新的阿洛酮糖-6-磷酸磷酸酶、包括所述酶的用于产生阿洛酮糖的组合物、利用所述酶产生阿洛酮糖的方法与流程

本申请涉及新的阿洛酮糖-6-磷酸磷酸酶、包括所述酶的用于产生d-阿洛酮糖的组合物，以及利用所述酶产生d-阿洛酮糖的方法。
背景技术：
：已知d-阿洛酮糖-3-表异构酶(ec5.1.3.30)和d-塔格糖-3-表异构酶(ec5.1.3.31)是催化d-果糖的3-差向异构化以产生d-阿洛酮糖的酶。当利用该酶通过单一酶促反应产生d-阿洛酮糖时，底物(即d-果糖)与产物(即d-阿洛酮糖)之间的反应平衡以恒定水平(产物/底物＝～20％-35％)存在。因此，高纯度d-阿洛酮糖的产生需要另外的处理以从酶反应产物中分离和去除相对高浓度的d-果糖。另一方面，chan等人.(2008.biochemistry.47:9608-9617)报道了来源于酿脓链球菌(streptococcuspyogenes)的d-核酮糖-5-磷酸-3-表异构酶(ec5.1.3.1)和来源于大肠杆菌(e.coli)的d-阿洛酮糖6-磷酸-3-表异构酶(ec5.1.3.-)，其能够催化d-果糖-6-磷酸和d-阿洛酮糖-6-磷酸的3-差向异构化。然而，由于较差的抗热性，这些酶在工业上不适用。在这样的情况下，本申请的发明人认真进行了研究以开发一种以经济的方式在工业规模上提高d-阿洛酮糖的转化率的方法。结果，本申请的发明人发现，在将作为廉价原料的蔗糖或淀粉(例如麦芽糖糊精)转化为d-阿洛酮糖-6-磷酸后，使用d-阿洛酮糖-6-磷酸特异性的阿洛酮糖-6-磷酸磷酸酶并参与不可逆的反应途径，使得能够通过利用d-阿洛酮糖产生途径中涉及到的两种或更多种酶的一锅法酶促转化来产生d-阿洛酮糖，并且可以显著提高至d-阿洛酮糖的转化率。基于该发现完成了本申请。公开内容技术问题本申请的一个目标是提供包含基序a和基序b的新的阿洛酮糖-6-磷酸磷酸酶。本申请的另一个目标是提供编码本文中所述的阿洛酮糖-6-磷酸磷酸酶的核酸和包含所述核酸的转化子。本申请的另一个目标是提供用于产生d-阿洛酮糖的组合物，其包含肌醇-单磷酸酶、表达肌醇-单磷酸酶的微生物或所述微生物的培养物。本申请的又一个目标是提供用于产生d-阿洛酮糖的方法，其包括使肌醇-单磷酸酶、表达肌醇-单磷酸酶的微生物或所述微生物的培养物与d-阿洛酮糖-6-磷酸接触，以将d-阿洛酮糖-6-磷酸转化为d-阿洛酮糖。技术方案现在将详细描述本申请。同时，本申请中公开的方面和实施方案的说明也可以应用于其他方面和实施方案的说明。另外，本申请中公开的各种要素的所有组合都落入本申请的范围内。此外，不应认为本申请的范围受到以下详细描述的限制。为了达到本申请的上述和其他目标，本申请的一个方面提供了阿洛酮糖-6-磷酸磷酸酶，其包含由xa1-xa2-xa3-dpldg-xa4表示的基序a，其中xa1是w、f、v、i或a，xa2是i、f、v、a或空位，xa3是v、i或l，且xa4是t或s；以及由ya1-d-ya2-wa1-ya3-wa2-ya4-wa3表示的基序b，其中ya1是w、y、t、l或v，ya2是v、i、c、f或a，wa1是aag、aas、sag、apg、apf、agg、apl或aga，ya3是w、i、p、m、v、y、f、r、l、t或s，wa2是llv、liv、lli、lii、ili、fia、alv、iia、vlv、vil、tig、nfc或pif，ya4是e、r、s、t、l、k或p，且wa3是eagg、eggg、eakg、kagg、aagg、yvdg、eaga或rlgv。具体地，在基序a中，xa1可以是w或f，xa2可以是i或v，xa3可以是v或i，且xa4可以是t。具体地，在基序b中，ya1可以是w，ya2可以是v或i，wa1可以是aag，ya3可以是w、i或v，wa2可以是llv、liv、lii或lli，ya4可以是e、r或s，且wa3可以是eagg或eggg。在基序a和基序b之间、在基序a的一端或在基序b的一端可以存在一个或多个氨基酸。可以从已知的肌醇-单磷酸酶的序列(例如，seqidno:1-20中所示的序列中除基序a和基序b外的序列)定义除基序a和基序b外的氨基酸序列。基序a和/或基序b是肌醇-单磷酸酶的序列中的活性位点。基序a和/或基序b也被称为作为酶的底物的肌醇磷酸的结合位点(参见federationofeuropeanbiochemicalsocieties，第294卷，期号1,2、16-18，1991年12月)。本申请的发明人已发现肌醇-单磷酸酶表现出阿洛酮糖-6-磷酸磷酸酶活性。本申请的发明人已发现肌醇-单磷酸酶的基序a和/或基序b可以是磷酸酶的底物结合位点。阿洛酮糖-6-磷酸磷酸酶可以包含例如seqidno:1-20中所示的氨基酸序列中的任一个。本申请的阿洛酮糖-6-磷酸磷酸酶可以是具有已知肌醇-单磷酸酶的功能的酶。本申请的阿洛酮糖-6-磷酸磷酸酶更加选择性地催化d-阿洛酮糖-6-磷酸的去磷酸化，并且可以是d-葡萄糖-1-磷酸、d-葡萄糖-6-磷酸或d-果糖-6-磷酸非特异性的。可以通过以下方式来产生本申请的阿洛酮糖-6-磷酸磷酸酶：用上述酶本身或表达上述酶的dna(例如，seqidno:21-40)转化菌株，培养转化的菌株，破碎培养物，然后纯化。可以通过柱色谱法进行纯化。菌株可以是例如，大肠杆菌(escherichiacoli)、谷氨酸棒杆菌(corynebacteriumglutamicum)、米曲霉(aspergillusoryzae)或枯草芽孢杆菌(bacillussubtilis)。根据本申请的一个实施方案，本申请的阿洛酮糖-6-磷酸磷酸酶可以包括与基序a和/或基序b的序列具有至少90％、至少95％、至少97％、至少99％或100％的同源性、相似性或同一性，并且表现出阿洛酮糖-6-磷酸磷酸酶活性的酶。可选地，本申请的阿洛酮糖-6-磷酸磷酸酶可以是与除基序a和/或基序b外的序列具有至少70％、至少75％、至少80％、至少85％、至少90％、至少95％、至少97％、至少99％或100％的同源性、相似性或同一性的酶。在一个实施方案中，本申请的阿洛酮糖-6-磷酸磷酸酶可以包括由以下序列组成的蛋白质，所述序列与seqidno:1-20中所示的序列中的基序a和/或基序b的序列具有至少90％、至少95％、至少97％、至少99％或100％的同源性、相似性或同一性，并且与seqidno:1-20中所示的序列中除基序a和/或基序b外的氨基酸序列具有至少70％、至少75％、至少80％、至少85％、至少90％、至少95％、至少97％、至少99％或100％的同源性、相似性或同一性，或者由任何两个上述值所限定的范围内的同源性、相似性或同一性。例如，本申请的范围还包括任何这样的氨基酸序列，其具有以上定义的同源性、相似性或同一性，并且表现出与阿洛酮糖-6-磷酸磷酸酶蛋白质，例如，由seqidno:1-20中所示的序列中的任一个组成的蛋白质相当的功效，并且为部分缺失的、修饰的、取代的或添加的。在基序a和基序b的序列的上游和下游添加不相干序列的蛋白质属于本申请的范围内，并且其天然存在的突变或沉默突变不排除在本申请的范围之外，只要它们具有与包含基序a和基序b的阿洛酮糖-6-磷酸磷酸酶相当的活性。特别地，在seqidno:1-20中所示的氨基酸序列中的任一个的上游和下游添加不相干序列的蛋白质，其天然存在的突变和沉默突变不排除在本申请的范围之外，只要其具有与包含基序a和基序b的阿洛酮糖-6-磷酸磷酸酶相当的活性。包含基序a和基序b的蛋白质或包含seqidno:1-20中所示的氨基酸序列的蛋白质也属于本申请的范围内，只要其具有与包含基序a和基序b的阿洛酮糖-6-磷酸磷酸酶相当的活性。本申请的另一个方面提供了编码阿洛酮糖-6-磷酸磷酸酶的核酸。如本文中所用的，术语“核酸”包括dna和rna分子，并且作为核酸的基本单元的核苷酸包括天然的核苷酸以及具有修饰的糖或碱基的类似物(参见scheit,nucleotideanalogs,johnwiley,newyork(1980)；uhlman和peyman,chemicalreviews,90:543-584(1990))。具体地，编码阿洛酮糖-6-磷酸磷酸酶的核酸可以包括包含可以被翻译成基序a和基序b的核苷酸的序列。在一个实施方案中，本申请的核酸可以由seqidno:21-40中所示的核苷酸序列中的任一个组成。更具体地，本申请的核酸可以包括这样的核酸，其与可以翻译成基序a和基序b的核苷酸具有至少90％、至少95％、至少99％或100％的同源性、相似性或同一性，并且在翻译后可以表现出所需的酶活性。具体地，本申请的核酸可以包括与可以被翻译成seqidno:21-40中所示的序列中的基序a和基序b的核苷酸具有至少90％、至少95％、至少99％或100％的同源性、相似性或同一性的核酸。可选地，本申请的核酸可以是与可以翻译成seqidno:21-40中所示的序列中除基序a和基序b外的基序的核苷酸具有至少80％、至少90％、至少95％、至少97％或至少99％的同源性、相似性或同一性的核酸。由于密码子简并性可以翻译成包括基序a和基序b的蛋白质的蛋白质，具体地，由seqidno:1-20中所示的氨基酸序列中的任一个组成的蛋白质，或者可以翻译成与由seqidno:1-20中所示的氨基酸序列中的任一个组成的蛋白质具有同源性、相似性或同一性的蛋白质的多核苷酸也属于本申请的范围内。根据本申请的包含基序a和基序b的酶可以来源于耐热或嗜热菌株。具体地，由seqidno:1-20中所示的氨基酸序列中的任一个组成的酶可以是来源于以下菌株的酶：海洋红嗜热盐菌(rhodothermusmarinus)、莱廷格热袍菌(thermotogalettingae)、红色亚栖热菌(meiothermusruber)、dictyoglomusturgidum、铁还原热棒菌(pyrobaculumferrireducens)、威吉利热厌氧杆菌(thermoanaerobacterwiegelii)、嗜热栖热菌(thermusthermophilus)、海滨嗜热球菌(thermococcuslitoralis)、嗜热脂肪地芽孢杆菌(geobacillusstearothermophilus)、嗜热厌氧绳菌(anaerolineathermophila)、嗜酸热硫化叶菌(sulfolobusacidocaldarius)、thermosulfidibactertakaii、强烈火球菌(pyrococcusfuriosus)、闪烁古生球菌(archaeoglobusfulgidus)、酸热脂环酸芽孢杆菌(alicyclobacillusacidocaldarius)、meiothermussilvanus、meiothermusrufus、台湾亚栖热菌(meiothermustaiwanensis)、meiothermuschliarophilus或meiothermuscerbereus。如本文中所用的，术语“同源性”或“同一性”表示两个给定的氨基酸序列或核苷酸序列之间的相关程度，其可以表示为百分比。术语“同源性”和“同一性”经常可互换使用。通过标准比对算法确定保守的多核苷酸或多肽的序列同源性或同一性，并且可以与所用程序建立的默认空位罚分一起使用。基本上同源或相同的序列通常均沿目标全长多核苷酸或多肽的至少约50％、60％、70％、80％或90％在中等严格性或高严格性下杂交。还考虑了在杂交多核苷酸中包含简并密码子以代替密码子的多核苷酸。可以利用已知的计算机算法如“fasta”程序，采用例如，如pearson等人.(1988),proc.natl.acad.sci.usa85:2444中的默认参数，来确定任何两个多核苷酸或多肽序列是否为同源的、相似的或相同的。可选地，可以利用needleman-wunsch算法(needleman和wunsch,1970,j.mol.biol.48:443-453)，如在emboss包(emboss:theeuropeanmolecularbiologyopensoftwaresuite,rice等人.,2000,trendsgenet.16:276-277)(版本5.0.0或更新的版本)的needle程序中实施的，来确定序列同源性、相似性或同一性。其他的程序包括gcg程序包(devereux,j.,等人,nucleicacidsresearch12:387(1984))、blastp、blastn、fasta(atschul,[s.][f.,][etal,jmolecbiol215]:403(1990)；guidetohugecomputers,martinj.bishop,[ed.,]academicpress,sandiego,1994，和[carillo等人.](1988)siamjappliedmath48:1073)。例如，可以使用ncbi数据库的blast工具来确定同源性、相似性或同一性。也可以使用其他可商购获得的或公开获得的程序如clustalw。可以例如，通过利用gap计算机程序(例如，needleman等人.,(1970)j.mol.biol.48:443，如由smith和waterman所修订的,adv.appl.math(1981)2:482)比较序列信息，来确定多核苷酸或多肽的百分比同源性、相似性或同一性。简言之，gap程序将相似性定义为相似的比对符号(即核苷酸或氨基酸)的数量除以两条序列中的较短一条中符号的总数。gap程序的默认参数可以包括：(1)一元比较矩阵(包含对于相同的值1和对于不同的值0)，和gribskov等人.(1986)nucl.acidsres.14:6745的加权的比较矩阵，如schwartz和dayhoff编辑,atlasofproteinsequenceandstructure,nationalbiomedicalresearchfoundation,pp.353-358(1979)所描述的；(2)对于每个空位的3.0的罚分，和对于每个空位中的每个符号的另外的0.10的罚分(或者空位开放罚分10.0，空位扩展罚分0.5)；以及(3)对于最后的空位无罚分。