu Trp

35 40 45

Gly Gly Glu Asp Glu Glu Asp Arg Asp Glu Leu Asp Arg Glu Glu Glu

50 55 60

Glu Glu Asp Gly Glu Glu Glu Glu Val Glu Ala Val Gly Ala Gly Ala

65 70 75 80

Gly Trp Asp Thr Pro Met Arg Asp Glu Asp Tyr Glu Gly Asp Met Glu

85 90 95

Glu Glu Val Glu Glu Glu Glu Glu Gly Val Phe Trp Thr Ser Gly Met

100 105 110

Ser Arg Ser Ser Trp Asp Asn Met Asp Tyr Val Trp Glu Glu Glu Asp

115 120 125

Glu Glu Glu Asp Leu Asp Tyr Tyr Leu Gly Asp Met Glu Glu Glu Asp

130 135 140

Leu Arg Gly Glu Asp Glu Glu Asp Glu Glu Glu Val Leu Glu Glu Val

145 150 155 160

Glu Glu Glu Asp Leu Asp Pro Val Thr Pro Leu Pro Pro Pro Pro Ala

165 170 175

Pro Arg Arg Cys Phe Thr Cys Pro Gln Cys Arg Lys Ser Phe Pro Arg

180 185 190

Arg Ser Phe Arg Pro Asn Leu Gln Leu Ala Asn Met Val Gln Val Ile

195 200 205

Arg Gln Met His Pro Thr Pro Gly Arg Gly Ser Arg Val Thr Asp Gln

210 215 220

Gly Ile Cys Pro Lys His Gln Glu Ala Leu Lys Leu Phe Cys Glu Val

225 230 235 240

Asp Glu Glu Ala Ile Cys Val Val Cys Arg Glu Ser Arg Ser His Lys

245 250 255

Gln His Ser Val Val Pro Leu Glu Glu Val Val Gln Glu Tyr Lys Ala

260 265 270

Lys Leu Gln Gly His Val Glu Pro Leu Arg Lys His Leu Glu Ala Val

275 280 285

Gln Lys Met Lys Ala Lys Glu Glu Arg Arg Val Thr Glu Leu Lys Ser

290 295 300

Gln Met Lys Ser Glu Leu Ala Ala Val Ala Ser Glu Phe Gly Arg Leu

305 310 315 320

Thr Arg Phe Leu Ala Glu Glu Gln Ala Gly Leu Glu Arg Arg Leu Arg

325 330 335

Glu Met His Glu Ala Gln Leu Gly Arg Ala Gly Ala Ala Ala Ser Arg

340 345 350

Leu Ala Glu Gln Ala Ala Gln Leu Ser Arg Leu Leu Ala Glu Ala Gln

355 360 365

Glu Arg Ser Gln Gln Gly Gly Leu Arg Leu Leu Gln Asp Ile Lys Glu

370 375 380

Thr Phe Asn Arg Cys Glu Glu Val Gln Leu Gln Pro Pro Glu Val Trp

385 390 395 400

Ser Pro Asp Pro Cys Gln Pro His Ser His Asp Phe Leu Thr Asp Ala

405 410 415

Ile Val Arg Lys Met Ser Arg Met Phe Cys Gln Ala Ala Arg Val Asp

420 425 430

Leu Thr Leu Asp Pro Asp Thr Ala His Pro Ala Leu Met Leu Ser Pro

435 440 445

Asp Arg Arg Gly Val Arg Leu Ala Glu Arg Arg Gln Glu Val Ala Asp

450 455 460

His Pro Lys Arg Phe Ser Ala Asp Cys Cys Val Leu Gly Ala Gln Gly

465 470 475 480

Phe Arg Ser Gly Arg His Tyr Trp Glu Val Glu Val Gly Gly Arg Arg

485 490 495

Gly Trp Ala Val Gly Ala Ala Arg Glu Ser Thr His His Lys Glu Lys

500 505 510

Val Gly Pro Gly Gly Ser Ser Val Gly Ser Gly Asp Ala Ser Ser Ser

515 520 525

Arg His His His Arg Arg Arg Arg Leu His Leu Pro Gln Gln Pro Leu

530 535 540

Leu Gln Arg Glu Val Trp Cys Val Gly Thr Asn Gly Lys Arg Tyr Gln

545 550 555 560

Ala Gln Ser Ser Thr Glu Gln Thr Leu Leu Ser Pro Ser Glu Lys Pro

565 570 575

Arg Arg Phe Gly Val Tyr Leu Asp Tyr Glu Ala Gly Arg Leu Gly Phe

580 585 590

Tyr Asn Ala Glu Thr Leu Ala His Val His Thr Phe Ser Ala Ala Phe

595 600 605

Leu Gly Glu Arg Val Phe Pro Phe Phe Arg Val Leu Ser Lys Gly Thr

610 615 620

Arg Ile Lys Leu Cys Pro

625 630

<210> 83

<211> 2172

<212> DNA

<213> 智人(Homo sapiens)

<400> 83

atggaaactg ctatgtgcgt ttgctgtcca tgttgtacat ggcagagatg ttgtcctcag 60

ttatgctcct gtctgtgctg caagttcatc ttcacctcag agcggaactg cacctgcttc 120

ccctgccctt acaaagatga gcggaactgc cagttctgcc actgcacctg ttctgagagc 180

cccaactgcc attggtgttg ctgctcttgg gccaatgatc ccaactgtaa gtgctgctgc 240

acagccagca gcaatctcaa ctgctactac tatgagagcc gctgctgccg caataccatc 300

atcactttcc acaagggccg cctcaggagc atccatacct cctccaagac tgccctgcgc 360

actgggagca gcgataccca ggtggatgaa gtaaagtcaa taccagccaa cagtcacctg 420

gtgaaccacc tcaattgccc catgtgcagc cggctgcgcc tgcactcatt catgctgccc 480

tgcaaccaca gcctgtgcga gaagtgcctg cggcagctgc agaagcacgc cgaggtcacc 540

gagaacttct tcatcctcat ctgcccagtg tgcgaccgct cgcactgcat gccctacagc 600

aacaagatgc agctgcccga gaactacctg cacgggcgtc tcaccaagcg ctacatgcag 660

gagcacggct acctcaagtg gcgctttgac cgctcctccg ggcccatcct ctgccaggtc 720

tgccgcaaca agcgcatcgc ttacaagcgc tgcatcacct gccgcctcaa cctgtgcaac 780

gactgcctca aggccttcca ctcggatgtg gccatgcaag accacgtctt tgtggacacc 840

agcgccgagg aacaggacga gaagatctgc atccaccacc catccagccg catcatcgag 900

tactgccgca atgacaacaa attgctctgc accttctgca agttctcttt ccacaatggc 960

cacgacacca ttagcctcat cgacgcctgc tccgagaggg ccgcctcact cttcagcgcc 1020

atcgccaagt tcaaagcagt ccgatatgaa attgataatg acctaatgga attcaacatc 1080

ttaaaaaaca gctttaaagc tgacaaggag gcaaagcgaa aagagatcag aaatggcttt 1140

ctcaagttgc gcagcattct tcaggagaaa gagaagatca tcatggagca gatagagaat 1200

ctagaagtgt ccaggcagaa ggaaattgaa aaatatgtgt atgttacaac catgaaagtg 1260

aacgagatgg atggtctgat cgcctactcc aaggaagccc tgaaggagac tggccaggtg 1320

gcattcctgc agtcagccaa gatcctggtg gaccagatcg aggacggcat ccagaccacc 1380

tacaggcctg acccacagct ccggctgcac tcaataaact acgtgccctt ggactttgtt 1440

gagctttcca gtgccatcca tgagctcttc cccacagggc ccaagaaggt acgctcctca 1500

ggggactccc tgccctcccc ctaccccgtg cactcagaaa caatgattgc caggaaggtc 1560

actttcagca cccacagcct cggcaaccag cacatatacc agcgaagctc ctccatgttg 1620

tccttcagca acactgacaa gaaggccaag gtgggtctgg aggcctgtgg gagagcccag 1680

tcagccaccc ccgccaaacc cacagacggc ctctacacct actggagtgc tggagcagac 1740

agccagtctg tacagaacag cagcagcttc cacaactggt actcattcaa cgatggctct 1800

gtgaagaccc caggcccaat tgttatctac cagactctgg tgtacccaag agctgccaag 1860

gtttactgga catgtccagc agaagacgtg gactcttttg agatggaatt ctatgaagtc 1920

attacttctc ctcctaacaa cgtacaaatg gagctctgtg gacaaattcg ggacataatg 1980

cagcaaaatc tggagctgca caacctgacc cccaacacag aatacgtgtt taaagttaga 2040

gccatcaatg ataatggtcc tgggcaatgg agtgatatct gcaaggtggt aacaccagat 2100

ggacatggga agaaccgagc taagtggggc ctgctgaaga atatccagtc tgccctccag 2160

aagcacttct ga 2172

<210> 84

<211> 723

<212> PRT

<213> 智人(Homo sapiens)

<400> 84

Met Glu Thr Ala Met Cys Val Cys Cys Pro Cys Cys Thr Trp Gln Arg

1 5 10 15

Cys Cys Pro Gln Leu Cys Ser Cys Leu Cys Cys Lys Phe Ile Phe Thr

20 25 30

Ser Glu Arg Asn Cys Thr Cys Phe Pro Cys Pro Tyr Lys Asp Glu Arg

35 40 45

Asn Cys Gln Phe Cys His Cys Thr Cys Ser Glu Ser Pro Asn Cys His

50 55 60

Trp Cys Cys Cys Ser Trp Ala Asn Asp Pro Asn Cys Lys Cys Cys Cys

65 70 75 80

Thr Ala Ser Ser Asn Leu Asn Cys Tyr Tyr Tyr Glu Ser Arg Cys Cys

85 90 95

Arg Asn Thr Ile Ile Thr Phe His Lys Gly Arg Leu Arg Ser Ile His

100 105 110

Thr Ser Ser Lys Thr Ala Leu Arg Thr Gly Ser Ser Asp Thr Gln Val

115 120 125

Asp Glu Val Lys Ser Ile Pro Ala Asn Ser His Leu Val Asn His Leu

130 135 140

Asn Cys Pro Met Cys Ser Arg Leu Arg Leu His Ser Phe Met Leu Pro

145 150 155 160

Cys Asn His Ser Leu Cys Glu Lys Cys Leu Arg Gln Leu Gln Lys His

165 170 175

Ala Glu Val Thr Glu Asn Phe Phe Ile Leu Ile Cys Pro Val Cys Asp

180 185 190

Arg Ser His Cys Met Pro Tyr Ser Asn Lys Met Gln Leu Pro Glu Asn

195 200 205

Tyr Leu His Gly Arg Leu Thr Lys Arg Tyr Met Gln Glu His Gly Tyr

210 215 220

Leu Lys Trp Arg Phe Asp Arg Ser Ser Gly Pro Ile Leu Cys Gln Val

225 230 235 240

Cys Arg Asn Lys Arg Ile Ala Tyr Lys Arg Cys Ile Thr Cys Arg Leu

245 250 255

Asn Leu Cys Asn Asp Cys Leu Lys Ala Phe His Ser Asp Val Ala Met

260 265 270

Gln Asp His Val Phe Val Asp Thr Ser Ala Glu Glu Gln Asp Glu Lys

275 280 285

Ile Cys Ile His His Pro Ser Ser Arg Ile Ile Glu Tyr Cys Arg Asn

290 295 300

Asp Asn Lys Leu Leu Cys Thr Phe Cys Lys Phe Ser Phe His Asn Gly

305 310 315 320

His Asp Thr Ile Ser Leu Ile Asp Ala Cys Ser Glu Arg Ala Ala Ser

325 330 335

Leu Phe Ser Ala Ile Ala Lys Phe Lys Ala Val Arg Tyr Glu Ile Asp

340 345 350

Asn Asp Leu Met Glu Phe Asn Ile Leu Lys Asn Ser Phe Lys Ala Asp

355 360 365

Lys Glu Ala Lys Arg Lys Glu Ile Arg Asn Gly Phe Leu Lys Leu Arg

370 375 380

Ser Ile Leu Gln Glu Lys Glu Lys Ile Ile Met Glu Gln Ile Glu Asn

385 390 395 400

Leu Glu Val Ser Arg Gln Lys Glu Ile Glu Lys Tyr Val Tyr Val Thr

405 410 415

Thr Met Lys Val Asn Glu Met Asp Gly Leu Ile Ala Tyr Ser Lys Glu

420 425 430

Ala Leu Lys Glu Thr Gly Gln Val Ala Phe Leu Gln Ser Ala Lys Ile

435 440 445

Leu Val Asp Gln Ile Glu Asp Gly Ile Gln Thr Thr Tyr Arg Pro Asp

450 455 460

Pro Gln Leu Arg Leu His Ser Ile Asn Tyr Val Pro Leu Asp Phe Val

465 470 475 480

Glu Leu Ser Ser Ala Ile His Glu Leu Phe Pro Thr Gly Pro Lys Lys

485 490 495

Val Arg Ser Ser Gly Asp Ser Leu Pro Ser Pro Tyr Pro Val His Ser

500 505 510

Glu Thr Met Ile Ala Arg Lys Val Thr Phe Ser Thr His Ser Leu Gly

515 520 525

Asn Gln His Ile Tyr Gln Arg Ser Ser Ser Met Leu Ser Phe Ser Asn

530 535 540

Thr Asp Lys Lys Ala Lys Val Gly Leu Glu Ala Cys Gly Arg Ala Gln

545 550 555 560

Ser Ala Thr Pro Ala Lys Pro Thr Asp Gly Leu Tyr Thr Tyr Trp Ser

565 570 575

Ala Gly Ala Asp Ser Gln Ser Val Gln Asn Ser Ser Ser Phe His Asn

580 585 590

Trp Tyr Ser Phe Asn Asp Gly Ser Val Lys Thr Pro Gly Pro Ile Val

595 600 605

Ile Tyr Gln Thr Leu Val Tyr Pro Arg Ala Ala Lys Val Tyr Trp Thr

610 615 620

Cys Pro Ala Glu Asp Val Asp Ser Phe Glu Met Glu Phe Tyr Glu Val

625 630 635 640

Ile Thr Ser Pro Pro Asn Asn Val Gln Met Glu Leu Cys Gly Gln Ile

645 650 655

Arg Asp Ile Met Gln Gln Asn Leu Glu Leu His Asn Leu Thr Pro Asn

660 665 670

Thr Glu Tyr Val Phe Lys Val Arg Ala Ile Asn Asp Asn Gly Pro Gly

675 680 685

Gln Trp Ser Asp Ile Cys Lys Val Val Thr Pro Asp Gly His Gly Lys

690 695 700

Asn Arg Ala Lys Trp Gly Leu Leu Lys Asn Ile Gln Ser Ala Leu Gln

705 710 715 720

Lys His Phe

<210> 85

<211> 1341

<212> DNA

<213> 智人(Homo sapiens)

<400> 85

atggactcag acttctcaca tgccttccag aaggaactca cctgcgtcat ctgtttgaac 60

tacctggtag accctgtcac catctgctgt gggcacagct tctgtaggcc ctgtctctgc 120

ctttcgtggg aggaagccca aagtcctgca aactgccctg catgcaggga accatcaccg 180

aaaatggact tcaaaaccaa tattcttctg aagaatttag tgaccattgc cagaaaagcc 240

agtctctggc aattcctgag ctctgagaaa caaatatgtg ggacccatag gcaaacaaag 300

aagatgttct gtgacatgga caagagtctc ctctgcttgc tgtgctccaa ctctcaggag 360

cacggggctc acaaacacca tcccatcgaa gaggcagctg aggaacaccg ggagaaactc 420

ttaaagcaaa tgaggatttt atggaaaaag attcaagaaa atcagagaaa tctatatgag 480

gagggaagaa cagccttcct ctggaggggc aatgtggttt tacgggcaca gatgatcagg 540

aatgagtata ggaagctgca tccggttctc cataaggaag aaaaacaaca tttagagaga 600

ctgaacaagg aataccaaga gatttttcag caactccaga gaagttgggt caaaatggat 660

caaaagagta aacacttgaa agaaatgtat caggaactaa tggaaatgtg tcataaacca 720

gatgtggagc tgctccagga tttgggagac atcgtggcaa ggagtgagtc cgtgctgctg 780

cacatgcccc agcctgtgaa tccagagctc actgcaggac ccatcactgg actggtgtac 840

aggctcaacc gcttccgagt ggaaatttcc ttccattttg aagtaaccaa tcacaatatc 900

aggctctttg aggatgtgag aagttggatg tttagacgtg gacctttgaa ttctgacaga 960

tctgactatt ttgctgcatg gggagccagg gtcttctcct ttgggaaaca ctactgggag 1020

ctggatgtgg acaactcttg tgactgggct ctgggagtct gtaacaactc ctggataagg 1080

aagaatagca caatggttaa ctctgaggac atatttcttc ttttgtgtct gaaggtggat 1140

aatcatttca atctcttgac cacctcccca gtgtttcctc actacataga gaaacctctg 1200

ggccgggttg gtgtgtttct tgattttgaa agtggaagtg tgagtttttt gaatgtcacc 1260

aagagttccc tcatatggag ttacccagct ggctccttaa cttttcctgt caggcctttc 1320

ttttacactg gccacagatg a 1341

<210> 86

<211> 446

<212> PRT

<213> 智人(Homo sapiens)

<400> 86

Met Asp Ser Asp Phe Ser His Ala Phe Gln Lys Glu Leu Thr Cys Val

1 5 10 15

Ile Cys Leu Asn Tyr Leu Val Asp Pro Val Thr Ile Cys Cys Gly His

20 25 30

Ser Phe Cys Arg Pro Cys Leu Cys Leu Ser Trp Glu Glu Ala Gln Ser

35 40 45

Pro Ala Asn Cys Pro Ala Cys Arg Glu Pro Ser Pro Lys Met Asp Phe

50 55 60

Lys Thr Asn Ile Leu Leu Lys Asn Leu Val Thr Ile Ala Arg Lys Ala

65 70 75 80

Ser Leu Trp Gln Phe Leu Ser Ser Glu Lys Gln Ile Cys Gly Thr His

85 90 95

Arg Gln Thr Lys Lys Met Phe Cys Asp Met Asp Lys Ser Leu Leu Cys

100 105 110

Leu Leu Cys Ser Asn Ser Gln Glu His Gly Ala His Lys His His Pro

115 120 125

Ile Glu Glu Ala Ala Glu Glu His Arg Glu Lys Leu Leu Lys Gln Met

130 135 140

Arg Ile Leu Trp Lys Lys Ile Gln Glu Asn Gln Arg Asn Leu Tyr Glu

145 150 155 160

Glu Gly Arg Thr Ala Phe Leu Trp Arg Gly Asn Val Val Leu Arg Ala

165 170 175

Gln Met Ile Arg Asn Glu Tyr Arg Lys Leu His Pro Val Leu His Lys

180 185 190

Glu Glu Lys Gln His Leu Glu Arg Leu Asn Lys Glu Tyr Gln Glu Ile

195 200 205

Phe Gln Gln Leu Gln Arg Ser Trp Val Lys Met Asp Gln Lys Ser Lys

210 215 220

His Leu Lys Glu Met Tyr Gln Glu Leu Met Glu Met Cys His Lys Pro

225 230 235 240

Asp Val Glu Leu Leu Gln Asp Leu Gly Asp Ile Val Ala Arg Ser Glu

245 250 255

Ser Val Leu Leu His Met Pro Gln Pro Val Asn Pro Glu Leu Thr Ala

260 265 270

Gly Pro Ile Thr Gly Leu Val Tyr Arg Leu Asn Arg Phe Arg Val Glu

275 280 285

Ile Ser Phe His Phe Glu Val Thr Asn His Asn Ile Arg Leu Phe Glu

290 295 300

Asp Val Arg Ser Trp Met Phe Arg Arg Gly Pro Leu Asn Ser Asp Arg

305 310 315 320

Ser Asp Tyr Phe Ala Ala Trp Gly Ala Arg Val Phe Ser Phe Gly Lys

325 330 335

His Tyr Trp Glu Leu Asp Val Asp Asn Ser Cys Asp Trp Ala Leu Gly

340 345 350

Val Cys Asn Asn Ser Trp Ile Arg Lys Asn Ser Thr Met Val Asn Ser

355 360 365

Glu Asp Ile Phe Leu Leu Leu Cys Leu Lys Val Asp Asn His Phe Asn

370 375 380

Leu Leu Thr Thr Ser Pro Val Phe Pro His Tyr Ile Glu Lys Pro Leu

385 390 395 400

Gly Arg Val Gly Val Phe Leu Asp Phe Glu Ser Gly Ser Val Ser Phe

405 410 415

Leu Asn Val Thr Lys Ser Ser Leu Ile Trp Ser Tyr Pro Ala Gly Ser

420 425 430

Leu Thr Phe Pro Val Arg Pro Phe Phe Tyr Thr Gly His Arg

435 440 445

<210> 87

<211> 1035

<212> DNA

<213> 智人(Homo sapiens)

<400> 87

atggcctctg gagtgggcgc ggccttcgag gaactgcctc acgacggcac gtgtgacgag 60

tgcgagcccg acgaggctcc gggggccgag gaagtgtgcc gagaatgcgg cttctgctac 120

tgccgccgcc atgccgaggc gcacaggcag aagttcctca gtcaccatct ggccgaatac 180

gtccacggct cccaggcctg gaccccgcca gctgacggag agggggcggg gaaggaagaa 240

gcggaggtca aggtggagca ggagagggag atagaaagcg aggcagggga agagagtgag 300

tcggaggaag agagcgagtc agaggaagag agcgagacag aggaagagag tgaggatgag 360

agcgatgagg agagtgaaga agacagcgag gaagaaatgg aggatgagca agaaagcgag 420

gccgaagaag acaaccaaga agaaggggaa tccgaggcgg agggagaaac tgaggcagaa 480

agtgaatttg acccagaaat agaaatggaa gcagagagag tggccaagag gaagtgtccg 540

gaccatgggc ttgatttgag tacctattgc caggaagata ggcagctcat ctgtgtcctg 600

tgtccagtca ttggggctca ccagggccac caactctcca ccctagacga agcctttgaa 660

gaattaagaa gcaaagactc aggtggactg aaggccgcta tgatcgaatt ggtggaaagg 720

ttgaagttca agagctcaga ccctaaagta actcgggacc aaatgaagat gtttatacag 780

caggaattta agaaagttca gaaagtgatt gctgatgagg agcagaaggc ccttcatcta 840

gtggacatcc aagaggcaat ggccacagct catgtgactg agatactggc agacatccaa 900

tcccacatgg ataggttgat gactcagatg gcccaagcca aggaacaact tgatacctct 960

aatgaatcag ctgagccaaa ggcagagggc gatgaggaag gacccagtgg tgccagtgaa 1020

gaagaggaca catga 1035

<210> 88

<211> 344

<212> PRT

<213> 智人(Homo sapiens)

<400> 88

Met Ala Ser Gly Val Gly Ala Ala Phe Glu Glu Leu Pro His Asp Gly

1 5 10 15

Thr Cys Asp Glu Cys Glu Pro Asp Glu Ala Pro Gly Ala Glu Glu Val

20 25 30

Cys Arg Glu Cys Gly Phe Cys Tyr Cys Arg Arg His Ala Glu Ala His

35 40 45

Arg Gln Lys Phe Leu Ser His His Leu Ala Glu Tyr Val His Gly Ser

50 55 60

Gln Ala Trp Thr Pro Pro Ala Asp Gly Glu Gly Ala Gly Lys Glu Glu

65 70 75 80

Ala Glu Val Lys Val Glu Gln Glu Arg Glu Ile Glu Ser Glu Ala Gly

85 90 95

Glu Glu Ser Glu Ser Glu Glu Glu Ser Glu Ser Glu Glu Glu Ser Glu

100 105 110

Thr Glu Glu Glu Ser Glu Asp Glu Ser Asp Glu Glu Ser Glu Glu Asp

115 120 125

Ser Glu Glu Glu Met Glu Asp Glu Gln Glu Ser Glu Ala Glu Glu Asp

130 135 140

Asn Gln Glu Glu Gly Glu Ser Glu Ala Glu Gly Glu Thr Glu Ala Glu

145 150 155 160

Ser Glu Phe Asp Pro Glu Ile Glu Met Glu Ala Glu Arg Val Ala Lys

165 170 175

Arg Lys Cys Pro Asp His Gly Leu Asp Leu Ser Thr Tyr Cys Gln Glu

180 185 190

Asp Arg Gln Leu Ile Cys Val Leu Cys Pro Val Ile Gly Ala His Gln

195 200 205

Gly His Gln Leu Ser Thr Leu Asp Glu Ala Phe Glu Glu Leu Arg Ser

210 215 220

Lys Asp Ser Gly Gly Leu Lys Ala Ala Met Ile Glu Leu Val Glu Arg

225 230 235 240

Leu Lys Phe Lys Ser Ser Asp Pro Lys Val Thr Arg Asp Gln Met Lys

245 250 255

Met Phe Ile Gln Gln Glu Phe Lys Lys Val Gln Lys Val Ile Ala Asp

260 265 270

Glu Glu Gln Lys Ala Leu His Leu Val Asp Ile Gln Glu Ala Met Ala

275 280 285

Thr Ala His Val Thr Glu Ile Leu Ala Asp Ile Gln Ser His Met Asp

290 295 300

Arg Leu Met Thr Gln Met Ala Gln Ala Lys Glu Gln Leu Asp Thr Ser

305 310 315 320

Asn Glu Ser Ala Glu Pro Lys Ala Glu Gly Asp Glu Glu Gly Pro Ser

325 330 335

Gly Ala Ser Glu Glu Glu Asp Thr

340

<210> 89

<211> 1743

<212> DNA

<213> 智人(Homo sapiens)

<400> 89

atgtcagaaa acagaaaacc gctgctgggc tttgtaagca aactcactag tgggactgca 60

cttgggaact caggcaagac tcactgcccc ctgtgcttgg ggcttttcaa agcccccagg 120

ctcttgcctt gtttgcatac agtttgcacc acgtgtctgg agcagctgga gcccttctca 180

gtagtggaca tccgaggggg agactctgac acaagctctg aggggtcaat attccaggaa 240

ctcaagccac gaagtctgca gtcgcagatc ggcatccttt gtcctgtatg tgatgctcag 300

gtggacctgc ccatgggtgg agtgaaggct ttaaccatag accacctggc cgtgaatgat 360

gtgatgctgg agagcctacg tggggaaggc cagggcctgg tgtgtgacct gtgcaacgac 420

agggaagtag agaagaggtg tcagacctgc aaagccaacc tctgccactt ctgctgccag 480

gctcataggc ggcagaagaa aacgacttac cacaccatgg tggacctaaa agacttgaaa 540

ggctacagcc ggattgggaa gcccatcctg tgtcctgttc accctgcaga ggaactgagg 600

ctgttctgtg agttctgtga ccggcccgtg tgccaggatt gtgtggtggg ggagcatcgg 660

gaacacccct gtgacttcac cagcaatgtc atccacaagc atggggactc tgtgtgggag 720

ctcctcaaag gtactcagcc ccacgtggag gccctggagg aagccctggc tcagatccac 780

ataataaaca gtgccctcca gaagcgagtg gaggcagtgg cagctgatgt ccggacattc 840

tcggagggct acattaaggc cattgaggag catcgggaca agctgctgaa gcagctggaa 900

gacatacggg cccagaagga aaattccctg cagctgcaga aggcccagct ggaacagtta 960

ctggcagaca tgcggactgg agtggagttc accgagcact tgctgaccag cggctcagac 1020

ttggagatcc tcatcaccaa gagggtggtg gtagaacggc tcaggaagct gaacaaagtt 1080

caatatagca cccgtcctgg agtaaatgat aagatacgct tctgtcctca ggagaaagca 1140

ggccagtgcc gtggctatga aatttatggt acgattaata ccaaagaggt tgatccagcc 1200

aaatgtgtcc tacaaggaga agacctccac agagcccggg agaaacagac ggcctctttc 1260

accctgcttt gtaaggatgc cgcaggagaa atcatgggca ggggaggaga caacgttcaa 1320

gttgccgttg tccctaaaga taagaaagac agcccagtca gaacaatggt ccaggataac 1380

aaggatggga catactacat ttcctacacc cccaaggaac ctggcgtcta tactgtgtgg 1440

gtctgcatca aagaacagca tgtgcagggc tcgccattca ctgtgatggt gaggagaaag 1500

caccgcccac actcaggcgt gtttcactgc tgcaccttct gctccagcgg gggccagaaa 1560

accgctcgct gcgcctgtgg aggcaccatg ccaggtgggt acctaggctg tggccatgga 1620

cacaaaggcc acccaggtca tccccactgg tcatgctgtg gaaaatttaa tgagaaatct 1680

gaatgcacat ggacaggtgg gcagagcgca ccgaggagtc tacttaggac tgtggctctc 1740

tga 1743

<210> 90

<211> 580

<212> PRT

<213> 智人(Homo sapiens)

<400> 90

Met Ser Glu Asn Arg Lys Pro Leu Leu Gly Phe Val Ser Lys Leu Thr

1 5 10 15

Ser Gly Thr Ala Leu Gly Asn Ser Gly Lys Thr His Cys Pro Leu Cys

20 25 30

Leu Gly Leu Phe Lys Ala Pro Arg Leu Leu Pro Cys Leu His Thr Val

35 40 45

Cys Thr Thr Cys Leu Glu Gln Leu Glu Pro Phe Ser Val Val Asp Ile

50 55 60

Arg Gly Gly Asp Ser Asp Thr Ser Ser Glu Gly Ser Ile Phe Gln Glu

65 70 75 80

Leu Lys Pro Arg Ser Leu Gln Ser Gln Ile Gly Ile Leu Cys Pro Val

85 90 95

Cys Asp Ala Gln Val Asp Leu Pro Met Gly Gly Val Lys Ala Leu Thr

100 105 110

Ile Asp His Leu Ala Val Asn Asp Val Met Leu Glu Ser Leu Arg Gly

115 120 125

Glu Gly Gln Gly Leu Val Cys Asp Leu Cys Asn Asp Arg Glu Val Glu

130 135 140

Lys Arg Cys Gln Thr Cys Lys Ala Asn Leu Cys His Phe Cys Cys Gln

145 150 155 160

Ala His Arg Arg Gln Lys Lys Thr Thr Tyr His Thr Met Val Asp Leu

165 170 175

Lys Asp Leu Lys Gly Tyr Ser Arg Ile Gly Lys Pro Ile Leu Cys Pro

180 185 190

Val His Pro Ala Glu Glu Leu Arg Leu Phe Cys Glu Phe Cys Asp Arg

195 200 205

Pro Val Cys Gln Asp Cys Val Val Gly Glu His Arg Glu His Pro Cys

210 215 220

Asp Phe Thr Ser Asn Val Ile His Lys His Gly Asp Ser Val Trp Glu

225 230 235 240

Leu Leu Lys Gly Thr Gln Pro His Val Glu Ala Leu Glu Glu Ala Leu

245 250 255

Ala Gln Ile His Ile Ile Asn Ser Ala Leu Gln Lys Arg Val Glu Ala

260 265 270

Val Ala Ala Asp Val Arg Thr Phe Ser Glu Gly Tyr Ile Lys Ala Ile

275 280 285

Glu Glu His Arg Asp Lys Leu Leu Lys Gln Leu Glu Asp Ile Arg Ala

290 295 300

Gln Lys Glu Asn Ser Leu Gln Leu Gln Lys Ala Gln Leu Glu Gln Leu

305 310 315 320

Leu Ala Asp Met Arg Thr Gly Val Glu Phe Thr Glu His Leu Leu Thr

325 330 335

Ser Gly Ser Asp Leu Glu Ile Leu Ile Thr Lys Arg Val Val Val Glu

340 345 350

Arg Leu Arg Lys Leu Asn Lys Val Gln Tyr Ser Thr Arg Pro Gly Val

355 360 365

Asn Asp Lys Ile Arg Phe Cys Pro Gln Glu Lys Ala Gly Gln Cys Arg

370 375 380

Gly Tyr Glu Ile Tyr Gly Thr Ile Asn Thr Lys Glu Val Asp Pro Ala

385 390 395 400

Lys Cys Val Leu Gln Gly Glu Asp Leu His Arg Ala Arg Glu Lys Gln

405 410 415

Thr Ala Ser Phe Thr Leu Leu Cys Lys Asp Ala Ala Gly Glu Ile Met

420 425 430

Gly Arg Gly Gly Asp Asn Val Gln Val Ala Val Val Pro Lys Asp Lys

435 440 445

Lys Asp Ser Pro Val Arg Thr Met Val Gln Asp Asn Lys Asp Gly Thr

450 455 460

Tyr Tyr Ile Ser Tyr Thr Pro Lys Glu Pro Gly Val Tyr Thr Val Trp

465 470 475 480

Val Cys Ile Lys Glu Gln His Val Gln Gly Ser Pro Phe Thr Val Met

485 490 495

Val Arg Arg Lys His Arg Pro His Ser Gly Val Phe His Cys Cys Thr

500 505 510

Phe Cys Ser Ser Gly Gly Gln Lys Thr Ala Arg Cys Ala Cys Gly Gly

515 520 525

Thr Met Pro Gly Gly Tyr Leu Gly Cys Gly His Gly His Lys Gly His

530 535 540

Pro Gly His Pro His Trp Ser Cys Cys Gly Lys Phe Asn Glu Lys Ser

545 550 555 560

Glu Cys Thr Trp Thr Gly Gly Gln Ser Ala Pro Arg Ser Leu Leu Arg

565 570 575

Thr Val Ala Leu

580

<210> 91

<211> 2280

<212> DNA

<213> 智人(Homo sapiens)

<400> 91

atggcagagg gtgaggatat gcagaccttc acttccatca tggacgcact ggtccgcatc 60

agtaccagca tgaagaacat ggagaaggaa ctgctgtgcc cagtgtgtca agagatgtac 120

aagcagccac tggtgctgcc ctgtacccac aacgtgtgcc aggcctgtgc ccgagaggtc 180

ttgggccagc agggctacat aggacatggt ggggacccca gctccgagcc cacctctcct 240

gcctccaccc cttccacccg cagcccccgc ctctcccgca gaactctccc caagccagac 300

cgcctggacc ggctgcttaa gtcaggcttt gggacatacc ctgggaggaa gcgaggtgct 360

ttgcaccccc aagtgatcat gttcccgtgc ccagcctgcc aaggtgatgt ggagcttggg 420

gagcggggtc tggcagggct tttccggaac ctgaccctgg agcgtgtggt ggagcggtac 480

cgccagagtg tgagtgtggg aggtgccatc ctgtgccagt tgtgcaagcc cccaccacta 540

gaggccacca agggctgcac agagtgccgc gccaccttct gcaatgagtg cttcaagctc 600

ttccacccct ggggcaccca gaaggcccag catgagccca ccctgcctac cctctccttc 660

cgacccaagg gccttatgtg cccagaccac aaggaagagg tgacccacta ctgcaagaca 720

tgccaacgcc tggtatgtca actctgccgg gtgcggcgca cccacagcgg gcacaagatc 780

acaccagtgc tcagtgccta ccaggccctc aaggacaagc tgacaaagag cctgacatac 840

atcctgggaa accaggacac ggtacagacc cagatctgtg agctggagga ggccgtgagg 900

cacaccgagg tgagtggtca gcaggccaag gaggaggtgt cgcagctggt gcgggggctg 960

ggggctgtgc tggaggagaa gcgggcatca ctgcttcagg ccattgaaga atgccagcag 1020

gagcggctgg cccgtctcag cgcccagatc caggagcacc ggagcctgct ggatggctca 1080

ggtctggtgg gctatgccca ggaagtactt aaggaaacag accagccttg ctttgtgcaa 1140

gccgccaagc agctgcacaa caggattgcc cgagccactg aagccctcca gacattccgg 1200

ccagctgcca gctcctcctt ccgccattgc cagctcgacg tgggacgtga gatgaagctg 1260

ctgacagagc ttaacttcct gcgagtgcct gaggcccccg tcattgacac ccagcgcacc 1320

tttgcctatg atcagatctt cctgtgctgg cggctgcccc cccattcacc acctgcctgg 1380

cactataccg ttgagttccg gcgcacggat gtgcctgctc agccaggccc cacccgctgg 1440

cagcggcggg aggaggtgag gggcaccagt gccctgcttg agaaccccga cacgggctct 1500

gtgtatgtgc tgcgtgtccg cggctgcaac aaggccggct acggcgaata cagtgaagat 1560

gtgcacctgc acacgccccc ggcacctgtc ctgcacttct tcctcgatag ccgctggggc 1620

gcaagccgag agcggctggc tatcagcaag gaccagcgag cagtacggag tgttccaggg 1680

ctgcccctgc tgctggctgc tgaccggctg ctgaccggct gccacctgag tgtggatgtg 1740

gtcctgggcg acgtggctgt gacccagggc cgcagctact gggcctgcgc cgtagaccca 1800

gcctcctact tggtcaaggt gggcgtcggg ctggagagca agcttcaaga aagtttccag 1860

ggtgcccccg atgtgatcag ccccaggtac gacccggaca gcgggcacga cagcggtgcc 1920

gaggatgcca cagtggaggc gtcgccaccc ttcgctttcc taaccattgg catgggcaag 1980

atcctgctgg ggtcgggggc aagctcaaac gcagggctga cagggaggga tggccccaca 2040

gccggctgca cagtgcccct gccaccccgc ctgggcatct gcctggacta tgagcggggc 2100

cgggtttcct tcctggatgc tgtttccttc cgtgggctct tggagtgccc cctggactgc 2160

tcagggcctg tgtgccctgc cttttgcttc atcgggggtg gcgcagtaca gctccaggag 2220

ccagtgggca ctaagcctga gaggaaagtc accattgggg gcttcgccaa gctggactga 2280

<210> 92

<211> 759

<212> PRT

<213> 智人(Homo sapiens)

<400> 92

Met Ala Glu Gly Glu Asp Met Gln Thr Phe Thr Ser Ile Met Asp Ala

1 5 10 15

Leu Val Arg Ile Ser Thr Ser Met Lys Asn Met Glu Lys Glu Leu Leu

20 25 30

Cys Pro Val Cys Gln Glu Met Tyr Lys Gln Pro Leu Val Leu Pro Cys

35 40 45

Thr His Asn Val Cys Gln Ala Cys Ala Arg Glu Val Leu Gly Gln Gln

50 55 60

Gly Tyr Ile Gly His Gly Gly Asp Pro Ser Ser Glu Pro Thr Ser Pro

65 70 75 80

Ala Ser Thr Pro Ser Thr Arg Ser Pro Arg Leu Ser Arg Arg Thr Leu

85 90 95

Pro Lys Pro Asp Arg Leu Asp Arg Leu Leu Lys Ser Gly Phe Gly Thr

100 105 110

Tyr Pro Gly Arg Lys Arg Gly Ala Leu His Pro Gln Val Ile Met Phe

115 120 125

Pro Cys Pro Ala Cys Gln Gly Asp Val Glu Leu Gly Glu Arg Gly Leu

130 135 140

Ala Gly Leu Phe Arg Asn Leu Thr Leu Glu Arg Val Val Glu Arg Tyr

145 150 155 160

Arg Gln Ser Val Ser Val Gly Gly Ala Ile Leu Cys Gln Leu Cys Lys

165 170 175

Pro Pro Pro Leu Glu Ala Thr Lys Gly Cys Thr Glu Cys Arg Ala Thr

180 185 190

Phe Cys Asn Glu Cys Phe Lys Leu Phe His Pro Trp Gly Thr Gln Lys

195 200 205

Ala Gln His Glu Pro Thr Leu Pro Thr Leu Ser Phe Arg Pro Lys Gly

210 215 220

Leu Met Cys Pro Asp His Lys Glu Glu Val Thr His Tyr Cys Lys Thr

225 230 235 240

Cys Gln Arg Leu Val Cys Gln Leu Cys Arg Val Arg Arg Thr His Ser

245 250 255

Gly His Lys Ile Thr Pro Val Leu Ser Ala Tyr Gln Ala Leu Lys Asp

260 265 270

Lys Leu Thr Lys Ser Leu Thr Tyr Ile Leu Gly Asn Gln Asp Thr Val

275 280 285

Gln Thr Gln Ile Cys Glu Leu Glu Glu Ala Val Arg His Thr Glu Val

290 295 300

Ser Gly Gln Gln Ala Lys Glu Glu Val Ser Gln Leu Val Arg Gly Leu

305 310 315 320

Gly Ala Val Leu Glu Glu Lys Arg Ala Ser Leu Leu Gln Ala Ile Glu

325 330 335

Glu Cys Gln Gln Glu Arg Leu Ala Arg Leu Ser Ala Gln Ile Gln Glu

340 345 350

His Arg Ser Leu Leu Asp Gly Ser Gly Leu Val Gly Tyr Ala Gln Glu

355 360 365

Val Leu Lys Glu Thr Asp Gln Pro Cys Phe Val Gln Ala Ala Lys Gln

370 375 380

Leu His Asn Arg Ile Ala Arg Ala Thr Glu Ala Leu Gln Thr Phe Arg

385 390 395 400

Pro Ala Ala Ser Ser Ser Phe Arg His Cys Gln Leu Asp Val Gly Arg

405 410 415

Glu Met Lys Leu Leu Thr Glu Leu Asn Phe Leu Arg Val Pro Glu Ala

420 425 430

Pro Val Ile Asp Thr Gln Arg Thr Phe Ala Tyr Asp Gln Ile Phe Leu

435 440 445

Cys Trp Arg Leu Pro Pro His Ser Pro Pro Ala Trp His Tyr Thr Val

450 455 460

Glu Phe Arg Arg Thr Asp Val Pro Ala Gln Pro Gly Pro Thr Arg Trp

465 470 475 480

Gln Arg Arg Glu Glu Val Arg Gly Thr Ser Ala Leu Leu Glu Asn Pro

485 490 495

Asp Thr Gly Ser Val Tyr Val Leu Arg Val Arg Gly Cys Asn Lys Ala

500 505 510

Gly Tyr Gly Glu Tyr Ser Glu Asp Val His Leu His Thr Pro Pro Ala

515 520 525

Pro Val Leu His Phe Phe Leu Asp Ser Arg Trp Gly Ala Ser Arg Glu

530 535 540

Arg Leu Ala Ile Ser Lys Asp Gln Arg Ala Val Arg Ser Val Pro Gly

545 550 555 560

Leu Pro Leu Leu Leu Ala Ala Asp Arg Leu Leu Thr Gly Cys His Leu

565 570 575

Ser Val Asp Val Val Leu Gly Asp Val Ala Val Thr Gln Gly Arg Ser

580 585 590

Tyr Trp Ala Cys Ala Val Asp Pro Ala Ser Tyr Leu Val Lys Val Gly

595 600 605

Val Gly Leu Glu Ser Lys Leu Gln Glu Ser Phe Gln Gly Ala Pro Asp

610 615 620

Val Ile Ser Pro Arg Tyr Asp Pro Asp Ser Gly His Asp Ser Gly Ala

625 630 635 640

Glu Asp Ala Thr Val Glu Ala Ser Pro Pro Phe Ala Phe Leu Thr Ile

645 650 655

Gly Met Gly Lys Ile Leu Leu Gly Ser Gly Ala Ser Ser Asn Ala Gly

660 665 670

Leu Thr Gly Arg Asp Gly Pro Thr Ala Gly Cys Thr Val Pro Leu Pro

675 680 685

Pro Arg Leu Gly Ile Cys Leu Asp Tyr Glu Arg Gly Arg Val Ser Phe

690 695 700

Leu Asp Ala Val Ser Phe Arg Gly Leu Leu Glu Cys Pro Leu Asp Cys

705 710 715 720

Ser Gly Pro Val Cys Pro Ala Phe Cys Phe Ile Gly Gly Gly Ala Val

725 730 735

Gln Leu Gln Glu Pro Val Gly Thr Lys Pro Glu Arg Lys Val Thr Ile

740 745 750

Gly Gly Phe Ala Lys Leu Asp

755

<210> 93

<211> 1917

<212> DNA

<213> 智人(Homo sapiens)

<400> 93

atggacggca gtggaccctt cagctgcccc atctgcctag agccactccg ggagccggtg 60

acgctgccct gcggccacaa cttctgtctc gcctgcctgg gcgcgctctg gccgcatcgt 120

ggcgcgagtg gagccggcgg acccggaggc gcggcccgct gcccgctgtg ccaggagccc 180

ttccccgacg gccttcagct ccgcaagaac cacacgctgt ccgagctgct gcagctccgc 240

cagggctcgg gccccgggtc cggccccggc ccggcccctg ccctggcccc ggagccctcg 300

gcacccagcg cgctgcccag tgtcccggag ccgtcggccc cctgcgctcc cgagccgtgg 360

cccgcgggcg aagagccagt gcgctgcgac gcgtgccccg agggcgcggc cctgcccgcc 420

gcgctgtcct gcctctcctg cctcgcctcc ttttgccccg cgcacctggg cccgcacgag 480

cgcagccccg ccctccgcgg acaccgcctg gtgccgccgc tgcgccggct agaggagagc 540

ctgtgcccgc gccacctacg gccgctcgag cgctactgcc gcgcggagcg cgtgtgtctg 600

tgcgaggcct gcgccgcaca ggagcaccgc ggccacgagc tggtgccgct ggagcaggag 660

cgcgcgcttc aggaggctga gcagtccaaa gtcctgagcg ccgtggagga ccgcatggac 720

gagctgggtg ctggcattgc acagtccagg cgcacagtgg ccctcatcaa gagtgcagcc 780

gtagcagagc gggagagggt gagccggctg tttgcagatg ctgcggccgc cctgcagggc 840

ttccagaccc aggtgctggg cttcatcgag gagggggaag ctgccatgct aggccgctcc 900

cagggtgacc tgcggcgaca ggaggaacag cgcagccgcc tgagccgagc ccgccagaat 960

ctcagccagg tccctgaagc tgactcagtc agcttcctgc aggagctgct ggcactaagg 1020

ctggccctgg aggatgggtg tggccctggg cctggacccc cgagggagct cagcttcacc 1080

aaatcatccc aagctgtccg tgcagtgaga gacatgctgg ccgtggcctg cgtcaaccag 1140

tgggagcagc tgagggggcc gggtggcaac gaggatgggc cacagaagct ggactcggaa 1200

gctgatgctg agccccaaga cctcgagagt acgaacctct tggagagtga agctcccagg 1260

gactatttcc tcaagtttgc ctatattgtg gatttggaca gcgacacagc agacaagttc 1320

ctgcagctgt ttggaaccaa aggtgtcaag agggtgctgt gtcctatcaa ctaccccttg 1380

tcgcccaccc gcttcaccca ttgtgagcag gtgctgggcg agggtgccct ggaccgaggc 1440

acctactact gggaggtgga gattatcgag ggctgggtca gcatgggggt catggccgaa 1500

gacttctccc cacaagagcc ctacgaccgc ggccggctgg gccgcaacgc ccactcctgc 1560

tgcctgcagt ggaatggacg cagcttctcc gtctggtttc atgggctgga ggctcccctg 1620

ccccacccct tctcgcccac ggttggggtc tgcctggaat acgctgaccg tgccttggcc 1680

ttctatgctg tacgggacgg caagatgagc ctcctgcgga ggctgaaggc ctcccggccc 1740

cgccggggtg gcatcccggc ctcccccatt gaccccttcc agagccgcct ggacagtcac 1800

tttgcggggc tcttcaccca cagactcaag cctgccttct tcctggagag tgtggacgcc 1860

cacttgcaga tcgggcccct caagaagtcc tgcatatccg tgctgaagag gaggtga 1917

<210> 94

<211> 638

<212> PRT

<213> 智人(Homo sapiens)

<400> 94

Met Asp Gly Ser Gly Pro Phe Ser Cys Pro Ile Cys Leu Glu Pro Leu

1 5 10 15

Arg Glu Pro Val Thr Leu Pro Cys Gly His Asn Phe Cys Leu Ala Cys

20 25 30

Leu Gly Ala Leu Trp Pro His Arg Gly Ala Ser Gly Ala Gly Gly Pro

35 40 45

Gly Gly Ala Ala Arg Cys Pro Leu Cys Gln Glu Pro Phe Pro Asp Gly

50 55 60

Leu Gln Leu Arg Lys Asn His Thr Leu Ser Glu Leu Leu Gln Leu Arg

65 70 75 80

Gln Gly Ser Gly Pro Gly Ser Gly Pro Gly Pro Ala Pro Ala Leu Ala

85 90 95

Pro Glu Pro Ser Ala Pro Ser Ala Leu Pro Ser Val Pro Glu Pro Ser

100 105 110

Ala Pro Cys Ala Pro Glu Pro Trp Pro Ala Gly Glu Glu Pro Val Arg

115 120 125

Cys Asp Ala Cys Pro Glu Gly Ala Ala Leu Pro Ala Ala Leu Ser Cys

130 135 140

Leu Ser Cys Leu Ala Ser Phe Cys Pro Ala His Leu Gly Pro His Glu

145 150 155 160

Arg Ser Pro Ala Leu Arg Gly His Arg Leu Val Pro Pro Leu Arg Arg

165 170 175

Leu Glu Glu Ser Leu Cys Pro Arg His Leu Arg Pro Leu Glu Arg Tyr

180 185 190

Cys Arg Ala Glu Arg Val Cys Leu Cys Glu Ala Cys Ala Ala Gln Glu

195 200 205

His Arg Gly His Glu Leu Val Pro Leu Glu Gln Glu Arg Ala Leu Gln

210 215 220

Glu Ala Glu Gln Ser Lys Val Leu Ser Ala Val Glu Asp Arg Met Asp

225 230 235 240

Glu Leu Gly Ala Gly Ile Ala Gln Ser Arg Arg Thr Val Ala Leu Ile

245 250 255

Lys Ser Ala Ala Val Ala Glu Arg Glu Arg Val Ser Arg Leu Phe Ala

260 265 270

Asp Ala Ala Ala Ala Leu Gln Gly Phe Gln Thr Gln Val Leu Gly Phe

275 280 285

Ile Glu Glu Gly Glu Ala Ala Met Leu Gly Arg Ser Gln Gly Asp Leu

290 295 300

Arg Arg Gln Glu Glu Gln Arg Ser Arg Leu Ser Arg Ala Arg Gln Asn

305 310 315 320

Leu Ser Gln Val Pro Glu Ala Asp Ser Val Ser Phe Leu Gln Glu Leu

325 330 335

Leu Ala Leu Arg Leu Ala Leu Glu Asp Gly Cys Gly Pro Gly Pro Gly

340 345 350

Pro Pro Arg Glu Leu Ser Phe Thr Lys Ser Ser Gln Ala Val Arg Ala

355 360 365

Val Arg Asp Met Leu Ala Val Ala Cys Val Asn Gln Trp Glu Gln Leu

370 375 380

Arg Gly Pro Gly Gly Asn Glu Asp Gly Pro Gln Lys Leu Asp Ser Glu

385 390 395 400

Ala Asp Ala Glu Pro Gln Asp Leu Glu Ser Thr Asn Leu Leu Glu Ser

405 410 415

Glu Ala Pro Arg Asp Tyr Phe Leu Lys Phe Ala Tyr Ile Val Asp Leu

420 425 430

Asp Ser Asp Thr Ala Asp Lys Phe Leu Gln Leu Phe Gly Thr Lys Gly

435 440 445

Val Lys Arg Val Leu Cys Pro Ile Asn Tyr Pro Leu Ser Pro Thr Arg

450 455 460

Phe Thr His Cys Glu Gln Val Leu Gly Glu Gly Ala Leu Asp Arg Gly

465 470 475 480

Thr Tyr Tyr Trp Glu Val Glu Ile Ile Glu Gly Trp Val Ser Met Gly

485 490 495

Val Met Ala Glu Asp Phe Ser Pro Gln Glu Pro Tyr Asp Arg Gly Arg

500 505 510

Leu Gly Arg Asn Ala His Ser Cys Cys Leu Gln Trp Asn Gly Arg Ser

515 520 525

Phe Ser Val Trp Phe His Gly Leu Glu Ala Pro Leu Pro His Pro Phe

530 535 540

Ser Pro Thr Val Gly Val Cys Leu Glu Tyr Ala Asp Arg Ala Leu Ala

545 550 555 560

Phe Tyr Ala Val Arg Asp Gly Lys Met Ser Leu Leu Arg Arg Leu Lys

565 570 575

Ala Ser Arg Pro Arg Arg Gly Gly Ile Pro Ala Ser Pro Ile Asp Pro

580 585 590

Phe Gln Ser Arg Leu Asp Ser His Phe Ala Gly Leu Phe Thr His Arg

595 600 605

Leu Lys Pro Ala Phe Phe Leu Glu Ser Val Asp Ala His Leu Gln Ile

610 615 620

Gly Pro Leu Lys Lys Ser Cys Ile Ser Val Leu Lys Arg Arg

625 630 635

<210> 95

<211> 675

<212> DNA

<213> 智人(Homo sapiens)

<400> 95

atgtctcgaa gaatcattgt gggaaccctt caaagaaccc agcgaaacat gaattctgga 60

atctcgcaag tcttccagag ggaactcacc tgccccatct gcatgaacta cttcatagac 120

ccggtcacca tagactgtgg gcacagcttt tgcaggccct gtttctacct caactggcaa 180

gacatcccaa ttcttactca gtgctttgaa tgcataaaga caatacagca gagaaacctc 240

aaaactaaca ttcgattgaa gaagatggct tcccttgcca gaaaagccag tctctggcta 300

ttcctgagct ctgaggagca aatgtgtggc attcacaggg agacaaagaa gatgttctgt 360

gaagtggaca ggagcctgct ctgtttgctg tgctccagct ctcaggagca ccggtatcac 420

agacactgtc ccgctgagtg ggctgctgag gaacactggg agaagctttt aaagaaaatg 480

cagtctttat gggaaaaagc ttgtgaaaat cagagaaacc tgaatgtgga aaccaccaga 540

atcagccact ggaaggcttt tggagacata ttatacagga gtgagtccgt gctgctgcac 600

atgccccagc ctctgaatct agcgctcagg gcagggccca tcactggact gagggacagg 660

ctcaaccaat tctga 675

<210> 96

<211> 224

<212> PRT

<213> 智人(Homo sapiens)

<400> 96

Met Ser Arg Arg Ile Ile Val Gly Thr Leu Gln Arg Thr Gln Arg Asn

1 5 10 15

Met Asn Ser Gly Ile Ser Gln Val Phe Gln Arg Glu Leu Thr Cys Pro

20 25 30

Ile Cys Met Asn Tyr Phe Ile Asp Pro Val Thr Ile Asp Cys Gly His

35 40 45

Ser Phe Cys Arg Pro Cys Phe Tyr Leu Asn Trp Gln Asp Ile Pro Ile

50 55 60

Leu Thr Gln Cys Phe Glu Cys Ile Lys Thr Ile Gln Gln Arg Asn Leu

65 70 75 80

Lys Thr Asn Ile Arg Leu Lys Lys Met Ala Ser Leu Ala Arg Lys Ala

85 90 95

Ser Leu Trp Leu Phe Leu Ser Ser Glu Glu Gln Met Cys Gly Ile His

100 105 110

Arg Glu Thr Lys Lys Met Phe Cys Glu Val Asp Arg Ser Leu Leu Cys

115 120 125

Leu Leu Cys Ser Ser Ser Gln Glu His Arg Tyr His Arg His Cys Pro

130 135 140

Ala Glu Trp Ala Ala Glu Glu His Trp Glu Lys Leu Leu Lys Lys Met

145 150 155 160

Gln Ser Leu Trp Glu Lys Ala Cys Glu Asn Gln Arg Asn Leu Asn Val

165 170 175

Glu Thr Thr Arg Ile Ser His Trp Lys Ala Phe Gly Asp Ile Leu Tyr

180 185 190

Arg Ser Glu Ser Val Leu Leu His Met Pro Gln Pro Leu Asn Leu Ala

195 200 205

Leu Arg Ala Gly Pro Ile Thr Gly Leu Arg Asp Arg Leu Asn Gln Phe

210 215 220

<210> 97

<211> 1359

<212> DNA

<213> 智人(Homo sapiens)

<400> 97

atgaattctg gaatcttaca ggtctttcag ggggaactca tctgccccct gtgcatgaac 60

tacttcatag acccggtcac catagactgt gggcacagct tttgcaggcc ttgtttctac 120

ctcaactggc aagacatccc atttcttgtc cagtgctctg aatgcacaaa gtcaaccgag 180

cagataaacc tcaaaaccaa cattcatttg aagaagatgg cttctcttgc cagaaaagtc 240

agtctctggc tattcctgag ctctgaggag caaatgtgtg gcactcacag ggagacaaag 300

aagatattct gtgaagtgga caggagcctg ctctgtttgc tgtgctccag ctctcaggag 360

caccggtatc acagacaccg tcccattgag tgggctgctg aggaacaccg ggagaagctt 420

ttacagaaaa tgcagtcttt gtgggaaaaa gcttgtgaaa atcacagaaa cctgaatgtg 480

gaaaccacca gaaccagatg ctggaaggat tatgtgaatt taaggctaga agcaattaga 540

gctgagtatc agaagatgcc tgcatttcat catgaagaag aaaaacataa tttggagatg 600

ctgaaaaaga aggggaaaga aatttttcat cgacttcatt taagtaaagc caaaatggct 660

cataggatgg agattttaag aggaatgtat gaggagctga acgaaatgtg ccataaacca 720

gatgtggagc tacttcaggc ttttggagac atattacaca ggagtgagtc cgtgctgctg 780

cacatgcccc agcctctgaa tccagagctc agtgcagggc ccatcactgg actgagggac 840

aggctcaacc aattccgagt gcatattact ctgcatcatg aagaagccaa caatgatatc 900

tttctgtatg aaattttgag aagcatgtgt attggatgtg accatcaaga tgtaccctat 960

ttcactgcaa cacctagaag ttttcttgca tggggtgttc agactttcac ctcgggcaaa 1020

tattactggg aggtccatgt aggggactcc tggaattggg cttttggtgt ctgtaatatg 1080

tatcggaaag agaagaatca gaatgagaag atagatggaa aggcgggact ctttcttctt 1140

gggtgtgtta agaatgacat tcaatgcagt ctctttacca cctccccact tatgctgcaa 1200

tatatcccaa aacctaccag ccgagtagga ttattcctgg attgtgaggc taagactgtg 1260

agctttgttg atgttaatca aagctcccta atatacacca tccctaattg ctctttctca 1320

cctcctctca ggcctatctt ttgctgtatt cacttctga 1359

<210> 98

<211> 452

<212> PRT

<213> 智人(Homo sapiens)

<400> 98

Met Asn Ser Gly Ile Leu Gln Val Phe Gln Gly Glu Leu Ile Cys Pro

1 5 10 15

Leu Cys Met Asn Tyr Phe Ile Asp Pro Val Thr Ile Asp Cys Gly His

20 25 30

Ser Phe Cys Arg Pro Cys Phe Tyr Leu Asn Trp Gln Asp Ile Pro Phe

35 40 45

Leu Val Gln Cys Ser Glu Cys Thr Lys Ser Thr Glu Gln Ile Asn Leu

50 55 60

Lys Thr Asn Ile His Leu Lys Lys Met Ala Ser Leu Ala Arg Lys Val

65 70 75 80

Ser Leu Trp Leu Phe Leu Ser Ser Glu Glu Gln Met Cys Gly Thr His

85 90 95

Arg Glu Thr Lys Lys Ile Phe Cys Glu Val Asp Arg Ser Leu Leu Cys

100 105 110

Leu Leu Cys Ser Ser Ser Gln Glu His Arg Tyr His Arg His Arg Pro

115 120 125

Ile Glu Trp Ala Ala Glu Glu His Arg Glu Lys Leu Leu Gln Lys Met

130 135 140

Gln Ser Leu Trp Glu Lys Ala Cys Glu Asn His Arg Asn Leu Asn Val

145 150 155 160

Glu Thr Thr Arg Thr Arg Cys Trp Lys Asp Tyr Val Asn Leu Arg Leu

165 170 175

Glu Ala Ile Arg Ala Glu Tyr Gln Lys Met Pro Ala Phe His His Glu

180 185 190

Glu Glu Lys His Asn Leu Glu Met Leu Lys Lys Lys Gly Lys Glu Ile

195 200 205

Phe His Arg Leu His Leu Ser Lys Ala Lys Met Ala His Arg Met Glu

210 215 220

Ile Leu Arg Gly Met Tyr Glu Glu Leu Asn Glu Met Cys His Lys Pro

225 230 235 240

Asp Val Glu Leu Leu Gln Ala Phe Gly Asp Ile Leu His Arg Ser Glu

245 250 255

Ser Val Leu Leu His Met Pro Gln Pro Leu Asn Pro Glu Leu Ser Ala

260 265 270

Gly Pro Ile Thr Gly Leu Arg Asp Arg Leu Asn Gln Phe Arg Val His

275 280 285

Ile Thr Leu His His Glu Glu Ala Asn Asn Asp Ile Phe Leu Tyr Glu

290 295 300

Ile Leu Arg Ser Met Cys Ile Gly Cys Asp His Gln Asp Val Pro Tyr

305 310 315 320

Phe Thr Ala Thr Pro Arg Ser Phe Leu Ala Trp Gly Val Gln Thr Phe

325 330 335

Thr Ser Gly Lys Tyr Tyr Trp Glu Val His Val Gly Asp Ser Trp Asn

340 345 350

Trp Ala Phe Gly Val Cys Asn Met Tyr Arg Lys Glu Lys Asn Gln Asn

355 360 365

Glu Lys Ile Asp Gly Lys Ala Gly Leu Phe Leu Leu Gly Cys Val Lys

370 375 380

Asn Asp Ile Gln Cys Ser Leu Phe Thr Thr Ser Pro Leu Met Leu Gln

385 390 395 400

Tyr Ile Pro Lys Pro Thr Ser Arg Val Gly Leu Phe Leu Asp Cys Glu

405 410 415

Ala Lys Thr Val Ser Phe Val Asp Val Asn Gln Ser Ser Leu Ile Tyr

420 425 430

Thr Ile Pro Asn Cys Ser Phe Ser Pro Pro Leu Arg Pro Ile Phe Cys

435 440 445

Cys Ile His Phe

450

<210> 99

<211> 1464

<212> DNA

<213> 智人(Homo sapiens)

<400> 99

atggcttggc aggtgagcct gccagagctg gaggaccggc ttcagtgtcc catctgcctg 60

gaggtcttca aggagcccct gatgctgcag tgtggccact cttactgcaa gggctgcctg 120

gtttccctgt cctgccacct ggatgccgag ctgcgctgcc ccgtgtgccg gcaggcggtg 180

gacggcagca gctccctgcc caacgtctcc ctggccaggg tgatcgaagc cctgaggctc 240

cctggggacc cggagcccaa ggtctgcgtg caccaccgga acccgctcag ccttttctgc 300

gagaaggacc aggagctcat ctgtggcctc tgcggtctgc tgggctccca ccaacaccac 360

ccggtcacgc ccgtctccac cgtctacagc cgcatgaagg aggagctcgc agccctcatc 420

tctgagctga agcaggagca gaaaaaggtg gatgagctca tcgccaaact ggtgaacaac 480

cggacccgaa tcgtcaatga gtcggatgtc ttcagctggg tgatccgccg cgagttccag 540

gagctgcacc acctggtgga tgaggagaag gcccgctgcc tggaggggat agggggtcac 600

acccgtggcc tggtggcctc cctggacatg cagctggagc aggcccaggg aacccgggag 660

cggctggccc aagccgagtg tgtgctggaa cagttcggca atgaggacca ccacaagttc 720

atccggaagt tccactccat ggcctccaga gcagagatgc cgcaggcccg gcccttagaa 780

ggcgcattca gccccatctc cttcaagcca ggcctccacc aggctgacat caagctgacc 840

gtgtggaaaa ggctcttccg gaaagttttg ccagccccgg agcctctcaa gttggaccct 900

gccactgccc acccactcct ggagctctcc aagggcaaca cggtggtgca gtgcgggctt 960

ctggcccagc ggcgagccag ccagcctgag cgcttcgact acagcacctg cgtcctggcc 1020

agccgcggct tctcctgcgg ccgccactac tgggaggtgg tggtgggcag caagagcgac 1080

tggcgcctgg gggtcatcaa gggcacagcc agccgtaagg gcaagctgaa caggtccccc 1140

gagcacggcg tgtggctgat cggcctgaag gagggccggg tgtacgaagc ctttgcctgc 1200

ccccgggtac ccctgcccgt ggccggccac ccccaccgca tcgggctcta cctgcactat 1260

gagcagggcg aactcacctt cttcgatgcc gaccgccccg atgacctgcg gccgctctac 1320

accttccagg ccgacttcca gggcaagctc taccccatcc tggacacctg ctggcacgag 1380

aggggcagca actcgctgcc catggtgctg cccccgccca gcgggcctgg ccccctcagc 1440

cccgagcagc ccaccaagct gtag 1464

<210> 100

<211> 487

<212> PRT

<213> 智人(Homo sapiens)

<400> 100

Met Ala Trp Gln Val Ser Leu Pro Glu Leu Glu Asp Arg Leu Gln Cys

1 5 10 15

Pro Ile Cys Leu Glu Val Phe Lys Glu Pro Leu Met Leu Gln Cys Gly

20 25 30

His Ser Tyr Cys Lys Gly Cys Leu Val Ser Leu Ser Cys His Leu Asp

35 40 45

Ala Glu Leu Arg Cys Pro Val Cys Arg Gln Ala Val Asp Gly Ser Ser

50 55 60

Ser Leu Pro Asn Val Ser Leu Ala Arg Val Ile Glu Ala Leu Arg Leu

65 70 75 80

Pro Gly Asp Pro Glu Pro Lys Val Cys Val His His Arg Asn Pro Leu

85 90 95

Ser Leu Phe Cys Glu Lys Asp Gln Glu Leu Ile Cys Gly Leu Cys Gly

100 105 110

Leu Leu Gly Ser His Gln His His Pro Val Thr Pro Val Ser Thr Val

115 120 125

Tyr Ser Arg Met Lys Glu Glu Leu Ala Ala Leu Ile Ser Glu Leu Lys

130 135 140

Gln Glu Gln Lys Lys Val Asp Glu Leu Ile Ala Lys Leu Val Asn Asn

145 150 155 160

Arg Thr Arg Ile Val Asn Glu Ser Asp Val Phe Ser Trp Val Ile Arg

165 170 175

Arg Glu Phe Gln Glu Leu His His Leu Val Asp Glu Glu Lys Ala Arg

180 185 190

Cys Leu Glu Gly Ile Gly Gly His Thr Arg Gly Leu Val Ala Ser Leu

195 200 205

Asp Met Gln Leu Glu Gln Ala Gln Gly Thr Arg Glu Arg Leu Ala Gln

210 215 220

Ala Glu Cys Val Leu Glu Gln Phe Gly Asn Glu Asp His His Lys Phe

225 230 235 240

Ile Arg Lys Phe His Ser Met Ala Ser Arg Ala Glu Met Pro Gln Ala

245 250 255

Arg Pro Leu Glu Gly Ala Phe Ser Pro Ile Ser Phe Lys Pro Gly Leu

260 265 270

His Gln Ala Asp Ile Lys Leu Thr Val Trp Lys Arg Leu Phe Arg Lys

275 280 285

Val Leu Pro Ala Pro Glu Pro Leu Lys Leu Asp Pro Ala Thr Ala His

290 295 300

Pro Leu Leu Glu Leu Ser Lys Gly Asn Thr Val Val Gln Cys Gly Leu

305 310 315 320

Leu Ala Gln Arg Arg Ala Ser Gln Pro Glu Arg Phe Asp Tyr Ser Thr

325 330 335

Cys Val Leu Ala Ser Arg Gly Phe Ser Cys Gly Arg His Tyr Trp Glu

340 345 350

Val Val Val Gly Ser Lys Ser Asp Trp Arg Leu Gly Val Ile Lys Gly

355 360 365

Thr Ala Ser Arg Lys Gly Lys Leu Asn Arg Ser Pro Glu His Gly Val

370 375 380

Trp Leu Ile Gly Leu Lys Glu Gly Arg Val Tyr Glu Ala Phe Ala Cys

385 390 395 400

Pro Arg Val Pro Leu Pro Val Ala Gly His Pro His Arg Ile Gly Leu

405 410 415

Tyr Leu His Tyr Glu Gln Gly Glu Leu Thr Phe Phe Asp Ala Asp Arg

420 425 430

Pro Asp Asp Leu Arg Pro Leu Tyr Thr Phe Gln Ala Asp Phe Gln Gly

435 440 445

Lys Leu Tyr Pro Ile Leu Asp Thr Cys Trp His Glu Arg Gly Ser Asn

450 455 460

Ser Leu Pro Met Val Leu Pro Pro Pro Ser Gly Pro Gly Pro Leu Ser

465 470 475 480

Pro Glu Gln Pro Thr Lys Leu

485

<210> 101

<211> 1359

<212> DNA

<213> 智人(Homo sapiens)

<400> 101

atgaattctg gaatcttgca agtcttccag agggcactca cctgtcccat ctgcatgaac 60

tacttcctag acccagtcac catagactgt gggcacagct tttgccggcc ctgtttgtac 120

ctcaactggc aagacacggc agttcttgct cagtgctctg aatgcaagaa gacaacgcgg 180

cagagaaacc tcaacactga catttgtttg aagaacatgg ctttcattgc cagaaaagcc 240

agcctccggc aattccttag ctctgaggag caaatatgtg ggatgcacag agagacaaag 300

aagatgttct gtgaagtgga caagagcctg ctctgtttgc cgtgctccaa ctctcaggag 360

caccggaatc acatacactg tcccattgag tgggctgctg aggaacgccg ggaggagctc 420

ctaaaaaaaa tgcagtcttt atgggaaaaa gcttgtgaaa atctcagaaa tctgaacatg 480

gaaaccacaa gaaccagatg ctggaaggat tatgtgagtt taaggataga agcaatcaga 540

gctgaatatc agaagatgcc tgcatttctc catgaagaag agcaacatca cttggaaagg 600

ctgcgaaagg agggcgagga catttttcag caactcaatg aaagcaaagc cagaatggaa 660

cattccaggg agcttttaag aggaatgtat gaggatctga agcaaatgtg ccataaagca 720

gatgtggagc tactccaggc ttttggagac atattacaca ggtatgagtc tctgctgctg 780

caagtgtctg agcctgtgaa tccagagctc agtgcagggc ccatcactgg actgctggac 840

agcctcagtg gattcagagt tgattttact ctgcagcctg aaagagccaa tagtcatatc 900

ttcctgtgtg gagatttgag aagcatgaat gttggatgtg accctcaaga tgatcccgat 960

atcactggaa aatctgaatg ttttcttgta tggggggctc aggctttcac atctggcaaa 1020

tattattggg aggttcatat gggggactct tggaattggg cttttggtgt ctgtaacaat 1080

tattggaaag agaagagaca gaatgacaag atagatggag aggagggact ctttcttctt 1140

ggatgtgtta aggaggacac tcactgcagt ctctttacca cctccccact tgtggtgcaa 1200

tatgttccaa gacctaccag cacagtagga ttattcctgg attgtgaagg tagaaccgtg 1260

agctttgttg atgttgatca aagttccctg atatacacca tccccaattg ctccttctca 1320

cctcctctca ggcctatctt ttgctgtagt cacttctga 1359

<210> 102

<211> 452

<212> PRT

<213> 智人(Homo sapiens)

<400> 102

Met Asn Ser Gly Ile Leu Gln Val Phe Gln Arg Ala Leu Thr Cys Pro

1 5 10 15

Ile Cys Met Asn Tyr Phe Leu Asp Pro Val Thr Ile Asp Cys Gly His

20 25 30

Ser Phe Cys Arg Pro Cys Leu Tyr Leu Asn Trp Gln Asp Thr Ala Val

35 40 45

Leu Ala Gln Cys Ser Glu Cys Lys Lys Thr Thr Arg Gln Arg Asn Leu

50 55 60

Asn Thr Asp Ile Cys Leu Lys Asn Met Ala Phe Ile Ala Arg Lys Ala

65 70 75 80

Ser Leu Arg Gln Phe Leu Ser Ser Glu Glu Gln Ile Cys Gly Met His

85 90 95

Arg Glu Thr Lys Lys Met Phe Cys Glu Val Asp Lys Ser Leu Leu Cys

100 105 110

Leu Pro Cys Ser Asn Ser Gln Glu His Arg Asn His Ile His Cys Pro

115 120 125

Ile Glu Trp Ala Ala Glu Glu Arg Arg Glu Glu Leu Leu Lys Lys Met

130 135 140

Gln Ser Leu Trp Glu Lys Ala Cys Glu Asn Leu Arg Asn Leu Asn Met

145 150 155 160

Glu Thr Thr Arg Thr Arg Cys Trp Lys Asp Tyr Val Ser Leu Arg Ile

165 170 175

Glu Ala Ile Arg Ala Glu Tyr Gln Lys Met Pro Ala Phe Leu His Glu

180 185 190

Glu Glu Gln His His Leu Glu Arg Leu Arg Lys Glu Gly Glu Asp Ile

195 200 205

Phe Gln Gln Leu Asn Glu Ser Lys Ala Arg Met Glu His Ser Arg Glu

210 215 220

Leu Leu Arg Gly Met Tyr Glu Asp Leu Lys Gln Met Cys His Lys Ala

225 230 235 240

Asp Val Glu Leu Leu Gln Ala Phe Gly Asp Ile Leu His Arg Tyr Glu

245 250 255

Ser Leu Leu Leu Gln Val Ser Glu Pro Val Asn Pro Glu Leu Ser Ala

260 265 270

Gly Pro Ile Thr Gly Leu Leu Asp Ser Leu Ser Gly Phe Arg Val Asp

275 280 285

Phe Thr Leu Gln Pro Glu Arg Ala Asn Ser His Ile Phe Leu Cys Gly

290 295 300

Asp Leu Arg Ser Met Asn Val Gly Cys Asp Pro Gln Asp Asp Pro Asp

305 310 315 320

Ile Thr Gly Lys Ser Glu Cys Phe Leu Val Trp Gly Ala Gln Ala Phe

325 330 335

Thr Ser Gly Lys Tyr Tyr Trp Glu Val His Met Gly Asp Ser Trp Asn

340 345 350

Trp Ala Phe Gly Val Cys Asn Asn Tyr Trp Lys Glu Lys Arg Gln Asn

355 360 365

Asp Lys Ile Asp Gly Glu Glu Gly Leu Phe Leu Leu Gly Cys Val Lys

370 375 380

Glu Asp Thr His Cys Ser Leu Phe Thr Thr Ser Pro Leu Val Val Gln

385 390 395 400

Tyr Val Pro Arg Pro Thr Ser Thr Val Gly Leu Phe Leu Asp Cys Glu

405 410 415

Gly Arg Thr Val Ser Phe Val Asp Val Asp Gln Ser Ser Leu Ile Tyr

420 425 430

Thr Ile Pro Asn Cys Ser Phe Ser Pro Pro Leu Arg Pro Ile Phe Cys

435 440 445

Cys Ser His Phe

450

<210> 103

<211> 894

<212> DNA

<213> 智人(Homo sapiens)