因此，如本文中所用的术语“同源性”或“同一性”指示多肽或多核苷酸之间的比较。本申请的另一方面提供了包含编码本文中所述的阿洛酮糖-6-磷酸磷酸酶的核酸的载体，或者包含编码阿洛酮糖-6-磷酸磷酸酶的核酸或包含编码阿洛酮糖-6-磷酸磷酸酶的核酸的载体的转化子。如本文中所用的，术语“载体”是指用于将碱基核酸克隆和/或转移到宿主细胞中的任何媒介物。载体可以是复制子，另一个dna区段可以连接至该复制子，以引起所连接区段的复制。“复制子”是指任何遗传元件(例如质粒、噬菌体、粘粒、染色体或病毒)，其作为体内dna复制的自主单元发挥功能，即能够在其自身的控制下复制。术语“载体”可以包括用于在体外、离体或体内将核酸引入宿主细胞中的病毒和非病毒媒介物。术语“载体”还可以包括微环dna。具体地，包含编码根据本申请的阿洛酮糖-6-磷酸磷酸酶的核酸的载体可以是pet21a-cj_rma、pet21a-cj_tle、pet21a-cj_mrub、pet21a-cj_dtu、pet21a-cj_msi、pet21a-cj_mruf、pet21a-cj_mta、pet21a-cj_mch、pet21a-cj_mce、pbt7-c-his-cj_pfe、pbt7-c-his-cj_twi、pbt7-c-his-cj_tth、pbt7-c-his-cj_tli、pbt7-c-his-cj_gst、pbt7-c-his-cj_ath、pbt7-c-his-cj_sac、pbt7-c-his-cj_tta、pbt7-c-his-cj_pfu、pbt7-c-his-cj_afu或pbt7-c-his-cj_aac。如本文中所用的，术语“转化”是指将包含编码目标蛋白质的核酸的载体引入宿主细胞中，以在宿主细胞中表达核酸编码的蛋白质。转化的核酸可以插入和位于宿主细胞的染色体中，或者可以在染色体外存在，只要其可以在宿主细胞中表达即可。核酸包括编码目标蛋白质的dna和rna。可以以任何形式引入核酸，只要其可以被引入宿主细胞中并在宿主细胞中表达即可。例如，可以将核酸以表达盒(其为包括其自主表达所需的所有元件的基因构建体)的形式引入宿主细胞中，但是其形式不限于此。通常，表达盒包括可操作地连接至核酸的启动子、转录终止信号、核糖体结合域和翻译终止信号。表达盒可以为可自我复制的表达载体的形式。可以将核酸原样引入宿主细胞中，并可操作地连接至在宿主细胞中表达所需的序列。如本文中所用的，术语“可操作地连接”是指起始和介导编码本申请的目标蛋白质的核酸转录的启动子序列与基因序列的之间的功能连接。对于在本申请中提及的氨基酸使用了以下缩写和名称。[表1]丙氨酸a精氨酸r天冬酰胺n天冬氨酸d半胱氨酸c谷氨酸e谷氨酰胺q甘氨酸g组氨酸h异亮氨酸i亮氨酸l赖氨酸k蛋氨酸m苯丙氨酸f脯氨酸p丝氨酸s苏氨酸t色氨酸w酪氨酸y缬氨酸v可以使用将核酸引入细胞中的任何转化方法。可以根据宿主细胞的类型，通过本领域已知的合适的标准技术进行转化方法。这样的转化方法的实例包括但不限于：电穿孔、磷酸钙(capo4)沉淀、氯化钙(cacl2)沉淀、逆转录病毒感染、显微注射、聚乙二醇(peg)方法、deae-葡聚糖方法、阳离子脂质体方法和醋酸锂-dmso方法。宿主细胞优选是高效引入dna且引入的dna以高水平表达的宿主细胞。宿主细胞的实例包括但不限于：属于棒杆菌属(corynebacterium)、埃希氏菌属(escherichia)和沙雷氏菌属(serratia)的微生物的细胞。具体地，宿主细胞可以是大肠杆菌细胞。本申请的转化子可以是：大肠杆菌bl21(de3)/pet21a-cj_rma(大肠杆菌_p1_cj_rma,kccm12057p)、大肠杆菌bl21(de3)/pet21a-cj_tle(大肠杆菌_p2_cj_tle,kccm12058p)、大肠杆菌bl21(de3)/pet21a-cj_mrub(大肠杆菌_p3_cj_mrub,kccm12059p)、大肠杆菌bl21(de3)/pet21a-cj_dtu(大肠杆菌_p4_cj_dtu,kccm12060p)、大肠杆菌bl21(de3)/pbt7-c-his-cj_pfe(大肠杆菌_p5_cj_pfe,kccm12061p)、大肠杆菌bl21(de3)/pbt7-c-his-cj_twi(大肠杆菌_p6_cj_twi,kccm12062p)、大肠杆菌bl21(de3)/pbt7-c-his-cj_tth(大肠杆菌_p7_cj_tth,kccm12063p)、大肠杆菌bl21(de3)/pbt7-c-his-cj_tli(大肠杆菌_p8_cj_tli,kccm12064p)、大肠杆菌bl21(de3)/pbt7-c-his-cj_gst(大肠杆菌_p9_cj_gst,kccm12065p)、大肠杆菌bl21(de3)/pbt7-c-his-cj_ath(大肠杆菌_p10_cj_ath,kccm12066p)、大肠杆菌bl21(de3)/pbt7-c-his-cj_sac(大肠杆菌_p11_cj_sac,kccm12067p)、大肠杆菌bl21(de3)/pbt7-c-his-cj_tta(大肠杆菌_p12_cj_tta,kccm12068p)、大肠杆菌bl21(de3)/pbt7-c-his-cj_pfu(大肠杆菌_p13_cj_pfu,kccm12069p)、大肠杆菌bl21(de3)/pbt7-c-his-cj_afu(大肠杆菌_p14_cj_afu,kccm12070p)、大肠杆菌bl21(de3)/pbt7-c-his-cj_aac(大肠杆菌_p15_cj_aac,kccm12071p)、大肠杆菌bl21(de3)/pet21a-cj_msi(大肠杆菌_p16_cj_msi,kccm12072p)、大肠杆菌bl21(de3)/pet21a-cj_mruf(大肠杆菌_p17_cj_mruf,kccm12073p)、大肠杆菌bl21(de3)/pet21a-cj_mta(大肠杆菌_p18_cj_mta,kccm12074p)、大肠杆菌bl21(de3)/pet21a-cj_mch(大肠杆菌_p19_cj_mch,kccm12075p)或大肠杆菌bl21(de3)/pet21a-cj_mce(大肠杆菌_p20_cj_mce,kccm12076p)。本申请的另一方面提供了用于产生d-阿洛酮糖的组合物，其包含肌醇-单磷酸酶、表达肌醇-单磷酸酶的微生物或表达肌醇-单磷酸酶的微生物的培养物。本申请的肌醇-单磷酸酶可以包含：由xa1-xa2-xa3-dpldg-xa4表示的基序a，其中xa1是w、f、v、i或a，xa2是i、f、v、a或空位，xa3是v、i或l，且xa4是t或s；以及由ya1-d-ya2-wa1-ya3-wa2-ya4-wa3表示的基序b，其中ya1是w、y、t、l或v，ya2是v、i、c、f或a，wa1是aag、aas、sag、apg、apf、agg、apl或aga，ya3是w、i、p、m、v、y、f、r、l、t或s，wa2是llv、liv、lli、lii、ili、fia、alv、iia、vlv、vil、tig、nfc或pif，ya4是e、r、s、t、l、k或p，且wa3是eagg、eggg、eakg、kagg、aagg、yvdg、eaga或rlgv。即，肌醇-单磷酸酶可以与阿洛酮糖-6-磷酸磷酸酶互换使用。因此，对阿洛酮糖-6-磷酸磷酸酶的说明可以适用于肌醇-单磷酸酶。在一个实施方案中，肌醇-单磷酸酶可以是由seqidno:1-20中所示的氨基酸序列中的任一个组成的酶。本申请的组合物还可包含d-阿洛酮糖产生途径(参见图1)中涉及到的酶和/或底物[(i)淀粉、麦芽糖糊精、蔗糖或以上的组合；(ii)磷酸；(iii)d-果糖-6-磷酸-3-表异构酶；(iv)d-葡萄糖-6-磷酸-异构酶；(v)葡萄糖磷酸变位酶或葡萄糖激酶；和/或(vi)α-葡聚糖磷酸化酶、淀粉磷酸化酶、麦芽糖糊精磷酸化酶、蔗糖磷酸化酶、α-淀粉酶、普鲁兰酶、异淀粉酶、葡糖淀粉酶或蔗糖酶]；表达d-阿洛酮糖产生途径中涉及到的酶的微生物；或者表达d-阿洛酮糖产生途径中涉及到的酶的微生物的培养物。另外的酶和底物仅是说明性的，并且不受限制，只要可以利用本申请的阿洛酮糖-6-磷酸磷酸酶产生d-阿洛酮糖即可。淀粉/麦芽糖糊精磷酸化酶(ec2.4.1.1)和α-葡聚糖磷酸化酶可以包括在磷酰基转移至葡萄糖以从淀粉或麦芽糖糊精产生d-葡萄糖-1-磷酸中具有活性的任何蛋白质。具体地，在从淀粉或麦芽糖糊精产生d-葡萄糖-1-磷酸中具有活性的蛋白质可以由seqidno:59中所示的氨基酸序列组成。可以通过seqidno:60中所示的序列编码在从淀粉或麦芽糖糊精产生d-葡萄糖-1-磷酸中具有活性的蛋白质。蔗糖磷酸化酶(ec2.4.1.7)可以包括在磷酰基转移至葡萄糖以从蔗糖产生d-葡萄糖-1-磷酸中具有活性的任何蛋白质。α-淀粉酶(ec3.2.1.1)、普鲁兰酶(ec3.2.1.41)、葡糖淀粉酶(ec3.2.1.3)和异淀粉酶为淀粉液化酶，并且可以包括在将淀粉或麦芽糖糊精转化为葡萄糖中具有活性的任何蛋白质。蔗糖酶(ec3.2.1.26)可以包括在将蔗糖转化为葡萄糖中具有活性的任何蛋白质。葡萄糖磷酸变位酶(ec5.4.2.2)可以包括在将d-葡萄糖-1-磷酸转化为d-葡萄糖-6-磷酸中具有活性的任何蛋白质。具体地，在将d-葡萄糖-1-磷酸转化为d-葡萄糖-6-磷酸中具有活性的蛋白质可以包含seqidno:61中所示的氨基酸序列。可以通过seqidno:62中所示的序列编码在将d-葡萄糖-1-磷酸转化为d-葡萄糖-6-磷酸中具有活性的蛋白质。葡萄糖激酶可以包括在磷酰基转移至葡萄糖以将葡萄糖转化为d-葡萄糖-6-磷酸中具有活性的任何蛋白质。具体地，葡萄糖激酶可以是多聚磷酸依赖型葡萄糖激酶。更具体地，葡萄糖激酶可以是由seqidno:77或78中所示的氨基酸序列组成的蛋白质。d-葡萄糖-6-磷酸-异构酶可以包括在将d-葡萄糖-6-磷酸转化为d-果糖-6-磷酸中具有活性的任何蛋白质。具体地，d-葡萄糖-6-磷酸-异构酶可以是包含seqidno:63中所示的氨基酸序列的蛋白质。可以通过seqidno:64中所示的序列编码d-葡萄糖-6-磷酸-异构酶。d-果糖-6-磷酸-3-表异构酶可以包括在将d-果糖-6-磷酸转化为d-阿洛酮糖-6-磷酸中具有活性的任何蛋白质。具体地，d-果糖-6-磷酸-3-表异构酶可以是包含seqidno:65中所示的氨基酸序列的蛋白质。可以通过seqidno:66中所示的序列编码d-果糖-6-磷酸-3-表异构酶。根据本申请的用于d-阿洛酮糖产生的组合物还可包括在将葡萄糖转化为淀粉、麦芽糖糊精或蔗糖中具有活性的蛋白质。蛋白质可以为例如，4-α-葡聚糖转移酶。在将葡萄糖转化为淀粉、麦芽糖糊精或蔗糖中具有活性的蛋白质可以是包含seqidno:67中所示的氨基酸序列的酶。具体地，可以通过seqidno:68中所示的核苷酸序列编码在将葡萄糖转化为淀粉、麦芽糖糊精或蔗糖中具有活性的蛋白质。根据本申请的用于d-阿洛酮糖产生的组合物还可包括本领域已知的任何合适的赋形剂。这样的赋形剂的实例包括但不限于：防腐剂、湿润剂、分散剂、助悬剂、缓冲剂、稳定剂和等渗剂。根据本申请的用于d-阿洛酮糖产生的组合物还可包括金属离子或金属盐。在一个实施方案中，金属离子可以是二价金属阳离子。具体地，金属离子可以选自：ni、mg、co、mn、fe和zn离子。更具体地，根据本申请的用于d-阿洛酮糖产生的组合物还可包括金属盐。甚至更具体地，金属盐可以选自：niso4、mgso4、mgcl2、nicl2、coso4、cocl2、mncl2、feso4、znso4和以上的混合物。本申请的另一方面提供了用于产生d-阿洛酮糖的方法，其包括使肌醇-单磷酸酶、表达所述酶的微生物或所述微生物的培养物与d-阿洛酮糖-6-磷酸接触，以将d-阿洛酮糖-6-磷酸转化为d-阿洛酮糖。肌醇-单磷酸酶可以包括由xa1-xa2-xa3-dpldg-xa4表示的基序a和由ya1-d-ya2-wa1-ya3-wa2-ya4-wa3表示的基序b。在本文中，肌醇-单磷酸酶可与阿洛酮糖-6-磷酸磷酸酶互换使用。本申请的方法还可包括在将d-阿洛酮糖-6-磷酸转化为d-阿洛酮糖之前，使d-果糖-6-磷酸-3-表异构酶、表达d-果糖-6-磷酸-3-表异构酶的微生物或表达d-果糖-6-磷酸-3-表异构酶的微生物的培养物与d-果糖-6-磷酸接触，以将d-果糖-6-磷酸转化为d-阿洛酮糖-6-磷酸。本申请的方法还可包括在将d-果糖-6-磷酸转化为d-阿洛酮糖-6-磷酸之前，使d-葡萄糖-6-磷酸-异构酶、表达d-葡萄糖-6-磷酸-异构酶的微生物或表达d-葡萄糖-6-磷酸-异构酶的微生物的培养物与d-葡萄糖-6-磷酸接触，以将d-葡萄糖-6-磷酸转化为d-果糖-6-磷酸。本申请的方法还可包括在将d-葡萄糖-6-磷酸转化为d-果糖-6-磷酸之前，使葡萄糖磷酸变位酶、表达葡萄糖磷酸变位酶的微生物或表达葡萄糖磷酸变位酶的微生物的培养物与d-葡萄糖-1-磷酸接触，以将d-葡萄糖-1-磷酸转化为d-葡萄糖-6-磷酸。本申请的方法还可包括在将d-葡萄糖-6-磷酸转化为d-果糖-6-磷酸之前，使葡萄糖激酶、表达葡萄糖激酶的微生物或表达葡萄糖激酶的微生物的培养物及磷酸与葡萄糖接触，以将葡萄糖转化为d-葡萄糖-6-磷酸。本申请的方法还可包括在将d-葡萄糖-1-磷酸转化为d-葡萄糖-6-磷酸之前，使α-葡聚糖磷酸化酶、淀粉磷酸化酶、麦芽糖糊精磷酸化酶或蔗糖磷酸化酶、表达磷酸化酶的微生物或表达磷酸化酶的微生物的培养物及磷酸与淀粉、麦芽糖糊精、蔗糖或以上的组合接触，以将淀粉、麦芽糖糊精、蔗糖或以上的组合转化为d-葡萄糖-1-磷酸。