<400> 103

atggctggtt atgccactac tcccagcccc atgcagaccc ttcaggagga agcggtgtgt 60

gccatctgct tggattactt caaggacccc gtgtccatca gctgtgggca caacttctgc 120

cgagggtgtg tgacccagct gtggagtaag gaggacgagg aggaccagaa cgaggaggaa 180

gatgaatggg aggaggagga ggacgaggaa gcggtggggg ccatggatgg atgggacggc 240

tccattcgag aggtgttgta tcgggggaat gctgacgaag agttgttcca agaccaagat 300

gacgatgaac tctggctcgg tgacagtggt ataactaatt gggacaacgt agactatatg 360

tgggacgagg aggaagaaga agaagaggaa gatcaggact attacctagg aggcttgaga 420

cctgacctga gaattgatgt ctaccgagaa gaagaaatac tggaagcata cgatgaggac 480

gaagatgaag agctgtatcc tgacatccac ccgcctcctt ccttgcccct tccagggcag 540

ttcacctgcc cccagtgccg aaagagcttt acacgtcgca gctttcgtcc caacttgcag 600

ctggccaaca tggtccagat aattcgccag atgtgcccca ctccttatcg gggaaaccgg 660

agtaatgatc agggcatgtg ctttaaacac caggaagccc tgaaactctt ctgtgaggtg 720

gacaaagagg ccatctgtgt ggtgtgccga gaatccagga gccacaaaca gcacagcgtg 780

ctgcctttgg aggaggtggt gcaggagtac caggaaataa agttggaaac aactctggtg 840

ggaatacttc agatagagca agaaagcatt cacagcaagg cctataatca gtaa 894

<210> 104

<211> 297

<212> PRT

<213> 智人(Homo sapiens)

<400> 104

Met Ala Gly Tyr Ala Thr Thr Pro Ser Pro Met Gln Thr Leu Gln Glu

1 5 10 15

Glu Ala Val Cys Ala Ile Cys Leu Asp Tyr Phe Lys Asp Pro Val Ser

20 25 30

Ile Ser Cys Gly His Asn Phe Cys Arg Gly Cys Val Thr Gln Leu Trp

35 40 45

Ser Lys Glu Asp Glu Glu Asp Gln Asn Glu Glu Glu Asp Glu Trp Glu

50 55 60

Glu Glu Glu Asp Glu Glu Ala Val Gly Ala Met Asp Gly Trp Asp Gly

65 70 75 80

Ser Ile Arg Glu Val Leu Tyr Arg Gly Asn Ala Asp Glu Glu Leu Phe

85 90 95

Gln Asp Gln Asp Asp Asp Glu Leu Trp Leu Gly Asp Ser Gly Ile Thr

100 105 110

Asn Trp Asp Asn Val Asp Tyr Met Trp Asp Glu Glu Glu Glu Glu Glu

115 120 125

Glu Glu Asp Gln Asp Tyr Tyr Leu Gly Gly Leu Arg Pro Asp Leu Arg

130 135 140

Ile Asp Val Tyr Arg Glu Glu Glu Ile Leu Glu Ala Tyr Asp Glu Asp

145 150 155 160

Glu Asp Glu Glu Leu Tyr Pro Asp Ile His Pro Pro Pro Ser Leu Pro

165 170 175

Leu Pro Gly Gln Phe Thr Cys Pro Gln Cys Arg Lys Ser Phe Thr Arg

180 185 190

Arg Ser Phe Arg Pro Asn Leu Gln Leu Ala Asn Met Val Gln Ile Ile

195 200 205

Arg Gln Met Cys Pro Thr Pro Tyr Arg Gly Asn Arg Ser Asn Asp Gln

210 215 220

Gly Met Cys Phe Lys His Gln Glu Ala Leu Lys Leu Phe Cys Glu Val

225 230 235 240

Asp Lys Glu Ala Ile Cys Val Val Cys Arg Glu Ser Arg Ser His Lys

245 250 255

Gln His Ser Val Leu Pro Leu Glu Glu Val Val Gln Glu Tyr Gln Glu

260 265 270

Ile Lys Leu Glu Thr Thr Leu Val Gly Ile Leu Gln Ile Glu Gln Glu

275 280 285

Ser Ile His Ser Lys Ala Tyr Asn Gln

290 295

<210> 105

<211> 1203

<212> DNA

<213> 智人(Homo sapiens)

<400> 105

atgaacttca cagtgggttt caagccgctg ctaggggatg cacacagcat ggacaacctg 60

gagaagcagc tcatctgccc catctgcctg gagatgttct ccaaaccagt ggtgatcctg 120

ccctgccaac acaacctgtg ccgcaaatgt gccaacgacg tcttccaggc ctcgaatcct 180

ctatggcagt cccggggctc caccactgtg tcttcaggag gccgtttccg ctgcccatcg 240

tgcaggcatg aggttgtcct ggacagacac ggtgtctacg gcctgcagcg aaacctgcta 300

gtggagaaca ttatcgacat ttacaagcag gagtcatcca ggccgctgca ctccaaggct 360

gagcagcacc tcatgtgcga ggagcatgaa gaagagaaga tcaatattta ctgcctgagc 420

tgtgaggtgc ccacctgctc tctctgcaag gtcttcggtg cccacaagga ctgtgaggtg 480

gccccactgc ccaccattta caaacgccag aagaaacagg atctcactct gttgcccagg 540

ctggagtgca gtggcacaaa cacaacttac tgcagccttg atctcccgag ctcaagtgat 600

cctcccatct tagcctcgca gaacactaag attatagata gtgagctcag cgatggcatc 660

gcgatgctgg tggcaggcaa tgaccgcgtg caagcagtga tcacacagat ggaggaggtg 720

tgccagacta tcgaggacaa tagccggagg cagaagcagt tgttaaacca gaggtttgag 780

agcctgtgcg cagtgctgga ggagcgcaag ggtgagctgc tgcaggcgct ggcccgggag 840

caagaggaga agctgcagcg cgtccgcggc ctcatccgtc agtatggcga ccacctggag 900

gcctcctcta agctggtgga gtctgccatc cagtccatgg aagagccaca aatggcgctg 960

tatctccagc aggccaagga gctgatcaat aaggtcgggg ccatgtcgaa ggtggagctg 1020

gcagggcggc cggagccagg ctatgagagc atggagcaat tcaccgtaag ggtggagcac 1080

gtggccgaaa tgctgcggac catcgacttc cagccaggcg cttccgggga ggaagaggag 1140

gtggccccag acggagagga gggcagcgcg gggccggagg aagagcggcc ggatgggcct 1200

taa 1203

<210> 106

<211> 400

<212> PRT

<213> 智人(Homo sapiens)

<400> 106

Met Asn Phe Thr Val Gly Phe Lys Pro Leu Leu Gly Asp Ala His Ser

1 5 10 15

Met Asp Asn Leu Glu Lys Gln Leu Ile Cys Pro Ile Cys Leu Glu Met

20 25 30

Phe Ser Lys Pro Val Val Ile Leu Pro Cys Gln His Asn Leu Cys Arg

35 40 45

Lys Cys Ala Asn Asp Val Phe Gln Ala Ser Asn Pro Leu Trp Gln Ser

50 55 60

Arg Gly Ser Thr Thr Val Ser Ser Gly Gly Arg Phe Arg Cys Pro Ser

65 70 75 80

Cys Arg His Glu Val Val Leu Asp Arg His Gly Val Tyr Gly Leu Gln

85 90 95

Arg Asn Leu Leu Val Glu Asn Ile Ile Asp Ile Tyr Lys Gln Glu Ser

100 105 110

Ser Arg Pro Leu His Ser Lys Ala Glu Gln His Leu Met Cys Glu Glu

115 120 125

His Glu Glu Glu Lys Ile Asn Ile Tyr Cys Leu Ser Cys Glu Val Pro

130 135 140

Thr Cys Ser Leu Cys Lys Val Phe Gly Ala His Lys Asp Cys Glu Val

145 150 155 160

Ala Pro Leu Pro Thr Ile Tyr Lys Arg Gln Lys Lys Gln Asp Leu Thr

165 170 175

Leu Leu Pro Arg Leu Glu Cys Ser Gly Thr Asn Thr Thr Tyr Cys Ser

180 185 190

Leu Asp Leu Pro Ser Ser Ser Asp Pro Pro Ile Leu Ala Ser Gln Asn

195 200 205

Thr Lys Ile Ile Asp Ser Glu Leu Ser Asp Gly Ile Ala Met Leu Val

210 215 220

Ala Gly Asn Asp Arg Val Gln Ala Val Ile Thr Gln Met Glu Glu Val

225 230 235 240

Cys Gln Thr Ile Glu Asp Asn Ser Arg Arg Gln Lys Gln Leu Leu Asn

245 250 255

Gln Arg Phe Glu Ser Leu Cys Ala Val Leu Glu Glu Arg Lys Gly Glu

260 265 270

Leu Leu Gln Ala Leu Ala Arg Glu Gln Glu Glu Lys Leu Gln Arg Val

275 280 285

Arg Gly Leu Ile Arg Gln Tyr Gly Asp His Leu Glu Ala Ser Ser Lys

290 295 300

Leu Val Glu Ser Ala Ile Gln Ser Met Glu Glu Pro Gln Met Ala Leu

305 310 315 320

Tyr Leu Gln Gln Ala Lys Glu Leu Ile Asn Lys Val Gly Ala Met Ser

325 330 335

Lys Val Glu Leu Ala Gly Arg Pro Glu Pro Gly Tyr Glu Ser Met Glu

340 345 350

Gln Phe Thr Val Arg Val Glu His Val Ala Glu Met Leu Arg Thr Ile

355 360 365

Asp Phe Gln Pro Gly Ala Ser Gly Glu Glu Glu Glu Val Ala Pro Asp

370 375 380

Gly Glu Glu Gly Ser Ala Gly Pro Glu Glu Glu Arg Pro Asp Gly Pro

385 390 395 400

<210> 107

<211> 1647

<212> DNA

<213> 智人(Homo sapiens)

<400> 107

atgagcgcat ctctgaatta caaatctttt tccaaagagc agcagaccat ggataactta 60

gagaagcaac tcatctgtcc catctgctta gagatgttca cgaaacctgt ggtgattctc 120

ccttgtcagc acaacctgtg taggaaatgt gccagtgata ttttccaggc ctctaacccg 180

tatttgccca caagaggagg taccaccatg gcatcagggg gccgattccg ctgcccatcc 240

tgtagacatg aagtggtttt ggatagacat ggggtatatg gacttcagag gaacctgctg 300

gtggaaaata tcattgacat ctacaagcag gagtccacca ggccagaaaa gaaatccgac 360

cagcccatgt gcgaggaaca tgaagaggag cgcatcaaca tctactgtct gaactgcgaa 420

gtacccacct gctctctgtg caaggtgttt ggtgcacaca aagactgcca ggtggctccc 480

ctcactcatg tgttccagag acagaagtct gagctcagtg atggcatcgc catcctcgtg 540

ggcagcaacg atcgagtcca gggagtgatc agccagctgg aagacacctg caaaactatc 600

gaggaatgtt gcagaaaaca gaaacaagag ctttgtgaga agtttgatta cctgtatggc 660

attttggagg agaggaagaa tgaaatgacc caagtcatta cccgaaccca agaggagaaa 720

ctggaacatg tccgtgctct gatcaaaaag tattctgatc atttggagaa cgtctcaaag 780

ttggttgagt caggaattca gtttatggat gagccagaaa tggcagtgtt tctgcagaat 840

gccaaaaccc tgctaaaaaa aatctcggaa gcatcaaagg catttcagat ggagaaaata 900

gaacatggct atgagaacat gaaccacttc acagtcaacc tcaatagaga agaaaagata 960

atacgtgaaa ttgactttta cagagaagat gaagatgaag aagaagaaga aggcggagaa 1020

ggagaaaaag aaggagaagg agaagtggga ggagaagcag tagaagtgga agaggtagaa 1080

aatgttcaaa cagagtttcc aggagaagat gaaaacccag aaaaagcttc agagctctct 1140

caggtggagc tgcaggctgc ccctggggca cttccagttt cctctccaga gccacctcca 1200

gccctgccac ctgctgcgga tgcccctgtg acacaggggg aggttgtacc cactggctct 1260

gagcagacca cagagtctga aactccagtc cctgcagcag cagaaactgc ggatcccttg 1320

ttttacccta gttggtataa aggccaaacc cggaaagcca ccaccaaccc accttgcacc 1380

ccagggagcg aaggtctggg gcaaataggg cctccaggtt ctgaggattc gaatgtacgg 1440

aaggcagaag tggcagcagc cgcagcgagt gagagggcag ctgtgagtgg taaggaaact 1500

agtgcacctg cagctacttc tcagattgga tttgaggctc ctcccctcca gggacaggct 1560

gcagctccag cgagtggcag tggagctgat tctgagccag ctcgccatat cttctccttt 1620

tcctggttga actccctaaa tgaatga 1647

<210> 108

<211> 548

<212> PRT

<213> 智人(Homo sapiens)

<400> 108

Met Ser Ala Ser Leu Asn Tyr Lys Ser Phe Ser Lys Glu Gln Gln Thr

1 5 10 15

Met Asp Asn Leu Glu Lys Gln Leu Ile Cys Pro Ile Cys Leu Glu Met

20 25 30

Phe Thr Lys Pro Val Val Ile Leu Pro Cys Gln His Asn Leu Cys Arg

35 40 45

Lys Cys Ala Ser Asp Ile Phe Gln Ala Ser Asn Pro Tyr Leu Pro Thr

50 55 60

Arg Gly Gly Thr Thr Met Ala Ser Gly Gly Arg Phe Arg Cys Pro Ser

65 70 75 80

Cys Arg His Glu Val Val Leu Asp Arg His Gly Val Tyr Gly Leu Gln

85 90 95

Arg Asn Leu Leu Val Glu Asn Ile Ile Asp Ile Tyr Lys Gln Glu Ser

100 105 110

Thr Arg Pro Glu Lys Lys Ser Asp Gln Pro Met Cys Glu Glu His Glu

115 120 125

Glu Glu Arg Ile Asn Ile Tyr Cys Leu Asn Cys Glu Val Pro Thr Cys

130 135 140

Ser Leu Cys Lys Val Phe Gly Ala His Lys Asp Cys Gln Val Ala Pro

145 150 155 160

Leu Thr His Val Phe Gln Arg Gln Lys Ser Glu Leu Ser Asp Gly Ile

165 170 175

Ala Ile Leu Val Gly Ser Asn Asp Arg Val Gln Gly Val Ile Ser Gln

180 185 190

Leu Glu Asp Thr Cys Lys Thr Ile Glu Glu Cys Cys Arg Lys Gln Lys

195 200 205

Gln Glu Leu Cys Glu Lys Phe Asp Tyr Leu Tyr Gly Ile Leu Glu Glu

210 215 220

Arg Lys Asn Glu Met Thr Gln Val Ile Thr Arg Thr Gln Glu Glu Lys

225 230 235 240

Leu Glu His Val Arg Ala Leu Ile Lys Lys Tyr Ser Asp His Leu Glu

245 250 255

Asn Val Ser Lys Leu Val Glu Ser Gly Ile Gln Phe Met Asp Glu Pro

260 265 270

Glu Met Ala Val Phe Leu Gln Asn Ala Lys Thr Leu Leu Lys Lys Ile

275 280 285

Ser Glu Ala Ser Lys Ala Phe Gln Met Glu Lys Ile Glu His Gly Tyr

290 295 300

Glu Asn Met Asn His Phe Thr Val Asn Leu Asn Arg Glu Glu Lys Ile

305 310 315 320

Ile Arg Glu Ile Asp Phe Tyr Arg Glu Asp Glu Asp Glu Glu Glu Glu

325 330 335

Glu Gly Gly Glu Gly Glu Lys Glu Gly Glu Gly Glu Val Gly Gly Glu

340 345 350

Ala Val Glu Val Glu Glu Val Glu Asn Val Gln Thr Glu Phe Pro Gly

355 360 365

Glu Asp Glu Asn Pro Glu Lys Ala Ser Glu Leu Ser Gln Val Glu Leu

370 375 380

Gln Ala Ala Pro Gly Ala Leu Pro Val Ser Ser Pro Glu Pro Pro Pro

385 390 395 400

Ala Leu Pro Pro Ala Ala Asp Ala Pro Val Thr Gln Gly Glu Val Val

405 410 415

Pro Thr Gly Ser Glu Gln Thr Thr Glu Ser Glu Thr Pro Val Pro Ala

420 425 430

Ala Ala Glu Thr Ala Asp Pro Leu Phe Tyr Pro Ser Trp Tyr Lys Gly

435 440 445

Gln Thr Arg Lys Ala Thr Thr Asn Pro Pro Cys Thr Pro Gly Ser Glu

450 455 460

Gly Leu Gly Gln Ile Gly Pro Pro Gly Ser Glu Asp Ser Asn Val Arg

465 470 475 480

Lys Ala Glu Val Ala Ala Ala Ala Ala Ser Glu Arg Ala Ala Val Ser

485 490 495

Gly Lys Glu Thr Ser Ala Pro Ala Ala Thr Ser Gln Ile Gly Phe Glu

500 505 510

Ala Pro Pro Leu Gln Gly Gln Ala Ala Ala Pro Ala Ser Gly Ser Gly

515 520 525

Ala Asp Ser Glu Pro Ala Arg His Ile Phe Ser Phe Ser Trp Leu Asn

530 535 540

Ser Leu Asn Glu

545

<210> 109

<211> 2225

<212> DNA

<213> 智人(Homo sapiens)

<400> 109

atggtttccc acgggtcctc gccctccctc ctggaggccc tgagcagcga cttcctggcc 60

tgtaaaatct gcctggagca gctgcgggca cccaagacac tgccctgcct gcatacctac 120

tgccaagact gcctggcaca gctggcggat ggcggccgcg tccgctgccc cgagtgccgc 180

gagacagtgc ctgtgccgcc cgagggtgtg gcctccttca agaccaactt cttcgtcaat 240

gggctgctgg acctggtgaa ggcccgggcc tgtggagacc tgcgtgccgg gaagccagcc 300

tgtgccctgt gtcccctggt gggtggcacc agcaccgggg ggccggccac ggcccggtgc 360

ctggactgtg ccgatgactt gtgccaggcc tgtgccgacg ggcaccgctg cacccgccag 420

acccacaccc accgcgtggt ggacctggtg ggctacaggg ccgggtggta tgatgaggag 480

gcccgggagc gccaagcggc ccagtgtccc cagcaccccg gggaggcact gcgcttcctg 540

tgccagccct gctcacagtt gctgtgcaga gagtgccgcc tagaccccca cctggaccac 600

ccctgcctgc ctctggctga agctgtgcgt gcccggaggc cgggcctgga gggactgctg 660

gccggtgtgg acaataacct ggtggagctg gaggcagcgc ggagggtgga gaaggaggcg 720

ctagcccggc tgcgggagca ggcggcccgg gtggggactc aggtggagga ggcggctgag 780

ggcgtcctcc gggccctgct ggcccagaag caggaggtgc tggggcagct acgagcccac 840

gtggaggctg ccgaagaagc tgctcgggag aggctggcgg agcttgaggg ccgggagcag 900

gtggccaggg ccgcagccgc cttcgcccgc cgggtactca gcctggggcg agaggccgag 960

atcctctccc tggaaggggc gatcgcacag cggctcaggc agctgcaggg ctgcccctgg 1020

gcaccaggcc cggccccctg cctgctccca cagctggagc tccatcctgg gctcctggac 1080

aagaactgcc accttcttcg gctgtccttt gaggagcagc agccccagaa ggatggtggg 1140

aaagacggag ctggtaccca gggaggtgag gagagccaga gccggaggga ggatgagccg 1200

aagactgaga gacagggtgg agtccagccc caggctggag atggagccca gaccccaaaa 1260

gaggaaaaag cccagacaac ccgagaagag ggagcccaga ccttggagga ggacagggcc 1320

cagacacccc acgaggatgg aggaccccag ccccacaggg gtggcagacc caacaagaag 1380

aaaaagttca aaggcaggct caagtcaatt tcccgggagc ccagcccagc cctggggccg 1440

aatctggacg gctctggcct cctccccaga cccatctttt actgcagttt ccccacgcgg 1500

atgcctggag acaagcggtc cccccggatc accgggctct gtcccttcgg tccccgggag 1560

atcctggtgg cggatgagca gaaccgggca ctgaaacgct tctccctcaa cggcgactac 1620

aagggcaccg tgccggtccc tgagggctgc tccccttgca gcgtggccgc cctgcagagc 1680

gcggtggcct tctccgctag cgcacggctc tatctcatca accccaacgg cgaagtgcag 1740

tggcgcaggg ccctgagcct ctcccaggcc agccacgcgg tggcggcact gcctagcggg 1800

gaccgcgtgg ctgtcagcgt ggcgggccac gtggaggtgt acaatatgga aggcagcctg 1860

gccacccggt tcattcctgg aggcaaggcc agccggggcc tgcgggcgct ggtgtttctg 1920

accaccagcc cccaggggca tttcgtgggg tcggactggc agcagaatag tgtggtaatc 1980

tgtgatgggc tgggccaggt ggttggggag tacaaggggc caggcctgca tggctgccag 2040

ccgggctccg tgtctgtgga taagaagggc tacatctttc tgacccttcg agaagtcaac 2100

aaggtggtga tcctggaccc gaaggggtcc ctccttggag acttcctgac agcctaccac 2160

ggcctggaaa agccccgggt taccaccatg gtggatggca ggtacctggt cgtgtccctc 2220

agtaa 2225

<210> 110

<211> 755

<212> PRT

<213> 智人(Homo sapiens)

<400> 110

Met Val Ser His Gly Ser Ser Pro Ser Leu Leu Glu Ala Leu Ser Ser

1 5 10 15

Asp Phe Leu Ala Cys Lys Ile Cys Leu Glu Gln Leu Arg Ala Pro Lys

20 25 30

Thr Leu Pro Cys Leu His Thr Tyr Cys Gln Asp Cys Leu Ala Gln Leu

35 40 45

Ala Asp Gly Gly Arg Val Arg Cys Pro Glu Cys Arg Glu Thr Val Pro

50 55 60

Val Pro Pro Glu Gly Val Ala Ser Phe Lys Thr Asn Phe Phe Val Asn

65 70 75 80

Gly Leu Leu Asp Leu Val Lys Ala Arg Ala Cys Gly Asp Leu Arg Ala

85 90 95

Gly Lys Pro Ala Cys Ala Leu Cys Pro Leu Val Gly Gly Thr Ser Thr

100 105 110

Gly Gly Pro Ala Thr Ala Arg Cys Leu Asp Cys Ala Asp Asp Leu Cys

115 120 125

Gln Ala Cys Ala Asp Gly His Arg Cys Thr Arg Gln Thr His Thr His

130 135 140

Arg Val Val Asp Leu Val Gly Tyr Arg Ala Gly Trp Tyr Asp Glu Glu

145 150 155 160

Ala Arg Glu Arg Gln Ala Ala Gln Cys Pro Gln His Pro Gly Glu Ala

165 170 175

Leu Arg Phe Leu Cys Gln Pro Cys Ser Gln Leu Leu Cys Arg Glu Cys

180 185 190

Arg Leu Asp Pro His Leu Asp His Pro Cys Leu Pro Leu Ala Glu Ala

195 200 205

Val Arg Ala Arg Arg Pro Gly Leu Glu Gly Leu Leu Ala Gly Val Asp

210 215 220

Asn Asn Leu Val Glu Leu Glu Ala Ala Arg Arg Val Glu Lys Glu Ala

225 230 235 240

Leu Ala Arg Leu Arg Glu Gln Ala Ala Arg Val Gly Thr Gln Val Glu

245 250 255

Glu Ala Ala Glu Gly Val Leu Arg Ala Leu Leu Ala Gln Lys Gln Glu

260 265 270

Val Leu Gly Gln Leu Arg Ala His Val Glu Ala Ala Glu Glu Ala Ala

275 280 285

Arg Glu Arg Leu Ala Glu Leu Glu Gly Arg Glu Gln Val Ala Arg Ala

290 295 300

Ala Ala Ala Phe Ala Arg Arg Val Leu Ser Leu Gly Arg Glu Ala Glu

305 310 315 320

Ile Leu Ser Leu Glu Gly Ala Ile Ala Gln Arg Leu Arg Gln Leu Gln

325 330 335

Gly Cys Pro Trp Ala Pro Gly Pro Ala Pro Cys Leu Leu Pro Gln Leu

340 345 350

Glu Leu His Pro Gly Leu Leu Asp Lys Asn Cys His Leu Leu Arg Leu

355 360 365

Ser Phe Glu Glu Gln Gln Pro Gln Lys Asp Gly Gly Lys Asp Gly Ala

370 375 380

Gly Thr Gln Gly Gly Glu Glu Ser Gln Ser Arg Arg Glu Asp Glu Pro

385 390 395 400

Lys Thr Glu Arg Gln Gly Gly Val Gln Pro Gln Ala Gly Asp Gly Ala

405 410 415

Gln Thr Pro Lys Glu Glu Lys Ala Gln Thr Thr Arg Glu Glu Gly Ala

420 425 430

Gln Thr Leu Glu Glu Asp Arg Ala Gln Thr Pro His Glu Asp Gly Gly

435 440 445

Pro Gln Pro His Arg Gly Gly Arg Pro Asn Lys Lys Lys Lys Phe Lys

450 455 460

Gly Arg Leu Lys Ser Ile Ser Arg Glu Pro Ser Pro Ala Leu Gly Pro

465 470 475 480

Asn Leu Asp Gly Ser Gly Leu Leu Pro Arg Pro Ile Phe Tyr Cys Ser

485 490 495

Phe Pro Thr Arg Met Pro Gly Asp Lys Arg Ser Pro Arg Ile Thr Gly

500 505 510

Leu Cys Pro Phe Gly Pro Arg Glu Ile Leu Val Ala Asp Glu Gln Asn

515 520 525

Arg Ala Leu Lys Arg Phe Ser Leu Asn Gly Asp Tyr Lys Gly Thr Val

530 535 540

Pro Val Pro Glu Gly Cys Ser Pro Cys Ser Val Ala Ala Leu Gln Ser

545 550 555 560

Ala Val Ala Phe Ser Ala Ser Ala Arg Leu Tyr Leu Ile Asn Pro Asn

565 570 575

Gly Glu Val Gln Trp Arg Arg Ala Leu Ser Leu Ser Gln Ala Ser His

580 585 590

Ala Val Ala Ala Leu Pro Ser Gly Asp Arg Val Ala Val Ser Val Ala

595 600 605

Gly His Val Glu Val Tyr Asn Met Glu Gly Ser Leu Ala Thr Arg Phe

610 615 620

Ile Pro Gly Gly Lys Ala Ser Arg Gly Leu Arg Ala Leu Val Phe Leu

625 630 635 640

Thr Thr Ser Pro Gln Gly His Phe Val Gly Ser Asp Trp Gln Gln Asn

645 650 655

Ser Val Val Ile Cys Asp Gly Leu Gly Gln Val Val Gly Glu Tyr Lys

660 665 670

Gly Pro Gly Leu His Gly Cys Gln Pro Gly Ser Val Ser Val Asp Lys

675 680 685

Lys Gly Tyr Ile Phe Leu Thr Leu Arg Glu Val Asn Lys Val Val Ile

690 695 700

Leu Asp Pro Lys Gly Ser Leu Leu Gly Asp Phe Leu Thr Ala Tyr His

705 710 715 720

Gly Leu Glu Lys Pro Arg Val Thr Thr Met Val Asp Gly Arg Tyr Leu

725 730 735

Val Val Ser Leu Ser Asn Gly Thr Ile His Ile Phe Arg Val Arg Ser

740 745 750

Pro Asp Ser

755

<210> 111

<211> 1461

<212> DNA

<213> 智人(Homo sapiens)

<400> 111

atggcctggg cgccgcccgg ggagcggctg cgcgaggatg cgcggtgccc ggtgtgcctg 60

gatttcctgc aggagccggt cagcgtggac tgcggccaca gcttctgcct caggtgcatc 120

tccgagttct gcgagaagtc ggacggcgcg cagggcggcg tctacgcctg tccgcagtgc 180

cggggcccct tccggccctc gggctttcgc cccaaccggc agctggcggg cctggtggag 240

agcgtgcggc ggctggggtt gggcgcgggg cccggggcgc ggcgatgcgc gcggcacggc 300

gaggacctga gccgcttctg cgaggaggac gaggcggcgc tgtgctgggt gtgcgacgcc 360

ggccccgagc acaggacgca ccgcacggcg ccgctgcagg aggccgccgg cagctaccag 420

gtaaagctcc agatggctct ggaacttatg aggaaagagt tggaggacgc cttgactcag 480

gaggccaacg tggggaaaaa gactgtcatt tggaaggaga aagtggaaat gcagaggcag 540

cgcttcagat tggagtttga gaagcatcgt ggctttctgg cccaggagga gcaacggcag 600

ctgaggcggc tggaggcgga ggagcgagcg acgctgcaga gactgcggga gagcaagagc 660

cggctggtcc agcagagcaa ggccctgaag gagctggcgg atgagctgca ggagaggtgc 720

cagcgcccgg ccctgggtct gctggagggt gtgagaggag tcctgagcag aagtaaggct 780

gtcacaaggc tggaagcaga gaacatcccc atggaactga agacagcatg ctgcatccct 840

gggaggaggg agctcttaag gaagttccaa gtggatgtaa agctggatcc cgccacggcg 900

cacccgagtc tgctcttgac cgccgacctg cgcagtgtgc aggatggaga accatggagg 960

gatgtcccca acaaccctga gcgatttgac acatggccct gcatcctggg tttgcagagc 1020

ttctcatcag ggaggcatta ctgggaggtt ctggtgggag aaggagcaga gtggggttta 1080

ggggtctgtc aagacacact gccaagaaag ggggaaacca cgccatctcc tgagaatggg 1140

gtctgggccc tgtggctgct gaaagggaat gagtacatgg tccttgcctc cccatcagtg 1200

cctcttctcc aactggaaag tcctcgctgc attgggattt tcttggacta tgaagccggt 1260

gaaatttcat tctacaatgt cacagatgga tcttatatct acacattcaa ccaactcttc 1320

tctggtcttc ttcggcctta ctttttcatc tgtgatgcaa ctcctcttat cttgccaccc 1380

acaacaatag cagggtcagg aaattgggca tccagggatc atttagatcc tgcttctgat 1440

gtaagagatg atcatctcta a 1461

<210> 112

<211> 486

<212> PRT

<213> 智人(Homo sapiens)

<400> 112

Met Ala Trp Ala Pro Pro Gly Glu Arg Leu Arg Glu Asp Ala Arg Cys

1 5 10 15

Pro Val Cys Leu Asp Phe Leu Gln Glu Pro Val Ser Val Asp Cys Gly

20 25 30

His Ser Phe Cys Leu Arg Cys Ile Ser Glu Phe Cys Glu Lys Ser Asp

35 40 45

Gly Ala Gln Gly Gly Val Tyr Ala Cys Pro Gln Cys Arg Gly Pro Phe

50 55 60

Arg Pro Ser Gly Phe Arg Pro Asn Arg Gln Leu Ala Gly Leu Val Glu

65 70 75 80

Ser Val Arg Arg Leu Gly Leu Gly Ala Gly Pro Gly Ala Arg Arg Cys

85 90 95

Ala Arg His Gly Glu Asp Leu Ser Arg Phe Cys Glu Glu Asp Glu Ala

100 105 110

Ala Leu Cys Trp Val Cys Asp Ala Gly Pro Glu His Arg Thr His Arg

115 120 125

Thr Ala Pro Leu Gln Glu Ala Ala Gly Ser Tyr Gln Val Lys Leu Gln

130 135 140

Met Ala Leu Glu Leu Met Arg Lys Glu Leu Glu Asp Ala Leu Thr Gln

145 150 155 160

Glu Ala Asn Val Gly Lys Lys Thr Val Ile Trp Lys Glu Lys Val Glu

165 170 175

Met Gln Arg Gln Arg Phe Arg Leu Glu Phe Glu Lys His Arg Gly Phe

180 185 190

Leu Ala Gln Glu Glu Gln Arg Gln Leu Arg Arg Leu Glu Ala Glu Glu

195 200 205

Arg Ala Thr Leu Gln Arg Leu Arg Glu Ser Lys Ser Arg Leu Val Gln

210 215 220

Gln Ser Lys Ala Leu Lys Glu Leu Ala Asp Glu Leu Gln Glu Arg Cys

225 230 235 240

Gln Arg Pro Ala Leu Gly Leu Leu Glu Gly Val Arg Gly Val Leu Ser

245 250 255

Arg Ser Lys Ala Val Thr Arg Leu Glu Ala Glu Asn Ile Pro Met Glu

260 265 270

Leu Lys Thr Ala Cys Cys Ile Pro Gly Arg Arg Glu Leu Leu Arg Lys

275 280 285

Phe Gln Val Asp Val Lys Leu Asp Pro Ala Thr Ala His Pro Ser Leu

290 295 300

Leu Leu Thr Ala Asp Leu Arg Ser Val Gln Asp Gly Glu Pro Trp Arg

305 310 315 320

Asp Val Pro Asn Asn Pro Glu Arg Phe Asp Thr Trp Pro Cys Ile Leu

325 330 335

Gly Leu Gln Ser Phe Ser Ser Gly Arg His Tyr Trp Glu Val Leu Val

340 345 350

Gly Glu Gly Ala Glu Trp Gly Leu Gly Val Cys Gln Asp Thr Leu Pro

355 360 365

Arg Lys Gly Glu Thr Thr Pro Ser Pro Glu Asn Gly Val Trp Ala Leu

370 375 380

Trp Leu Leu Lys Gly Asn Glu Tyr Met Val Leu Ala Ser Pro Ser Val

385 390 395 400

Pro Leu Leu Gln Leu Glu Ser Pro Arg Cys Ile Gly Ile Phe Leu Asp

405 410 415

Tyr Glu Ala Gly Glu Ile Ser Phe Tyr Asn Val Thr Asp Gly Ser Tyr

420 425 430

Ile Tyr Thr Phe Asn Gln Leu Phe Ser Gly Leu Leu Arg Pro Tyr Phe

435 440 445

Phe Ile Cys Asp Ala Thr Pro Leu Ile Leu Pro Pro Thr Thr Ile Ala

450 455 460

Gly Ser Gly Asn Trp Ala Ser Arg Asp His Leu Asp Pro Ala Ser Asp

465 470 475 480

Val Arg Asp Asp His Leu

485

<210> 113

<211> 1212

<212> DNA

<213> 智人(Homo sapiens)

<400> 113

atgcacaatt ttgaggaaga gttaacttgt cccatatgtt atagtatttt tgaagatcct 60

cgtgtactgc catgctctca tacattttgt agaaattgtt tggaaaacat tcttcaggca 120

tctggtaact tttatatatg gagaccttta cgaattccac tcaagtgccc taattgcaga 180

agtattactg aaattgctcc aactggcatt gaatctttac ctgttaattt tgcactaagg 240

gctattattg aaaagtacca gcaagaagac catccagata ttgtcacctg ccctgaacat 300

tacaggcaac cattaaatgt ttactgtcta ttagataaaa aattagtttg tggtcattgc 360

cttaccatag gtcaacatca tggtcatcct atagatgacc ttcaaagtgc ctatttgaaa 420

gaaaaggaca ctcctcaaaa actgcttgaa cagttgactg acacacactg gacagatctt 480

acccatctta ttgaaaagct gaaagaacaa aaatctcatt ctgagaaaat gatccaaggc 540

gataaggaag ctgttctcca gtattttaag gagcttaatg atacattaga acagaaaaaa 600

aaaagtttcc taacggctct ctgtgatgtt ggcaatctaa ttaatcaaga atatactcca 660

caaattgaaa gaatgaagga aatacgagag cagcagcttg aattaatggc actgacaata 720

tctttacaag aagagtctcc acttaaattt cttgaaaaag ttgatgatgt acgccagcat 780

gtacagatct tgaaacaaag accacttcct gaggttcaac ccgttgaaat ttatcctcga 840

gtaagcaaaa tattgaaaga agaatggagc agaacagaaa ttggacaaat taagaacgtt 900

ctcattccca aaatgaaaat ttctccaaaa aggatgtcat gttcctggcc tggtaaggat 960

gaaaaggaag ttgaattttt aaaaatttta aacattgttg tagttacatt aatttcagta 1020

atactgatgt cgatactctt tttcaaccaa cacatcataa cctttttaag tgaaatcact 1080

ttaatatggt tttctgaagc ctctctatct gtttaccaaa gtttatctaa cagtctgcat 1140

aaggtaaaga atatactgtg tcacattttc tatttgttga aggaatttgt gtggaaaata 1200

gtttcccatt ga 1212

<210> 114

<211> 403

<212> PRT

<213> 智人(Homo sapiens)

<400> 114

Met His Asn Phe Glu Glu Glu Leu Thr Cys Pro Ile Cys Tyr Ser Ile

1 5 10 15

Phe Glu Asp Pro Arg Val Leu Pro Cys Ser His Thr Phe Cys Arg Asn

20 25 30

Cys Leu Glu Asn Ile Leu Gln Ala Ser Gly Asn Phe Tyr Ile Trp Arg

35 40 45

Pro Leu Arg Ile Pro Leu Lys Cys Pro Asn Cys Arg Ser Ile Thr Glu

50 55 60

Ile Ala Pro Thr Gly Ile Glu Ser Leu Pro Val Asn Phe Ala Leu Arg

65 70 75 80

Ala Ile Ile Glu Lys Tyr Gln Gln Glu Asp His Pro Asp Ile Val Thr

85 90 95

Cys Pro Glu His Tyr Arg Gln Pro Leu Asn Val Tyr Cys Leu Leu Asp

100 105 110

Lys Lys Leu Val Cys Gly His Cys Leu Thr Ile Gly Gln His His Gly

115 120 125

His Pro Ile Asp Asp Leu Gln Ser Ala Tyr Leu Lys Glu Lys Asp Thr

130 135 140

Pro Gln Lys Leu Leu Glu Gln Leu Thr Asp Thr His Trp Thr Asp Leu

145 150 155 160

Thr His Leu Ile Glu Lys Leu Lys Glu Gln Lys Ser His Ser Glu Lys

165 170 175

Met Ile Gln Gly Asp Lys Glu Ala Val Leu Gln Tyr Phe Lys Glu Leu

180 185 190

Asn Asp Thr Leu Glu Gln Lys Lys Lys Ser Phe Leu Thr Ala Leu Cys

195 200 205

Asp Val Gly Asn Leu Ile Asn Gln Glu Tyr Thr Pro Gln Ile Glu Arg

210 215 220

Met Lys Glu Ile Arg Glu Gln Gln Leu Glu Leu Met Ala Leu Thr Ile

225 230 235 240

Ser Leu Gln Glu Glu Ser Pro Leu Lys Phe Leu Glu Lys Val Asp Asp

245 250 255

Val Arg Gln His Val Gln Ile Leu Lys Gln Arg Pro Leu Pro Glu Val

260 265 270

Gln Pro Val Glu Ile Tyr Pro Arg Val Ser Lys Ile Leu Lys Glu Glu

275 280 285

Trp Ser Arg Thr Glu Ile Gly Gln Ile Lys Asn Val Leu Ile Pro Lys

290 295 300

Met Lys Ile Ser Pro Lys Arg Met Ser Cys Ser Trp Pro Gly Lys Asp

305 310 315 320

Glu Lys Glu Val Glu Phe Leu Lys Ile Leu Asn Ile Val Val Val Thr

325 330 335

Leu Ile Ser Val Ile Leu Met Ser Ile Leu Phe Phe Asn Gln His Ile

340 345 350

Ile Thr Phe Leu Ser Glu Ile Thr Leu Ile Trp Phe Ser Glu Ala Ser

355 360 365

Leu Ser Val Tyr Gln Ser Leu Ser Asn Ser Leu His Lys Val Lys Asn

370 375 380

Ile Leu Cys His Ile Phe Tyr Leu Leu Lys Glu Phe Val Trp Lys Ile

385 390 395 400

Val Ser His

<210> 115

<211> 1416

<212> DNA

<213> 智人(Homo sapiens)

<400> 115

atggagtttg tgacagccct ggtgaacctc caagaggagt ctagctgtcc catctgtctg 60

gagtacttga aagacccagt gaccatcaac tgtgggcaca acttctgtcg ctcctgcctc 120

agtgtatcct ggaaggatct agatgatacc tttccctgtc ctgtctgccg tttttgcttt 180

ccatacaaga gcttcaggag gaacccccag ctccgtaatt tgactgaaat tgctaaacaa 240

ctccagatta ggaggagcaa gagaaagagg cagaaagaga atgccatgtg tgaaaaacac 300

aaccagtttc tgaccctctt ctgtgttaaa gatctagaga tcttatgtac acagtgcagt 360

ttctccacta aacaccagaa gcactacatt tgccctatta agaaagctgc ctcttatcac 420

agagaaattc tagaaggtag ccttgagccc ttgaggaata atatagaacg agttgaaaaa 480

gtgataattc tgcaaggcag caaatcagtg gagctgaaaa agaaggtaga atataagagg 540

gaagaaataa attctgagtt tgagcaaata agattgtttt tacagaatga acaagagatg 600

attcttaggc agatacaaga tgaagagatg aacattttag caaaactaaa tgaaaacctt 660

gtagaacttt cagattatgt ttccacatta aaacatctac tgagggaggt agagggcaag 720

tctgtgcagt caaacctgga attactgaca caagctaaga gtatgcacca caagtatcaa 780

aacctaaaat gccctgaact cttttcattt agattaacaa aatatggttt cagtcttcct 840

cctcaatatt ctggcttgga cagaattatc aagccatttc aagtagatgt gattctagat 900

ctcaacacag cacatcctca acttcttgtc tctgaggata gaaaagctgt gcgatatgaa 960

agaaaaaaac gaaacatttg ttatgaccca aggagatttt atgtctgccc tgctgtccta 1020

ggctctcaga gatttagttc tggccgacat tactgggaag tagaagtggg aaacaaacct 1080

aaatggatat tgggtgtgtg tcaagactgt cttcttagga actggcagga tcagccatca 1140

gttctgggcg gattctgggc aattgggcga tacatgaaga gtggttatgt tgcgtcaggt 1200

cctaagacaa cccagcttct gccagtagta aaacccagta aaattggtat ttttctggac 1260

tatgaattgg gtgatctttc cttttataat atgaatgata ggtctattct ctatactttt 1320

aacgattgtt tcacagaagc cgtttggcct tatttctata ctggaacaga ttccgaacct 1380

cttaaaatct gctcagtatc agattctgaa agataa 1416

<210> 116

<211> 471

<212> PRT

<213> 智人(Homo sapiens)

<400> 116

Met Glu Phe Val Thr Ala Leu Val Asn Leu Gln Glu Glu Ser Ser Cys

1 5 10 15

Pro Ile Cys Leu Glu Tyr Leu Lys Asp Pro Val Thr Ile Asn Cys Gly

20 25 30

His Asn Phe Cys Arg Ser Cys Leu Ser Val Ser Trp Lys Asp Leu Asp

35 40 45

Asp Thr Phe Pro Cys Pro Val Cys Arg Phe Cys Phe Pro Tyr Lys Ser

50 55 60

Phe Arg Arg Asn Pro Gln Leu Arg Asn Leu Thr Glu Ile Ala Lys Gln

65 70 75 80

Leu Gln Ile Arg Arg Ser Lys Arg Lys Arg Gln Lys Glu Asn Ala Met

85 90 95

Cys Glu Lys His Asn Gln Phe Leu Thr Leu Phe Cys Val Lys Asp Leu

100 105 110

Glu Ile Leu Cys Thr Gln Cys Ser Phe Ser Thr Lys His Gln Lys His

115 120 125

Tyr Ile Cys Pro Ile Lys Lys Ala Ala Ser Tyr His Arg Glu Ile Leu

130 135 140

Glu Gly Ser Leu Glu Pro Leu Arg Asn Asn Ile Glu Arg Val Glu Lys

145 150 155 160

Val Ile Ile Leu Gln Gly Ser Lys Ser Val Glu Leu Lys Lys Lys Val

165 170 175

Glu Tyr Lys Arg Glu Glu Ile Asn Ser Glu Phe Glu Gln Ile Arg Leu

180 185 190

Phe Leu Gln Asn Glu Gln Glu Met Ile Leu Arg Gln Ile Gln Asp Glu

195 200 205

Glu Met Asn Ile Leu Ala Lys Leu Asn Glu Asn Leu Val Glu Leu Ser

210 215 220

Asp Tyr Val Ser Thr Leu Lys His Leu Leu Arg Glu Val Glu Gly Lys

225 230 235 240

Ser Val Gln Ser Asn Leu Glu Leu Leu Thr Gln Ala Lys Ser Met His

245 250 255

His Lys Tyr Gln Asn Leu Lys Cys Pro Glu Leu Phe Ser Phe Arg Leu

260 265 270

Thr Lys Tyr Gly Phe Ser Leu Pro Pro Gln Tyr Ser Gly Leu Asp Arg

275 280 285

Ile Ile Lys Pro Phe Gln Val Asp Val Ile Leu Asp Leu Asn Thr Ala

290 295 300

His Pro Gln Leu Leu Val Ser Glu Asp Arg Lys Ala Val Arg Tyr Glu

305 310 315 320

Arg Lys Lys Arg Asn Ile Cys Tyr Asp Pro Arg Arg Phe Tyr Val Cys

325 330 335

Pro Ala Val Leu Gly Ser Gln Arg Phe Ser Ser Gly Arg His Tyr Trp

340 345 350

Glu Val Glu Val Gly Asn Lys Pro Lys Trp Ile Leu Gly Val Cys Gln

355 360 365

Asp Cys Leu Leu Arg Asn Trp Gln Asp Gln Pro Ser Val Leu Gly Gly

370 375 380

Phe Trp Ala Ile Gly Arg Tyr Met Lys Ser Gly Tyr Val Ala Ser Gly

385 390 395 400

Pro Lys Thr Thr Gln Leu Leu Pro Val Val Lys Pro Ser Lys Ile Gly

405 410 415

Ile Phe Leu Asp Tyr Glu Leu Gly Asp Leu Ser Phe Tyr Asn Met Asn

420 425 430

Asp Arg Ser Ile Leu Tyr Thr Phe Asn Asp Cys Phe Thr Glu Ala Val

435 440 445

Trp Pro Tyr Phe Tyr Thr Gly Thr Asp Ser Glu Pro Leu Lys Ile Cys

450 455 460

Ser Val Ser Asp Ser Glu Arg

465 470

<210> 117

<211> 630

<212> DNA

<213> 智人(Homo sapiens)

<400> 117

atggaatttg ttacggccct ggctgacctc cgagcagagg ctagctgtcc catctgtctg 60

gactacttga aagacccagt gaccatcagc tgtgggcata acttctgtct ctcctgcatc 120

attatgtcct ggaaggatct acatgatagt ttcccctgcc ccttttgcca cttttgctgt 180

ccagaaagga aatttataag caatccccag ctgggtagtt tgactgaaat tgctaagcaa 240

ctccagataa gaagcaagaa gaggaagagg caggaagaga agcatgtgtg taagaagcat 300

aatcaggttt tgactttctt ctgtcagaaa gacctagagc ttttatgtcc aaggtgcagt 360

ttgtccactg atcaccagca tcactgtgtt tggcccataa agaaggctgc ctcctatcat 420

aggaaaaaac tggaggaata caatgcaccg tggaaggaga gagtggaact aattgaaaaa 480

gtcataacta tgcaaaccag gaaatcactg gaactgaaga aaaagatgga gtctccttct 540

gtcaccaggc tggagtgcag ttgcacgatc tcggctcact tcaacctccg cctcccggga 600

tcaagcgatt cttctgcctc tggctcctga 630

<210> 118

<211> 209

<212> PRT

<213> 智人(Homo sapiens)

<400> 118

Met Glu Phe Val Thr Ala Leu Ala Asp Leu Arg Ala Glu Ala Ser Cys

1 5 10 15

Pro Ile Cys Leu Asp Tyr Leu Lys Asp Pro Val Thr Ile Ser Cys Gly

20 25 30

His Asn Phe Cys Leu Ser Cys Ile Ile Met Ser Trp Lys Asp Leu His

35 40 45

Asp Ser Phe Pro Cys Pro Phe Cys His Phe Cys Cys Pro Glu Arg Lys

50 55 60

Phe Ile Ser Asn Pro Gln Leu Gly Ser Leu Thr Glu Ile Ala Lys Gln

65 70 75 80

Leu Gln Ile Arg Ser Lys Lys Arg Lys Arg Gln Glu Glu Lys His Val

85 90 95

Cys Lys Lys His Asn Gln Val Leu Thr Phe Phe Cys Gln Lys Asp Leu

100 105 110

Glu Leu Leu Cys Pro Arg Cys Ser Leu Ser Thr Asp His Gln His His

115 120 125

Cys Val Trp Pro Ile Lys Lys Ala Ala Ser Tyr His Arg Lys Lys Leu

130 135 140

Glu Glu Tyr Asn Ala Pro Trp Lys Glu Arg Val Glu Leu Ile Glu Lys

145 150 155 160

Val Ile Thr Met Gln Thr Arg Lys Ser Leu Glu Leu Lys Lys Lys Met

165 170 175

Glu Ser Pro Ser Val Thr Arg Leu Glu Cys Ser Cys Thr Ile Ser Ala

180 185 190

His Phe Asn Leu Arg Leu Pro Gly Ser Ser Asp Ser Ser Ala Ser Gly

195 200 205

Ser

<210> 119

<211> 1428

<212> DNA

<213> 智人(Homo sapiens)

<400> 119

atggcgtgca gcctcaagga cgagctgctg tgctccatct gcctgagcat ctaccaggac 60

ccggtgagcc tgggctgcga gcattacttc tgccgccgct gcatcacgga gcactgggtg 120

cggcaggagg cgcagggcgc ccgcgactgc cccgagtgcc ggcgcacgtt cgccgagccc 180

gcgctggcgc ccagcctcaa gctggccaac atcgtggagc gctacagctc cttcccgctg 240

gacgccatcc tcaacgcgcg ccgcgccgcg cgaccctgcc aggcgcacga caaggtcaag 300

ctcttctgcc tcacggaccg cgcgcttctc tgcttcttct gcgacgagcc tgcactgcac 360

gagcagcatc aggtcaccgg catcgacgac gccttcgacg agctgcagag ggagctgaag 420

gaccaacttc aggcccttca agacagcgag cgggaacaca ccgaagcgct gcagctgctc 480

aagcgacaac tggcggagac caagtcttcc accaagagcc tgcggaccac tatcggcgag 540

gccttcgagc ggctgcaccg gctgctgcgt gaacgccaga aggccatgct agaggagctg 600

gaggcggaca cggcccgcac gctgaccgac atcgagcaga aagtccagcg ctacagccag 660

cagctgcgca aggtccagga gggagcccag atcctgcagg agcggctggc tgaaaccgac 720

cggcacacct tcctggctgg ggtggcctca ctgtccgagc ggctcaaggg aaaaatccat 780

gagaccaacc tcacatatga agacttcccg acctccaagt acacaggccc cctgcagtac 840

accatctgga agtccctgtt ccaggacatc cacccagtgc cagccgccct aaccctggac 900

ccgggcacag cccaccagcg cctgatcctg tcggacgact gcaccattgt ggcttacggc 960

aacttgcacc cacagccact gcaggactcg ccaaagcgct tcgatgtgga ggtgtcggtg 1020

ctgggttctg aagccttcag tagtggcgtc cactactggg aggtggtggt ggcggagaag 1080

acccagtggg tgatcgggct ggcacacgaa gccgcaagcc gcaagggcag catccagatc 1140

cagcccagcc gcggcttcta ctgcatcgtg atgcacgatg gcaaccagta cagcgcctgc 1200

acggagccct ggacgcggct taacgtccgg gacaagcttg acaaggtggg tgtcttcctg 1260

gactatgacc aaggcttgct catcttctac aatgctgatg acatgtcctg gctctacacc 1320

ttccgcgaga agttccctgg caagctctgc tcttacttca gccctggcca gagccacgcc 1380

aatggcaaga acgttcagcc gctgcggatc aacaccgtcc gcatctag 1428

<210> 120

<211> 475

<212> PRT

<213> 智人(Homo sapiens)

<400> 120

Met Ala Cys Ser Leu Lys Asp Glu Leu Leu Cys Ser Ile Cys Leu Ser

1 5 10 15

Ile Tyr Gln Asp Pro Val Ser Leu Gly Cys Glu His Tyr Phe Cys Arg

20 25 30

Arg Cys Ile Thr Glu His Trp Val Arg Gln Glu Ala Gln Gly Ala Arg

35 40 45

Asp Cys Pro Glu Cys Arg Arg Thr Phe Ala Glu Pro Ala Leu Ala Pro

50 55 60

Ser Leu Lys Leu Ala Asn Ile Val Glu Arg Tyr Ser Ser Phe Pro Leu

65 70 75 80

Asp Ala Ile Leu Asn Ala Arg Arg Ala Ala Arg Pro Cys Gln Ala His

85 90 95

Asp Lys Val Lys Leu Phe Cys Leu Thr Asp Arg Ala Leu Leu Cys Phe

100 105 110

Phe Cys Asp Glu Pro Ala Leu His Glu Gln His Gln Val Thr Gly Ile

115 120 125

Asp Asp Ala Phe Asp Glu Leu Gln Arg Glu Leu Lys Asp Gln Leu Gln

130 135 140

Ala Leu Gln Asp Ser Glu Arg Glu His Thr Glu Ala Leu Gln Leu Leu

145 150 155 160

Lys Arg Gln Leu Ala Glu Thr Lys Ser Ser Thr Lys Ser Leu Arg Thr

165 170 175

Thr Ile Gly Glu Ala Phe Glu Arg Leu His Arg Leu Leu Arg Glu Arg

180 185 190

Gln Lys Ala Met Leu Glu Glu Leu Glu Ala Asp Thr Ala Arg Thr Leu

195 200 205

Thr Asp Ile Glu Gln Lys Val Gln Arg Tyr Ser Gln Gln Leu Arg Lys

210 215 220

Val Gln Glu Gly Ala Gln Ile Leu Gln Glu Arg Leu Ala Glu Thr Asp

225 230 235 240

Arg His Thr Phe Leu Ala Gly Val Ala Ser Leu Ser Glu Arg Leu Lys

245 250 255

Gly Lys Ile His Glu Thr Asn Leu Thr Tyr Glu Asp Phe Pro Thr Ser

260 265 270

Lys Tyr Thr Gly Pro Leu Gln Tyr Thr Ile Trp Lys Ser Leu Phe Gln

275 280 285

Asp Ile His Pro Val Pro Ala Ala Leu Thr Leu Asp Pro Gly Thr Ala

290 295 300

His Gln Arg Leu Ile Leu Ser Asp Asp Cys Thr Ile Val Ala Tyr Gly

305 310 315 320

Asn Leu His Pro Gln Pro Leu Gln Asp Ser Pro Lys Arg Phe Asp Val

325 330 335

Glu Val Ser Val Leu Gly Ser Glu Ala Phe Ser Ser Gly Val His Tyr

340 345 350

Trp Glu Val Val Val Ala Glu Lys Thr Gln Trp Val Ile Gly Leu Ala

355 360 365

His Glu Ala Ala Ser Arg Lys Gly Ser Ile Gln Ile Gln Pro Ser Arg

370 375 380

Gly Phe Tyr Cys Ile Val Met His Asp Gly Asn Gln Tyr Ser Ala Cys

385 390 395 400

Thr Glu Pro Trp Thr Arg Leu Asn Val Arg Asp Lys Leu Asp Lys Val

405 410 415

Gly Val Phe Leu Asp Tyr Asp Gln Gly Leu Leu Ile Phe Tyr Asn Ala

420 425 430

Asp Asp Met Ser Trp Leu Tyr Thr Phe Arg Glu Lys Phe Pro Gly Lys

435 440 445

Leu Cys Ser Tyr Phe Ser Pro Gly Gln Ser His Ala Asn Gly Lys Asn

450 455 460

Val Gln Pro Leu Arg Ile Asn Thr Val Arg Ile

465 470 475

<210> 121

<211> 1062

<212> DNA

<213> 智人(Homo sapiens)

<400> 121

atggattata agtcgagcct gatccaggat gggaatccca tggagaactt ggagaagcag 60

ctgatctgcc ctatctgcct ggagatgttt accaagccag tggtcatctt gccgtgccag 120

cacaacctgt gccggaagtg tgccaatgac atcttccagg ctgcaaatcc ctactggacc 180

agccggggca gctcagtgtc catgtctgga ggccgtttcc gctgccccac ctgccgccac 240

gaggtgatca tggatcgtca cggagtgtac ggcctgcaga ggaacctgct ggtggagaac 300

atcatcgaca tctacaaaca ggagtgctcc agtcggccgc tgcagaaggg cagtcacccc 360

atgtgcaagg agcacgaaga tgagaaaatc aacatctact gtctcacgtg tgaggtgccc 420

acctgctcca tgtgcaaggt gtttgggatc cacaaggcct gcgaggtggc cccattgcag 480

agtgtcttcc agggacaaaa gactgaactg aataactgta tctccatgct ggtggcgggg 540

aatgaccgtg tgcagaccat catcactcag ctggaggatt cccgtcgagt gaccaaggag 600

aacagtcacc aggtaaagga agagctgagc cagaagtttg acacgttgta tgccatcctg 660

gatgagaaga aaagtgagtt gctgcagcgg atcacgcagg agcaggagaa aaagcttagc 720

ttcatcgagg ccctcatcca gcagtaccag gagcagctgg acaagtccac aaagctggtg 780

gaaactgcca tccagtccct ggacgagcct gggggagcca ccttcctctt gactgccaag 840

caactcatca aaagcattgt ggaagcttcc aagggctgcc agctggggaa gacagagcag 900

ggctttgaga acatggactt ctttactttg gatttagagc acatagcaga cgccctgaga 960

gccattgact ttgggacaga tgaggaagag gaagaattca ttgaagaaga agatcaggaa 1020

gaggaagagt ccacagaagg gaaggaagaa ggacaccagt aa 1062

<210> 122

<211> 353

<212> PRT

<213> 智人(Homo sapiens)

<400> 122

Met Asp Tyr Lys Ser Ser Leu Ile Gln Asp Gly Asn Pro Met Glu Asn

1 5 10 15

Leu Glu Lys Gln Leu Ile Cys Pro Ile Cys Leu Glu Met Phe Thr Lys

20 25 30

Pro Val Val Ile Leu Pro Cys Gln His Asn Leu Cys Arg Lys Cys Ala

35 40 45

Asn Asp Ile Phe Gln Ala Ala Asn Pro Tyr Trp Thr Ser Arg Gly Ser

50 55 60

Ser Val Ser Met Ser Gly Gly Arg Phe Arg Cys Pro Thr Cys Arg His

65 70 75 80

Glu Val Ile Met Asp Arg His Gly Val Tyr Gly Leu Gln Arg Asn Leu

85 90 95

Leu Val Glu Asn Ile Ile Asp Ile Tyr Lys Gln Glu Cys Ser Ser Arg

100 105 110

Pro Leu Gln Lys Gly Ser His Pro Met Cys Lys Glu His Glu Asp Glu

115 120 125

Lys Ile Asn Ile Tyr Cys Leu Thr Cys Glu Val Pro Thr Cys Ser Met

130 135 140

Cys Lys Val Phe Gly Ile His Lys Ala Cys Glu Val Ala Pro Leu Gln

145 150 155 160

Ser Val Phe Gln Gly Gln Lys Thr Glu Leu Asn Asn Cys Ile Ser Met

165 170 175

Leu Val Ala Gly Asn Asp Arg Val Gln Thr Ile Ile Thr Gln Leu Glu

180 185 190

Asp Ser Arg Arg Val Thr Lys Glu Asn Ser His Gln Val Lys Glu Glu

195 200 205

Leu Ser Gln Lys Phe Asp Thr Leu Tyr Ala Ile Leu Asp Glu Lys Lys

210 215 220

Ser Glu Leu Leu Gln Arg Ile Thr Gln Glu Gln Glu Lys Lys Leu Ser

225 230 235 240

Phe Ile Glu Ala Leu Ile Gln Gln Tyr Gln Glu Gln Leu Asp Lys Ser

245 250 255

Thr Lys Leu Val Glu Thr Ala Ile Gln Ser Leu Asp Glu Pro Gly Gly

260 265 270

Ala Thr Phe Leu Leu Thr Ala Lys Gln Leu Ile Lys Ser Ile Val Glu

275 280 285

Ala Ser Lys Gly Cys Gln Leu Gly Lys Thr Glu Gln Gly Phe Glu Asn

290 295 300

Met Asp Phe Phe Thr Leu Asp Leu Glu His Ile Ala Asp Ala Leu Arg

305 310 315 320

Ala Ile Asp Phe Gly Thr Asp Glu Glu Glu Glu Glu Phe Ile Glu Glu

325 330 335

Glu Asp Gln Glu Glu Glu Glu Ser Thr Glu Gly Lys Glu Glu Gly His

340 345 350

Gln

<210> 123

<211> 1350

<212> DNA

<213> 智人(Homo sapiens)

<400> 123

atggattcag acgacctgca agtcttccag aatgagctca tttgctgcat ttgcgtgaac 60

tacttcatag atccggtcac cattgactgt gggcacagct tttgcaggcc ctgcctctgc 120

ctctgctcag aagaaggcag agcaccaatg cgctgccctt cgtgcagaaa aatctcagag 180

aagcccaact tcaacaccaa tgtggtactc aaaaagctgt cttccctagc cagacagacc 240

agacctcaga acatcaacag ctcagacaat atctgtgtgc tccatgagga gactaaggag 300

ctcttctgtg aggctgacaa gagattgctc tgtgggccct gctctgagtc accagagcac 360

atggctcaca gccacagccc aataggatgg gctgctgagg aatgcaggga gaaacttata 420

aaggaaatgg actatttatg ggaaatcaat caagagacaa gaaacaatct aaatcaggaa 480

actagaacat ttcattcgtt aaaggactat gtgtcagtaa ggaagaggat aatcactatt 540

caatatcaaa agatgcctat atttctcgat gaggaggagc aacggcatct gcaggcactg 600

gaaagagaag cagaagagct tttccaacaa ctacaagaca gtcaagtgag aatgacccaa 660

catttagaaa ggatgaaaga catgtacaga gagctgtggg agacatgcca cgtgcctgac 720

gtggagctgc tccaggatgt gagaaatgta tcagcaagga ctgatttggc acagatgcaa 780

aagccccagc cagtgaaccc agagctcact tcatggtgca taactggagt cctagacatg 840

ctcaacaact tcagagtgga tagtgctctg agcacggaaa tgattccttg ctatataagc 900

ctttctgagg atgtgagata tgtgatattt ggagatgacc atctcagtgc tcccacggat 960

ccccagggag tggacagctt tgctgtgtgg ggagcgcaag cattcacctc cggcaagcat 1020

tactgggagg tggatgtgac cctctcctcc aactggattc tgggagtctg tcaagattcc 1080

aggactgcag atgccaattt cgttattgat tctgatgaaa gatttttttt aatttcctca 1140

aagaggagca atcactatag tctctccacc aactctccac ctttaattca gtatgtgcaa 1200

aggcctctgg gtcaagttgg ggtgtttctg gattatgata atggatctgt gagttttttt 1260

gatgtttcta aaggttctct tatctatggt tttcctcctt cctccttctc ttcccctctg 1320

aggcctttct tttgctttgg ttgtacatga 1350

<210> 124

<211> 449

<212> PRT

<213> 智人(Homo sapiens)

<400> 124

Met Asp Ser Asp Asp Leu Gln Val Phe Gln Asn Glu Leu Ile Cys Cys

1 5 10 15

Ile Cys Val Asn Tyr Phe Ile Asp Pro Val Thr Ile Asp Cys Gly His

20 25 30

Ser Phe Cys Arg Pro Cys Leu Cys Leu Cys Ser Glu Glu Gly Arg Ala

35 40 45

Pro Met Arg Cys Pro Ser Cys Arg Lys Ile Ser Glu Lys Pro Asn Phe

50 55 60

Asn Thr Asn Val Val Leu Lys Lys Leu Ser Ser Leu Ala Arg Gln Thr

65 70 75 80

Arg Pro Gln Asn Ile Asn Ser Ser Asp Asn Ile Cys Val Leu His Glu

85 90 95

Glu Thr Lys Glu Leu Phe Cys Glu Ala Asp Lys Arg Leu Leu Cys Gly

100 105 110

Pro Cys Ser Glu Ser Pro Glu His Met Ala His Ser His Ser Pro Ile

115 120 125

Gly Trp Ala Ala Glu Glu Cys Arg Glu Lys Leu Ile Lys Glu Met Asp

130 135 140

Tyr Leu Trp Glu Ile Asn Gln Glu Thr Arg Asn Asn Leu Asn Gln Glu

145 150 155 160

Thr Arg Thr Phe His Ser Leu Lys Asp Tyr Val Ser Val Arg Lys Arg

165 170 175

Ile Ile Thr Ile Gln Tyr Gln Lys Met Pro Ile Phe Leu Asp Glu Glu

180 185 190

Glu Gln Arg His Leu Gln Ala Leu Glu Arg Glu Ala Glu Glu Leu Phe

195 200 205

Gln Gln Leu Gln Asp Ser Gln Val Arg Met Thr Gln His Leu Glu Arg

210 215 220

Met Lys Asp Met Tyr Arg Glu Leu Trp Glu Thr Cys His Val Pro Asp

225 230 235 240

Val Glu Leu Leu Gln Asp Val Arg Asn Val Ser Ala Arg Thr Asp Leu

245 250 255

Ala Gln Met Gln Lys Pro Gln Pro Val Asn Pro Glu Leu Thr Ser Trp

260 265 270

Cys Ile Thr Gly Val Leu Asp Met Leu Asn Asn Phe Arg Val Asp Ser

275 280 285

Ala Leu Ser Thr Glu Met Ile Pro Cys Tyr Ile Ser Leu Ser Glu Asp

290 295 300

Val Arg Tyr Val Ile Phe Gly Asp Asp His Leu Ser Ala Pro Thr Asp

305 310 315 320

Pro Gln Gly Val Asp Ser Phe Ala Val Trp Gly Ala Gln Ala Phe Thr

325 330 335

Ser Gly Lys His Tyr Trp Glu Val Asp Val Thr Leu Ser Ser Asn Trp

340 345 350

Ile Leu Gly Val Cys Gln Asp Ser Arg Thr Ala Asp Ala Asn Phe Val

355 360 365

Ile Asp Ser Asp Glu Arg Phe Phe Leu Ile Ser Ser Lys Arg Ser Asn

370 375 380

His Tyr Ser Leu Ser Thr Asn Ser Pro Pro Leu Ile Gln Tyr Val Gln

385 390 395 400

Arg Pro Leu Gly Gln Val Gly Val Phe Leu Asp Tyr Asp Asn Gly Ser

405 410 415

Val Ser Phe Phe Asp Val Ser Lys Gly Ser Leu Ile Tyr Gly Phe Pro

420 425 430

Pro Ser Ser Phe Ser Ser Pro Leu Arg Pro Phe Phe Cys Phe Gly Cys

435 440 445

Thr

<210> 125

<211> 1554

<212> DNA

<213> 智人(Homo sapiens)

<400> 125

atggccgcgc agctgctgga ggagaagctg acctgcgcca tctgcctggg gctctaccag 60

gacccagtga cgctgccctg cggccacaac ttctgcgggg cctgcatccg ggactggtgg 120

gaccgctgcg gaaaggcgtg ccccgagtgc cgggagccct ttcccgacgg cgccgagctg 180

cgccgcaacg tggccctcag cggcgtgctg gaggtggtgc gcgccgggcc cgcccgggat 240

cccggccccg atcccggccc cggccccgac cctgccgcgc gctgcccccg ccacgggcgg 300

ccgctggagc tcttctgccg gaccgagggc cgctgtgtgt gcagcgtgtg caccgtgcgc 360

gagtgtcgcc tccacgagcg ggcgctgctg gatgccgagc gcctcaagcg cgaggcccag 420

ctgagagcca gcctggaggt tacccagcag caggccaccc aggccgaagg ccagctacta 480

gagctgcgca agcaaagcag ccagatccag aactcggcct gcatcttggc ctcctgggtc 540

tccggcaagt tcagcagcct gctacaggcc ctggaaatac agcacacgac agcactgagg 600

agcatcgagg tggccaagac gcaggcgctg gcacaggctc gagacgagga gcagcggctg 660

cgggtccatt tggaggctgt ggctcgccat ggctgcagga tccgggagct cctggagcag 720

gtggatgagc agaccttcct gcaggaatcg cagctcctcc agcccccagg gcctcttggg 780

ccactgaccc ctctgcagtg ggatgaagac caacagctgg gtgacctgaa gcagttgcta 840

agccggctgt gtggcctcct cttggaagag gggagccacc ctggggcacc agccaagcct 900

gtggacttag cccccgtgga ggccccaggt cccctggcac cggtcccaag cacagtttgt 960

ccactgagga ggaaactctg gcagaattat cgcaatctga cctttgatcc agtcagcgcc 1020

aaccgtcact tctatctgtc gcgccaggac cagcaggtga agcactgtcg tcagtcccgg 1080

ggcccaggcg ggcccggcag ctttgagctc tggcaggtgc aatgtgccca gagcttccag 1140

gccgggcacc actactggga ggtgcgcgcg tcagaccact cggtgacact gggcgtctcc 1200

tacccgcaac tgccacggtg caggctgggg ccccacacag acaacattgg ccggggaccc 1260

tgctcctggg ggctctgcgt ccaggaggac agcctccagg cctggcacaa cggggaagcc 1320

cagcgcctcc caggggtgtc agggcggctc ctgggcatgg atttggacct ggcctcaggc 1380

tgcctcacct tctacagcct ggagccccag acccagcccc tgtacacctt ccatgccctc 1440

ttcaaccagc ccctcacccc cgtcttctgg ctcctcgagg gtaggaccct gaccctgtgc 1500

catcagccag gggctgtgtt ccctctgggg ccccaggaag aggtgctcag ctga 1554

<210> 126

<211> 517

<212> PRT

<213> 智人(Homo sapiens)

<400> 126

Met Ala Ala Gln Leu Leu Glu Glu Lys Leu Thr Cys Ala Ile Cys Leu

1 5 10 15

Gly Leu Tyr Gln Asp Pro Val Thr Leu Pro Cys Gly His Asn Phe Cys

20 25 30

Gly Ala Cys Ile Arg Asp Trp Trp Asp Arg Cys Gly Lys Ala Cys Pro

35 40 45

Glu Cys Arg Glu Pro Phe Pro Asp Gly Ala Glu Leu Arg Arg Asn Val

50 55 60

Ala Leu Ser Gly Val Leu Glu Val Val Arg Ala Gly Pro Ala Arg Asp

65 70 75 80

Pro Gly Pro Asp Pro Gly Pro Gly Pro Asp Pro Ala Ala Arg Cys Pro

85 90 95

Arg His Gly Arg Pro Leu Glu Leu Phe Cys Arg Thr Glu Gly Arg Cys

100 105 110

Val Cys Ser Val Cys Thr Val Arg Glu Cys Arg Leu His Glu Arg Ala

115 120 125

Leu Leu Asp Ala Glu Arg Leu Lys Arg Glu Ala Gln Leu Arg Ala Ser

130 135 140

Leu Glu Val Thr Gln Gln Gln Ala Thr Gln Ala Glu Gly Gln Leu Leu

145 150 155 160

Glu Leu Arg Lys Gln Ser Ser Gln Ile Gln Asn Ser Ala Cys Ile Leu

165 170 175

Ala Ser Trp Val Ser Gly Lys Phe Ser Ser Leu Leu Gln Ala Leu Glu

180 185 190

Ile Gln His Thr Thr Ala Leu Arg Ser Ile Glu Val Ala Lys Thr Gln

195 200 205

Ala Leu Ala Gln Ala Arg Asp Glu Glu Gln Arg Leu Arg Val His Leu

210 215 220

Glu Ala Val Ala Arg His Gly Cys Arg Ile Arg Glu Leu Leu Glu Gln

225 230 235 240

Val Asp Glu Gln Thr Phe Leu Gln Glu Ser Gln Leu Leu Gln Pro Pro

245 250 255

Gly Pro Leu Gly Pro Leu Thr Pro Leu Gln Trp Asp Glu Asp Gln Gln

260 265 270

Leu Gly Asp Leu Lys Gln Leu Leu Ser Arg Leu Cys Gly Leu Leu Leu

275 280 285

Glu Glu Gly Ser His Pro Gly Ala Pro Ala Lys Pro Val Asp Leu Ala

290 295 300

Pro Val Glu Ala Pro Gly Pro Leu Ala Pro Val Pro Ser Thr Val Cys

305 310 315 320

Pro Leu Arg Arg Lys Leu Trp Gln Asn Tyr Arg Asn Leu Thr Phe Asp

325 330 335

Pro Val Ser Ala Asn Arg His Phe Tyr Leu Ser Arg Gln Asp Gln Gln

340 345 350

Val Lys His Cys Arg Gln Ser Arg Gly Pro Gly Gly Pro Gly Ser Phe

355 360 365

Glu Leu Trp Gln Val Gln Cys Ala Gln Ser Phe Gln Ala Gly His His

370 375 380

Tyr Trp Glu Val Arg Ala Ser Asp His Ser Val Thr Leu Gly Val Ser

385 390 395 400

Tyr Pro Gln Leu Pro Arg Cys Arg Leu Gly Pro His Thr Asp Asn Ile

405 410 415

Gly Arg Gly Pro Cys Ser Trp Gly Leu Cys Val Gln Glu Asp Ser Leu

420 425 430

Gln Ala Trp His Asn Gly Glu Ala Gln Arg Leu Pro Gly Val Ser Gly

435 440 445

Arg Leu Leu Gly Met Asp Leu Asp Leu Ala Ser Gly Cys Leu Thr Phe

450 455 460

Tyr Ser Leu Glu Pro Gln Thr Gln Pro Leu Tyr Thr Phe His Ala Leu

465 470 475 480

Phe Asn Gln Pro Leu Thr Pro Val Phe Trp Leu Leu Glu Gly Arg Thr

485 490 495

Leu Thr Leu Cys His Gln Pro Gly Ala Val Phe Pro Leu Gly Pro Gln

500 505 510

Glu Glu Val Leu Ser

515

<210> 127

<211> 3651

<212> DNA

<213> 智人(Homo sapiens)