本申请的方法还可包括在将葡萄糖转化为d-葡萄糖-6-磷酸之前，使α-淀粉酶、普鲁兰酶、葡糖淀粉酶、蔗糖酶或异淀粉酶、表达α-淀粉酶、普鲁兰酶、葡糖淀粉酶、蔗糖酶或异淀粉酶的微生物，或者表达α-淀粉酶、普鲁兰酶、葡糖淀粉酶、蔗糖酶或异淀粉酶的微生物的培养物与淀粉、麦芽糖糊精、蔗糖或以上的组合接触，以将淀粉、麦芽糖糊精、蔗糖或以上的组合转化为葡萄糖。本申请的又一个方面提供了用于产生d-阿洛酮糖的方法，其包括使(a)肌醇-单磷酸酶、d-果糖-6-磷酸-3-表异构酶、d-葡萄糖-6-磷酸-异构酶、葡萄糖磷酸变位酶或葡萄糖激酶和α-葡聚糖磷酸化酶、淀粉磷酸化酶、麦芽糖糊精磷酸化酶、蔗糖磷酸化酶、α-淀粉酶、普鲁兰酶、异淀粉酶、葡糖淀粉酶或蔗糖酶，或(b)表达酶(a)的微生物或所述微生物的培养物与淀粉、麦芽糖糊精、蔗糖或以上的组合及磷酸接触。在本申请的方法中，可以在5.0-9.0的ph，具体地6.0-8.0的ph下进行接触反应。在本申请的方法中，可以在40℃-80℃的温度，具体地40℃-60℃或50℃-60℃的温度下进行接触反应。在本申请的方法中，接触反应可以进行2小时至24小时，具体地6-24小时。在本申请的方法中，接触反应可以在5.0-9.0的ph下、在40℃-80℃的温度下进行，和/或进行2小时至24小时。具体地，接触反应可以在6.0-8.0的ph下、在40℃-60℃或50℃-60℃的温度下进行，和/或进行6小时至24小时。本申请的方法还可包括纯化反应产物d-阿洛酮糖。对纯化d-阿洛酮糖的方法没有特别限制。可以通过本领域已知的任何合适的方法纯化d-阿洛酮糖。这样的纯化方法的非限制性实例包括：色谱法、分步结晶和离子纯化，其可以单独或组合进行。例如，可以通过色谱法纯化d-阿洛酮糖。在该情形下，可以基于附着在离子树脂上的糖与金属离子之间的结合力的微小差异来分离目标糖。根据本申请的每种方法还可包括在纯化步骤之前或之后漂白和/或脱矿质。漂白和/或脱矿质使d-阿洛酮糖更纯净而没有杂质。有利效应本申请的新的阿洛酮糖-6-磷酸磷酸酶为耐热的肌醇-单磷酸酶。由于耐热性，本申请的酶可以参与d-阿洛酮糖-6-磷酸转化为d-阿洛酮糖的途径中，使得能够在工业规模上生产d-阿洛酮糖。根据本申请的酶的使用允许进行从作为廉价的原料的葡萄糖或淀粉(例如，麦芽糖糊精)合成d-阿洛酮糖的途径。另外，当使用本申请的酶时，可以通过d-阿洛酮糖-6-磷酸的不可逆的去磷酸化产生d-阿洛酮糖。因此，根据本申请的酶的使用极大地提高了d-阿洛酮糖的转化率。此外，本申请的基于使用肌醇-单磷酸酶的方法能够以高转化率产生高浓度的d-阿洛酮糖，简化或省略了反应产物的分离和纯化。因此，本申请的方法可以以简单的方式进行并且从经济的角度来看是有利的。附图说明图1以图示示出了用于从淀粉(例如，麦芽糖糊精)或葡萄糖产生d-阿洛酮糖的酶促反应途径。图2a、2b和2c显示了用以确定分子量的尺寸标志物(m)、表达的辅酶(s)和纯化的重组酶(e)(pet21a-cj_rma、pet21a-cj_tle、pet21a-cj_mrub、pet21a-cj_dtu、pbt7-c-his-cj_pfe、pbt7-c-his-cj_twi、pbt7-c-his-cj_tth、pbt7-c-his-cj_tli、pbt7-c-his-cj_gst、pbt7-c-his-cj_ath、pbt7-c-his-cj_sac、pbt7-c-his-cj_tta、pbt7-c-his-cj_pfu、pbt7-c-his-cj_afu、pbt7-c-his-cj_aac、pet21a-cj_msi、pet21a-cj_mruf、pet21a-cj_mta、pet21a-cj_mch、pet21a-cj_mce)的sds-page凝胶图像，其在蛋白质电泳后采集。图3a显示了用于去磷酸化的肌醇-单磷酸酶(pet21a-cj_rma、pet21a-cj_tle、pet21a-cj_mrub、pet21a-cj_dtu、pbt7-c-his-cj_pfe、pbt7-c-his-cj_twi、pbt7-c-his-cj_tth、pbt7-c-his-cj_tli、pbt7-c-his-cj_gst、pbt7-c-his-cj_ath、pbt7-c-his-cj_sac、pbt7-c-his-cj_tta、pbt7-c-his-cj_pfu、pbt7-c-his-cj_afu、pbt7-c-his-cj_aac)的相对活性(％，左侧y轴，柱状图)，和肌醇-单磷酸酶在包含d-葡萄糖-6-磷酸、d-葡萄糖-1-磷酸、d-果糖-6-磷酸和d-阿洛酮糖-6-磷酸的去磷酸化混合物中的选择性去磷酸化速率(％，右侧y轴，方形点)。图3b针对去磷酸化比较了肌醇-单磷酸酶(pet21a-cj_mrub、pet21a-cj_msi、pet21a-cj_mruf、pet21a-cj_mta、pet21a-cj_mch、pet21a-cj_mce)的相对活性(％)。图4a显示了hplc色谱图，其证实了通过利用α-葡聚糖磷酸化酶、葡萄糖磷酸变位酶、d-葡萄糖-6-磷酸-异构酶、4-α-葡聚糖转移酶、d-果糖-6-磷酸-3-表异构酶和阿洛酮糖-6-磷酸磷酸酶的多个酶促反应，从麦芽糖糊精产生了d-阿洛酮糖。图4b显示了hplc色谱图，其证实了在根据本申请的肌醇-单磷酸酶存在下，d-阿洛酮糖-6-磷酸在包含d-葡萄糖-1-磷酸、d-葡萄糖-6-磷酸、d-果糖-6-磷酸和d-阿洛酮糖-6-磷酸的反应溶液中通过选择性去磷酸化，产生了d-阿洛酮糖。发明模式将参考以下实施例详细说明本申请。然而，提供这些实施例是为了帮助理解本申请，并不限制本申请的范围。实施例实施例1：肌醇-单磷酸酶的重组表达载体和转化的微生物的产生为了提供d-阿洛酮糖产生途径所必需的阿洛酮糖-6-磷酸磷酸酶，筛选了耐热的肌醇-单磷酸酶基因。具体地，从genbank中登记的以下微生物的基因组序列筛选了肌醇-单磷酸酶基因(rma、tle、mrub、dtu、msi、mruf、mta、mch和mce)：海洋红嗜热盐菌、莱廷格热袍菌、红色亚栖热菌、dictyoglomusturgidum、铁还原热棒菌、威吉利热厌氧杆菌、嗜热栖热菌、海滨嗜热球菌、嗜热脂肪地芽孢杆菌、嗜热厌氧绳菌、嗜酸热硫化叶菌、thermosulfidibactertakaii、强烈火球菌、闪烁古生球菌、酸热脂环酸芽孢杆菌、meiothermussilvanus、meiothermusrufus、台湾亚栖热菌、meiothermuschliarophilus和meiothermuscerbereus。基于筛选的基因的核苷酸序列(seqidno:21、22、23、24、36、37、38、39和40，按基因的顺序)和氨基酸序列(seqidno:1、2、3、4、16、17、18、19和20，按基因的顺序)的信息，设计出了正向引物(seqidno:41、43、45、47、49、51、53、55和57)和反向引物(seqidno:42、44、46、48、50、52、54、56和58)。通过聚合酶链式反应(pcr)，利用合成的引物从海洋红嗜热盐菌、莱廷格热袍菌、红色亚栖热菌、dictyoglomusturgidum、meiothermussilvanus、meiothermusrufus、台湾亚栖热菌、meiothermuschliarophilus和meiothermuscerbereus的基因组dna扩增基因。利用限制酶ndei和xhoi或sa1i，将扩增的肌醇-单磷酸酶基因插入质粒载体pet21a(novagen)中用于大肠杆菌表达，以构建重组表达载体，其分别命名为pet21a-cj_rma(ndei/xhoi)、pet21a-cj_tle(ndei/xhoi)、pet21a-cj_mrub(ndei/xhoi)、pet21a-cj_dtu(ndei/xhoi)、pet21a-cj_msi(ndei/sali)、pet21a-cj_mruf(ndei/sali)、pet21a-cj_mta(ndei/sali)、pet21a-cj_mch(ndei/sali)和pet21a-cj_mce(ndei/sali)。另外，筛选了来源于铁还原热棒菌、威吉利热厌氧杆菌、嗜热栖热菌、海滨嗜热球菌、嗜热脂肪地芽孢杆菌、嗜热厌氧绳菌、嗜酸热硫化叶菌、thermosulfidibactertakaii、强烈火球菌、闪烁古生球菌和酸热脂环酸芽孢杆菌的肌醇-单磷酸酶基因(pfe、twi、tth、tli、gst、ath、sac、tta、pfu、afu和aac)。基于筛选的基因的核苷酸序列(seqidno:25、26、27、28、29、30、31、32、33、34和35，按基因的顺序)和氨基酸序列(seqidno:5、6、7、8、9、10、11、12、13、14和15，按基因的顺序)的信息，请求bioneer(korea)进行了dna合成。将dna插入载体pbt7-c-his(bioneer)中，以构建重组表达载体，其被命名为pbt7-c-his-cj_pfe、pbt7-c-his-cj_twi、pbt7-c-his-cj_tth、pbt7-c-his-cj_tli、pbt7-c-his-cj_gst、pbt7-c-his-cj_ath、pbt7-c-his-cj_sac、pbt7-c-his-cj_tta、pbt7-c-his-cj_pfu、pbt7-c-his-cj_afu和pbt7-c-his-cj_aac。通过一般的转化技术(参见sambrook等人.1989)将表达载体转化至大肠杆菌bl21(de3)菌株中，以产生转化的微生物，其被命名为大肠杆菌bl21(de3)/pet21a-cj_rma(大肠杆菌_p1_cj_rma,kccm12057p)、大肠杆菌bl21(de3)/pet21a-cj_tle(大肠杆菌_p2_cj_tle,kccm12058p)、大肠杆菌bl21(de3)/pet21a-cj_mrub(大肠杆菌_p3_cj_mrub,kccm12059p)、大肠杆菌bl21(de3)/pet21a-cj_dtu(大肠杆菌_p4_cj_dtu,kccm12060p)、大肠杆菌bl21(de3)/pbt7-c-his-cj_pfe(大肠杆菌_p5_cj_pfe,kccm12061p)、大肠杆菌bl21(de3)/pbt7-c-his-cj_twi(大肠杆菌_p6_cj_twi,kccm12062p)、大肠杆菌bl21(de3)/pbt7-c-his-cj_tth(大肠杆菌_p7_cj_tth,kccm12063p)、大肠杆菌bl21(de3)/pbt7-c-his-cj_tli(大肠杆菌_p8_cj_tli,kccm12064p)、大肠杆菌bl21(de3)/pbt7-c-his-cj_gst(大肠杆菌_p9_cj_gst,kccm12065p)、大肠杆菌bl21(de3)/pbt7-c-his-cj_ath(大肠杆菌_p10_cj_ath,kccm12066p)、大肠杆菌bl21(de3)/pbt7-c-his-cj_sac(大肠杆菌_p11_cj_sac,kccm12067p)、大肠杆菌bl21(de3)/pbt7-c-his-cj_tta(大肠杆菌_p12_cj_tta,kccm12068p)、大肠杆菌bl21(de3)/pbt7-c-his-cj_pfu(大肠杆菌_p13_cj_pfu,kccm12069p)、大肠杆菌bl21(de3)/pbt7-c-his-cj_afu(大肠杆菌_p14_cj_afu,kccm12070p)、大肠杆菌bl21(de3)/pbt7-c-his-cj_aac(大肠杆菌_p15_cj_aac,kccm12071p)、大肠杆菌bl21(de3)/pet21a-cj_msi(大肠杆菌_p16_cj_msi,kccm12072p)、大肠杆菌bl21(de3)/pet21a-cj_mruf(大肠杆菌_p17_cj_mruf,kccm12073p)、大肠杆菌bl21(de3)/pet21a-cj_mta(大肠杆菌_p18_cj_mta,kccm12074p)、大肠杆菌bl21(de3)/pet21a-cj_mch(大肠杆菌_p19_cj_mch,kccm12075p)和大肠杆菌bl21(de3)/pet21a-cj_mce(大肠杆菌_p20_cj_mce,kccm12076p)。于2017年7月10日，根据布达佩斯条约(登陆号：kccm12057p-kccm12076p)将转化的菌株保藏在韩国微生物保藏中心(koreanculturecenterofmicroorganisms,kccm)。实施例2：d-阿洛酮糖产生途径所必需的酶的产生为了提供作为d-阿洛酮糖产生途径所必需的耐热酶的来源于新阿波罗栖热袍菌(thermotoganeapolitana)的α-葡聚糖磷酸化酶、葡萄糖磷酸变位酶、d-葡萄糖-6-磷酸-异构酶和d-果糖-6-磷酸-3-表异构酶，筛选了对应于酶的基因(ct1、ct2、tn1和fp3e，按酶的顺序)。基于筛选的基因的核苷酸序列(seqidno:60、62、64和66，按酶的顺序)和氨基酸序列(seqidno:59、61、63和65，按酶的顺序)，设计了正向引物(seqidno:69、71、73和75)和反向引物(seqidno:70、72、74和76)。通过聚合酶链式反应(pcr)，利用引物从作为模板的新阿波罗栖热袍菌的基因组dna扩增酶基因。pcr采用以下条件进行总共25个循环：95℃下变性30sec，55℃下退火30sec，和68℃下聚合2min。利用限制酶ndei和xhoi将扩增的酶基因插入质粒载体pet21a(novagen)中用于大肠杆菌表达，以构建重组表达载体，其被命名为pet21a-cj_ct1、pet21a-cj_ct2、pet21a-cj_tn1和pet21a-cj_fp3e。