<400> 127

atggccagga actgctctga gtgcaaggag aagagggcag cacatatcct ctgcacctac 60

tgcaatcgct ggctgtgcag ctcttgcaca gaggaacacc gacacagccc tgtccccggg 120

ggcccattct ttcctcgggc ccagaaggga tctccaggag tgaatggtgg tcccggagac 180

ttcaccttgt attgtcctct acacacacag gaagtactca agctattctg tgagacatgt 240

gatatgctca cttgccatag ctgcctagtg gtggaacaca aagaacacag gtgcagacat 300

gttgaagaag ttttgcaaaa ccagaggatg cttctggaag gtgtgactac acaggtggca 360

cataagaaat ccagtctaca gacatctgca aagcaaattg aggacaggat ttttgaagtg 420

aagcatcagc ataggaaggt ggaaaaccag atcaaaatgg ccaagatggt tctgatgaat 480

gagctgaaca aacaggccaa tgggctaata gaggaattag aggggattac taatgagaga 540

aagcggaagc tggaacagca gttacagagc atcatggttc tcaaccgtca gtttgagcat 600

gtgcagaatt tcatcaactg ggctgtctgc agcaaaacca gtgtcccttt tcttttcagc 660

aaagagctga ttgtgtttca gatgcagcga ttgctggaga caagttgtaa cacagatcct 720

ggctcccctt ggagtatcag attcacctgg gagcctaact tctggaccaa gcagctagct 780

tctcttggct gcataactac tgaaggtgga caaatgtcca gggcagatgc tcctgcttat 840

ggaggcttac aggggtcatc acccttttat caaagccacc agtctccagt ggctcagcaa 900

gaggctctta gccacccctc acacaagttc cagtctccag cagtgtgctc ctcatctgtg 960

tgctgctccc actgctcccc agtctcgcct tccctcaaag gccaggtccc cccacccagc 1020

atacacccag cccacagctt caggcagccc cctgagatgg tgccccagca gctggggtct 1080

ctgcagtgct ctgccctgct gcccagggag aaagagctgg cctgcagccc tcatccacca 1140

aagctgctgc agccctggct ggaaacccag ccccccgtgg agcaggagag cacatcccag 1200

cggctggggc agcagctgac ttcccagccc gtgtgcattg tccccccaca ggatgttcag 1260

caaggagccc atgcccagcc caccttacag acaccctcta tccaagtcca gtttggccac 1320

caccagaagc tgaagctcag tcactttcag cagcagccac agcagcagct accacctcca 1380

ccaccacccc tcccccatcc cccacctccc cttccccctc ccccacagca gccacaccca 1440

cctcttcctc catcccagca tctggcttct agtcagcacg agagccctcc tggccctgcc 1500

tgttctcaga acatggacat aatgcatcac aagtttgagc tggaggaaat gcagaaggac 1560

ttggagcttc ttctccaggc tcaacagccc agcctgcaac tgagtcagac caaatctcct 1620

cagcatcttc agcaaaccat tgtggggcag atcaactaca tcgtgaggca gccagcacct 1680

gtccagtccc agagccagga ggagaccctg caggctacag atgagccccc agcatctcag 1740

ggctcaaagc cggctctccc tcttgacaag aatactgctg ctgccttgcc ccaggcgtct 1800

ggggaagaaa cccctctcag tgtcccccca gtggacagca ccatccagca ctcctctcca 1860

aatgtggtga gaaagcactc cacctcgctg agcatcatgg gcttttccaa cactctggag 1920

atggagttgt catctaccag gttggagagg cccctagagc cacagatcca gagtgtgagc 1980

aacctgacag ctggtgcccc ccaggcagta ccaagcctgc tgagtgctcc ccccaaaatg 2040

gtgtccagcc tgacaagtgt tcaaaaccag gccatgccca gcctgacaac cagtcaccta 2100

cagactgtgc ccagccttgt gcatagcaca ttccagtcca tgcccaacct gataagtgac 2160

tcccctcagg ctatggcaag cctggcaagt gatcaccctc aggctgggcc cagcctaatg 2220

tctggtcaca cccaggctgt gccgagtctg gcaacttgtc ctctgcagag catccctcca 2280

gtttctgaca tgcagccaga aactgggtcc agctccagtt ctggccgaac ttcagggagc 2340

ctgtgtccca gagatggggc tgatccctcc ctggagaatg ctctgtgtaa ggtaaaactg 2400

gaggagccaa ttaacctctc tgtgaagaaa cctccactgg cgccagtggt cagcacgtct 2460

acagctctgc agcagtacca gaacccaaaa gagtgtgaga attttgaaca aggagcccta 2520

gagctggatg caaaagagaa ccagagcatc agagccttca atagtgagca taagattccc 2580

tatgtgcgac tggagcgact caagatctgt gctgcctcct caggagagat gcctgtgttc 2640

aaactgaagc cacagaagaa tgatcaggat gggagcttcc tgctgatcat cgagtgtggc 2700

actgagtcct ccagcatgtc cattaaggtc agccaggaca gactgtctga ggccacccag 2760

gccccaggtc tggagggaag aaaggtcact gtcacttctt tggctgggca gcggccacca 2820

gaagtggagg gcacatctcc tgaagaacac agactcattc ctcgaacccc aggagccaag 2880

aagggccccc cagccccaat agagaatgag gacttctgtg ctgtttgcct caatggcgga 2940

gagttactgt gctgtgaccg ctgccccaaa gtgttccacc tctcctgcca tgtgccagcc 3000

ttgctcagct tcccaggggg agagtgggtg tgtaccttgt gccgcagcct gacccagccc 3060

gagatggagt acgactgtga gaatgcctgc tataaccagc ctggaatgcg ggcatctcct 3120

ggcctaagca tgtatgacca gaagaagtgt gagaagctgg tattgtcctt gtgctgcaat 3180

aacctcagcc tgcccttcca tgaacctgtc agccccctgg cccggcatta ttaccagatt 3240

atcaagaggc ccatggacct gtcaatcatc cggaggaagc tgcaaaagaa ggacccagct 3300

cactatacca ccccagagga ggtggtatca gatgtgcgcc tcatgttctg gaactgtgct 3360

aagttcaatt atcctgactc cgaggttgca gaggctggcc gctgcctgga agtgttcttt 3420

gagggctggt tgaaggagat ctacccggag aaacggtttg cccagccaag gcaggaggac 3480

tcagactccg aggaggtgtc tagtgagagt ggatgttcca ctccccaggg cttcccgtgg 3540

cctccctaca tgcaggaggg catccaaccc aagaggcggc gacgacatat ggagaatgaa 3600

agagcaaaaa gaatgtcatt tcgcctggcc aacagcatct ctcaggtgtg a 3651

<210> 128

<211> 1216

<212> PRT

<213> 智人(Homo sapiens)

<400> 128

Met Ala Arg Asn Cys Ser Glu Cys Lys Glu Lys Arg Ala Ala His Ile

1 5 10 15

Leu Cys Thr Tyr Cys Asn Arg Trp Leu Cys Ser Ser Cys Thr Glu Glu

20 25 30

His Arg His Ser Pro Val Pro Gly Gly Pro Phe Phe Pro Arg Ala Gln

35 40 45

Lys Gly Ser Pro Gly Val Asn Gly Gly Pro Gly Asp Phe Thr Leu Tyr

50 55 60

Cys Pro Leu His Thr Gln Glu Val Leu Lys Leu Phe Cys Glu Thr Cys

65 70 75 80

Asp Met Leu Thr Cys His Ser Cys Leu Val Val Glu His Lys Glu His

85 90 95

Arg Cys Arg His Val Glu Glu Val Leu Gln Asn Gln Arg Met Leu Leu

100 105 110

Glu Gly Val Thr Thr Gln Val Ala His Lys Lys Ser Ser Leu Gln Thr

115 120 125

Ser Ala Lys Gln Ile Glu Asp Arg Ile Phe Glu Val Lys His Gln His

130 135 140

Arg Lys Val Glu Asn Gln Ile Lys Met Ala Lys Met Val Leu Met Asn

145 150 155 160

Glu Leu Asn Lys Gln Ala Asn Gly Leu Ile Glu Glu Leu Glu Gly Ile

165 170 175

Thr Asn Glu Arg Lys Arg Lys Leu Glu Gln Gln Leu Gln Ser Ile Met

180 185 190

Val Leu Asn Arg Gln Phe Glu His Val Gln Asn Phe Ile Asn Trp Ala

195 200 205

Val Cys Ser Lys Thr Ser Val Pro Phe Leu Phe Ser Lys Glu Leu Ile

210 215 220

Val Phe Gln Met Gln Arg Leu Leu Glu Thr Ser Cys Asn Thr Asp Pro

225 230 235 240

Gly Ser Pro Trp Ser Ile Arg Phe Thr Trp Glu Pro Asn Phe Trp Thr

245 250 255

Lys Gln Leu Ala Ser Leu Gly Cys Ile Thr Thr Glu Gly Gly Gln Met

260 265 270

Ser Arg Ala Asp Ala Pro Ala Tyr Gly Gly Leu Gln Gly Ser Ser Pro

275 280 285

Phe Tyr Gln Ser His Gln Ser Pro Val Ala Gln Gln Glu Ala Leu Ser

290 295 300

His Pro Ser His Lys Phe Gln Ser Pro Ala Val Cys Ser Ser Ser Val

305 310 315 320

Cys Cys Ser His Cys Ser Pro Val Ser Pro Ser Leu Lys Gly Gln Val

325 330 335

Pro Pro Pro Ser Ile His Pro Ala His Ser Phe Arg Gln Pro Pro Glu

340 345 350

Met Val Pro Gln Gln Leu Gly Ser Leu Gln Cys Ser Ala Leu Leu Pro

355 360 365

Arg Glu Lys Glu Leu Ala Cys Ser Pro His Pro Pro Lys Leu Leu Gln

370 375 380

Pro Trp Leu Glu Thr Gln Pro Pro Val Glu Gln Glu Ser Thr Ser Gln

385 390 395 400

Arg Leu Gly Gln Gln Leu Thr Ser Gln Pro Val Cys Ile Val Pro Pro

405 410 415

Gln Asp Val Gln Gln Gly Ala His Ala Gln Pro Thr Leu Gln Thr Pro

420 425 430

Ser Ile Gln Val Gln Phe Gly His His Gln Lys Leu Lys Leu Ser His

435 440 445

Phe Gln Gln Gln Pro Gln Gln Gln Leu Pro Pro Pro Pro Pro Pro Leu

450 455 460

Pro His Pro Pro Pro Pro Leu Pro Pro Pro Pro Gln Gln Pro His Pro

465 470 475 480

Pro Leu Pro Pro Ser Gln His Leu Ala Ser Ser Gln His Glu Ser Pro

485 490 495

Pro Gly Pro Ala Cys Ser Gln Asn Met Asp Ile Met His His Lys Phe

500 505 510

Glu Leu Glu Glu Met Gln Lys Asp Leu Glu Leu Leu Leu Gln Ala Gln

515 520 525

Gln Pro Ser Leu Gln Leu Ser Gln Thr Lys Ser Pro Gln His Leu Gln

530 535 540

Gln Thr Ile Val Gly Gln Ile Asn Tyr Ile Val Arg Gln Pro Ala Pro

545 550 555 560

Val Gln Ser Gln Ser Gln Glu Glu Thr Leu Gln Ala Thr Asp Glu Pro

565 570 575

Pro Ala Ser Gln Gly Ser Lys Pro Ala Leu Pro Leu Asp Lys Asn Thr

580 585 590

Ala Ala Ala Leu Pro Gln Ala Ser Gly Glu Glu Thr Pro Leu Ser Val

595 600 605

Pro Pro Val Asp Ser Thr Ile Gln His Ser Ser Pro Asn Val Val Arg

610 615 620

Lys His Ser Thr Ser Leu Ser Ile Met Gly Phe Ser Asn Thr Leu Glu

625 630 635 640

Met Glu Leu Ser Ser Thr Arg Leu Glu Arg Pro Leu Glu Pro Gln Ile

645 650 655

Gln Ser Val Ser Asn Leu Thr Ala Gly Ala Pro Gln Ala Val Pro Ser

660 665 670

Leu Leu Ser Ala Pro Pro Lys Met Val Ser Ser Leu Thr Ser Val Gln

675 680 685

Asn Gln Ala Met Pro Ser Leu Thr Thr Ser His Leu Gln Thr Val Pro

690 695 700

Ser Leu Val His Ser Thr Phe Gln Ser Met Pro Asn Leu Ile Ser Asp

705 710 715 720

Ser Pro Gln Ala Met Ala Ser Leu Ala Ser Asp His Pro Gln Ala Gly

725 730 735

Pro Ser Leu Met Ser Gly His Thr Gln Ala Val Pro Ser Leu Ala Thr

740 745 750

Cys Pro Leu Gln Ser Ile Pro Pro Val Ser Asp Met Gln Pro Glu Thr

755 760 765

Gly Ser Ser Ser Ser Ser Gly Arg Thr Ser Gly Ser Leu Cys Pro Arg

770 775 780

Asp Gly Ala Asp Pro Ser Leu Glu Asn Ala Leu Cys Lys Val Lys Leu

785 790 795 800

Glu Glu Pro Ile Asn Leu Ser Val Lys Lys Pro Pro Leu Ala Pro Val

805 810 815

Val Ser Thr Ser Thr Ala Leu Gln Gln Tyr Gln Asn Pro Lys Glu Cys

820 825 830

Glu Asn Phe Glu Gln Gly Ala Leu Glu Leu Asp Ala Lys Glu Asn Gln

835 840 845

Ser Ile Arg Ala Phe Asn Ser Glu His Lys Ile Pro Tyr Val Arg Leu

850 855 860

Glu Arg Leu Lys Ile Cys Ala Ala Ser Ser Gly Glu Met Pro Val Phe

865 870 875 880

Lys Leu Lys Pro Gln Lys Asn Asp Gln Asp Gly Ser Phe Leu Leu Ile

885 890 895

Ile Glu Cys Gly Thr Glu Ser Ser Ser Met Ser Ile Lys Val Ser Gln

900 905 910

Asp Arg Leu Ser Glu Ala Thr Gln Ala Pro Gly Leu Glu Gly Arg Lys

915 920 925

Val Thr Val Thr Ser Leu Ala Gly Gln Arg Pro Pro Glu Val Glu Gly

930 935 940

Thr Ser Pro Glu Glu His Arg Leu Ile Pro Arg Thr Pro Gly Ala Lys

945 950 955 960

Lys Gly Pro Pro Ala Pro Ile Glu Asn Glu Asp Phe Cys Ala Val Cys

965 970 975

Leu Asn Gly Gly Glu Leu Leu Cys Cys Asp Arg Cys Pro Lys Val Phe

980 985 990

His Leu Ser Cys His Val Pro Ala Leu Leu Ser Phe Pro Gly Gly Glu

995 1000 1005

Trp Val Cys Thr Leu Cys Arg Ser Leu Thr Gln Pro Glu Met Glu

1010 1015 1020

Tyr Asp Cys Glu Asn Ala Cys Tyr Asn Gln Pro Gly Met Arg Ala

1025 1030 1035

Ser Pro Gly Leu Ser Met Tyr Asp Gln Lys Lys Cys Glu Lys Leu

1040 1045 1050

Val Leu Ser Leu Cys Cys Asn Asn Leu Ser Leu Pro Phe His Glu

1055 1060 1065

Pro Val Ser Pro Leu Ala Arg His Tyr Tyr Gln Ile Ile Lys Arg

1070 1075 1080

Pro Met Asp Leu Ser Ile Ile Arg Arg Lys Leu Gln Lys Lys Asp

1085 1090 1095

Pro Ala His Tyr Thr Thr Pro Glu Glu Val Val Ser Asp Val Arg

1100 1105 1110

Leu Met Phe Trp Asn Cys Ala Lys Phe Asn Tyr Pro Asp Ser Glu

1115 1120 1125

Val Ala Glu Ala Gly Arg Cys Leu Glu Val Phe Phe Glu Gly Trp

1130 1135 1140

Leu Lys Glu Ile Tyr Pro Glu Lys Arg Phe Ala Gln Pro Arg Gln

1145 1150 1155

Glu Asp Ser Asp Ser Glu Glu Val Ser Ser Glu Ser Gly Cys Ser

1160 1165 1170

Thr Pro Gln Gly Phe Pro Trp Pro Pro Tyr Met Gln Glu Gly Ile

1175 1180 1185

Gln Pro Lys Arg Arg Arg Arg His Met Glu Asn Glu Arg Ala Lys

1190 1195 1200

Arg Met Ser Phe Arg Leu Ala Asn Ser Ile Ser Gln Val

1205 1210 1215

<210> 129

<211> 2352

<212> DNA

<213> 智人(Homo sapiens)

<400> 129

atggaggaag agctgaagtg tcccgtgtgc ggctctctgt ttcgggagcc tatcatcctg 60

ccctgttccc acaatgtctg cctgccttgc gctcgcacca tcgcggtgca gaccccggac 120

ggtgagcagc acctgcccca gccgctcctg ctttcccggg gatcggggct gcaggcgggc 180

gccgccgccg ctgcctctct ggagcacgac gctgcggctg gcccggcctg cggcggtgca 240

ggcgggagtg cagctggcgg cctcggcggc ggtgcgggag gtggcggaga ccacgcggac 300

aagctcagct tgtacagcga gacagacagc ggctacgggt cctacacccc gagcctcaag 360

tcccccaacg gggttcgcgt gctgcccatg gtgcccgcac cacccggctc ctcggctgcg 420

gcggctcggg gtgccgcctg ctcctcgctg tcctcgtctt cgagctccat cacgtgcccg 480

cagtgccacc gcagcgcatc cctggaccac cgcggcctgc gcggcttcca gcgcaaccgg 540

ctgctcgagg ccatcgtgca gcggtaccag cagggccgcg gggccgtgcc ggggacgtct 600

gcagccgcgg cggtggccat ctgccagctg tgcgaccgca ccccgccaga gccagcagcc 660

acgctctgcg agcagtgcga cgtcctctac tgctctgcct gccagctcaa gtgccatcca 720

tcccggggac ccttcgccaa gcatcgcctg gtgcagccgc cgccgccgcc gccgccgccc 780

gccgaggcag cctccgggcc cactggcacc gcccagggcg cccccagcgg aggcggcggc 840

tgcaagagcc cgggaggcgc gggggcgggg gcgactgggg gcagcacggc ccgcaagttc 900

cccacgtgtc ccgagcatga aatggagaac tacagcatgt actgcgtgag ctgtcgaacc 960

ccggtgtgtt atctgtgcct ggaggagggc cggcacgcca agcacgaggt gaagccgctg 1020

ggggccatgt ggaagcagca caaggcacaa ctatctcagg ccttaaatgg agtttcagat 1080

aaggcaaagg aagcaaagga gtttctggtt cagctaaaga acatattgca gcagatccag 1140

gaaaacggac tggactacga agcctgcctc gttgctcagt gtgatgccct tgtggatgct 1200

ttaactcgtc agaaagccaa gctgctcacc aaggtgacta aagagaggga acacaagttg 1260

aagatggttt gggaccagat caatcactgc acattgaagc tgcgtcagtc caccggactg 1320

atggagtact gcctggaggt gatcaaggag aacgacccct ccgggttctt acagatctca 1380

gatgctctga tcaagcgcgt ccaggtgtct caggagcagt gggtcaaagg cgccctggag 1440

ccgaaagtgt ctgcggagtt tgatctgact ttggacagcg agccgctgct gcaggccatc 1500

caccagctgg acttcattca gatgaaatgt agggtgccac ccgtccccct actgcagctg 1560

gagaaatgct gcacccgtaa caacagcgtc acgctggcct ggaggatgcc acccttcacc 1620

cacagccccg tggacggcta catcctggag ctggacgacg gtgccggggg acagttccgg 1680

gaagtgtacg tcggtaagga gactttgtgt accatcgacg gtcttcactt caacagcacc 1740

tacaacgccc gagtcaaagc tttcaactct tctggtgtcg ggccttacag taaaactgtc 1800

gtcctgcaga catccgatgt ggcctggttc acatttgacc ccaactctgg gcatcgggac 1860

atcattttat ccaatgacaa ccagacagcc acctgcagca gctatgacga ccgggtggtg 1920

ctgggcacag ctgcgttctc caagggcgtg cactactggg agctgcacgt ggaccggtac 1980

gacaaccacc cagaccccgc cttcggggtg gccagggcca gcgtggtcaa ggacatgatg 2040

ctgggcaagg atgacaaggc ctgggccatg tatgtggaca acaaccgcag ctggttcatg 2100

cactgcaact cccacaccaa caggacggaa ggtggcgtgt gcaagggggc caccgtgggc 2160

gtgctgctgg acctgaataa gcacactctc accttcttca tcaacgggca gcagcagggc 2220

cccacagcct tcagccacgt ggacggggtc ttcatgccag ccctcagcct caaccgcaac 2280

gtgcaggtca ccctgcacac aggattggaa gtgccgacta acctggggcg gccaaagctg 2340

tcaggcaatt ag 2352

<210> 130

<211> 783

<212> PRT

<213> 智人(Homo sapiens)

<400> 130

Met Glu Glu Glu Leu Lys Cys Pro Val Cys Gly Ser Leu Phe Arg Glu

1 5 10 15

Pro Ile Ile Leu Pro Cys Ser His Asn Val Cys Leu Pro Cys Ala Arg

20 25 30

Thr Ile Ala Val Gln Thr Pro Asp Gly Glu Gln His Leu Pro Gln Pro

35 40 45

Leu Leu Leu Ser Arg Gly Ser Gly Leu Gln Ala Gly Ala Ala Ala Ala

50 55 60

Ala Ser Leu Glu His Asp Ala Ala Ala Gly Pro Ala Cys Gly Gly Ala

65 70 75 80

Gly Gly Ser Ala Ala Gly Gly Leu Gly Gly Gly Ala Gly Gly Gly Gly

85 90 95

Asp His Ala Asp Lys Leu Ser Leu Tyr Ser Glu Thr Asp Ser Gly Tyr

100 105 110

Gly Ser Tyr Thr Pro Ser Leu Lys Ser Pro Asn Gly Val Arg Val Leu

115 120 125

Pro Met Val Pro Ala Pro Pro Gly Ser Ser Ala Ala Ala Ala Arg Gly

130 135 140

Ala Ala Cys Ser Ser Leu Ser Ser Ser Ser Ser Ser Ile Thr Cys Pro

145 150 155 160

Gln Cys His Arg Ser Ala Ser Leu Asp His Arg Gly Leu Arg Gly Phe

165 170 175

Gln Arg Asn Arg Leu Leu Glu Ala Ile Val Gln Arg Tyr Gln Gln Gly

180 185 190

Arg Gly Ala Val Pro Gly Thr Ser Ala Ala Ala Ala Val Ala Ile Cys

195 200 205

Gln Leu Cys Asp Arg Thr Pro Pro Glu Pro Ala Ala Thr Leu Cys Glu

210 215 220

Gln Cys Asp Val Leu Tyr Cys Ser Ala Cys Gln Leu Lys Cys His Pro

225 230 235 240

Ser Arg Gly Pro Phe Ala Lys His Arg Leu Val Gln Pro Pro Pro Pro

245 250 255

Pro Pro Pro Pro Ala Glu Ala Ala Ser Gly Pro Thr Gly Thr Ala Gln

260 265 270

Gly Ala Pro Ser Gly Gly Gly Gly Cys Lys Ser Pro Gly Gly Ala Gly

275 280 285

Ala Gly Ala Thr Gly Gly Ser Thr Ala Arg Lys Phe Pro Thr Cys Pro

290 295 300

Glu His Glu Met Glu Asn Tyr Ser Met Tyr Cys Val Ser Cys Arg Thr

305 310 315 320

Pro Val Cys Tyr Leu Cys Leu Glu Glu Gly Arg His Ala Lys His Glu

325 330 335

Val Lys Pro Leu Gly Ala Met Trp Lys Gln His Lys Ala Gln Leu Ser

340 345 350

Gln Ala Leu Asn Gly Val Ser Asp Lys Ala Lys Glu Ala Lys Glu Phe

355 360 365

Leu Val Gln Leu Lys Asn Ile Leu Gln Gln Ile Gln Glu Asn Gly Leu

370 375 380

Asp Tyr Glu Ala Cys Leu Val Ala Gln Cys Asp Ala Leu Val Asp Ala

385 390 395 400

Leu Thr Arg Gln Lys Ala Lys Leu Leu Thr Lys Val Thr Lys Glu Arg

405 410 415

Glu His Lys Leu Lys Met Val Trp Asp Gln Ile Asn His Cys Thr Leu

420 425 430

Lys Leu Arg Gln Ser Thr Gly Leu Met Glu Tyr Cys Leu Glu Val Ile

435 440 445

Lys Glu Asn Asp Pro Ser Gly Phe Leu Gln Ile Ser Asp Ala Leu Ile

450 455 460

Lys Arg Val Gln Val Ser Gln Glu Gln Trp Val Lys Gly Ala Leu Glu

465 470 475 480

Pro Lys Val Ser Ala Glu Phe Asp Leu Thr Leu Asp Ser Glu Pro Leu

485 490 495

Leu Gln Ala Ile His Gln Leu Asp Phe Ile Gln Met Lys Cys Arg Val

500 505 510

Pro Pro Val Pro Leu Leu Gln Leu Glu Lys Cys Cys Thr Arg Asn Asn

515 520 525

Ser Val Thr Leu Ala Trp Arg Met Pro Pro Phe Thr His Ser Pro Val

530 535 540

Asp Gly Tyr Ile Leu Glu Leu Asp Asp Gly Ala Gly Gly Gln Phe Arg

545 550 555 560

Glu Val Tyr Val Gly Lys Glu Thr Leu Cys Thr Ile Asp Gly Leu His

565 570 575

Phe Asn Ser Thr Tyr Asn Ala Arg Val Lys Ala Phe Asn Ser Ser Gly

580 585 590

Val Gly Pro Tyr Ser Lys Thr Val Val Leu Gln Thr Ser Asp Val Ala

595 600 605

Trp Phe Thr Phe Asp Pro Asn Ser Gly His Arg Asp Ile Ile Leu Ser

610 615 620

Asn Asp Asn Gln Thr Ala Thr Cys Ser Ser Tyr Asp Asp Arg Val Val

625 630 635 640

Leu Gly Thr Ala Ala Phe Ser Lys Gly Val His Tyr Trp Glu Leu His

645 650 655

Val Asp Arg Tyr Asp Asn His Pro Asp Pro Ala Phe Gly Val Ala Arg

660 665 670

Ala Ser Val Val Lys Asp Met Met Leu Gly Lys Asp Asp Lys Ala Trp

675 680 685

Ala Met Tyr Val Asp Asn Asn Arg Ser Trp Phe Met His Cys Asn Ser

690 695 700

His Thr Asn Arg Thr Glu Gly Gly Val Cys Lys Gly Ala Thr Val Gly

705 710 715 720

Val Leu Leu Asp Leu Asn Lys His Thr Leu Thr Phe Phe Ile Asn Gly

725 730 735

Gln Gln Gln Gly Pro Thr Ala Phe Ser His Val Asp Gly Val Phe Met

740 745 750

Pro Ala Leu Ser Leu Asn Arg Asn Val Gln Val Thr Leu His Thr Gly

755 760 765

Leu Glu Val Pro Thr Asn Leu Gly Arg Pro Lys Leu Ser Gly Asn

770 775 780

<210> 131

<211> 1458

<212> DNA

<213> 智人(Homo sapiens)

<400> 131

atggatccca cagccttggt ggaagccatt gtggaagaag tggcctgtcc catctgtatg 60

accttcctga gggagcccat gagcattgac tgtggccaca gcttctgcca cagctgtctc 120

tctggactct gggagatccc aggagaatcc cagaactggg gttacacctg tcccctctgt 180

cgagctcctg tccagccaag gaacctgcgg cctaattggc agctggccaa tgttgtagaa 240

aaagtccgtc tgctaaggct acatccagga atggggctga agggtgacct gtgtgagcgc 300

catggggaaa agctgaagat gttctgcaaa gaggatgtct tgataatgtg tgaggcctgc 360

agccagtccc cagagcatga ggcccacagt gttgtgccaa tggaggatgt tgcctgggag 420

tacaagtggg aacttcatga ggccctcgaa catctgaaga aagagcaaga agaggcctgg 480

aagcttgaag ttggtgaaag gaaacgaact gccacctgga agatacaggt ggaaacccga 540

aaacagagta ttgtatggga gtttgaaaaa taccagcgat tactagagaa aaagcagcca 600

ccacatcggc agctgggggc agaggtagca gcagctctgg ccagcctaca gcgggaggca 660

gcggagacca tgcagaaact ggagttgaac catagcgagc tcatccagca gagccaggtc 720

ctgtggagga tgattgcaga gttgaaagag aggtcgcaga ggcctgtccg ctggatgttg 780

caggatattc aggaagtgtt aaacaggagc aaatcttgga gcttgcagca gccagaacca 840

atctccctgg agttgaagac agattgccgt gtgctggggc taagagagat cctgaagact 900

tatgcagctg atgtgcgctt ggatccagat actgcttact cccgtctcat cgtgtctgag 960

gacagaaaac gtgtgcacta tggagacacc aaccagaaac tgccagacaa tcctgagaga 1020

ttttaccgct ataatatcgt cctgggaagc cagtgcatct cctcaggccg gcactactgg 1080

gaggtggagg tgggagacag gtctgagtgg ggcctgggag tatgtaagca aaatgtagac 1140

cggaaggagg tggtctactt atccccccac tatggattct gggtgataag gctgaggaag 1200

ggaaatgagt accgagcagg caccgatgag tacccaatcc tgtccttgcc ggtccctcct 1260

cgccgggtgg gaatcttcgt ggattatgag gcccatgaca tttctttcta caatgtgact 1320

gactgtggct cccacatctt cactttcccc cgctatccct tccctgggcg cctcctgccc 1380

tattttagtc cttgctacag cattggaacc aacaacactg ctcctctggc catctgctcc 1440

ctggatgggg aggactaa 1458

<210> 132

<211> 485

<212> PRT

<213> 智人(Homo sapiens)

<400> 132

Met Asp Pro Thr Ala Leu Val Glu Ala Ile Val Glu Glu Val Ala Cys

1 5 10 15

Pro Ile Cys Met Thr Phe Leu Arg Glu Pro Met Ser Ile Asp Cys Gly

20 25 30

His Ser Phe Cys His Ser Cys Leu Ser Gly Leu Trp Glu Ile Pro Gly

35 40 45

Glu Ser Gln Asn Trp Gly Tyr Thr Cys Pro Leu Cys Arg Ala Pro Val

50 55 60

Gln Pro Arg Asn Leu Arg Pro Asn Trp Gln Leu Ala Asn Val Val Glu

65 70 75 80

Lys Val Arg Leu Leu Arg Leu His Pro Gly Met Gly Leu Lys Gly Asp

85 90 95

Leu Cys Glu Arg His Gly Glu Lys Leu Lys Met Phe Cys Lys Glu Asp

100 105 110

Val Leu Ile Met Cys Glu Ala Cys Ser Gln Ser Pro Glu His Glu Ala

115 120 125

His Ser Val Val Pro Met Glu Asp Val Ala Trp Glu Tyr Lys Trp Glu

130 135 140

Leu His Glu Ala Leu Glu His Leu Lys Lys Glu Gln Glu Glu Ala Trp

145 150 155 160

Lys Leu Glu Val Gly Glu Arg Lys Arg Thr Ala Thr Trp Lys Ile Gln

165 170 175

Val Glu Thr Arg Lys Gln Ser Ile Val Trp Glu Phe Glu Lys Tyr Gln

180 185 190

Arg Leu Leu Glu Lys Lys Gln Pro Pro His Arg Gln Leu Gly Ala Glu

195 200 205

Val Ala Ala Ala Leu Ala Ser Leu Gln Arg Glu Ala Ala Glu Thr Met

210 215 220

Gln Lys Leu Glu Leu Asn His Ser Glu Leu Ile Gln Gln Ser Gln Val

225 230 235 240

Leu Trp Arg Met Ile Ala Glu Leu Lys Glu Arg Ser Gln Arg Pro Val

245 250 255

Arg Trp Met Leu Gln Asp Ile Gln Glu Val Leu Asn Arg Ser Lys Ser

260 265 270

Trp Ser Leu Gln Gln Pro Glu Pro Ile Ser Leu Glu Leu Lys Thr Asp

275 280 285

Cys Arg Val Leu Gly Leu Arg Glu Ile Leu Lys Thr Tyr Ala Ala Asp

290 295 300

Val Arg Leu Asp Pro Asp Thr Ala Tyr Ser Arg Leu Ile Val Ser Glu

305 310 315 320

Asp Arg Lys Arg Val His Tyr Gly Asp Thr Asn Gln Lys Leu Pro Asp

325 330 335

Asn Pro Glu Arg Phe Tyr Arg Tyr Asn Ile Val Leu Gly Ser Gln Cys

340 345 350

Ile Ser Ser Gly Arg His Tyr Trp Glu Val Glu Val Gly Asp Arg Ser

355 360 365

Glu Trp Gly Leu Gly Val Cys Lys Gln Asn Val Asp Arg Lys Glu Val

370 375 380

Val Tyr Leu Ser Pro His Tyr Gly Phe Trp Val Ile Arg Leu Arg Lys

385 390 395 400

Gly Asn Glu Tyr Arg Ala Gly Thr Asp Glu Tyr Pro Ile Leu Ser Leu

405 410 415

Pro Val Pro Pro Arg Arg Val Gly Ile Phe Val Asp Tyr Glu Ala His

420 425 430

Asp Ile Ser Phe Tyr Asn Val Thr Asp Cys Gly Ser His Ile Phe Thr

435 440 445

Phe Pro Arg Tyr Pro Phe Pro Gly Arg Leu Leu Pro Tyr Phe Ser Pro

450 455 460

Cys Tyr Ser Ile Gly Thr Asn Asn Thr Ala Pro Leu Ala Ile Cys Ser

465 470 475 480

Leu Asp Gly Glu Asp

485

<210> 133

<211> 1026

<212> DNA

<213> 智人(Homo sapiens)

<400> 133

atggaggagg agcttgccat ccaacagggt caactggaga caactctgaa ggagcttcag 60

accctgagga acatgcagaa ggaagctatt gctgctcaca aggaaaacaa gctacatctg 120

cagcaacatg tgtccatgga gtttctaaag ctgcatcagt tcctgcacag caaagaaaag 180

gacattttaa ctgagctccg ggaagagggg aaagccttga atgaggagat ggagttgaat 240

ctgagccagc ttcaggagca atgtctctta gccaaggata tgttggtgag cattcaggca 300

aagacggaac aacagaactc cttcgacttt ctcaaagaca tcacaactct cttacatagc 360

ttggagcaag gaatgaaggt gctggcaacc agagagctta tttccagaaa gctgaacctg 420

ggccagtaca aaggtcctat ccagtacatg gtatggaggg aaatgcagga cactctctgc 480

ccaggcctgt ctccactaac tctggaccct aaaacagctc acccaaatct ggtgctctcc 540

aaaagccaaa ccagcgtctg gcatggtgac attaagaaga taatgcctga tgatcctgag 600

aggtttgact caagtgtggc tgtactgggc tcaagaggct tcacctctgg aaagtggtac 660

tgggaagtag aagtagcaaa gaagacaaaa tggacagttg gagttgtcag agaatccatc 720

attcggaagg gcagctgtcc tctaactcct gagcaaggat tctggctttt aagactaagg 780

aaccaaactg atctaaaggc tctggatttg ccttctttca gtctgacact gactaacaac 840

ctcgacaagg tgggcatata cctggattat gaaggaggac agttgtcctt ctacaatgct 900

aaaaccatga ctcacattta caccttcagt aacactttca tggagaaact ttatccctac 960

ttctgcccct gccttaatga tggtggagag aataaagaac cattgcacat cttacatcca 1020

cagtaa 1026

<210> 134

<211> 341

<212> PRT

<213> 智人(Homo sapiens)

<400> 134

Met Glu Glu Glu Leu Ala Ile Gln Gln Gly Gln Leu Glu Thr Thr Leu

1 5 10 15

Lys Glu Leu Gln Thr Leu Arg Asn Met Gln Lys Glu Ala Ile Ala Ala

20 25 30

His Lys Glu Asn Lys Leu His Leu Gln Gln His Val Ser Met Glu Phe

35 40 45

Leu Lys Leu His Gln Phe Leu His Ser Lys Glu Lys Asp Ile Leu Thr

50 55 60

Glu Leu Arg Glu Glu Gly Lys Ala Leu Asn Glu Glu Met Glu Leu Asn

65 70 75 80

Leu Ser Gln Leu Gln Glu Gln Cys Leu Leu Ala Lys Asp Met Leu Val

85 90 95

Ser Ile Gln Ala Lys Thr Glu Gln Gln Asn Ser Phe Asp Phe Leu Lys

100 105 110

Asp Ile Thr Thr Leu Leu His Ser Leu Glu Gln Gly Met Lys Val Leu

115 120 125

Ala Thr Arg Glu Leu Ile Ser Arg Lys Leu Asn Leu Gly Gln Tyr Lys

130 135 140

Gly Pro Ile Gln Tyr Met Val Trp Arg Glu Met Gln Asp Thr Leu Cys

145 150 155 160

Pro Gly Leu Ser Pro Leu Thr Leu Asp Pro Lys Thr Ala His Pro Asn

165 170 175

Leu Val Leu Ser Lys Ser Gln Thr Ser Val Trp His Gly Asp Ile Lys

180 185 190

Lys Ile Met Pro Asp Asp Pro Glu Arg Phe Asp Ser Ser Val Ala Val

195 200 205

Leu Gly Ser Arg Gly Phe Thr Ser Gly Lys Trp Tyr Trp Glu Val Glu

210 215 220

Val Ala Lys Lys Thr Lys Trp Thr Val Gly Val Val Arg Glu Ser Ile

225 230 235 240

Ile Arg Lys Gly Ser Cys Pro Leu Thr Pro Glu Gln Gly Phe Trp Leu

245 250 255

Leu Arg Leu Arg Asn Gln Thr Asp Leu Lys Ala Leu Asp Leu Pro Ser

260 265 270

Phe Ser Leu Thr Leu Thr Asn Asn Leu Asp Lys Val Gly Ile Tyr Leu

275 280 285

Asp Tyr Glu Gly Gly Gln Leu Ser Phe Tyr Asn Ala Lys Thr Met Thr

290 295 300

His Ile Tyr Thr Phe Ser Asn Thr Phe Met Glu Lys Leu Tyr Pro Tyr

305 310 315 320

Phe Cys Pro Cys Leu Asn Asp Gly Gly Glu Asn Lys Glu Pro Leu His

325 330 335

Ile Leu His Pro Gln

340

<210> 135

<211> 1047

<212> DNA

<213> 智人(Homo sapiens)

<400> 135

atgcagtttg gggaactcct tgctgctgtg aggaaggccc aggccaatgt gatgctcttc 60

ttagaggaga aggagcaagc tgcgctgagc caggccaacg gtatcaaggc ccacctggag 120

tacaggagtg ccgagatgga gaagagtaag caggagctgg agacgatggc ggccatcagc 180

aacactgtcc agttcttgga ggagtactgc aagtttaaga acactgaaga catcaccttc 240

cctagtgttt acatagggct gaaggataaa ctctcgggca tccgcaaagt tatcacggaa 300

tccactgtac acttaatcca gttgctggag aactataaga aaaagctcca ggagttttcc 360

aaggaagagg agtatgacat cagaactcaa gtgtctgcca ttgttcagcg caaatattgg 420

acttccaaac ctgagcccag caccagggaa cagttcctcc aatatgtgca tgacatcacg 480

ttcgacccgg acacagcaca caagtatctc cggctgcagg aggagaaccg caaggtcacc 540

aacaccacgc cctgggagca tccctacccg gacctcccca gcaggttcct gcactggcgg 600

caggtgctgt cccagcagag tctgtacctg cacaggtact attttgaggt ggagatcttc 660

ggggcaggca cctatgttgg cctgacctgc aaaggcatcg accagaaagg ggaggagcgc 720

agcagttgca tttccggaaa caacttctcc tggagcctcc aatggaacgg gaaggagttc 780

acggcctggt acagtgacat ggagacccca ctcaaagctg gccctttctg gaggctcggg 840

gtctatattg acttcccagg agggatcctt tccttctatg gcgtagagta tgattccatg 900

actctggttc acaagtttgc ctgcaagttt tcagaaccag tctatgctgc cttctggctt 960

tccaagaagg aaaacgccat ccggattgta gatctgggag aggaacccga gaagccagca 1020

ccgtccttgg tggggactgc tccctag 1047

<210> 136

<211> 348

<212> PRT

<213> 智人(Homo sapiens)

<400> 136

Met Gln Phe Gly Glu Leu Leu Ala Ala Val Arg Lys Ala Gln Ala Asn

1 5 10 15

Val Met Leu Phe Leu Glu Glu Lys Glu Gln Ala Ala Leu Ser Gln Ala

20 25 30

Asn Gly Ile Lys Ala His Leu Glu Tyr Arg Ser Ala Glu Met Glu Lys

35 40 45

Ser Lys Gln Glu Leu Glu Thr Met Ala Ala Ile Ser Asn Thr Val Gln

50 55 60

Phe Leu Glu Glu Tyr Cys Lys Phe Lys Asn Thr Glu Asp Ile Thr Phe

65 70 75 80

Pro Ser Val Tyr Ile Gly Leu Lys Asp Lys Leu Ser Gly Ile Arg Lys

85 90 95

Val Ile Thr Glu Ser Thr Val His Leu Ile Gln Leu Leu Glu Asn Tyr

100 105 110

Lys Lys Lys Leu Gln Glu Phe Ser Lys Glu Glu Glu Tyr Asp Ile Arg

115 120 125

Thr Gln Val Ser Ala Ile Val Gln Arg Lys Tyr Trp Thr Ser Lys Pro

130 135 140

Glu Pro Ser Thr Arg Glu Gln Phe Leu Gln Tyr Val His Asp Ile Thr

145 150 155 160

Phe Asp Pro Asp Thr Ala His Lys Tyr Leu Arg Leu Gln Glu Glu Asn

165 170 175

Arg Lys Val Thr Asn Thr Thr Pro Trp Glu His Pro Tyr Pro Asp Leu

180 185 190

Pro Ser Arg Phe Leu His Trp Arg Gln Val Leu Ser Gln Gln Ser Leu

195 200 205

Tyr Leu His Arg Tyr Tyr Phe Glu Val Glu Ile Phe Gly Ala Gly Thr

210 215 220

Tyr Val Gly Leu Thr Cys Lys Gly Ile Asp Gln Lys Gly Glu Glu Arg

225 230 235 240

Ser Ser Cys Ile Ser Gly Asn Asn Phe Ser Trp Ser Leu Gln Trp Asn

245 250 255

Gly Lys Glu Phe Thr Ala Trp Tyr Ser Asp Met Glu Thr Pro Leu Lys

260 265 270

Ala Gly Pro Phe Trp Arg Leu Gly Val Tyr Ile Asp Phe Pro Gly Gly

275 280 285

Ile Leu Ser Phe Tyr Gly Val Glu Tyr Asp Ser Met Thr Leu Val His

290 295 300

Lys Phe Ala Cys Lys Phe Ser Glu Pro Val Tyr Ala Ala Phe Trp Leu

305 310 315 320

Ser Lys Lys Glu Asn Ala Ile Arg Ile Val Asp Leu Gly Glu Glu Pro

325 330 335

Glu Lys Pro Ala Pro Ser Leu Val Gly Thr Ala Pro

340 345

<210> 137

<211> 2607

<212> DNA

<213> 智人(Homo sapiens)

<400> 137

atggcttcgt tccccgagac cgatttccag atctgcttgc tgtgcaagga gatgtgcggc 60

tcgccggcgc cgctctcctc caactcgtcc gcgtcgtcgt cctcctcgca gacgtccacg 120

tcgtcggggg gcggcggcgg gggccctggg gcggcggcgc gccgcctaca cgtcctgccc 180

tgcctgcacg ccttctgccg cccctgcctc gaggcgcacc ggctgccggc ggcgggcggc 240

ggcgcggcgg gagagccgct caagctgcgc tgccccgtgt gcgaccagaa agtagtgcta 300

gccgaggcgg cgggtatgga cgcgctgcct tcgtccgcct tcctgcttag caacctgctc 360

gacgcggtgg tggccactgc cgacgagccg ccgcccaaga acgggcgcgc cggcgctccg 420

gcgggagcgg gcggccacag caaccaccgg caccacgctc accacgcgca cccgcgcgcg 480

tccgcctccg cgccgccact cccgcaggcg ccgcagccgc ccgcgccttc ccgctcggca 540

cccggcggcc ctgccgcttc cccgtcggcg ctgctgctcc gccgtcctca cggctgcagc 600

tcgtgcgatg agggcaacgc agcttcttcg cgctgcctcg actgccagga gcacctgtgc 660

gacaactgcg tccgagcgca ccagcgcgtg cgcctcacca aggaccacta catcgagcgc 720

ggcccgccgg gtcccggtgc cgcagcagcg gcgcagcagc tcgggctcgg gccgcccttt 780

cccggcccgc ccttctccat cctctcagtg tttcccgagc gcctcggctt ctgccagcac 840

cacgacgacg aggtgctgca cctgtactgt gacacttgct ctgtacccat ctgtcgtgag 900

tgcacaatgg gccggcatgg gggccacagc ttcatctacc tccaggaggc actgcaggac 960

tcacgggcac tcaccatcca gctgctggca gatgcccagc agggacgaca ggcaatccag 1020

ctgagcatcg agcaggccca gacggtggcg gaacaggtgg agatgaaggc gaaggttgtg 1080

cagtcggagg tcaaagccgt gacggcgagg cataagaaag ccctggagga acgcgagtgt 1140

gagctgctgt ggaaggtaga aaagatccgc caggtgaaag ccaagtctct gtacctgcag 1200

gtggagaagc tgcggcaaaa cctcaacaag cttgagagca ccatcagtgc cgtgcagcag 1260

gtcctggagg agggtagagc gctagacatc ctactggccc gagaccggat gctggcccag 1320

gtgcaggagc tgaagaccgt gcggagcctc ctgcagcccc aggaagacga ccgagtcatg 1380

ttcacacccc ccgatcaggc actgtacctt gccatcaagt cttttggctt tgttagcagc 1440

ggggcctttg ccccactcac caaggccaca ggcgatggcc tcaagcgtgc cctccagggt 1500

aaggtggcct ccttcacagt cattggttat gaccacgatg gtgagccccg cctctcagga 1560

ggcgacctga tgtcggctgt ggtcctgggc cctgatggca acctgtttgg tgcagaggtg 1620

agtgatcagc agaatgggac atacgtggtg agttaccgac cccagctgga gggtgagcac 1680

ctggtatctg tgacactgtg caaccagcac attgagaaca gccctttcaa ggtggtggtc 1740

aagtcaggcc gcagctacgt gggcattggg ctcccgggcc tgagcttcgg cagtgagggt 1800

gacagcgatg gcaagctctg ccgcccttgg ggtgtgagtg tagacaagga gggctacatc 1860

attgtcgccg accgcagcaa caaccgcatc caggtgttca agccctgcgg cgccttccac 1920

cacaaattcg gcaccctggg ctcccggcct gggcagttcg accgaccagc cggcgtggcc 1980

tgtgacgcct cacgcaggat cgtggtggct gacaaggaca atcatcgcat ccagatcttc 2040

acgttcgagg gccagttcct cctcaagttt ggtgagaaag gaaccaagaa tgggcagttc 2100

aactaccctt gggatgtggc ggtgaattct gagggcaaga tcctggtctc agacacgagg 2160

aaccaccgga tccagctgtt tgggcctgat ggtgtcttcc taaacaagta tggcttcgag 2220

ggggctctct ggaagcactt tgactcccca cggggtgtgg ccttcaacca tgagggccac 2280

ttggtggtca ctgacttcaa caaccaccgg ctcctggtta ttcaccccga ctgccagtcg 2340

gcacgctttc tgggctcgga gggcacaggc aatgggcagt tcctgcgccc acaaggggta 2400

gctgtggacc aggaagggcg catcattgtg gcggattcca ggaaccatcg ggtacagatg 2460

tttgaatcca acggcagctt cctgtgcaag tttggtgctc aaggcagcgg ctttgggcag 2520

atggaccgcc cttccggcat cgccatcacc cccgacggaa tgatcgttgt ggtggacttt 2580

ggcaacaatc gaatcctcgt cttctaa 2607

<210> 138

<211> 868

<212> PRT

<213> 智人(Homo sapiens)

<400> 138

Met Ala Ser Phe Pro Glu Thr Asp Phe Gln Ile Cys Leu Leu Cys Lys

1 5 10 15

Glu Met Cys Gly Ser Pro Ala Pro Leu Ser Ser Asn Ser Ser Ala Ser

20 25 30

Ser Ser Ser Ser Gln Thr Ser Thr Ser Ser Gly Gly Gly Gly Gly Gly

35 40 45

Pro Gly Ala Ala Ala Arg Arg Leu His Val Leu Pro Cys Leu His Ala

50 55 60

Phe Cys Arg Pro Cys Leu Glu Ala His Arg Leu Pro Ala Ala Gly Gly

65 70 75 80

Gly Ala Ala Gly Glu Pro Leu Lys Leu Arg Cys Pro Val Cys Asp Gln

85 90 95

Lys Val Val Leu Ala Glu Ala Ala Gly Met Asp Ala Leu Pro Ser Ser

100 105 110

Ala Phe Leu Leu Ser Asn Leu Leu Asp Ala Val Val Ala Thr Ala Asp

115 120 125

Glu Pro Pro Pro Lys Asn Gly Arg Ala Gly Ala Pro Ala Gly Ala Gly

130 135 140

Gly His Ser Asn His Arg His His Ala His His Ala His Pro Arg Ala

145 150 155 160

Ser Ala Ser Ala Pro Pro Leu Pro Gln Ala Pro Gln Pro Pro Ala Pro

165 170 175

Ser Arg Ser Ala Pro Gly Gly Pro Ala Ala Ser Pro Ser Ala Leu Leu

180 185 190

Leu Arg Arg Pro His Gly Cys Ser Ser Cys Asp Glu Gly Asn Ala Ala

195 200 205

Ser Ser Arg Cys Leu Asp Cys Gln Glu His Leu Cys Asp Asn Cys Val

210 215 220

Arg Ala His Gln Arg Val Arg Leu Thr Lys Asp His Tyr Ile Glu Arg

225 230 235 240

Gly Pro Pro Gly Pro Gly Ala Ala Ala Ala Ala Gln Gln Leu Gly Leu

245 250 255

Gly Pro Pro Phe Pro Gly Pro Pro Phe Ser Ile Leu Ser Val Phe Pro

260 265 270

Glu Arg Leu Gly Phe Cys Gln His His Asp Asp Glu Val Leu His Leu

275 280 285

Tyr Cys Asp Thr Cys Ser Val Pro Ile Cys Arg Glu Cys Thr Met Gly

290 295 300

Arg His Gly Gly His Ser Phe Ile Tyr Leu Gln Glu Ala Leu Gln Asp

305 310 315 320

Ser Arg Ala Leu Thr Ile Gln Leu Leu Ala Asp Ala Gln Gln Gly Arg

325 330 335

Gln Ala Ile Gln Leu Ser Ile Glu Gln Ala Gln Thr Val Ala Glu Gln

340 345 350

Val Glu Met Lys Ala Lys Val Val Gln Ser Glu Val Lys Ala Val Thr

355 360 365

Ala Arg His Lys Lys Ala Leu Glu Glu Arg Glu Cys Glu Leu Leu Trp

370 375 380

Lys Val Glu Lys Ile Arg Gln Val Lys Ala Lys Ser Leu Tyr Leu Gln

385 390 395 400

Val Glu Lys Leu Arg Gln Asn Leu Asn Lys Leu Glu Ser Thr Ile Ser

405 410 415

Ala Val Gln Gln Val Leu Glu Glu Gly Arg Ala Leu Asp Ile Leu Leu

420 425 430

Ala Arg Asp Arg Met Leu Ala Gln Val Gln Glu Leu Lys Thr Val Arg

435 440 445

Ser Leu Leu Gln Pro Gln Glu Asp Asp Arg Val Met Phe Thr Pro Pro

450 455 460

Asp Gln Ala Leu Tyr Leu Ala Ile Lys Ser Phe Gly Phe Val Ser Ser

465 470 475 480

Gly Ala Phe Ala Pro Leu Thr Lys Ala Thr Gly Asp Gly Leu Lys Arg

485 490 495

Ala Leu Gln Gly Lys Val Ala Ser Phe Thr Val Ile Gly Tyr Asp His

500 505 510

Asp Gly Glu Pro Arg Leu Ser Gly Gly Asp Leu Met Ser Ala Val Val

515 520 525

Leu Gly Pro Asp Gly Asn Leu Phe Gly Ala Glu Val Ser Asp Gln Gln

530 535 540

Asn Gly Thr Tyr Val Val Ser Tyr Arg Pro Gln Leu Glu Gly Glu His

545 550 555 560

Leu Val Ser Val Thr Leu Cys Asn Gln His Ile Glu Asn Ser Pro Phe

565 570 575

Lys Val Val Val Lys Ser Gly Arg Ser Tyr Val Gly Ile Gly Leu Pro

580 585 590

Gly Leu Ser Phe Gly Ser Glu Gly Asp Ser Asp Gly Lys Leu Cys Arg

595 600 605

Pro Trp Gly Val Ser Val Asp Lys Glu Gly Tyr Ile Ile Val Ala Asp

610 615 620

Arg Ser Asn Asn Arg Ile Gln Val Phe Lys Pro Cys Gly Ala Phe His

625 630 635 640

His Lys Phe Gly Thr Leu Gly Ser Arg Pro Gly Gln Phe Asp Arg Pro

645 650 655

Ala Gly Val Ala Cys Asp Ala Ser Arg Arg Ile Val Val Ala Asp Lys

660 665 670

Asp Asn His Arg Ile Gln Ile Phe Thr Phe Glu Gly Gln Phe Leu Leu

675 680 685

Lys Phe Gly Glu Lys Gly Thr Lys Asn Gly Gln Phe Asn Tyr Pro Trp

690 695 700

Asp Val Ala Val Asn Ser Glu Gly Lys Ile Leu Val Ser Asp Thr Arg

705 710 715 720

Asn His Arg Ile Gln Leu Phe Gly Pro Asp Gly Val Phe Leu Asn Lys

725 730 735

Tyr Gly Phe Glu Gly Ala Leu Trp Lys His Phe Asp Ser Pro Arg Gly

740 745 750

Val Ala Phe Asn His Glu Gly His Leu Val Val Thr Asp Phe Asn Asn

755 760 765

His Arg Leu Leu Val Ile His Pro Asp Cys Gln Ser Ala Arg Phe Leu

770 775 780

Gly Ser Glu Gly Thr Gly Asn Gly Gln Phe Leu Arg Pro Gln Gly Val

785 790 795 800

Ala Val Asp Gln Glu Gly Arg Ile Ile Val Ala Asp Ser Arg Asn His

805 810 815

Arg Val Gln Met Phe Glu Ser Asn Gly Ser Phe Leu Cys Lys Phe Gly

820 825 830

Ala Gln Gly Ser Gly Phe Gly Gln Met Asp Arg Pro Ser Gly Ile Ala

835 840 845

Ile Thr Pro Asp Gly Met Ile Val Val Val Asp Phe Gly Asn Asn Arg

850 855 860

Ile Leu Val Phe

865

<210> 139

<211> 1434

<212> DNA

<213> 智人(Homo sapiens)

<400> 139

atgtcggctg cgcccggcct cctgcaccag gagctgtcct gcccgctgtg cctgcagctg 60

ttcgacgcgc ccgtgacagc cgagtgcggc cacagtttct gccgcgcctg cctaggccgc 120

gtggccgggg agccggcggc ggatggcacc gttctctgcc cctgctgcca ggcccccacg 180

cggccgcagg cactcagcac caacctgcag ctggcgcgcc tggtggaggg gctggcccag 240

gtgccgcagg gccactgcga ggagcacctg gacccgctga gcatctactg cgagcaggac 300

cgcgcgctgg tgtgcggagt gtgcgcctca ctcggctcgc accgcggtca tcgcctcctg 360

cctgccgccg aggcccacgc acgcctcaag acacagctgc cacagcagaa actgcagctg 420

caggaggcat gcatgcgcaa ggagaagagt gtggctgtgc tggagcatca gctggtggag 480

gtggaggaga cagtgcgtca gttccggggg gccgtggggg agcagctggg caagatgcgg 540

gtgttcctgg ctgcactgga gggctccttg gaccgcgagg cagagcgtgt acggggtgag 600

gcaggggtcg ccttgcgccg ggagctgggg agcctgaact cttacctgga gcagctgcgg 660

cagatggaga aggtcctgga ggaggtggcg gacaagccgc agactgagtt cctcatgaaa 720

tactgcctgg tgaccagcag gctgcagaag atcctggcag agtctccccc acccgcccgt 780

ctggacatcc agctgccaat tatctcagat gacttcaaat tccaggtgtg gaggaagatg 840

ttccgggctc tgatgccagc gctggaggag ctgacctttg acccgagctc tgcgcacccg 900

agcctggtgg tgtcttcctc tggccgccgc gtggagtgct cggagcagaa ggcgccgccg 960

gccggggagg acccgcgcca gttcgacaag gcggtggcgg tggtggcgca ccagcagctc 1020

tccgagggcg agcactactg ggaggtggat gttggcgaca agccgcgctg ggcgctgggc 1080

gtgatcgcgg ccgaggcccc ccgccgcggg cgcctgcacg cggtgccctc gcagggcctg 1140

tggctgctgg ggctgcgcga gggcaagatc ctggaggcac acgtggaggc caaggagccg 1200

cgcgctctgc gcagccccga gaggcggccc acgcgcattg gcctttacct gagcttcggc 1260

gacggcgtcc tctccttcta cgatgccagc gacgccgacg cgctcgtgcc gctttttgcc 1320

ttccacgagc gcctgcccag gcccgtgtac cccttcttcg acgtgtgctg gcacgacaag 1380

ggcaagaatg cccagccgct gctgctcgtg ggtcccgaag gcgccgaggc ctga 1434

<210> 140

<211> 477

<212> PRT

<213> 智人(Homo sapiens)

<400> 140

Met Ser Ala Ala Pro Gly Leu Leu His Gln Glu Leu Ser Cys Pro Leu

1 5 10 15

Cys Leu Gln Leu Phe Asp Ala Pro Val Thr Ala Glu Cys Gly His Ser

20 25 30

Phe Cys Arg Ala Cys Leu Gly Arg Val Ala Gly Glu Pro Ala Ala Asp

35 40 45

Gly Thr Val Leu Cys Pro Cys Cys Gln Ala Pro Thr Arg Pro Gln Ala

50 55 60

Leu Ser Thr Asn Leu Gln Leu Ala Arg Leu Val Glu Gly Leu Ala Gln

65 70 75 80

Val Pro Gln Gly His Cys Glu Glu His Leu Asp Pro Leu Ser Ile Tyr

85 90 95

Cys Glu Gln Asp Arg Ala Leu Val Cys Gly Val Cys Ala Ser Leu Gly

100 105 110

Ser His Arg Gly His Arg Leu Leu Pro Ala Ala Glu Ala His Ala Arg

115 120 125

Leu Lys Thr Gln Leu Pro Gln Gln Lys Leu Gln Leu Gln Glu Ala Cys

130 135 140

Met Arg Lys Glu Lys Ser Val Ala Val Leu Glu His Gln Leu Val Glu

145 150 155 160

Val Glu Glu Thr Val Arg Gln Phe Arg Gly Ala Val Gly Glu Gln Leu

165 170 175

Gly Lys Met Arg Val Phe Leu Ala Ala Leu Glu Gly Ser Leu Asp Arg

180 185 190

Glu Ala Glu Arg Val Arg Gly Glu Ala Gly Val Ala Leu Arg Arg Glu

195 200 205

Leu Gly Ser Leu Asn Ser Tyr Leu Glu Gln Leu Arg Gln Met Glu Lys

210 215 220

Val Leu Glu Glu Val Ala Asp Lys Pro Gln Thr Glu Phe Leu Met Lys

225 230 235 240

Tyr Cys Leu Val Thr Ser Arg Leu Gln Lys Ile Leu Ala Glu Ser Pro

245 250 255

Pro Pro Ala Arg Leu Asp Ile Gln Leu Pro Ile Ile Ser Asp Asp Phe

260 265 270

Lys Phe Gln Val Trp Arg Lys Met Phe Arg Ala Leu Met Pro Ala Leu

275 280 285

Glu Glu Leu Thr Phe Asp Pro Ser Ser Ala His Pro Ser Leu Val Val

290 295 300

Ser Ser Ser Gly Arg Arg Val Glu Cys Ser Glu Gln Lys Ala Pro Pro

305 310 315 320

Ala Gly Glu Asp Pro Arg Gln Phe Asp Lys Ala Val Ala Val Val Ala

325 330 335

His Gln Gln Leu Ser Glu Gly Glu His Tyr Trp Glu Val Asp Val Gly

340 345 350

Asp Lys Pro Arg Trp Ala Leu Gly Val Ile Ala Ala Glu Ala Pro Arg

355 360 365

Arg Gly Arg Leu His Ala Val Pro Ser Gln Gly Leu Trp Leu Leu Gly

370 375 380

Leu Arg Glu Gly Lys Ile Leu Glu Ala His Val Glu Ala Lys Glu Pro

385 390 395 400

Arg Ala Leu Arg Ser Pro Glu Arg Arg Pro Thr Arg Ile Gly Leu Tyr

405 410 415

Leu Ser Phe Gly Asp Gly Val Leu Ser Phe Tyr Asp Ala Ser Asp Ala

420 425 430

Asp Ala Leu Val Pro Leu Phe Ala Phe His Glu Arg Leu Pro Arg Pro

435 440 445

Val Tyr Pro Phe Phe Asp Val Cys Trp His Asp Lys Gly Lys Asn Ala

450 455 460

Gln Pro Leu Leu Leu Val Gly Pro Glu Gly Ala Glu Ala

465 470 475

<210> 141

<211> 753

<212> DNA

<213> 智人(Homo sapiens)

<400> 141

atggcttggc aggtgagcct gctggagctg gaggaccggc ttcagtgtcc catctgcctg 60

gaggtcttca aggagtccct aatgctacag tgcggccact cctactgcaa gggctgcctg 120

gtttccctgt cctaccacct ggacaccaag gtgcgctgcc ccatgtgctg gcaggtggtg 180

gacggcagca gctccttgcc caacgtctcc ctggcctggg tgatcgaagc cctgaggctc 240

cctggggacc cggagcccaa ggtctgcgtg caccaccgga acccgctcag ccttttctgc 300

gagaaggacc aggagctcat ctgtggcctc tgcggtctgc tgggctccca ccaacaccac 360

ccggtcacgc ccgtctccac cgtctgcagc cgcatgaagg aggagctcgc agccctcttc 420

tctgagctga agcaggagca gaagaaggtg gatgagctca tcgccaaact ggtgaaaaac 480

cggacccgaa tcgtcaatga gtcggatgtc ttcagctggg tgatccgccg cgagttccag 540

gagctgcgcc acccggtgga cgaggagaag gcccgctgcc tggaggggat agggggtcac 600

acccgtggcc tggtggcctc cctggacatg cagctggagc aggcccaggg aacccgggag 660

cggctggccc aagccgagtg tgtgctggaa cagttcggca atgaggacca ccatgagttc 720

atctggaagt tccactccat ggcctccagg taa 753

<210> 142

<211> 250

<212> PRT

<213> 智人(Homo sapiens)

<400> 142

Met Ala Trp Gln Val Ser Leu Leu Glu Leu Glu Asp Arg Leu Gln Cys

1 5 10 15

Pro Ile Cys Leu Glu Val Phe Lys Glu Ser Leu Met Leu Gln Cys Gly

20 25 30

His Ser Tyr Cys Lys Gly Cys Leu Val Ser Leu Ser Tyr His Leu Asp

35 40 45

Thr Lys Val Arg Cys Pro Met Cys Trp Gln Val Val Asp Gly Ser Ser

50 55 60

Ser Leu Pro Asn Val Ser Leu Ala Trp Val Ile Glu Ala Leu Arg Leu

65 70 75 80

Pro Gly Asp Pro Glu Pro Lys Val Cys Val His His Arg Asn Pro Leu

85 90 95

Ser Leu Phe Cys Glu Lys Asp Gln Glu Leu Ile Cys Gly Leu Cys Gly

100 105 110

Leu Leu Gly Ser His Gln His His Pro Val Thr Pro Val Ser Thr Val

115 120 125

Cys Ser Arg Met Lys Glu Glu Leu Ala Ala Leu Phe Ser Glu Leu Lys

130 135 140

Gln Glu Gln Lys Lys Val Asp Glu Leu Ile Ala Lys Leu Val Lys Asn

145 150 155 160

Arg Thr Arg Ile Val Asn Glu Ser Asp Val Phe Ser Trp Val Ile Arg

165 170 175

Arg Glu Phe Gln Glu Leu Arg His Pro Val Asp Glu Glu Lys Ala Arg

180 185 190

Cys Leu Glu Gly Ile Gly Gly His Thr Arg Gly Leu Val Ala Ser Leu

195 200 205

Asp Met Gln Leu Glu Gln Ala Gln Gly Thr Arg Glu Arg Leu Ala Gln

210 215 220

Ala Glu Cys Val Leu Glu Gln Phe Gly Asn Glu Asp His His Glu Phe

225 230 235 240

Ile Trp Lys Phe His Ser Met Ala Ser Arg

245 250

<210> 143

<211> 753

<212> DNA

<213> 智人(Homo sapiens)

<400> 143

atggcttggc aggtgagcct gctggagctg gaggactggc ttcagtgtcc catctgcctg 60

gaggtcttca aggagtccct aatgctacag tgcggccact cctactgcaa gggctgcctg 120

gtttccctgt cctaccacct ggacaccaag gtgcgctgcc ccatgtgctg gcaggtggtg 180

gacggcagca gctccttgcc caacgtctcc ctggcctggg tgatcgaagc cctgaggctc 240

cctggggacc cggagcccaa ggtctgcgtg caccaccgga acccgctcag ccttttctgc 300

gagaaggacc aggagctcat ctgtggcctc tgcggtctgc tgggctccca ccaacaccac 360

ccggtcacgc ccgtctccac cgtctgcagc cgcatgaagg aggagctcgc agccctcttc 420

tctgagctga agcaggagca gaagaaggtg gatgagctca tcgccaaact ggtgaaaaac 480

cggacccgaa tcgtcaatga gtcggatgtc ttcagctggg tgatccgccg cgagttccag 540

gagctgcgcc acccggtgga cgaggagaag gcccgctgcc tggaggggat agggggtcac 600

acccgtggcc tggtggcctc cctggacatg cagctggagc aggcccaggg aacccgggag 660

cggctggccc aagccgagtg tgtgctggaa cagttcggaa atgaggacca ccatgagttc 720

atctggaagt tccactccat ggcctccagg taa 753

<210> 144

<211> 250

<212> PRT

<213> 智人(Homo sapiens)

<400> 144

Met Ala Trp Gln Val Ser Leu Leu Glu Leu Glu Asp Trp Leu Gln Cys

1 5 10 15

Pro Ile Cys Leu Glu Val Phe Lys Glu Ser Leu Met Leu Gln Cys Gly

20 25 30

His Ser Tyr Cys Lys Gly Cys Leu Val Ser Leu Ser Tyr His Leu Asp

35 40 45

Thr Lys Val Arg Cys Pro Met Cys Trp Gln Val Val Asp Gly Ser Ser

50 55 60

Ser Leu Pro Asn Val Ser Leu Ala Trp Val Ile Glu Ala Leu Arg Leu

65 70 75 80

Pro Gly Asp Pro Glu Pro Lys Val Cys Val His His Arg Asn Pro Leu

85 90 95

Ser Leu Phe Cys Glu Lys Asp Gln Glu Leu Ile Cys Gly Leu Cys Gly

100 105 110

Leu Leu Gly Ser His Gln His His Pro Val Thr Pro Val Ser Thr Val

115 120 125

Cys Ser Arg Met Lys Glu Glu Leu Ala Ala Leu Phe Ser Glu Leu Lys

130 135 140

Gln Glu Gln Lys Lys Val Asp Glu Leu Ile Ala Lys Leu Val Lys Asn

145 150 155 160

Arg Thr Arg Ile Val Asn Glu Ser Asp Val Phe Ser Trp Val Ile Arg

165 170 175

Arg Glu Phe Gln Glu Leu Arg His Pro Val Asp Glu Glu Lys Ala Arg

180 185 190

Cys Leu Glu Gly Ile Gly Gly His Thr Arg Gly Leu Val Ala Ser Leu

195 200 205

Asp Met Gln Leu Glu Gln Ala Gln Gly Thr Arg Glu Arg Leu Ala Gln

210 215 220

Ala Glu Cys Val Leu Glu Gln Phe Gly Asn Glu Asp His His Glu Phe

225 230 235 240

Ile Trp Lys Phe His Ser Met Ala Ser Arg

245 250

<210> 145

<211> 1404

<212> DNA

<213> 智人(Homo sapiens)

<400> 145

atggcacatg tcgaggtctt ggccagactc cagaaagaga ccaagtgccc catctgtttg 60

gacgatctga cagatcccgt caccgtcgag tgcggacaca acttctgtcg ttcctgcatc 120

aaagacttct gggcagggca gcaagccacg tcctcctgtc ctgtctgccg acaccagtgc 180

caacacagga acctcagaag caatgcccag ctgggaaaca tgattgaaac tgcccagctg 240

ctccaaggca tggagaacaa gaggcacgag agcagcacca gttgcgagag gcacaaccag 300

gccctgaccc tcttctgtga ggatgacctg cagttgctct gtgaccagtg cgtggagcct 360

gagagccacg ggcgccacca ggtgctgtcc attacagagg ctgcctctct ccacaggaaa 420

caccttcagg actatagcaa gctcctgaag tgggaagtga aagagattca agggctaatg 480

agcgcactaa acaaaagaac cgtgaccctg agggagcaag cagaggcaca gaggtcacag 540

ctaacctctg agtgtgagaa gctcatgcgg tttctagacc aggaagagcg ggcagctttc 600

tccaggttag aagacgagga gatgaggctc gagaagagac tgcttgataa catagcagca 660

ttagaacacc acggttccag cctcagagac ctcctgagac acttgatgct gaccggagag 720

ctctccgaag ccaagatgtt gtccacggtc aaggattttt acctgaattg taggcgtcag 780

ctcatcagcc caagcatttt cccagttcag ttgagaagag tggagtacag ctttcctctc 840

cagtattcgg ccctacagaa agttatacag cattttacag acaacgtcac tctagacctg 900

aagacagccc atccaaacct gctcatttct aaggatagaa cctgcgtaac atttacaaag 960

aaaagacaac gtattcctgg ttcttcaagt tttaccaaga gccctgttgt gttggggatt 1020

ccacatttta attctgggag acacttctgg gaggtgcagg taggaaagaa gccaaagtgg 1080

gctattggca tttgcaaagc tgattcctct atcggggaaa ggcagtcccc taacccctgg 1140

gggtactgga ggattgtttg gcagggggac agcttcaacg tctcaggagc tgatccagac 1200

tctcggctga aggccgcaag agctacaagc attggtgttt tcctggacta tgaactggga 1260

gaggtttctt tctatggcat gcctgagaaa tgtcacctct atactttcag ggacactttt 1320

tctggtcctg tctgccctta tttctacata gggcctcagt cagaacctct tagactctgt 1380

tctgccactg attcagaatg ctaa 1404

<210> 146

<211> 467

<212> PRT

<213> 智人(Homo sapiens)