通过一般的转化技术(参见sambrook等人.1989)将重组表达载体转化至大肠杆菌bl21(de3)菌株中，以产生转化的微生物，其被命名为大肠杆菌bl21(de3)/pet21a-cj_ct1(kccm11990p)、大肠杆菌bl21(de3)/pet21a-cj_ct2(kccm11991p)、大肠杆菌bl21(de3)/pet21a-cj_tn1(kccm11992p)和大肠杆菌bl21(de3)/cj_tn_fp3e(kccm11848p)。于2016年6月23日，根据布达佩斯条约将菌株保藏在韩国微生物保藏中心(kccm)。实施例3：重组酶的产生在本实施例中，产生了重组酶。首先，用实施例1和2中产生的每种转化的微生物接种含有5mllb液体培养基的培养管。将接种物在37℃下于振荡孵化器中培养直至在600nm处的吸光度达到2.0。将培养肉汤添加到培养烧瓶中的lb液体培养基中，然后加入主要培养物。当培养物在600nm处的吸光度达到2.0时，加入1mmiptg以诱导重组酶的表达和产生。培养温度维持在37℃，并以180rpm搅拌。将培养肉汤在8,000×g和4℃下离心20min以收集细菌细胞。将收集的细菌细胞用50mmtris-hcl缓冲液(ph8.0)洗涤两次，并悬浮在相同的缓冲液中。然后，使用超声均质器破碎细胞。将细胞裂解物在13,000×g和4℃下离心20min。通过his标签亲和层析从上清液中纯化重组酶。用50mmtris-hcl缓冲液(ph8.0)对纯化的重组酶进行透析，然后用于后续反应。通过sds-page测定纯化的重组酶的分子量。利用转化的微生物产生的纯化的酶的名称和分子量如下所示(图2a、2b和2c)：从大肠杆菌bl21(de3)/pet21a-cj_rma(大肠杆菌_p1_cj_rma)产生的酶(rma)为30.3kda；从大肠杆菌bl21(de3)/pet21a-cj_tle(大肠杆菌_p2_cj_tle)产生的酶(tle)为28.5kda；从大肠杆菌bl21(de3)/pet21a-cj_mrub(大肠杆菌_p3_cj_mrub)产生的酶(mrub)为28kda；从大肠杆菌bl21(de3)/pet21a-cj_dtu(大肠杆菌_p4_cj_dtu)产生的酶(dtu)为30.2kda；从大肠杆菌bl21(de3)/pbt7-c-his-cj_pfe(大肠杆菌_p5_cj_pfe)产生的酶(pfe)为27.2kda；从大肠杆菌bl21(de3)/pbt7-c-his-cj_twi(大肠杆菌_p6_cj_twi)产生的酶(twi)为28.8kda；从大肠杆菌bl21(de3)/pbt7-c-his-cj_tth(大肠杆菌_p7_cj_tth)产生的酶(tth)为28.6kda；从大肠杆菌bl21(de3)/pbt7-c-his-cj_tli(大肠杆菌_p8_cj_tli)产生的酶(tli)为28kda；从大肠杆菌bl21(de3)/pbt7-c-his-cj_gst(大肠杆菌_p9_cj_gst)产生的酶(gst)为29.6kda；从大肠杆菌bl21(de3)/pbt7-c-his-cj_ath(大肠杆菌_p10_cj_ath)产生的酶(ath)为28.7kda；从大肠杆菌bl21(de3)/pbt7-c-his-cj_sac(大肠杆菌_p11_cj_sac)产生的酶(sac)为29.9kda；从大肠杆菌bl21(de3)/pbt7-c-his-cj_tta(大肠杆菌_p12_cj_tta)产生的酶(tta)为28.6kda；从大肠杆菌bl21(de3)/pbt7-c-his-cj_pfu(大肠杆菌_p13_cj_pfu)产生的酶(pfu)为27.9kda；从大肠杆菌bl21(de3)/pbt7-c-his-cj_afu(大肠杆菌_p14_cj_afu)产生的酶(afu)为28kda；从大肠杆菌bl21(de3)/pbt7-c-his-cj_aac(大肠杆菌_p15_cj_aac)产生的酶(aac)为29kda；从大肠杆菌bl21(de3)/pet21a-cj_msi(大肠杆菌_p16_cj_msi)产生的酶(msi)为28.1kda；从大肠杆菌bl21(de3)/pet21a-cj_mruf(大肠杆菌_p17_cj_mruf)产生的酶(mruf)为28kda；从大肠杆菌bl21(de3)/pet21a-cj_mta(大肠杆菌_p18_cj_mta)产生的酶(mta)为28.1kda；从大肠杆菌bl21(de3)/pet21a-cj_mch(大肠杆菌_p19_cj_mch)产生的酶(mch)为28.4kda；从大肠杆菌bl21(de3)/pet21a-cj_mce(大肠杆菌_p20_cj_mce)产生的酶(mce)为28.1kda；从大肠杆菌bl21(de3)/pet21a-cj_ct1(kccm11990p)产生的酶(ct1)；从大肠杆菌bl21(de3)/pet21a-cj_ct2(kccm11991p)产生的酶(ct2)；从大肠杆菌bl21(de3)/pet21a-cj_tn1(kccm11992p)产生的酶(tn1)；和从大肠杆菌bl21(de3)/cj_tn_fp3e(kccm11848p)产生的酶(fp3e)。实施例4：肌醇-单磷酸酶的活性分析4-1.阿洛酮糖-6-磷酸磷酸酶的活性分析阿洛酮糖-6-磷酸很难购买。因此，发明人直接从d-果糖-6-磷酸产生了d-阿洛酮糖-6-磷酸，并研究了用于d-阿洛酮糖产生的肌醇-单磷酸酶的活性。具体地，将50mmd-果糖-6-磷酸悬浮在50mmtris-hcl(ph7.0)中，然后向其中加入实施例3中产生的d-果糖-6-磷酸-3-表异构酶(fp3e)和0.1单位/ml的20种肌醇-单磷酸酶中的每种。允许混合物在70℃下反应1h。通过hplc(sp_0810柱(shodex)，aminexhpx-87c柱(bio-rad)，80℃，流动相流速0.6ml/min，折光率检测器)证实了d-阿洛酮糖的产生。研究了所有20种肌醇-单磷酸酶对d-阿洛酮糖-6-磷酸的去磷酸化效能(图3a和3b)。4-2.肌醇-单磷酸酶对d-阿洛酮糖-6-磷酸的特异性去磷酸化的活性分析测量了在肌醇-单磷酸酶的存在下，包含d-葡萄糖-6-磷酸、d-葡萄糖-1-磷酸、d-果糖-6-磷酸和d-阿洛酮糖-6-磷酸的混合物中d-阿洛酮糖-6-磷酸的特异性去磷酸化速率。具体地，向1％(w/v)d-葡萄糖-6-磷酸、d-葡萄糖-1-磷酸、d-果糖-6-磷酸和d-阿洛酮糖-6-磷酸的混合物中加入0.1单位/ml的每种肌醇-单磷酸酶和5mmmgcl2。允许反应在50℃下进行12h。通过hplc(aminexhpx-87c柱(bio-rad)，80℃，流动相流速0.6ml/min)分析反应产物。使用折光率检测器检测d-阿洛酮糖和其他糖类(果糖和葡萄糖)的产生。结果，酶mrub显示最高的d-阿洛酮糖-6-磷酸的特异性去磷酸化速率(图3a)。实施例5：通过多个酶促反应分析酶的活性对于从麦芽糖糊精产生d-阿洛酮糖，允许酶ct1、ct2、tn1、fp3e和mrub同时与麦芽糖糊精反应。向0.1单位/ml的每种酶、5mmmgcl2和20mm磷酸钠(ph7.0)中添加5％(w/v)麦芽糖糊精。允许混合物在50℃的温度下反应12h。通过hplc(aminexhpx-87c柱(bio-rad)，80℃，流动相流速0.6ml/min，折光率检测器)分析反应产物。结果，证实了通过多个酶促反应从麦芽糖糊精产生d-阿洛酮糖(图4a)。尽管已经详细描述了本申请的实施方案，但是本领域技术人员将理解，可以以其他特定形式来实现本申请，而不改变本申请的精神或基本特征。因此，应当注意，前述实施方案在所有方面仅是说明性的，并且不应被解释为限制本申请。本申请的范围由所附的权利要求而不是本申请的详细描述来界定。在权利要求的含义和范围内所做的所有改变或修改或其等效形式都应解释为落入本申请的范围内。<110>cj第一制糖株式会社(cjcheiljedangcorporation)<120>新的阿洛酮糖-6-磷酸磷酸酶、包括所述磷酸酶的用于产生阿洛酮糖的组合物和利用所述磷酸酶产生阿洛酮糖的方法<130>p18-6064<150>kr2017/0097334<151>2017-07-31<160>78<170>kopatentin3.0<210>1<211>272<212>prt<213>人工序列(artificialsequence)<220><223>rma的氨基酸序列<400>1metmetaspalatyrthrleutyrgluglualaarggluvalalaala151015argleualaargaspalaglyglnilealaargtyrtyralaglyarg202530valthrvalargglulysglytyrasngluleuvalthrglnalaasp354045glugluvalglnargpheleuilegluglnilehisarghisphepro505560gluhisalaileleualaglugluasnleuseraspmetglnaspgly65707580arggluglyalaserpheargtrpileileaspproileaspglythr859095thrasnphethrhisglyvalproprotyrglyileserleualaleu100105110glnhisgluglyargthrvalvalglyvalvaltyraspvalprohis115120125aspgluleuphethralavalargglyglyglyleutyrvalasngly130135140valargalaargvalserglnthrgluthrleuargglualaleuile145150155160thrthrglypheprotyrarggluvalvalhisleugluglutyrleu165170175glualaleuglyargvalileargalathrargglyvalargargpro180185190glyalaalaservalaspleualatrpvalalacysglyargpheasp195200205glyphephegluthrglyleuserprotrpaspvalalaalaglyile210215220leuleuvalglugluglyglyglyargvalthraspphehisglyarg225230235240proaspproilephealaargglnmetleualathrasnglyargile245250255hisglualaleucysgluleuvalalaproleuhishisvaltyrala260265270<210>2<211>255<212>prt<213>人工序列(artificialsequence)<220><223>tle的氨基酸序列<400>2metaspargmetasppheserilelysilealaarglysvalglyleu151015tyrleumetgluhistrpglyasnalagluasnvalargglnlysser202530serpheglnaspleuvalseraspcysasplysglnalaglnlysmet354045ilevalglnlysilelysasphispheproaspaspalaileleuala505560glugluglyleupheglulysglyaspargmettrpileileasppro65707580ileaspglythrmetasntyrvalhisglyleuproserphealaile859095glyilealatyrvalglulysgluglnvalileleuglyvalalahis100105110aspprovalleuasngluthrtyrtyralailelysglyglnglyala115120125tyrlysasnglygluargileasnvalsergluasnserleuleulys130135140aspserileglyasnthrglyphetyrthraspphethrglyilephe145150155160ileseralaileglulyslysvalargargvalargmetthrglyser165170175alaileleualaglyalatyrvalalacysglyargpheaspphephe180185190ilealalysargalaasnsertrpaspvalalaproleuphevalleu195200205valproglualaglyglyilevalthraspleuserglyasnglnala210215220hisleuasnthrglyasnpheleupheserasnglyleuleuhisasp225230235240glnvalleugluvalilearggluvalasnlyslysvalarglys245250255<210>3<211>258<212>prt<213>人工序列(artificialsequence)<220><223>mrub的氨基酸序列<400>3metaspleuargalatyrleuglnthralaleuaspalaalatyrleu151015alalysglyilehisglntyrtyrglnglulysglyphethrglnser202530thrlysserthrprothraspleuvalthrglnalaasphisgluser354045glualaalailearggluleuilealaserarghisproasphisval505560valleuglyglugluglnglyglnasplysgluglyalapheargtrp65707580ilevalaspproleuaspglythrvalasntyralahisglyphepro