<400> 146

Met Ala His Val Glu Val Leu Ala Arg Leu Gln Lys Glu Thr Lys Cys

1 5 10 15

Pro Ile Cys Leu Asp Asp Leu Thr Asp Pro Val Thr Val Glu Cys Gly

20 25 30

His Asn Phe Cys Arg Ser Cys Ile Lys Asp Phe Trp Ala Gly Gln Gln

35 40 45

Ala Thr Ser Ser Cys Pro Val Cys Arg His Gln Cys Gln His Arg Asn

50 55 60

Leu Arg Ser Asn Ala Gln Leu Gly Asn Met Ile Glu Thr Ala Gln Leu

65 70 75 80

Leu Gln Gly Met Glu Asn Lys Arg His Glu Ser Ser Thr Ser Cys Glu

85 90 95

Arg His Asn Gln Ala Leu Thr Leu Phe Cys Glu Asp Asp Leu Gln Leu

100 105 110

Leu Cys Asp Gln Cys Val Glu Pro Glu Ser His Gly Arg His Gln Val

115 120 125

Leu Ser Ile Thr Glu Ala Ala Ser Leu His Arg Lys His Leu Gln Asp

130 135 140

Tyr Ser Lys Leu Leu Lys Trp Glu Val Lys Glu Ile Gln Gly Leu Met

145 150 155 160

Ser Ala Leu Asn Lys Arg Thr Val Thr Leu Arg Glu Gln Ala Glu Ala

165 170 175

Gln Arg Ser Gln Leu Thr Ser Glu Cys Glu Lys Leu Met Arg Phe Leu

180 185 190

Asp Gln Glu Glu Arg Ala Ala Phe Ser Arg Leu Glu Asp Glu Glu Met

195 200 205

Arg Leu Glu Lys Arg Leu Leu Asp Asn Ile Ala Ala Leu Glu His His

210 215 220

Gly Ser Ser Leu Arg Asp Leu Leu Arg His Leu Met Leu Thr Gly Glu

225 230 235 240

Leu Ser Glu Ala Lys Met Leu Ser Thr Val Lys Asp Phe Tyr Leu Asn

245 250 255

Cys Arg Arg Gln Leu Ile Ser Pro Ser Ile Phe Pro Val Gln Leu Arg

260 265 270

Arg Val Glu Tyr Ser Phe Pro Leu Gln Tyr Ser Ala Leu Gln Lys Val

275 280 285

Ile Gln His Phe Thr Asp Asn Val Thr Leu Asp Leu Lys Thr Ala His

290 295 300

Pro Asn Leu Leu Ile Ser Lys Asp Arg Thr Cys Val Thr Phe Thr Lys

305 310 315 320

Lys Arg Gln Arg Ile Pro Gly Ser Ser Ser Phe Thr Lys Ser Pro Val

325 330 335

Val Leu Gly Ile Pro His Phe Asn Ser Gly Arg His Phe Trp Glu Val

340 345 350

Gln Val Gly Lys Lys Pro Lys Trp Ala Ile Gly Ile Cys Lys Ala Asp

355 360 365

Ser Ser Ile Gly Glu Arg Gln Ser Pro Asn Pro Trp Gly Tyr Trp Arg

370 375 380

Ile Val Trp Gln Gly Asp Ser Phe Asn Val Ser Gly Ala Asp Pro Asp

385 390 395 400

Ser Arg Leu Lys Ala Ala Arg Ala Thr Ser Ile Gly Val Phe Leu Asp

405 410 415

Tyr Glu Leu Gly Glu Val Ser Phe Tyr Gly Met Pro Glu Lys Cys His

420 425 430

Leu Tyr Thr Phe Arg Asp Thr Phe Ser Gly Pro Val Cys Pro Tyr Phe

435 440 445

Tyr Ile Gly Pro Gln Ser Glu Pro Leu Arg Leu Cys Ser Ala Thr Asp

450 455 460

Ser Glu Cys

465

<210> 147

<211> 7

<212> PRT

<213> 人工序列(Artificial Sequence)

<220>

<221> SIGNAL

<223> SV40细胞核定位信号(SV40 nuclear localization signal)

<400> 147

Pro Lys Lys Lys Arg Lys Val

1 5

<210> 148

<211> 21

<212> DNA

<213> 人工序列(Artificial Sequence)

<220>

<221> attenuator

<223> PML#4

<400> 148

ctccaagatc taaaccgaga a 21

<210> 149

<211> 21

<212> DNA

<213> 人工序列(Artificial Sequence)

<220>

<221> attenuator

<223> PML#9

<400> 149

cacccgcaag accaacaaca t 21

<210> 150

<211> 21

<212> DNA

<213> 人工序列(Artificial Sequence)

<220>

<221> attenuator

<223> RNF4#5

<400> 150

ccctgtttcc taagaacgaa a 21

<210> 151

<211> 21

<212> DNA

<213> 人工序列(Artificial Sequence)

<220>

<221> attenuator

<223> RNF4#6

<400> 151

taggccgagc tttgcgggaa a 21

<210> 152

<211> 21

<212> DNA

<213> 人工序列(Artificial Sequence)

<220>

<221> attenuator

<223> RNF4#8

<400> 152

aagactgttt cgaaaccaac a 21

<210> 153

<211> 19

<212> DNA/RNA

<213> 人工序列(Artificial Sequence)

<220>

<221> misc_feature

<223> SUMO1 siRNA集合有义链1(SUMO1 siRNA pooled sense strand 1)

<400> 153

tcaagaaacu caaagaatc 19

<210> 154

<211> 19

<212> DNA

<213> 人工序列(Artificial Sequence)

<220>

<221> misc_RNA

<223> SUMO1 siRNA集合有义链2(SUMO1 siRNA pooled sense strand 2)

<400> 154

gacagggtgt tccaatgaa 19

<210> 155

<211> 19

<212> DNA

<213> 人工序列(Artificial Sequence)

<220>

<221> misc_RNA

<223> SUMO1 siRNA集合有义链3(SUMO1 siRNA pooled sense strand 3)

<400> 155

ggtttctctt tgagggtca 19

<210> 156

<211> 19

<212> DNA

<213> 人工序列(Artificial Sequence)

<220>

<221> misc_RNA

<223> SUMO1 siRNA集合有义链4(SUMO1 siRNA pooled sense strand 4)

<400> 156

gaataaatgg gcatgccaa 19

<210> 157

<211> 19

<212> RNA

<213> 人工序列(Artificial Sequence)

<220>

<221> misc_RNA

<223> SUMO2/3 siRNA集合有义链1(SUMO2/3 siRNA pooled sense strand 1)

<400> 157

cccauuccuu uauuguaca 19

<210> 158

<211> 19

<212> RNA

<213> 人工序列(Artificial Sequence)

<220>

<221> misc_RNA

<223> SUMO2/3 siRNA集合有义链2(SUMO2/3 siRNA pooled sense strand 2)

<400> 158

cagagaauga ccacaucaa 19

<210> 159

<211> 19

<212> RNA

<213> 人工序列(Artificial Sequence)

<220>

<221> misc_RNA

<223> SUMO2/3 siRNA集合有义链3(SUMO2/3 siRNA pooled sense strand 3)

<400> 159

caguuauguu gucguguau 19

<210> 160

<211> 21

<212> DNA

<213> 人工序列(Artificial Sequence)

<220>

<221> misc_RNA

<223> shRNF4的反义序列(antisense sequence of shRNF4)

<400> 160

tggcgtttct gggagtatgg g 21

<210> 161

<211> 10

<212> PRT

<213> 人工序列(Artificial Sequence)

<220>

<221> PEPTIDE

<223> 抗原肽(antigen peptide)

<400> 161

Asp Leu Thr His Asn Asp Ser Val Val Ile

1 5 10

<210> 162

<211> 21

<212> DNA

<213> 人工序列(Artificial Sequence)

<220>

<221> misc_RNA

<223> TRIM27 siRNA有义链(TRIM27 siRNA sense strand)

<400> 162

aactcttagg cctaacccag a 21

<210> 163

<211> 11

<212> PRT

<213> 人工序列(Artificial Sequence)

<220>

<221> DOMAIN

<223> HIV Tat的蛋白转导域(protein transduction domain of HIV Tat)

<400> 163

Tyr Gly Arg Lys Lys Arg Arg Gln Arg Arg Arg

1 5 10

<210> 164

<211> 12207

<212> DNA

<213> 智人(Homo sapiens)

<400> 164

atggcgagcc gcgatagcaa ccacgctggc gagagctttc tcggctccga cggggacgag 60

gaggcgaccc gggagctgga gaccgaggag gagtcggagg gcgaggagga cgagacggcg 120

gcggagtcgg aggaggagcc ggactccagg ttatcagacc aggatgaaga gggaaagatc 180

aagcaggagt atatcatatc tgacccctcc ttttccatgg tgacagtcca aagggaagat 240

agtgggataa cctgggaaac caattcaagt agatcttcta ctccttgggc ttcagaagaa 300

agtcagactt ctggtgtgtg tagtcgggaa gggtcaactg tgaattctcc tcctggaaat 360

gtttccttta ttgtggatga agtgaaaaag gttcggaaaa ggactcataa gtcaaagcat 420

ggttcaccat cattacgccg gaaaggcaac agaaaaagaa attcttttga atcccaagat 480

gttccaacaa acaaaaaagg cagtccttta acttcagcaa gccaggtact aaccacggag 540

aaagagaagt catatactgg catttatgat aaagcaagaa aaaagaagac cacttcaaat 600

acacctccga ttactggggc aatatacaaa gaacacaagc cattagtgtt aagaccagtc 660

tacataggaa cagtacaata taaaattaag atgtttaatt cggttaaaga agaattaatt 720

cctctacaat tttatggaac attgccaaag ggttatgtaa ttaaagaaat acattatagg 780

aaagggaaag atgcatccat tagtctagag ccagatttgg acaatagtgg ttctaataca 840

gtgtccaaaa cacgcaaatt agtagcccaa agcatagagg ataaagtaaa agaggttttt 900

ccaccctgga gaggcgcact ctccaaagga tcagagtccc taaccttaat gttcagtcat 960

gaagatcaaa agaaaattta tgctgattct cccctaaatg ccacatctgc attggagcac 1020

acagttccct cttattcaag tagtggcaga gcagaacaag gaatacagct caggcattca 1080

cagtcagtgc cacaacagcc agaagatgaa gcaaaaccac atgaagtgga acctccatct 1140

gtgacacccg acacacctgc aactatgttc ctgagaacaa caaaggaaga atgtgagctt 1200

gcttcaccag gaactgcagc ttcagagaat gactcttcag tctcaccatc atttgctaat 1260

gaggtaaaga aggaagatgt gtattctgct caccattcca tttctctgga ggcagcgtca 1320

ccaggtctgg cagcatctac ccaggatggt ttggacccag accaagaaca gccggacctg 1380

acttcaatag aaagggcaga accagtctcc gcaaaactga cccctaccca tcccagtgtc 1440

aaaggagaga aggaggaaaa catgcttgag ccatccattt ctctttctga acctctaatg 1500

ttagaagaac cagagaaaga agaaatagaa acttccctac ccatagctat tacccctgaa 1560

cctgaagatt ctaatttagt agaagaagag atcgtagaac ttgattaccc agaaagccca 1620

ttggtttccg agaagccctt cccaccacat atgtcccctg aagtggagca caaagaagaa 1680

gagcttattc taccattatt ggcagcatca tctcctgaac atgttgcttt gtctgaggaa 1740

gaaagagagg aaattgcatc tgtttctact ggttctgctt ttgtatcaga gtattcagta 1800

ccacaggatt tgaaccatga attacaggag caagaaggtg agccagttcc cccatccaat 1860

gtagaagcta tagctgaaca tgcagttttg tcagaagaag agaatgagga atttgaggct 1920

tattccccag ctgcagcccc tacatctgag agctctctct caccatccac aactgagaag 1980

acttcagaga accagtctcc actgttttca acagttacac cagaatacat ggtcctatca 2040

ggagacgagg cctcagaaag tgggtgttac acaccagact ccacatctgc ttctgaatat 2100

tcagttccat cactggcaac aaaagagtca ctgaagaaaa caattgaccg taagtccccg 2160

ttaatattga aaggtgtttc tgagtacatg attccatcag aagagaagga agacactgga 2220

tcgtttactc cagctgtggc ccctgcttct gagccctctc tctcaccatc cacaaccgaa 2280

aagacttctg aatgccagtc accactgcct tctactgcca catcagaaca cgtggtccca 2340

tcagaaggag aggacctagg aagtgaacgt ttcacaccgg attcaaagtt gatctccaaa 2400

tatgcagccc cactcaatgc aacacaggaa tctcaaaaga aaataatcaa tgaggcatcc 2460

caattcaaac caaaaggtat ttctgagcac acagttctgt cagtagacgg caaggaggtc 2520

attggaccat cttccccaga tttggttgtt gcatctgaac actctttccc accacacaca 2580

accgagatga cttctgaatg ccaggcccca ccactttcag ccaccccatc tgaatatgtt 2640

gttctatcag acgaagaggc agtcgagttg gaacgataca caccctcttc tacatctgct 2700

tctgaatttt cagtaccacc atatgcaaca ccggaggcac aggaggaaga aattgtccat 2760

agatctctaa atctaaaagg tgcatcctca cccatgaatt tatcagaaga agatcaagaa 2820

gacattggac ctttttctcc agattctgca tttgtgtcag aattctcatt tccaccgtat 2880

gcaacccagg aagcagagaa aagagaattt gagtgcgatt ctccaatatg tttaacatca 2940

ccatctgagc acactatttt gtcagatgaa gacactgaag aagcggaact gttctctcca 3000

gactcagcat cacaagtttc aatccctccc tttagaatct cagaaacaga gaaaaatgaa 3060

cttgagcctg attcactatt aactgcagtg tctgcttcag gttattcctg cttttcagaa 3120

gcagatgagg aagacattgg atccacagct gctacacctg tatctgagca gttcagttca 3180

tcacagaagc aaaaagctga aactttccct ttgatgtctc cgcttgaaga cttaagtctg 3240

ccgccttcaa cagataaatc agagaaagca gaaattaagc cagagattcc aacaacctca 3300

acatctgtat ctgaatatct cattttggca cagaagcaga aaactcaagc atatttagag 3360

cctgagtctg aagacttgat tccttcacat ttaaccagtg aagtggagaa gggagaaagg 3420

gaggcaagtt catcagtagc tgcaatacct gctgctttac ctgcacaatc atctatagta 3480

aaggaagaaa ccaaacctgc atctccacat tcagttttac ctgattcagt ccctgcaatc 3540

aagaaagaac aggaacccac agcagcactc actctaaaag ctgcagatga acagatggct 3600

ttgtcaaaag tcagaaagga agaaattgtg cctgattctc aagaagctac agcacatgta 3660

tcacaggatc aaaaaatgga gcctcagcct ccaaatgttc cagagtctga gatgaaatat 3720

tcagttttgc ctgacatggt agatgagcca aagaagggtg tcaagcccaa attagttcta 3780

aatgtgactt ctgaactaga acagagaaag ttgtccaaga atgagcctga agtaataaaa 3840

ccatattcac ctctaaagga aacatcttta tctggacctg aggctttatc agcagtgaaa 3900

atggagatga aacatgattc caaaataaca actacaccta tagtgcttca ttcagcttcc 3960

tcaggagtgg aaaagcaagt tgaacatggt ccacctgcac tagcattttc agctttgtca 4020

gaagaaatta aaaaagaaat tgaacccagt tcctcaacaa ctacagcatc tgtaactaag 4080

cttgattcaa acttaaccag agcagtaaaa gaagaaatcc caacagattc atctcttatc 4140

actcctgtag atcgtccagt cttaacaaaa gtaggaaagg gtgaattagg aagtggtttg 4200

ccaccactgg taacatctgc agatgaacat tcagttcttg cagaagaaga caaggtggca 4260

attaaaggtg cttctcccat tgaaacttca tccaaacatt tagcttggtc agaagcagag 4320

aaggaaatta aatttgattc acttccaagt gtctcctcta tagcagagca ttctgttttg 4380

tcagaagtag aagccaaaga agttaaagct gggttgccag taatcaaaac atcatcttct 4440

cagcattcag ataaatctga ggaagcaagg gtagaagaca aacaagatct tttattttct 4500

acagtctgtg actctgaacg tttggtttca tcacagaaga agagcttgat gtctacctca 4560

gaggtgttag agcctgaaca tgagcttcca ctcagcctat ggggtgagat aaagaagaaa 4620

gaaactgaac ttccttcatc acaaaatgtg tcacctgcat ccaaacatat aatcccaaaa 4680

ggcaaagatg aggaaacagc aagttcatct cctgagttgg aaaatttagc atcaggttta 4740

gccccaacat tactgctcct cagtgatgat aagaacaaac cggcagtgga ggtatcttct 4800

acagctcagg gagacttccc atcagaaaaa caagatgttg ctttggcaga gctgtctttg 4860

gaacctgaga agaaagacaa gccacaccaa ccgttggaat taccaaatgc tgggtcagaa 4920

ttttctagtg atttaggtag acaaagtgga tccataggta caaaacaagc aaagtctccc 4980

ataactgaaa cagaggattc tgttttagaa aaaggcccag ctgagcttag gagcagagaa 5040

ggaaaagaag aaaatagaga gctttgtgca tcttctacga tgcctgcaat ttcagagctt 5100

tcatcattgc ttagggagga atctcagaat gaagaaatta aacctttctc tcccaagatc 5160

atcagcctag agtcgaaaga accacctgcc tctgtagctg aaggaggcaa cccagaagaa 5220

tttcagccat ttactttttc tttaaaagga ttatcagagg aggttagcca tccagccgac 5280

tttaaaaagg gaggaaatca agaaataggc ccattaccac caactggaaa tttgaaggca 5340

caagtcatgg gagatatttt agataagcta agtgaagaaa caggccaccc aaattcatcc 5400

caggtactcc agagtataac agaaccatca aagattgctc cttctgacct ccttgtagaa 5460

caaaaaaaga cagaaaaagc acttcattca gatcaaactg ttaaattacc tgatgtaagc 5520

acctcttctg aagataaaca agatctgggt attaagcagt tttcacttat gagagagaat 5580

ttgcctttgg aacaatcaaa atcatttatg acaaccaagc ctgcggatgt caaagaaaca 5640

aaaatggaag aattctttat ttctccaaag gatgaaaact ggatgttggg aaagccagaa 5700

aatgtggcta gtcaacacga acagagaata gcaggatctg tgcagctgga ttcctctagc 5760

agcaatgagc tgaggccagg gcagctcaag gctgctgtgt ccagtaagga ccatacatgt 5820

gaagtgagaa agcaggtcct gccgcattct gctgaagaat ctcatttgtc atcacaagaa 5880

gcagtatctg ctcttgatac ttccagtggt aatacagaga ccttatcaag taaaagttac 5940

tcttctgaag aagtaaagct ggctgaagaa ccaaagtctt tagtcctagc tggaaatgta 6000

gagagaaaca tagcagaggg gaaggagatt cattctttga tggagagtga aagtttgcta 6060

ttggagaaag caaacacaga gctttcctgg ccttccaaag aagatagcca ggaaaaaatt 6120

aaactacctc ctgaaagatt cttccagaaa ccagtgtctg gcctatcagt ggaacaggtg 6180

aagtcagaaa caatctcctc ttctgtcaaa acagcccatt tcccggcaga aggtgtggaa 6240

cctgcattgg gcaatgaaaa agaagcacac aggagcacac ctccttttcc tgaagagaag 6300

ccattggaag aatcaaaaat ggttcagtca aaggttattg atgatgctga tgagggaaag 6360

aaaccatcac ctgaagtaaa aatacccaca caaagaaaac ccatctcctc aatccatgca 6420

agagagcctc aatccccaga gtcacctgag gtgacacaaa atccacctac acaaccaaag 6480

gtggctaagc cggaccttcc tgaggaaaag ggaaagaaag gaatttcatc tttcaaatcg 6540

tggatgtcca gcttgttttt tggatcgagc actccagata acaaagttgc tgaacaagaa 6600

gacttagaaa cacagccaag tccatccgta gaaaaagcag tgactgtgat agatcctgaa 6660

ggtacaattc ccaccaattt taatgtagct gagaaaccag ctgatcattc attatcagag 6720

gtaaaactta aaactgctga tgaacccaga ggtactttag taaaatctgg tgacggtcaa 6780

aacgttaaag aaaaatccat gattttatca aatgtagaag atttacaaca gccaaaattc 6840

atttctgagg tgtctaggga agattatgga aaaaaagaaa tctcaggcga ttcagaggaa 6900

atgaacataa actcagtagt tacttctgct gatggtgaga accttgaaat tcaatcttat 6960

tcactaatcg gtgagaaatt ggttatggaa gaagccaaaa ctattgttcc tcctcatgtt 7020

actgatagta aaagagtcca gaagccagca atcgctcctc catctaaatg gaatatttct 7080

atttttaagg aagagccaag aagtgatcaa aaacaaaaat cactcctttc atttgatgta 7140

gtagataagg tgccacaaca gccaaaatca gcttcctcca actttgcaag taaaaatatc 7200

acaaaggaat cagagaaacc agagtcaatt attttgccag tagaagaatc aaaaggcagt 7260

ttaattgatt tcagtgaaga cagactcaag aaagaaatgc aaaatcctac ttccttgaaa 7320

atttctgaag aggaaacaaa actcaggtct gttagtccaa ctgagaagaa agataatttg 7380

gaaaacagat catatacctt ggcagaaaag aaggtgctgg cagaaaaaca aaactctgtg 7440

gccccattag agcttagaga tagtaatgaa atagggaaga cacaaattac acttggatct 7500

agatctactg aactgaaaga atcaaaagcc gatgctatgc cacagcactt ctatcaaaat 7560

gaagactaca atgaaagacc caaaatcatt gttggttctg aaaaggagaa aggtgaagaa 7620

aaagaaaatc aggtatatgt gctttcagaa ggaaagaagc agcaggaaca tcagccttat 7680

tctgtgaatg tagccgagtc tatgagtaga gaatcagata tctctttagg tcattctttg 7740

ggtgaaactc aatcattttc attagttaaa gctacatcag ttactgaaaa atcagaagcc 7800

atgctcgcag aggctcaccc agaaatcaga gaagcaaagg cagtaggaac ccaaccacat 7860

cctttagaag aaagtaaagt tttggtggag aaaaccaaga ctttcctgcc ggtggctctt 7920

tcttgtcgtg atgaaataga gaaccactct ttatctcagg aaggaaatct agtattagaa 7980

aagtcaagca gagatatgcc agatcacagt gaagaaaaag aacagttcag agagtcagag 8040

ctatcgaaag gcggttcagt agatatcaca aaagaaactg tgaaacaagg atttcaagaa 8100

aaggcagtag gaacccaacc acgtccttta gaagaaagta aagttttggt ggagaaaact 8160

aagactttcc tgccagtggt tctttcttgt catgatgaaa tagagaacca ctctttgtct 8220

caggaaggaa atctagtgtt agaaaagtca agcagagata tgccagatca cagtgaagaa 8280

aaagaacagt tcaaagagtc agagctatgg aaaggtggtt cagtagatat cacaaaagaa 8340

agtatgaaag aaggatttcc atctaaagaa tccgaaagga ctttagctcg tccttttgat 8400

gaaactaaga gctcagaaac accgccatat ttgctgtcac ctgtaaaacc acaaactctt 8460

gcttcaggag cttctccaga aattaacgca gtgaagaaaa aagaaatgcc acgatcagaa 8520

ttgactccag aaaggcatac agttcatact attcagacat ctaaagatga cacatccgat 8580

gtgcctaaac aatctgttct tgtttcaaag caccacttgg aggctgcgga agatacccgt 8640

gtaaaggaac cactgtcttc agcaaaaagc aactatgctc aatttatatc taatacatca 8700

gcaagcaatg ctgataaaat ggtttctaat aaagaaatgc ccaaggaacc tgaagacaca 8760

tatgcaaaag gtgaagactt tacagtgact agtaagccag ccggactttc agaagatcag 8820

aagactgcct ttagtatcat ttctgaaggc tgtgagatat tgaatattca tgctccggcc 8880

tttatttctt caatcgatca ggaagaaagt gaacaaatgc aagataaatt agaatatttg 8940

gaagagaaag cctcatttaa aaccatacca ctccctgatg atagtgaaac agttgcttgt 9000

cataaaacat taaagagcag gttagaagat gaaaaagtta ccccattgaa agaaaataaa 9060

caaaaggaaa ctcataagac aaaagaagag atatccacag attcagaaac tgatttatca 9120

tttattcagc ccacaattcc cagtgaagag gattattttg aaaaatatac tttgattgat 9180

tataacatct ccccagaccc agaaaaacag aaagctccac agaaattaaa tgttgaagag 9240

aaactctcaa aggaagttac agaagaaact atctctttcc cagtaagttc agtggaaagt 9300

gcactagaac atgaatatga cttggtgaaa ttagatgaaa gtttttatgg accagaaaag 9360

ggccacaaca tattatctca tccagagacc caaagccaaa actcagctga caggaatgtt 9420

tcaaaggaca caaagagaga tgtggactca aagtcaccgg ggatgccttt atttgaagca 9480

gaggaaggag ttctatcacg aacccagata tttcctacca ctattaaagt cattgatcca 9540

gaatttctgg aggagccacc tgcacttgca tttttatata aggatctgta tgaagaagca 9600

gttggagaga aaaagaagga agaggagaca gcttctgaag gtgacagtgt gaattctgag 9660

gcatcatttc ccagcagaaa ttctgacact gatgatggaa caggaatata ttttgagaag 9720

tacatactca aagatgacat tctccatgac acatctctaa ctcaaaagga ccagggccaa 9780

ggtctggaag aaaaacgagt tggtaaggat gattcatacc aaccgatagc tgcagaaggg 9840

gaaatttggg gaaagtttgg aactatttgc agggagaaga gtctggaaga acagaaaggt 9900

gtttatgggg aaggagaatc agtagaccat gtggagaccg ttggtaacgt agcgatgcag 9960

aagaaagctc ccatcacaga ggacgtcaga gtggctaccc agaaaataag ttatgcggtt 10020

ccatttgaag acacccatca tgttctggag cgtgcagatg aagcaggcag tcacggtaat 10080

gaagtcggaa atgcaagtcc agaggtcaat ctgaatgtcc cagtacaagt gtccttcccg 10140

gaggaagaat ttgcatctgg tgcaactcat gttcaagaaa catcactaga agaacctaaa 10200

atcctggtcc cacctgagcc aagtgaagag aggctccgta atagccctgt tcaggatgag 10260

tatgaattta cagaatccct gcataatgaa gtggttcctc aagacatatt atcagaagaa 10320

ctgtcttcag aatccacacc tgaagatgtc ttatctcaag gaaaggaatc ctttgagcac 10380

atcagtgaaa atgaatttgc gagtgaggca gaacaaagta cacctgctga acaaaaagag 10440

ttgggcagcg agaggaaaga agaagaccaa ttatcatctg aggtagtaac tgaaaaggca 10500

caaaaagagc tgaaaaagtc ccagattgac acatactgtt acacctgcaa atgtccaatt 10560

tctgccactg acaaggtgtt tggcacccac aaagaccatg aagtttcaac gcttgacaca 10620

gctataagtg ctgtaaaggt tcaattagca gaatttctag aaaatttaca agaaaagtcc 10680

ttgaggattg aagcctttgt tagtgagata gaatcctttt ttaataccat tgaggaaaac 10740

tgtagtaaaa atgagaaaag gctagaagaa cagaatgagg aaatgatgaa gaaggtttta 10800

gcacagtatg atgagaaagc ccagagcttt gaggaagtga agaagaagaa gatggagttc 10860

ctgcatgagc agatggtcca ctttctgcag agcatggaca ctgccaaaga caccctggag 10920

accatcgtga gagaagcaga ggagcttgat gaggccgtct tcctgacttc gtttgaggaa 10980

atcaatgaaa ggttgctttc tgcaatggag agcactgctt ctttagagaa aatgcctgct 11040

gcgttttccc ttttcgaaca ttatgatgac agctcggcaa gaagtgacca gatgttaaaa 11100

caagtggctg ttccacagcc tcctagatta gaacctcagg aaccaaattc tgccaccagc 11160

acaacaattg cagtttactg gagcatgaac aaggaagatg tcattgattc atttcaggtt 11220

tactgcatgg aggagccaca agatgatcaa gaagtaaatg agttggtaga agaatacaga 11280

ctgacagtga aagaaagcta ctgcattttt gaagatctgg aacctgaccg atgctatcaa 11340

gtgtgggtga tggctgtgaa cttcactgga tgtagcctgc ccagtgaaag ggccatcttt 11400

aggacagcac cctccacccc tgtgatccgc gctgaggact gtactgtgtg ttggaacaca 11460

gccactatcc gatggcggcc caccacccca gaggccacgg agacctacac tctggagtac 11520

tgcagacagc actctcctga gggagagggc ctcagatctt tctctggaat caaaggactc 11580

cagctgaaag ttaacctcca acccaatgat aactactttt tctatgtgag ggccatcaat 11640

gcatttggga caagtgaaca gagtgaagct gctctcatct ccaccagagg aaccagattt 11700

ctcttgttga gagaaacagc tcatcctgct ctacacattt cctcaagtgg gacagtgatc 11760

agctttggtg agaggagacg gctgacggaa atcccgtcag tgctgggtga ggagctgcct 11820

tcctgtggcc agcattactg ggaaaccaca gtcacagact gcccagcata tcgactcggc 11880

atctgctcca gctcggctgt gcaggcaggt gccctaggac aaggggagac ctcatggtac 11940

atgcactgct ctgagccaca gagatacaca tttttctaca gtggtattgt gagtgatgtt 12000

catgtgactg agcgtccagc cagagtgggc atcctgctgg actacaacaa ccagagactt 12060

atcttcatca acgcagagag cgagcagttg ctcttcatca tcaggcacag gtttaatgag 12120

ggtgtccacc ctgcctttgc cctggagaaa cctggaaaat gtactttgca cctggggata 12180

gagcccccgg attctgtaag gcacaag 12207

<210> 165

<211> 4069

<212> PRT

<213> 智人(Homo sapiens)

<400> 165

Met Ala Ser Arg Asp Ser Asn His Ala Gly Glu Ser Phe Leu Gly Ser

1 5 10 15

Asp Gly Asp Glu Glu Ala Thr Arg Glu Leu Glu Thr Glu Glu Glu Ser

20 25 30

Glu Gly Glu Glu Asp Glu Thr Ala Ala Glu Ser Glu Glu Glu Pro Asp

35 40 45

Ser Arg Leu Ser Asp Gln Asp Glu Glu Gly Lys Ile Lys Gln Glu Tyr

50 55 60

Ile Ile Ser Asp Pro Ser Phe Ser Met Val Thr Val Gln Arg Glu Asp

65 70 75 80

Ser Gly Ile Thr Trp Glu Thr Asn Ser Ser Arg Ser Ser Thr Pro Trp

85 90 95

Ala Ser Glu Glu Ser Gln Thr Ser Gly Val Cys Ser Arg Glu Gly Ser

100 105 110

Thr Val Asn Ser Pro Pro Gly Asn Val Ser Phe Ile Val Asp Glu Val

115 120 125

Lys Lys Val Arg Lys Arg Thr His Lys Ser Lys His Gly Ser Pro Ser

130 135 140

Leu Arg Arg Lys Gly Asn Arg Lys Arg Asn Ser Phe Glu Ser Gln Asp

145 150 155 160

Val Pro Thr Asn Lys Lys Gly Ser Pro Leu Thr Ser Ala Ser Gln Val

165 170 175

Leu Thr Thr Glu Lys Glu Lys Ser Tyr Thr Gly Ile Tyr Asp Lys Ala

180 185 190

Arg Lys Lys Lys Thr Thr Ser Asn Thr Pro Pro Ile Thr Gly Ala Ile

195 200 205

Tyr Lys Glu His Lys Pro Leu Val Leu Arg Pro Val Tyr Ile Gly Thr

210 215 220

Val Gln Tyr Lys Ile Lys Met Phe Asn Ser Val Lys Glu Glu Leu Ile

225 230 235 240

Pro Leu Gln Phe Tyr Gly Thr Leu Pro Lys Gly Tyr Val Ile Lys Glu

245 250 255

Ile His Tyr Arg Lys Gly Lys Asp Ala Ser Ile Ser Leu Glu Pro Asp

260 265 270

Leu Asp Asn Ser Gly Ser Asn Thr Val Ser Lys Thr Arg Lys Leu Val

275 280 285

Ala Gln Ser Ile Glu Asp Lys Val Lys Glu Val Phe Pro Pro Trp Arg

290 295 300

Gly Ala Leu Ser Lys Gly Ser Glu Ser Leu Thr Leu Met Phe Ser His

305 310 315 320

Glu Asp Gln Lys Lys Ile Tyr Ala Asp Ser Pro Leu Asn Ala Thr Ser

325 330 335

Ala Leu Glu His Thr Val Pro Ser Tyr Ser Ser Ser Gly Arg Ala Glu

340 345 350

Gln Gly Ile Gln Leu Arg His Ser Gln Ser Val Pro Gln Gln Pro Glu

355 360 365

Asp Glu Ala Lys Pro His Glu Val Glu Pro Pro Ser Val Thr Pro Asp

370 375 380

Thr Pro Ala Thr Met Phe Leu Arg Thr Thr Lys Glu Glu Cys Glu Leu

385 390 395 400

Ala Ser Pro Gly Thr Ala Ala Ser Glu Asn Asp Ser Ser Val Ser Pro

405 410 415

Ser Phe Ala Asn Glu Val Lys Lys Glu Asp Val Tyr Ser Ala His His

420 425 430

Ser Ile Ser Leu Glu Ala Ala Ser Pro Gly Leu Ala Ala Ser Thr Gln

435 440 445

Asp Gly Leu Asp Pro Asp Gln Glu Gln Pro Asp Leu Thr Ser Ile Glu

450 455 460

Arg Ala Glu Pro Val Ser Ala Lys Leu Thr Pro Thr His Pro Ser Val

465 470 475 480

Lys Gly Glu Lys Glu Glu Asn Met Leu Glu Pro Ser Ile Ser Leu Ser

485 490 495

Glu Pro Leu Met Leu Glu Glu Pro Glu Lys Glu Glu Ile Glu Thr Ser

500 505 510

Leu Pro Ile Ala Ile Thr Pro Glu Pro Glu Asp Ser Asn Leu Val Glu

515 520 525

Glu Glu Ile Val Glu Leu Asp Tyr Pro Glu Ser Pro Leu Val Ser Glu

530 535 540

Lys Pro Phe Pro Pro His Met Ser Pro Glu Val Glu His Lys Glu Glu

545 550 555 560

Glu Leu Ile Leu Pro Leu Leu Ala Ala Ser Ser Pro Glu His Val Ala

565 570 575

Leu Ser Glu Glu Glu Arg Glu Glu Ile Ala Ser Val Ser Thr Gly Ser

580 585 590

Ala Phe Val Ser Glu Tyr Ser Val Pro Gln Asp Leu Asn His Glu Leu

595 600 605

Gln Glu Gln Glu Gly Glu Pro Val Pro Pro Ser Asn Val Glu Ala Ile

610 615 620

Ala Glu His Ala Val Leu Ser Glu Glu Glu Asn Glu Glu Phe Glu Ala

625 630 635 640

Tyr Ser Pro Ala Ala Ala Pro Thr Ser Glu Ser Ser Leu Ser Pro Ser

645 650 655

Thr Thr Glu Lys Thr Ser Glu Asn Gln Ser Pro Leu Phe Ser Thr Val

660 665 670

Thr Pro Glu Tyr Met Val Leu Ser Gly Asp Glu Ala Ser Glu Ser Gly

675 680 685

Cys Tyr Thr Pro Asp Ser Thr Ser Ala Ser Glu Tyr Ser Val Pro Ser

690 695 700

Leu Ala Thr Lys Glu Ser Leu Lys Lys Thr Ile Asp Arg Lys Ser Pro

705 710 715 720

Leu Ile Leu Lys Gly Val Ser Glu Tyr Met Ile Pro Ser Glu Glu Lys

725 730 735

Glu Asp Thr Gly Ser Phe Thr Pro Ala Val Ala Pro Ala Ser Glu Pro

740 745 750

Ser Leu Ser Pro Ser Thr Thr Glu Lys Thr Ser Glu Cys Gln Ser Pro

755 760 765

Leu Pro Ser Thr Ala Thr Ser Glu His Val Val Pro Ser Glu Gly Glu

770 775 780

Asp Leu Gly Ser Glu Arg Phe Thr Pro Asp Ser Lys Leu Ile Ser Lys

785 790 795 800

Tyr Ala Ala Pro Leu Asn Ala Thr Gln Glu Ser Gln Lys Lys Ile Ile

805 810 815

Asn Glu Ala Ser Gln Phe Lys Pro Lys Gly Ile Ser Glu His Thr Val

820 825 830

Leu Ser Val Asp Gly Lys Glu Val Ile Gly Pro Ser Ser Pro Asp Leu

835 840 845

Val Val Ala Ser Glu His Ser Phe Pro Pro His Thr Thr Glu Met Thr

850 855 860

Ser Glu Cys Gln Ala Pro Pro Leu Ser Ala Thr Pro Ser Glu Tyr Val

865 870 875 880

Val Leu Ser Asp Glu Glu Ala Val Glu Leu Glu Arg Tyr Thr Pro Ser

885 890 895

Ser Thr Ser Ala Ser Glu Phe Ser Val Pro Pro Tyr Ala Thr Pro Glu

900 905 910

Ala Gln Glu Glu Glu Ile Val His Arg Ser Leu Asn Leu Lys Gly Ala

915 920 925

Ser Ser Pro Met Asn Leu Ser Glu Glu Asp Gln Glu Asp Ile Gly Pro

930 935 940

Phe Ser Pro Asp Ser Ala Phe Val Ser Glu Phe Ser Phe Pro Pro Tyr

945 950 955 960

Ala Thr Gln Glu Ala Glu Lys Arg Glu Phe Glu Cys Asp Ser Pro Ile

965 970 975

Cys Leu Thr Ser Pro Ser Glu His Thr Ile Leu Ser Asp Glu Asp Thr

980 985 990

Glu Glu Ala Glu Leu Phe Ser Pro Asp Ser Ala Ser Gln Val Ser Ile

995 1000 1005

Pro Pro Phe Arg Ile Ser Glu Thr Glu Lys Asn Glu Leu Glu Pro

1010 1015 1020

Asp Ser Leu Leu Thr Ala Val Ser Ala Ser Gly Tyr Ser Cys Phe

1025 1030 1035

Ser Glu Ala Asp Glu Glu Asp Ile Gly Ser Thr Ala Ala Thr Pro

1040 1045 1050

Val Ser Glu Gln Phe Ser Ser Ser Gln Lys Gln Lys Ala Glu Thr

1055 1060 1065

Phe Pro Leu Met Ser Pro Leu Glu Asp Leu Ser Leu Pro Pro Ser

1070 1075 1080

Thr Asp Lys Ser Glu Lys Ala Glu Ile Lys Pro Glu Ile Pro Thr

1085 1090 1095

Thr Ser Thr Ser Val Ser Glu Tyr Leu Ile Leu Ala Gln Lys Gln

1100 1105 1110

Lys Thr Gln Ala Tyr Leu Glu Pro Glu Ser Glu Asp Leu Ile Pro

1115 1120 1125

Ser His Leu Thr Ser Glu Val Glu Lys Gly Glu Arg Glu Ala Ser

1130 1135 1140

Ser Ser Val Ala Ala Ile Pro Ala Ala Leu Pro Ala Gln Ser Ser

1145 1150 1155

Ile Val Lys Glu Glu Thr Lys Pro Ala Ser Pro His Ser Val Leu

1160 1165 1170

Pro Asp Ser Val Pro Ala Ile Lys Lys Glu Gln Glu Pro Thr Ala

1175 1180 1185

Ala Leu Thr Leu Lys Ala Ala Asp Glu Gln Met Ala Leu Ser Lys

1190 1195 1200

Val Arg Lys Glu Glu Ile Val Pro Asp Ser Gln Glu Ala Thr Ala

1205 1210 1215

His Val Ser Gln Asp Gln Lys Met Glu Pro Gln Pro Pro Asn Val

1220 1225 1230

Pro Glu Ser Glu Met Lys Tyr Ser Val Leu Pro Asp Met Val Asp

1235 1240 1245

Glu Pro Lys Lys Gly Val Lys Pro Lys Leu Val Leu Asn Val Thr

1250 1255 1260

Ser Glu Leu Glu Gln Arg Lys Leu Ser Lys Asn Glu Pro Glu Val

1265 1270 1275

Ile Lys Pro Tyr Ser Pro Leu Lys Glu Thr Ser Leu Ser Gly Pro

1280 1285 1290

Glu Ala Leu Ser Ala Val Lys Met Glu Met Lys His Asp Ser Lys

1295 1300 1305

Ile Thr Thr Thr Pro Ile Val Leu His Ser Ala Ser Ser Gly Val

1310 1315 1320

Glu Lys Gln Val Glu His Gly Pro Pro Ala Leu Ala Phe Ser Ala

1325 1330 1335

Leu Ser Glu Glu Ile Lys Lys Glu Ile Glu Pro Ser Ser Ser Thr

1340 1345 1350

Thr Thr Ala Ser Val Thr Lys Leu Asp Ser Asn Leu Thr Arg Ala

1355 1360 1365

Val Lys Glu Glu Ile Pro Thr Asp Ser Ser Leu Ile Thr Pro Val

1370 1375 1380

Asp Arg Pro Val Leu Thr Lys Val Gly Lys Gly Glu Leu Gly Ser

1385 1390 1395

Gly Leu Pro Pro Leu Val Thr Ser Ala Asp Glu His Ser Val Leu

1400 1405 1410

Ala Glu Glu Asp Lys Val Ala Ile Lys Gly Ala Ser Pro Ile Glu

1415 1420 1425

Thr Ser Ser Lys His Leu Ala Trp Ser Glu Ala Glu Lys Glu Ile

1430 1435 1440

Lys Phe Asp Ser Leu Pro Ser Val Ser Ser Ile Ala Glu His Ser

1445 1450 1455

Val Leu Ser Glu Val Glu Ala Lys Glu Val Lys Ala Gly Leu Pro

1460 1465 1470

Val Ile Lys Thr Ser Ser Ser Gln His Ser Asp Lys Ser Glu Glu

1475 1480 1485

Ala Arg Val Glu Asp Lys Gln Asp Leu Leu Phe Ser Thr Val Cys

1490 1495 1500

Asp Ser Glu Arg Leu Val Ser Ser Gln Lys Lys Ser Leu Met Ser

1505 1510 1515

Thr Ser Glu Val Leu Glu Pro Glu His Glu Leu Pro Leu Ser Leu

1520 1525 1530

Trp Gly Glu Ile Lys Lys Lys Glu Thr Glu Leu Pro Ser Ser Gln

1535 1540 1545

Asn Val Ser Pro Ala Ser Lys His Ile Ile Pro Lys Gly Lys Asp

1550 1555 1560

Glu Glu Thr Ala Ser Ser Ser Pro Glu Leu Glu Asn Leu Ala Ser

1565 1570 1575

Gly Leu Ala Pro Thr Leu Leu Leu Leu Ser Asp Asp Lys Asn Lys

1580 1585 1590

Pro Ala Val Glu Val Ser Ser Thr Ala Gln Gly Asp Phe Pro Ser

1595 1600 1605

Glu Lys Gln Asp Val Ala Leu Ala Glu Leu Ser Leu Glu Pro Glu

1610 1615 1620

Lys Lys Asp Lys Pro His Gln Pro Leu Glu Leu Pro Asn Ala Gly

1625 1630 1635

Ser Glu Phe Ser Ser Asp Leu Gly Arg Gln Ser Gly Ser Ile Gly

1640 1645 1650

Thr Lys Gln Ala Lys Ser Pro Ile Thr Glu Thr Glu Asp Ser Val

1655 1660 1665

Leu Glu Lys Gly Pro Ala Glu Leu Arg Ser Arg Glu Gly Lys Glu

1670 1675 1680

Glu Asn Arg Glu Leu Cys Ala Ser Ser Thr Met Pro Ala Ile Ser

1685 1690 1695

Glu Leu Ser Ser Leu Leu Arg Glu Glu Ser Gln Asn Glu Glu Ile

1700 1705 1710

Lys Pro Phe Ser Pro Lys Ile Ile Ser Leu Glu Ser Lys Glu Pro

1715 1720 1725

Pro Ala Ser Val Ala Glu Gly Gly Asn Pro Glu Glu Phe Gln Pro

1730 1735 1740

Phe Thr Phe Ser Leu Lys Gly Leu Ser Glu Glu Val Ser His Pro

1745 1750 1755

Ala Asp Phe Lys Lys Gly Gly Asn Gln Glu Ile Gly Pro Leu Pro

1760 1765 1770

Pro Thr Gly Asn Leu Lys Ala Gln Val Met Gly Asp Ile Leu Asp

1775 1780 1785

Lys Leu Ser Glu Glu Thr Gly His Pro Asn Ser Ser Gln Val Leu

1790 1795 1800

Gln Ser Ile Thr Glu Pro Ser Lys Ile Ala Pro Ser Asp Leu Leu

1805 1810 1815

Val Glu Gln Lys Lys Thr Glu Lys Ala Leu His Ser Asp Gln Thr

1820 1825 1830

Val Lys Leu Pro Asp Val Ser Thr Ser Ser Glu Asp Lys Gln Asp

1835 1840 1845

Leu Gly Ile Lys Gln Phe Ser Leu Met Arg Glu Asn Leu Pro Leu

1850 1855 1860

Glu Gln Ser Lys Ser Phe Met Thr Thr Lys Pro Ala Asp Val Lys

1865 1870 1875

Glu Thr Lys Met Glu Glu Phe Phe Ile Ser Pro Lys Asp Glu Asn

1880 1885 1890

Trp Met Leu Gly Lys Pro Glu Asn Val Ala Ser Gln His Glu Gln

1895 1900 1905

Arg Ile Ala Gly Ser Val Gln Leu Asp Ser Ser Ser Ser Asn Glu

1910 1915 1920

Leu Arg Pro Gly Gln Leu Lys Ala Ala Val Ser Ser Lys Asp His

1925 1930 1935

Thr Cys Glu Val Arg Lys Gln Val Leu Pro His Ser Ala Glu Glu

1940 1945 1950

Ser His Leu Ser Ser Gln Glu Ala Val Ser Ala Leu Asp Thr Ser

1955 1960 1965

Ser Gly Asn Thr Glu Thr Leu Ser Ser Lys Ser Tyr Ser Ser Glu

1970 1975 1980

Glu Val Lys Leu Ala Glu Glu Pro Lys Ser Leu Val Leu Ala Gly

1985 1990 1995

Asn Val Glu Arg Asn Ile Ala Glu Gly Lys Glu Ile His Ser Leu

2000 2005 2010

Met Glu Ser Glu Ser Leu Leu Leu Glu Lys Ala Asn Thr Glu Leu

2015 2020 2025

Ser Trp Pro Ser Lys Glu Asp Ser Gln Glu Lys Ile Lys Leu Pro

2030 2035 2040

Pro Glu Arg Phe Phe Gln Lys Pro Val Ser Gly Leu Ser Val Glu

2045 2050 2055

Gln Val Lys Ser Glu Thr Ile Ser Ser Ser Val Lys Thr Ala His

2060 2065 2070

Phe Pro Ala Glu Gly Val Glu Pro Ala Leu Gly Asn Glu Lys Glu

2075 2080 2085

Ala His Arg Ser Thr Pro Pro Phe Pro Glu Glu Lys Pro Leu Glu

2090 2095 2100

Glu Ser Lys Met Val Gln Ser Lys Val Ile Asp Asp Ala Asp Glu

2105 2110 2115

Gly Lys Lys Pro Ser Pro Glu Val Lys Ile Pro Thr Gln Arg Lys

2120 2125 2130

Pro Ile Ser Ser Ile His Ala Arg Glu Pro Gln Ser Pro Glu Ser

2135 2140 2145

Pro Glu Val Thr Gln Asn Pro Pro Thr Gln Pro Lys Val Ala Lys

2150 2155 2160

Pro Asp Leu Pro Glu Glu Lys Gly Lys Lys Gly Ile Ser Ser Phe

2165 2170 2175

Lys Ser Trp Met Ser Ser Leu Phe Phe Gly Ser Ser Thr Pro Asp

2180 2185 2190

Asn Lys Val Ala Glu Gln Glu Asp Leu Glu Thr Gln Pro Ser Pro

2195 2200 2205

Ser Val Glu Lys Ala Val Thr Val Ile Asp Pro Glu Gly Thr Ile

2210 2215 2220

Pro Thr Asn Phe Asn Val Ala Glu Lys Pro Ala Asp His Ser Leu

2225 2230 2235

Ser Glu Val Lys Leu Lys Thr Ala Asp Glu Pro Arg Gly Thr Leu

2240 2245 2250

Val Lys Ser Gly Asp Gly Gln Asn Val Lys Glu Lys Ser Met Ile

2255 2260 2265

Leu Ser Asn Val Glu Asp Leu Gln Gln Pro Lys Phe Ile Ser Glu

2270 2275 2280

Val Ser Arg Glu Asp Tyr Gly Lys Lys Glu Ile Ser Gly Asp Ser

2285 2290 2295

Glu Glu Met Asn Ile Asn Ser Val Val Thr Ser Ala Asp Gly Glu

2300 2305 2310

Asn Leu Glu Ile Gln Ser Tyr Ser Leu Ile Gly Glu Lys Leu Val

2315 2320 2325

Met Glu Glu Ala Lys Thr Ile Val Pro Pro His Val Thr Asp Ser

2330 2335 2340

Lys Arg Val Gln Lys Pro Ala Ile Ala Pro Pro Ser Lys Trp Asn

2345 2350 2355

Ile Ser Ile Phe Lys Glu Glu Pro Arg Ser Asp Gln Lys Gln Lys

2360 2365 2370

Ser Leu Leu Ser Phe Asp Val Val Asp Lys Val Pro Gln Gln Pro

2375 2380 2385

Lys Ser Ala Ser Ser Asn Phe Ala Ser Lys Asn Ile Thr Lys Glu

2390 2395 2400

Ser Glu Lys Pro Glu Ser Ile Ile Leu Pro Val Glu Glu Ser Lys

2405 2410 2415

Gly Ser Leu Ile Asp Phe Ser Glu Asp Arg Leu Lys Lys Glu Met

2420 2425 2430

Gln Asn Pro Thr Ser Leu Lys Ile Ser Glu Glu Glu Thr Lys Leu

2435 2440 2445

Arg Ser Val Ser Pro Thr Glu Lys Lys Asp Asn Leu Glu Asn Arg

2450 2455 2460

Ser Tyr Thr Leu Ala Glu Lys Lys Val Leu Ala Glu Lys Gln Asn

2465 2470 2475

Ser Val Ala Pro Leu Glu Leu Arg Asp Ser Asn Glu Ile Gly Lys

2480 2485 2490

Thr Gln Ile Thr Leu Gly Ser Arg Ser Thr Glu Leu Lys Glu Ser

2495 2500 2505

Lys Ala Asp Ala Met Pro Gln His Phe Tyr Gln Asn Glu Asp Tyr

2510 2515 2520

Asn Glu Arg Pro Lys Ile Ile Val Gly Ser Glu Lys Glu Lys Gly

2525 2530 2535

Glu Glu Lys Glu Asn Gln Val Tyr Val Leu Ser Glu Gly Lys Lys

2540 2545 2550

Gln Gln Glu His Gln Pro Tyr Ser Val Asn Val Ala Glu Ser Met

2555 2560 2565

Ser Arg Glu Ser Asp Ile Ser Leu Gly His Ser Leu Gly Glu Thr

2570 2575 2580

Gln Ser Phe Ser Leu Val Lys Ala Thr Ser Val Thr Glu Lys Ser

2585 2590 2595

Glu Ala Met Leu Ala Glu Ala His Pro Glu Ile Arg Glu Ala Lys

2600 2605 2610

Ala Val Gly Thr Gln Pro His Pro Leu Glu Glu Ser Lys Val Leu

2615 2620 2625

Val Glu Lys Thr Lys Thr Phe Leu Pro Val Ala Leu Ser Cys Arg

2630 2635 2640

Asp Glu Ile Glu Asn His Ser Leu Ser Gln Glu Gly Asn Leu Val

2645 2650 2655

Leu Glu Lys Ser Ser Arg Asp Met Pro Asp His Ser Glu Glu Lys

2660 2665 2670

Glu Gln Phe Arg Glu Ser Glu Leu Ser Lys Gly Gly Ser Val Asp

2675 2680 2685

Ile Thr Lys Glu Thr Val Lys Gln Gly Phe Gln Glu Lys Ala Val

2690 2695 2700

Gly Thr Gln Pro Arg Pro Leu Glu Glu Ser Lys Val Leu Val Glu

2705 2710 2715

Lys Thr Lys Thr Phe Leu Pro Val Val Leu Ser Cys His Asp Glu

2720 2725 2730

Ile Glu Asn His Ser Leu Ser Gln Glu Gly Asn Leu Val Leu Glu

2735 2740 2745

Lys Ser Ser Arg Asp Met Pro Asp His Ser Glu Glu Lys Glu Gln

2750 2755 2760

Phe Lys Glu Ser Glu Leu Trp Lys Gly Gly Ser Val Asp Ile Thr

2765 2770 2775

Lys Glu Ser Met Lys Glu Gly Phe Pro Ser Lys Glu Ser Glu Arg

2780 2785 2790

Thr Leu Ala Arg Pro Phe Asp Glu Thr Lys Ser Ser Glu Thr Pro

2795 2800 2805

Pro Tyr Leu Leu Ser Pro Val Lys Pro Gln Thr Leu Ala Ser Gly

2810 2815 2820

Ala Ser Pro Glu Ile Asn Ala Val Lys Lys Lys Glu Met Pro Arg

2825 2830 2835

Ser Glu Leu Thr Pro Glu Arg His Thr Val His Thr Ile Gln Thr

2840 2845 2850

Ser Lys Asp Asp Thr Ser Asp Val Pro Lys Gln Ser Val Leu Val

2855 2860 2865

Ser Lys His His Leu Glu Ala Ala Glu Asp Thr Arg Val Lys Glu

2870 2875 2880

Pro Leu Ser Ser Ala Lys Ser Asn Tyr Ala Gln Phe Ile Ser Asn

2885 2890 2895

Thr Ser Ala Ser Asn Ala Asp Lys Met Val Ser Asn Lys Glu Met

2900 2905 2910

Pro Lys Glu Pro Glu Asp Thr Tyr Ala Lys Gly Glu Asp Phe Thr

2915 2920 2925

Val Thr Ser Lys Pro Ala Gly Leu Ser Glu Asp Gln Lys Thr Ala

2930 2935 2940

Phe Ser Ile Ile Ser Glu Gly Cys Glu Ile Leu Asn Ile His Ala

2945 2950 2955

Pro Ala Phe Ile Ser Ser Ile Asp Gln Glu Glu Ser Glu Gln Met

2960 2965 2970

Gln Asp Lys Leu Glu Tyr Leu Glu Glu Lys Ala Ser Phe Lys Thr

2975 2980 2985

Ile Pro Leu Pro Asp Asp Ser Glu Thr Val Ala Cys His Lys Thr

2990 2995 3000

Leu Lys Ser Arg Leu Glu Asp Glu Lys Val Thr Pro Leu Lys Glu

3005 3010 3015

Asn Lys Gln Lys Glu Thr His Lys Thr Lys Glu Glu Ile Ser Thr

3020 3025 3030

Asp Ser Glu Thr Asp Leu Ser Phe Ile Gln Pro Thr Ile Pro Ser

3035 3040 3045

Glu Glu Asp Tyr Phe Glu Lys Tyr Thr Leu Ile Asp Tyr Asn Ile

3050 3055 3060

Ser Pro Asp Pro Glu Lys Gln Lys Ala Pro Gln Lys Leu Asn Val

3065 3070 3075

Glu Glu Lys Leu Ser Lys Glu Val Thr Glu Glu Thr Ile Ser Phe

3080 3085 3090

Pro Val Ser Ser Val Glu Ser Ala Leu Glu His Glu Tyr Asp Leu

3095 3100 3105

Val Lys Leu Asp Glu Ser Phe Tyr Gly Pro Glu Lys Gly His Asn

3110 3115 3120

Ile Leu Ser His Pro Glu Thr Gln Ser Gln Asn Ser Ala Asp Arg

3125 3130 3135

Asn Val Ser Lys Asp Thr Lys Arg Asp Val Asp Ser Lys Ser Pro

3140 3145 3150

Gly Met Pro Leu Phe Glu Ala Glu Glu Gly Val Leu Ser Arg Thr

3155 3160 3165

Gln Ile Phe Pro Thr Thr Ile Lys Val Ile Asp Pro Glu Phe Leu

3170 3175 3180

Glu Glu Pro Pro Ala Leu Ala Phe Leu Tyr Lys Asp Leu Tyr Glu

3185 3190 3195

Glu Ala Val Gly Glu Lys Lys Lys Glu Glu Glu Thr Ala Ser Glu

3200 3205 3210

Gly Asp Ser Val Asn Ser Glu Ala Ser Phe Pro Ser Arg Asn Ser

3215 3220 3225

Asp Thr Asp Asp Gly Thr Gly Ile Tyr Phe Glu Lys Tyr Ile Leu

3230 3235 3240

Lys Asp Asp Ile Leu His Asp Thr Ser Leu Thr Gln Lys Asp Gln

3245 3250 3255

Gly Gln Gly Leu Glu Glu Lys Arg Val Gly Lys Asp Asp Ser Tyr

3260 3265 3270

Gln Pro Ile Ala Ala Glu Gly Glu Ile Trp Gly Lys Phe Gly Thr

3275 3280 3285

Ile Cys Arg Glu Lys Ser Leu Glu Glu Gln Lys Gly Val Tyr Gly

3290 3295 3300

Glu Gly Glu Ser Val Asp His Val Glu Thr Val Gly Asn Val Ala

3305 3310 3315

Met Gln Lys Lys Ala Pro Ile Thr Glu Asp Val Arg Val Ala Thr

3320 3325 3330

Gln Lys Ile Ser Tyr Ala Val Pro Phe Glu Asp Thr His His Val

3335 3340 3345

Leu Glu Arg Ala Asp Glu Ala Gly Ser His Gly Asn Glu Val Gly

3350 3355 3360

Asn Ala Ser Pro Glu Val Asn Leu Asn Val Pro Val Gln Val Ser

3365 3370 3375

Phe Pro Glu Glu Glu Phe Ala Ser Gly Ala Thr His Val Gln Glu

3380 3385 3390

Thr Ser Leu Glu Glu Pro Lys Ile Leu Val Pro Pro Glu Pro Ser

3395 3400 3405

Glu Glu Arg Leu Arg Asn Ser Pro Val Gln Asp Glu Tyr Glu Phe

3410 3415 3420

Thr Glu Ser Leu His Asn Glu Val Val Pro Gln Asp Ile Leu Ser

3425 3430 3435

Glu Glu Leu Ser Ser Glu Ser Thr Pro Glu Asp Val Leu Ser Gln

3440 3445 3450

Gly Lys Glu Ser Phe Glu His Ile Ser Glu Asn Glu Phe Ala Ser

3455 3460 3465

Glu Ala Glu Gln Ser Thr Pro Ala Glu Gln Lys Glu Leu Gly Ser

3470 3475 3480

Glu Arg Lys Glu Glu Asp Gln Leu Ser Ser Glu Val Val Thr Glu

3485 3490 3495

Lys Ala Gln Lys Glu Leu Lys Lys Ser Gln Ile Asp Thr Tyr Cys

3500 3505 3510

Tyr Thr Cys Lys Cys Pro Ile Ser Ala Thr Asp Lys Val Phe Gly

3515 3520 3525

Thr His Lys Asp His Glu Val Ser Thr Leu Asp Thr Ala Ile Ser

3530 3535 3540

Ala Val Lys Val Gln Leu Ala Glu Phe Leu Glu Asn Leu Gln Glu

3545 3550 3555

Lys Ser Leu Arg Ile Glu Ala Phe Val Ser Glu Ile Glu Ser Phe

3560 3565 3570

Phe Asn Thr Ile Glu Glu Asn Cys Ser Lys Asn Glu Lys Arg Leu

3575 3580 3585

Glu Glu Gln Asn Glu Glu Met Met Lys Lys Val Leu Ala Gln Tyr

3590 3595 3600

Asp Glu Lys Ala Gln Ser Phe Glu Glu Val Lys Lys Lys Lys Met

3605 3610 3615

Glu Phe Leu His Glu Gln Met Val His Phe Leu Gln Ser Met Asp

3620 3625 3630

Thr Ala Lys Asp Thr Leu Glu Thr Ile Val Arg Glu Ala Glu Glu

3635 3640 3645

Leu Asp Glu Ala Val Phe Leu Thr Ser Phe Glu Glu Ile Asn Glu

3650 3655 3660

Arg Leu Leu Ser Ala Met Glu Ser Thr Ala Ser Leu Glu Lys Met

3665 3670 3675

Pro Ala Ala Phe Ser Leu Phe Glu His Tyr Asp Asp Ser Ser Ala

3680 3685 3690

Arg Ser Asp Gln Met Leu Lys Gln Val Ala Val Pro Gln Pro Pro

3695 3700 3705

Arg Leu Glu Pro Gln Glu Pro Asn Ser Ala Thr Ser Thr Thr Ile

3710 3715 3720

Ala Val Tyr Trp Ser Met Asn Lys Glu Asp Val Ile Asp Ser Phe

3725 3730 3735

Gln Val Tyr Cys Met Glu Glu Pro Gln Asp Asp Gln Glu Val Asn

3740 3745 3750

Glu Leu Val Glu Glu Tyr Arg Leu Thr Val Lys Glu Ser Tyr Cys

3755 3760 3765

Ile Phe Glu Asp Leu Glu Pro Asp Arg Cys Tyr Gln Val Trp Val

3770 3775 3780

Met Ala Val Asn Phe Thr Gly Cys Ser Leu Pro Ser Glu Arg Ala

3785 3790 3795

Ile Phe Arg Thr Ala Pro Ser Thr Pro Val Ile Arg Ala Glu Asp

3800 3805 3810

Cys Thr Val Cys Trp Asn Thr Ala Thr Ile Arg Trp Arg Pro Thr

3815 3820 3825

Thr Pro Glu Ala Thr Glu Thr Tyr Thr Leu Glu Tyr Cys Arg Gln

3830 3835 3840

His Ser Pro Glu Gly Glu Gly Leu Arg Ser Phe Ser Gly Ile Lys

3845 3850 3855

Gly Leu Gln Leu Lys Val Asn Leu Gln Pro Asn Asp Asn Tyr Phe

3860 3865 3870

Phe Tyr Val Arg Ala Ile Asn Ala Phe Gly Thr Ser Glu Gln Ser

3875 3880 3885

Glu Ala Ala Leu Ile Ser Thr Arg Gly Thr Arg Phe Leu Leu Leu

3890 3895 3900

Arg Glu Thr Ala His Pro Ala Leu His Ile Ser Ser Ser Gly Thr

3905 3910 3915

Val Ile Ser Phe Gly Glu Arg Arg Arg Leu Thr Glu Ile Pro Ser

3920 3925 3930

Val Leu Gly Glu Glu Leu Pro Ser Cys Gly Gln His Tyr Trp Glu

3935 3940 3945

Thr Thr Val Thr Asp Cys Pro Ala Tyr Arg Leu Gly Ile Cys Ser

3950 3955 3960

Ser Ser Ala Val Gln Ala Gly Ala Leu Gly Gln Gly Glu Thr Ser

3965 3970 3975

Trp Tyr Met His Cys Ser Glu Pro Gln Arg Tyr Thr Phe Phe Tyr

3980 3985 3990

Ser Gly Ile Val Ser Asp Val His Val Thr Glu Arg Pro Ala Arg

3995 4000 4005

Val Gly Ile Leu Leu Asp Tyr Asn Asn Gln Arg Leu Ile Phe Ile

4010 4015 4020

Asn Ala Glu Ser Glu Gln Leu Leu Phe Ile Ile Arg His Arg Phe

4025 4030 4035

Asn Glu Gly Val His Pro Ala Phe Ala Leu Glu Lys Pro Gly Lys

4040 4045 4050

Cys Thr Leu His Leu Gly Ile Glu Pro Pro Asp Ser Val Arg His

4055 4060 4065

Lys

<210> 166

<211> 1350

<212> DNA

<213> 智人(Homo sapiens)

<400> 166

atggcttctg ctatcacgca gtgttctacc agtgagctca cctgctcgat ctgcacagac 60

tatttgacag accctgtcac catttgttgt gggcacagat tttgtagtcc ctgtctctgc 120

ctcttgtggg aagatacact aactcctaat tgctgccctg tgtgcaggga aatatcacag 180

caaatgtact tcaaacgcat tatttttgct gagaaacaag ttattcctac aagagaatca 240

gtcccctgcc agttatcaag ctctgccatg ctgatctgta ggagacacca ggaaatcaag 300

aacctcatct gtgaaactga taggagcctg ctgtgttttc tatgctctca atccccaagg 360

catgctactc acaaacacta tatgacaagg gaggctgatg aatactatag gaagaaactc 420

ctgattcaaa tgaagtccat ctggaaaaaa aaacagaaaa atcaaagaaa tctaaacaga 480

gagaccaaca taatcggaac atgggaagtt tttataaatt tgcggagcat gatgatcagt 540

gctgaatatc ctaaggtgtg tcaatacctc cgtgaagaag agcaaaagca cgtagagagc 600

ctggcaagag aaggcaggat aatttttcag caactcaaga gaagtcaaac tagaatggct 660

aagatgggta tactcctgag agaaatgtat gagaaactga aggaaatgag ctgtaaagca 720

gatgtgaacc tgcctcagga tttgggagac gtaatgaaaa ggaatgagtt tctgaggctg 780

gccatgcctc agcctgtgaa cccacagctc agtgcatgga ccatcactgg ggtgtcagaa 840

aggcttaact tcttcagagt gtatatcacc ttggatcgta agatatgcag taatcacaag 900

cttctgttcg aagacctaag gcatttgcag tgcagccttg atgatacaga catgtcctgt 960

aatccaacaa gtacacagta tacttcttca tggggagctc agatcctcag ctctggcaaa 1020

cattactggg aggtggatgt gaaagactct tgtaattggg ttataggact ttgcagagaa 1080

gcctggacaa agaggaatga catgcgactt gactctgagg gtatctttct actcctgtgt 1140

ctcaaagtgg atgaccattt cagtctcttc tctacctccc cactgttacc tcactatatt 1200

ccaaggcccc aaggctggct aggggtgttc ctggattatg aatgtgggat agtgagcttt 1260

gttaatgttg cccaaagttc cctcatttgt agtttcctct cacgcatctt ctattttcct 1320

ctcagacctt tcatttgcca cggatctaaa 1350

<210> 167

<211> 450

<212> PRT

<213> 智人(Homo sapiens)

<400> 167

Met Ala Ser Ala Ile Thr Gln Cys Ser Thr Ser Glu Leu Thr Cys Ser

1 5 10 15

Ile Cys Thr Asp Tyr Leu Thr Asp Pro Val Thr Ile Cys Cys Gly His

20 25 30

Arg Phe Cys Ser Pro Cys Leu Cys Leu Leu Trp Glu Asp Thr Leu Thr

35 40 45

Pro Asn Cys Cys Pro Val Cys Arg Glu Ile Ser Gln Gln Met Tyr Phe

50 55 60

Lys Arg Ile Ile Phe Ala Glu Lys Gln Val Ile Pro Thr Arg Glu Ser

65 70 75 80

Val Pro Cys Gln Leu Ser Ser Ser Ala Met Leu Ile Cys Arg Arg His

85 90 95

Gln Glu Ile Lys Asn Leu Ile Cys Glu Thr Asp Arg Ser Leu Leu Cys

100 105 110

Phe Leu Cys Ser Gln Ser Pro Arg His Ala Thr His Lys His Tyr Met

115 120 125

Thr Arg Glu Ala Asp Glu Tyr Tyr Arg Lys Lys Leu Leu Ile Gln Met

130 135 140

Lys Ser Ile Trp Lys Lys Lys Gln Lys Asn Gln Arg Asn Leu Asn Arg

145 150 155 160

Glu Thr Asn Ile Ile Gly Thr Trp Glu Val Phe Ile Asn Leu Arg Ser

165 170 175

Met Met Ile Ser Ala Glu Tyr Pro Lys Val Cys Gln Tyr Leu Arg Glu

180 185 190

Glu Glu Gln Lys His Val Glu Ser Leu Ala Arg Glu Gly Arg Ile Ile

195 200 205

Phe Gln Gln Leu Lys Arg Ser Gln Thr Arg Met Ala Lys Met Gly Ile

210 215 220

Leu Leu Arg Glu Met Tyr Glu Lys Leu Lys Glu Met Ser Cys Lys Ala

225 230 235 240

Asp Val Asn Leu Pro Gln Asp Leu Gly Asp Val Met Lys Arg Asn Glu

245 250 255

Phe Leu Arg Leu Ala Met Pro Gln Pro Val Asn Pro Gln Leu Ser Ala

260 265 270

Trp Thr Ile Thr Gly Val Ser Glu Arg Leu Asn Phe Phe Arg Val Tyr

275 280 285

Ile Thr Leu Asp Arg Lys Ile Cys Ser Asn His Lys Leu Leu Phe Glu

290 295 300

Asp Leu Arg His Leu Gln Cys Ser Leu Asp Asp Thr Asp Met Ser Cys

305 310 315 320

Asn Pro Thr Ser Thr Gln Tyr Thr Ser Ser Trp Gly Ala Gln Ile Leu

325 330 335

Ser Ser Gly Lys His Tyr Trp Glu Val Asp Val Lys Asp Ser Cys Asn

340 345 350

Trp Val Ile Gly Leu Cys Arg Glu Ala Trp Thr Lys Arg Asn Asp Met

355 360 365

Arg Leu Asp Ser Glu Gly Ile Phe Leu Leu Leu Cys Leu Lys Val Asp

370 375 380

Asp His Phe Ser Leu Phe Ser Thr Ser Pro Leu Leu Pro His Tyr Ile

385 390 395 400

Pro Arg Pro Gln Gly Trp Leu Gly Val Phe Leu Asp Tyr Glu Cys Gly

405 410 415

Ile Val Ser Phe Val Asn Val Ala Gln Ser Ser Leu Ile Cys Ser Phe

420 425 430

Leu Ser Arg Ile Phe Tyr Phe Pro Leu Arg Pro Phe Ile Cys His Gly

435 440 445

Ser Lys

450

<210> 168

<211> 68

<212> PRT

<213> 人工序列(Artificial Sequence)

<220>

<221> SIGNAL

<223> 前导序列(Leader Sequence)

<220>

<221> PHOSPHORYLATION

<222> (5)..(11)

<223> 6His标签(6His Tag)

<220>

<221> TRANSMEM

<222> (40)..(51)

<223> TAT肽(TAT peptide)

<220>

<221> PHOSPHORYLATION

<222> (58)..(67)

<223> HA标签(HA tag)

<400> 168

Met Arg Gly Ser His His His His His His Gly Met Ala Ser Met Thr

1 5 10 15

Gly Gly Gln Gln Met Gly Arg Asp Leu Tyr Asp Asp Asp Asp Lys Asp

20 25 30

Arg Trp Gly Ser Lys Leu Gly Tyr Gly Arg Lys Lys Arg Arg Gln Arg

35 40 45

Arg Arg Gly Gly Ser Thr Met Ser Gly Tyr Pro Tyr Asp Val Pro Asp

50 55 60

Tyr Ala Gly Ser

65

<210> 169

<211> 536

<212> PRT

<213> 人工序列(Artificial Sequence)

<220>

<221> SIGNAL

<223> 前导序列和TRIM11(Leader sequence and TRIM11)

<400> 169

Met Arg Gly Ser His His His His His His Gly Met Ala Ser Met Thr

1 5 10 15

Gly Gly Gln Gln Met Gly Arg Asp Leu Tyr Asp Asp Asp Asp Lys Asp

20 25 30

Arg Trp Gly Ser Lys Leu Gly Tyr Gly Arg Lys Lys Arg Arg Gln Arg

35 40 45

Arg Arg Gly Gly Ser Thr Met Ser Gly Tyr Pro Tyr Asp Val Pro Asp

50 55 60

Tyr Ala Gly Ser Met Ala Ala Pro Asp Leu Ser Thr Asn Leu Gln Glu

65 70 75 80

Glu Ala Thr Cys Ala Ile Cys Leu Asp Tyr Phe Thr Asp Pro Val Met

85 90 95

Thr Asp Cys Gly His Asn Phe Cys Arg Glu Cys Ile Arg Arg Cys Trp

100 105 110

Gly Gln Pro Glu Gly Pro Tyr Ala Cys Pro Glu Cys Arg Glu Leu Ser

115 120 125

Pro Gln Arg Asn Leu Arg Pro Asn Arg Pro Leu Ala Lys Met Ala Glu

130 135 140

Met Ala Arg Arg Leu His Pro Pro Ser Pro Val Pro Gln Gly Val Cys

145 150 155 160

Pro Ala His Arg Glu Pro Leu Ala Ala Phe Cys Gly Asp Glu Leu Arg

165 170 175

Leu Leu Cys Ala Ala Cys Glu Arg Ser Gly Glu His Trp Ala His Arg

180 185 190

Val Arg Pro Leu Gln Asp Ala Ala Glu Asp Leu Lys Ala Lys Leu Glu

195 200 205

Lys Ser Leu Glu His Leu Arg Lys Gln Met Gln Asp Ala Leu Leu Phe

210 215 220

Gln Ala Gln Ala Asp Glu Thr Cys Val Leu Trp Gln Lys Met Val Glu

225 230 235 240

Ser Gln Arg Gln Asn Val Leu Gly Glu Phe Glu Arg Leu Arg Arg Leu

245 250 255

Leu Ala Glu Glu Glu Gln Gln Leu Leu Gln Arg Leu Glu Glu Glu Glu

260 265 270

Leu Glu Val Leu Pro Arg Leu Arg Glu Gly Ala Ala His Leu Gly Gln

275 280 285

Gln Ser Ala His Leu Ala Glu Leu Ile Ala Glu Leu Glu Gly Arg Cys

290 295 300

Gln Leu Pro Ala Leu Gly Leu Leu Gln Asp Ile Lys Asp Ala Leu Arg

305 310 315 320

Arg Val Gln Asp Val Lys Leu Gln Pro Pro Glu Val Val Pro Met Glu

325 330 335

Leu Arg Thr Val Cys Arg Val Pro Gly Leu Val Glu Thr Leu Arg Arg

340 345 350

Phe Arg Gly Asp Val Thr Leu Asp Pro Asp Thr Ala Asn Pro Glu Leu

355 360 365

Ile Leu Ser Glu Asp Arg Arg Ser Val Gln Arg Gly Asp Leu Arg Gln

370 375 380

Ala Leu Pro Asp Ser Pro Glu Arg Phe Asp Pro Gly Pro Cys Val Leu

385 390 395 400

Gly Gln Glu Arg Phe Thr Ser Gly Arg His Tyr Trp Glu Val Glu Val

405 410 415

Gly Asp Arg Thr Ser Trp Ala Leu Gly Val Cys Arg Glu Asn Val Asn

420 425 430

Arg Lys Glu Lys Gly Glu Leu Ser Ala Gly Asn Gly Phe Trp Ile Leu

435 440 445

Val Phe Leu Gly Ser Tyr Tyr Asn Ser Ser Glu Arg Ala Leu Ala Pro

450 455 460

Leu Arg Asp Pro Pro Arg Arg Val Gly Ile Phe Leu Asp Tyr Glu Ala

465 470 475 480

Gly His Leu Ser Phe Tyr Ser Ala Thr Asp Gly Ser Leu Leu Phe Ile

485 490 495

Phe Pro Glu Ile Pro Phe Ser Gly Thr Leu Arg Pro Leu Phe Ser Pro

500 505 510

Leu Ser Ser Ser Pro Thr Pro Met Thr Ile Cys Arg Pro Lys Gly Gly

515 520 525

Ser Gly Asp Thr Leu Ala Pro Gln

530 535

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