859095phetyralavalserileglyleuglualahisglygluvalvalleu100105110glyvalvalleuaspthralaargglygluleuphethralathrlys115120125glyglyglyalatyrleuasnglyargproileargvalserthrarg130135140serthrleuvalglyserleuleualathrglypheprotyraspval145150155160serlysaspthrgluasnleuvaltyrpheglnargalaleuthrlys165170175glyleumetvalargargproglyalaalaalaleuaspleualatyr180185190valalaalaglyargleuaspglyphetrpgluvallysleuasnpro195200205trpaspvalalaalaglytrpleuilevalserglualaglyglyarg210215220ilethrglyleuglnglygluasptyrargleuglyasnargtyrleu225230235240valalaserasnglyleuilehisglyproleuleuaspthrilehis245250255glyarg<210>4<211>265<212>prt<213>人工序列(artificialsequence)<220><223>dtu的氨基酸序列<400>4metlysargileleugluvalalailelysthrilelysglusergly151015asnileleuleuasntyrileglygluglulysgluilegluleulys202530glyileserasnleuvalthrglnvalasplysleusergluarghis354045ileleulysserileglugluasnpheproasphisserileleuthr505560glugluthrglypheileasnlysasnserglutyrthrtrpileval65707580aspproleuaspglythrthrasntyralahisasnpheprophephe859095glyileserilealaleuilelysasnlysgluileileleuglyleu100105110iletyraspproileargaspgluleuphetyralailelysasnglu115120125glyalatyrleuasnaspargargilegluvalserlysthrgluser130135140leugluasnserleuileserphealapheprotyrgluleuserleu145150155160gluglulysasnpheilepropheileasnpheserserargthrhis165170175glyileargargthrglyseralaalailegluilealatyrvalgly180185190cysglyargleuaspglyphetrpalalyslysleulysprotrpasp195200205ileseralaglyileleuilevalgluglualalysglylysvalthr210215220asppheserglyasnasnileaspilehisthraspasnileleuphe225230235240serasnglylysilehisglnglumetilelysileleuasnleugly245250255lysilepheileargasnglulysphe260265<210>5<211>251<212>prt<213>人工序列(artificialsequence)<220><223>pfe的氨基酸序列<400>5metleuglyvalleuglualavalalavalargalaserhispheleu151015metglutyrpheargalaglyargglyvalaspvalvalserarglys202530gluaspaspvalthrarggluvalaspilealaalagluthrleuile354045tyrlysmetleuargglualaphelysgluglyglyvalleutyrala505560glugluglyglyiletyrargtrpglyaspgluargtyrilepheval65707580leuaspproleuaspglyserleuasntyralavalglyvalprophe859095phealavalserilealaalaglylyshisarggluglythrleuala100105110aspleuglutyralavalvalalaileproprothrglyaspvaltyr115120125thralaalaproglyvalglyalaarglysasnglylysproleuarg130135140argthrproargserasnilevalphevalalavalserasnserphe145150155160proprolysthrcysgluvalvalargargleuglyleuargglyarg165170175serleuglyserseralaalagluleualatyrthrvalgluglyile180185190alaargglypheleuaspleuargglylysleuargleuleuaspval195200205alaglyalaleuthrileglylystyrvalaspglypheargtyrval210215220valmetglyaspthrlysprohisserlysvalserleuvalalagly225230235240aspvalaspphevalasnalaalathrthrasp245250<210>6<211>257<212>prt<213>人工序列(artificialsequence)<220><223>twi的氨基酸序列<400>6metlysaspglulysglyilevalvalaspileilelyslysalagly151015gluileleuglnaspglytrpasnlyslysasnphelysiletyrarg202530lysglythrileasnleuvalthrgluileasplyslysilegluphe354045leuileileglnleuleulysglntyrpheproasptyrglyileleu505560thrglugluserlysgluileasnserlysalaasnvalargtrpile65707580ileaspproleuaspglythrthrasntyrilelysglntyrprophe859095valalaileserilealaleuglulysgluglygluleuileleugly100105110valvaltyrasnproileleuasnglumetpheilealaglnlysgly115120125cysglyalathrtyrasnglylysserilehisvalserlysilelys130135140gluleuglyseralavalleualaserglypheprotyraspalatrp145150155160gluasnproaspasnasnalalysglntrpargglnpheleuthrarg165170175serleuserleuargcysaspglyseralaalaleuaspleucysarg180185190valalacysglyglnleuaspglytyrtrpglulysglyileserpro195200205trpaspvalalaalaglyilevalileleuargglualaglyglyile210215220ilethrasptyrleuglyglugluasnphephelysargglygluval225230235240valalaalaasnprovalleuhisalaglnmetleulysvalleuasn245250255asn<210>7<211>264<212>prt<213>人工序列(artificialsequence)<220><223>tth的氨基酸序列<400>7metileglyargarghisprotyrtyrprotyrleuglualaalaleu151015glualaalaserleualaargglyilehisleutyrtyrleuglulys202530glyphethrgluglythrlysserglyprothraspleuvalthrgln354045alaaspargglualagluglualavallysglyleuleuleuserarg505560pheproglualaglypheleuglyglugluglyglysergluglygly65707580lysalaleuargpheilevalaspproleuaspglythrvalasntyr859095alahisglypheprophephealavalserilealaleuglualaglu100105110glyalaileglnmetglyvalvalmetaspthralaargglygluval115120125phetyralaleuargglygluglyalatyrleuasnglyargproile130135140argvalthrglyarggluserleuvalglyserleuleualathrgly145150155160pheprotyraspvalalalysaspprogluasnleuthrtyrpheglu165170175argalaleuglylysglyleuleuvalargargproglyalaalaala180185190leuaspleualatyrvalalaalaglyargleugluglyphetrpglu195200205vallysleuasnprotrpaspvalalaalaglytrpleuleuvalglu210215220glualaglyglyargvalthraspleugluglyasnprotyrargleu225230235240glyserargtyrileleualathrasnglyargvalhisglualaleu245250255argargthrleuleuglyleuasp260<210>8<211>251<212>prt<213>人工序列(artificialsequence)<220><223>tli的氨基酸序列<400>8mettyrglutrpasngluilealaleuasnleualalysaspileglu151015arggluvalmetproleupheglythrlyslysalaglyglupheile202530glypheserproserglyasplysthrlysleuvalasplysvalala354045gluaspvalvalleuglutyrleuargproleuglyvalasnvalval505560serglugluileglyasnileglualaglyserglutyrthrileval65707580valaspproileaspglyserpheasnpheileglnglyileproile859095pheglypheserphealavalphelysasnglulysprovaltyrala100105110metiletyrglupheilethrlysasnvaltyrgluglyileprogly115120125gluglyalatyrleuasnglygluargileargvalarghisleuasn130135140glulysserileserileserphetyrthrargglyargglyalaarg145150155160leuvalglulysvallysargthrargvalleuglyalailealaval165170175gluleualatyrleualaargglyserleuaspglyvalileaspile180185190argasntyrvalargprothraspilealaalaglytyrileileala195200205lysglualaglyalaileilethraspaspserglyglugluilelys210215220pheargleuaspalaargglulysleuasnileilealavalasnasp225230235240lysargleuleulysleuileleugluvalile245250<210>9<211>264<212>prt<213>人工序列(artificialsequence)<220><223>gst的氨基酸序列<400>9metalaglulystrpglugluileaspargtyralaargglntrpile151015aspglualaglylysargileargalaserphealalysglnleuthr202530valglualalysgluasnproasnaspleuvalthrasnvalasparg354045alailegluglnphephealagluhisileargargglnpheproser505560hisargleuleuglyglugluglypheglyaspargileaspalaleu65707580aspglyvalvaltrpvalileaspproileaspglythrmetasnphe859095valhisglnargarghisphealavalserileglyilephegluasp100105110glyileglyglnleuglytyrvaltyraspvalvalpheaspgluleu115120125tyralaalaglnlysglyargglyvalpheleuasnglygluproleu130135140glyleuleuglnproalaprovalalagluserileilealaileasn145150155160glythrtrpleumetgluasnlysargleuasphisargproleumet165170175argleualalysglualaargglythrargsertyrglyseralaala180185190leugluleualatyrvalalaalaglyargleuaspalatyrileser195200205proargleuserprotrpaspphealaglyglymetileleuileglu210215220glualaglyglymetvalthrthrleuaspglylysproleuaspleu225230235240leuglyargasnservalleualaalalysproglyvalhisgluglu245250255ileleuargargtyrleuhisasp260<210>10<211>259<212>prt<213>人工序列(artificialsequence)<220><223>ath的氨基酸序列<400>10metthrthrserleuargaspleuleuaspphealavalgluthrala151015tyrleualaglyargthrthrleualatyrpheglnthrglyvalgln202530alagluphelysalaaspserserprovalthrleualaaspargala354045alagluglnleuileargglnargileglulysargpheprohishis505560alailevalglygluglupheglyvalglnglyseralaaspalathr65707580hisargtrppheileaspproileaspglythrlysserpheleuarg859095glyileproleutyralavalleuleuglyleugluilegluglyarg100105110valglnvalglyvalalatyrtyrproalametaspglumetleuser115120125alaalaaspglygluglycystrptrpasnglyargargalaargval130135140serthralaserargleualaglualatrpvalthrserthrasppro145150155160tyrasnpheglnlysthrglylysaspalaalatrpglnargilegln165170175alavalsertyrhisargglyglytrpglyaspalatyrglytyrleu180185190leuvalalathrglyargalagluvalmetleuaspproilemetasn195200205glutrpaspcysalapropheproproilepheargglualaglygly210215220phepheglyasptrpglnglyasngluthriletyrglyglygluala225230235240leualathrthrglnvalleuleuprogluvalleuglucysleuhis245250255serserleu<210>11<211>267<212>prt<213>人工序列(artificialsequence)<220><223>sac的氨基酸序列<400>11metlysarggluaspvalglulysvalalaserglualaserlystyr151015iletyrglugluarggluasnlysaspvalaspargvalileasnval202530hisglyasnaspvalthrargileileasplysargsergluaspphe354045ilevalaspargleulysserleuglytyrasnileleuilevalthr505560glugluserglyvalileaspsertyrglylysasntyrasptyrile65707580alailevalaspproleuaspglyserthrasnphevalserglyile859095protrpserservalserilealailetyrasnargaspglugluasp100105110ileleuserserasnvalglyalavalserserilephethrprotyr115120125thrphesertyraspgluglyseralatyrvalasnglyvallysile130135140alagluilelyslysproglulysileleuleuleualatyrpheser145150155160argserlysleuproasnleulysleuphepheglulysileglygln165170175glytyrlysileargserleuglyseralaserleuaspmetileleu180185190valcysthrglyargalathrmetphepheaspileargglylysleu195200205argasnvalaspilealaalaserserasnphecysserargleugly210215220valileprotyraspileglyleuarglysilelysserserleuthr225230235240gluvalservalvallysaspleuvalileserleuaspgluserleu245250255leuargserpheserleualaleuglnthrval260265<210>12<211>258<212>prt<213>人工序列(artificialsequence)<220><223>tta的氨基酸序列<400>12metleugluleuleulysaspthralalyslysalaglyargleuleu151015lysileargpheleugluleuleuasnglymetgluthrgluvalarg202530glulysglylysseraspphevalthrargvalaspvalgluvalglu354045asntyrilelysgluleuleualaasnthrasnileservalvalgly505560glugluserphelysglygluileproglythrcysilepheileasp65707580proileaspglythrargasnphemetarglysasnprohispheala859095ileasnleualatyrglnglulysglylysleuleualaglyvalthr100105110tyraspprometlysasnglumetpheseralaalapheserlysgly115120125alapheleuasnglygluargiletyralaserthrasnlysaspile130135140glylysalaileilealaileglyleuprotyrargglyarggluleu145150155160ileaspileglnthrasnleutyrargasnilepheleuasnglyala165170175alathrarghisthrglyseralaalaleuaspleualatyrileser180185190cysglyargtyraspalaalailetyrphetyrleuserprotrpasp195200205valalaproglyileleuleuvalgluglualaglyglygluvalglu210215220glythrmetglyarggluproileglnglytrpileilealaserasn225230235240lysvalilehisprogluvallysaspileleugluglyserleulys245250255alaval<210>13<211>254<212>prt<213>人工序列(artificialsequence)<220><223>pfu的氨基酸序列<400>13metlysleulysphetrparggluvalalaileaspileileserasp151015phegluthrthrilemetprophepheglyasnproaspglyglylys202530leuvallysileserproserglyaspgluthrlysleuvalasplys354045leualagluaspleuileleuserargilethrgluleuglyvalasn505560valvalserglugluvalglyvalileaspasngluserglutyrthr65707580valilevalaspproleuaspglysertyrasnpheilealaglyile859095prophephealaleuserleualavalphelyslysasplysproile100105110tyralaileiletyrgluprometthrgluargphephegluglyile115120125proglygluglyalapheleuasnglylysargilelysvalarglys130135140thrproaspglulysproserileserphetyrserargglylysgly145150155160hisgluilevallyshisvallysargthrargthrleuglyalaile165170175alaleugluleualatyrleualametglyalaleuaspglyvalval180185190aspvalarglystyrvalargprothraspilealaalaglythrile195200205ilealalysglualaglyalaleuilelysaspseralaglylysasp210215220ileaspileserpheasnalathraspargleuaspvalilealaval225230235240asnserglugluleuleulysthrileleuserleuleuglu245250<210>14<211>252<212>prt<213>人工序列(artificialsequence)<220><223>afu的氨基酸序列<400>14metaspgluargaspalaleuargileserarggluilealaglyglu151015valarglysalailealasermetproleuarggluargvallysasp202530valglymetglylysaspglythrprothrlysalaalaaspargval354045alagluaspalaalaleugluileleuarglysgluargvalthrval505560valthrglugluserglyvalleuglygluglyaspvalphevalala65707580leuaspproleuaspglythrpheasnalathrargglyileproval859095tyrservalserleucysphesertyrserasplysleulysaspala100105110phepheglytyrvaltyrasnleualathrglyaspglutyrtyrala115120125aspserserglyalatyrargasnglygluargilegluvalserasp130135140alaglugluleutyrcysasnalaileiletyrtyrproasparglys145150155160pheprophelysargmetargilepheglyseralaalathrgluleu165170175cysphephealaaspglyserpheaspcyspheleuaspileargpro180185190glylysmetleuargiletyraspalaalaalaglyvalpheileala195200205glulysalaglyglylysvalthrgluleuaspglygluserleugly210215220asnlyslyspheaspmetglngluargleuasnilevalalaalaasn225230235240glulysleuhisprolysleuleugluleuilelys245250<210>15<211>265<212>prt<213>人工序列(artificialsequence)<220><223>aac的氨基酸序列<400>15metasplysalatrpleuaspglyleualagluvalvalarggluala151015glyargleuvalglugluilealaargglnglypheaspthrglnphe202530lyshisproglugluargargaspprovalthrthralaaspleuala354045cysaspalapheleulysgluargleuleuthrleuleuprogluala505560glytrpleuserglugluthrlysaspargproaspargleuglulys65707580argtrpvaltrpilevalaspproileaspglythrargglupheval859095argargileproglutyralaileservalalaleualaargaspgly100105110gluprovalalaglyalavalvalasnproalathrglyaspleuphe115120125leuglyalavalglyvalglyalatrpargasnglythrprometval130135140cysserargileargglygluargleuthrileleuglyserargser145150155160glumetasnargglygluphegluprophealaglyileleugluval165170175argalavalglyserilealatyrlysleualaleuvalalaalagly180185190glualaaspglythrpheserleuglyprolyshisglutrpaspile195200205alaalaglyvalalaleuvalleualaalaglyglyargvalhisasp210215220glyalaglyargpropheargpheasnglnprohisthrleuthrarg225230235240glyilevalalaalathrargglualatyrglyaspleualaleuleu245250255ilegluarghisalaproargargala260265<210>16<211>258<212>prt<213>人工序列(artificialsequence)<220><223>msi的氨基酸序列<400>16metaspleuglnargtyrleuglualaalaleuaspalaalatyrleu151015alaargglyilehisleutyrtyrargglulysglyphealaleuglu202530serlysthrserprothraspleuvalthrglnalaasparggluser354045gluglualaileargserleuleuleugluargpheproasphisval505560valleuglyglugluglyglyglngluglyhisserasppheargtrp65707580ilevalaspproleuaspglythrvalasntyralahisglyphepro859095phetyralavalserilealaleugluvalglyglygluvalvalval100105110glyalavalleuaspthralaargglygluleuphethralaleulys115120125glygluglyalapheglnasnglyargproileargvalserserthr130135140alathrleuileglyserleuleualathrglypheprotyraspval145150155160alalysaspalagluasnleuthrtyrpheglnargalaleualalys165170175glyleuthrvalargargproglyalaalaalaleuaspleualatyr180185190valalaalaglyargleugluglyphetrpgluvallysleuasnpro195200205trpaspvalalaalaglytrpleuleuilethrglualaglyglythr210215220valserglyileglnglygluprotyrargleuglyasnargtyrleu225230235240valalaserasnglylysilehisgluglnleuleuaspthrleuhis245250255glyarg<210>17<211>258<212>prt<213>人工序列(artificialsequence)<220><223>mruf的氨基酸序列<400>17metaspleuargprotyrleuglualaalaleuaspalaalatyrleu151015alalysglyilehisglntyrtyrglnglulysglyphethrhisser202530thrlysserthrprothraspvalvalthrglnalaasphisgluala354045glualaalaileargalaleuilealagluargpheproasphisval505560valleuglyglugluglnglyglngluglygluglyglupheargtrp65707580ilevalaspproleuaspglythrvalasntyralahisglyphepro859095phetyrglyvalserileglyleugluvalargglygluvalvalleu100105110glyvalvalleuaspthralaargglyaspleuphethralathrlys115120125glyglyglyalatyrleuasnglyargproleuargvalserglnarg130135140thrthrleuvalglyserleuleualathrglypheprotyraspval145150155160alaargaspprogluasnleuthrtyrpheglnargalaleualalys165170175glyleuthrvalargargproglyalaalaalaleuaspleualaasn180185190valalaalaglyargleuaspglyphetrpgluvallysleuasnpro195200205trpaspvalalaalaglytrpleuleuilethrglualaglyglygln210215220valthrglypheglnglyglualatyrargleuglyasnargtyrleu225230235240valalaserasnglyargilehisglnalaleuleuglualaleugln245250255glyarg<210>18<211>258<212>prt<213>人工序列(artificialsequence)<220><223>mta的氨基酸序列<400>18metaspleuargalatyrleuglnthralaleuaspalaalatyrleu151015alalysglyilehisglntyrtyrglnglulysglyphethrglnser202530serlysserthrprothraspleuvalthrglnalaasphisgluser354045glualaalailearggluleuilealaalaarghisproasphisval505560valleuglyglugluglnglyglnasplysgluglyalapheargtrp65707580ilevalaspproleuaspglythrvalasntyralahisglyphepro859095phetyralavalserileglyleuglualahisglygluvalvalleu100105110glyvalvalleuaspthralaargglygluleuphethralathrlys115120125glyglyglyalatyrleuasnglyargproileargvalserthrarg130135140serthrleuvalglyserleuleualathrglypheprotyraspval145150155160serlysaspthrgluasnleuthrtyrpheglnargalaleuthrlys165170175glyleumetvalargargproglyalaalaalaleuaspleualatyr180185190valalaalaglyargleuaspglyphetrpgluvallysleuasnpro195200205trpaspvalalaalaglytrpleuilevalserglualaglyglyarg210215220valserglyleuglnglygluasptyrargleuglyasnargtyrleu225230235240valalaserasnglyleuilehisglnproleuleuaspthrleuhis245250255glyarg<210>19<211>258<212>prt<213>人工序列(artificialsequence)<220><223>mch的氨基酸序列<400>19metaspleuargpropheleugluthralaleualaalaalatyrval151015alaargglyilehisglnleutyrglnasplysglyphethrglnser202530thrlysserthrprothraspvalvalthrglnalaasplysgluala354045glualaalaileargalaleuilegluglnarghisproglyhisval505560valleuglygluglugluglyglnglnglygluglyglutyrargtrp65707580ilevalaspproleuaspglythrvalasntyralahisglyphepro859095phetyrcysvalserileglyleugluvalargglygluvalmetval100105110glyvalvalleuaspthralaargglyaspleuphethralathrarg115120125glyglyglyalatyrpheasnglyargprometargvalserargser130135140prolysleuleuglyserleuilealathrglypheprotyraspval145150155160alaargasparggluasnleuthrtyrleugluargvalleuphelys165170175glyilethrvalargargproglyalaalaalaleuaspleualatyr180185190valalacysglyargleuaspglyphetrpgluvallysleuasnpro195200205trpaspvalalaalaglytrpleuleuvalgluglualaglyglyarg210215220ilethrglyilehisglygluprotyrargmetglyasnargtyrleu225230235240metalaserasnglyhisilehisglugluleuleualathrilehis245250255glyarg<210>20<211>258<212>prt<213>人工序列(artificialsequence)<220><223>mce的氨基酸序列<400>20metaspleuargvaltyrleuglnthralaleuaspalaalatyrleu151015alalysglyilehisglntyrtyrglnglulysglyphethrleuser202530serlysserthrprothraspleuvalthrglnalaasphisgluser354045glualaalailearggluleuilealaalaarghisproasphisval505560valleuglyglugluglnglyglnasparggluglyalapheargtrp65707580ilevalaspproleuaspglythrvalasntyralahisglyphepro859095phetyralavalserileglyleuglualahisglygluvalvalleu100105110glyvalileleuaspthralaargglyaspleuphethralathrlys115120125glyglyglyalaphemetasnglyargproileargvalserproarg130135140alathrleuvalglyserleuleualathrglypheprotyraspval145150155160serlysaspthrgluasnleuvaltyrpheglnargalaleuthrarg165170175glyleumetvalargargproglyalaalaalaleuaspleualatyr180185190valalaalaglyargleuaspglyphetrpgluvallysleuasnpro195200205trpaspvalalaalaglytrpleuileileargglualaglyglythr210215220valthrglymetgluglyglualatyrargleuglyasnargtyrleu225230235240valalaserasnglyleuilehisglnproleuleuaspthrilehis245250255glyarg<210>21<211>819<212>dna<213>人工序列(artificialsequence)<220><223>rma的核苷酸序列<400>21atgatggatgcctacacgctttacgaagaagcccgtgaggtggcggcccggctggcccgc60gatgccggacagatcgcccgctactatgccggtcgggtgaccgtccgcgaaaaaggctat120aacgagctggtcacgcaggccgacgaagaagtacagcgcttcctgatcgagcagatccac180cggcatttccccgagcacgcgattctggccgaggagaacctgtccgacatgcaggacggc240agggaaggggcgtcgtttcgatggatcatcgatccgatcgacggcacgacgaacttcacg300cacggtgtgccgccctatggcatcagtctggcactccagcatgaaggccggacggtcgtg360ggggtcgtctacgacgtgccccacgacgagctgttcaccgcagtgcggggcggcgggctg420tacgtcaacggggtgcgcgcccgggttagccagaccgaaacgctccgggaggcgctcatc480acgaccggcttcccctaccgggaagtcgtgcatctggaagaatatctggaggcgctcggg540cgtgtgattcgagcgacgcgaggggtgcgtcggccgggcgcggcttcggtcgatctggcc600tgggtggcctgcggacgcttcgacggattcttcgagacgggcctgagtccctgggatgta660gcggccggcatcctgctggtcgaagaaggcggaggacgggtgacggactttcacgggcgg720cccgatccgatctttgcccgccagatgctggccacgaacgggcgcatccacgaggcgctc780tgcgagctggtcgcgccgctgcaccacgtctacgcctga819<210>22<211>768<212>dna<213>人工序列(artificialsequence)<220><223>tle的核苷酸序列<400>22atggacagaatggatttttcaataaagatcgccagaaaagtcggtctctacctgatggaa60cactggggcaatgcagaaaatgttcggcaaaaaagctcttttcaggacctggtaagtgac120tgcgacaagcaagcacagaaaatgattgttcagaagataaaagaccattttccagatgac180gctattttagccgaggaagggctattcgaaaaaggagacagaatgtggataattgaccca240atagatggcacaatgaactacgtccatggtctaccttcttttgcgattggaatcgcctac300gttgaaaaagagcaagttatccttggcgtagcacacgatccagttctcaacgagacatac360tatgcaatcaaagggcagggtgcgtataaaaatggggaaagaataaatgtttcagaaaat420tcgcttttgaaagactctattggaaatactggcttttatacagattttacagggatcttc480atcagcgcaatcgaaaagaaagtcaggcgggtgagaatgacaggcagtgctatacttgct540ggcgcttatgttgcctgtggaaggtttgatttctttatagcaaaaagagccaattcttgg600gatgtagctcctttattcgtcctcgtcccagaagctggtggaatcgtaacggacctgtcc660ggaaaccaggcacatctcaacaccggtaattttcttttcagtaacggtctattacacgat720caggtcttagaagtcataagagaggtgaataaaaaagtaagaaaatga768<210>23<211>777<212>dna<213>人工序列(artificialsequence)<220><223>mrub的核苷酸序列<400>23atggacttgcgtgcctatctgcaaaccgccctcgacgccgcctatttagccaaaggcatc60caccagtactaccaggaaaaaggctttacccagagcaccaagtccacccccaccgatcta120gtcacccaggccgaccacgagtcggaggcggccatccgagaactcattgcttcgcgccac180cccgaccacgtggtgctgggcgaggaacaaggccaggacaaagagggggccttccgctgg240attgtagaccccctggacggcaccgtaaactacgcccatggcttccctttttatgcggtg300agcatcggcctggaagcccacggcgaggtggtgctgggggtggttctggataccgcccgg360ggcgagctatttaccgctaccaaggggggcggggcctacctgaatggtcgtcccattcgg420gtatccacccgttcaaccctggtgggcagccttctagccaccggattcccctacgatgtg480agcaaggacaccgaaaacctggtctactttcagcgggccctgaccaaggggctcatggtg540cggcgaccgggcgcggcagccctcgacctggcttatgtggccgctgggcgcctggacggc600ttctgggaagtgaagctcaacccctgggatgtggccgcgggctggctcatcgttagcgag660gccggagggcggataaccggccttcaaggggaggattaccggctgggcaaccgctaccta720gtggcctccaacggcctgatccacgggccgctgctggacaccatccacgggcgatag777<210>24<211>798<212>dna<213>人工序列(artificialsequence)<220><223>dtu的核苷酸序列<400>24atgaagagaatacttgaagtagctataaaaacaattaaagaatctggtaatatacttttg60aactacataggcgaggaaaaagaaatagaacttaaaggaatttccaatttagtcactcaa120gtagataaactctcagaaagacatatactaaaatccatagaagaaaattttccggatcat180tcaatcttgacagaagaaacaggctttataaataaaaattctgaatatacctggattgta240gatcccttagatggaaccacaaattatgcccataatttccctttttttggaattagtatt300gccctcataaaaaataaagaaataatacttggattaatctatgatcccataagagacgag360ttattttatgccataaaaaatgaaggtgcatacctaaatgataggagaatagaggtatca420aaaacagaaagtcttgaaaattctctcattagttttgcctttccttacgaattaagtctt480gaggagaaaaattttattccttttataaatttttcctctagaactcatggtataagaagg540acaggttcggcagcaatagaaatagcctatgtagggtgtggaagacttgatggattttgg600gcaaagaaactaaaaccatgggatattagtgcaggcattctaatagtggaagaggcaaaa660ggtaaagttacagattttagtggaaacaatattgatattcatacagacaatattttgttt720tcaaacggtaaaatacaccaagagatgataaaaatcttaaatttaggtaaaatattcatt780agaaatgaaaaattttga798<210>25<211>756<212>dna<213>人工序列(artificialsequence)<220><223>pfe的核苷酸序列<400>25atgctcggcgtcttggaggccgtggcggtccgcgcctcgcattttttaatggagtacttc60agagccggcagaggcgtagacgtcgtgtcgcgtaaagaggacgacgtcaccagggaggtg120gacatcgccgcggagacgctcatatacaaaatgctcagagaggcctttaaagaaggcggc180gtcctatacgcagaggaaggcggcatctacaggtggggagacgagcgctacatcttcgtc240ctagacccactcgacgggtcgctgaactacgccgtgggcgtgcccttcttcgccgtatct300atcgcggcagggaaacatagagagggcaccctggccgatttagaatacgccgtcgtggcg360ataccccccacgggagacgtctacacggcggcccccggcgtcggcgcccgcaaaaacgga420aagccgttgaggaggaccccgaggagcaatatagtattcgtagcggtgagcaacagcttc480cccccaaagacctgcgaagtcgtgaggcggctggggctcagggggaggagcctcggcagc540tccgccgccgagctggcgtacacggtggaaggcatagcccgcggcttcctagacctccgg600ggcaagctcagactgctcgacgtggcgggggcattgacgataggcaaatacgtagacggc660ttccggtacgtggtgatgggcgacaccaagccgcactccaaggtgtctctagtggcgggc720gacgtagatttcgtaaatgccgcaaccacagattga756<210>26<211>774<212>dna<213>人工序列(artificialsequence)<220><223>twi的核苷酸序列<400>26atgaaagatgagaaaggaattgtagtagatattataaaaaaagcaggagagattcttcaa60gatgggtggaataaaaaaaattttaaaatttatcggaaaggaacaattaatttagttaca120gaaatagataagaaaattgaatttcttataatacaacttttaaaacaatattttcctgat180tatggtattcttacagaagaaagcaaagaaattaacagcaaagcaaacgtgcgatggatt240atagatcccctagatggtactactaattatataaaacaatatccgtttgtagccatatca300attgctttagagaaagaaggagaattaattttgggagttgtttataatccgattttaaat360gaaatgtttatagctcaaaaaggatgtggtgcgacttataatgggaagtcaattcatgtt420tctaaaataaaagaattgggatcggcagttctagcatcgggctttccttatgatgcgtgg480gaaaatcctgataataatgctaaacaatggaggcaatttcttactcgtagtttatctctt540cgatgcgatggatcagcagctctcgatctttgtagagtagcttgtggacaattggatggt600tattgggaaaagggaattagcccgtgggatgttgcagctggtattgtaatattgcgcgaa660gcaggaggaataattactgattatttgggtgaggaaaatttctttaaacgtggagaagtt720gttgctgcaaatcctgttctccacgcacaaatgttgaaagttttaaataattag774<210>27<211>795<212>dna<213>人工序列(artificialsequence)<220><223>tth的核苷酸序列<400>27atgatcgggaggcgccacccctactacccgtacctggaggcggccctggaggcggcgagc60ctcgcccggggcatccacctctactacctggaaaagggcttcaccgaggggaccaagtcc120ggccccaccgacctggtgacccaggcggaccgggaggcggaagaggcggtgaagggcctc180ctcctctcccgcttccccgaggcggggtttctgggggaggaagggggaagcgagggcggg240aaggccctccgcttcatcgtggacccgctggacgggacggtgaactacgcccacggcttc300cccttcttcgccgtatccatcgccctcgaggccgagggcgccatccagatgggcgtggtg360atggacaccgcccggggggaggtcttctacgccctgaggggggaaggggcctacctcaac420ggccgccccatccgggtcacggggcgggagagcctcgtggggagcctcctcgccacgggc480ttcccctacgacgtggccaaggacccggaaaacctcacctactttgaacgggccctgggc540aagggcctgctcgtccgcaggcctggggcggcggccctggacctggcctacgtggccgcg600gggcggctggagggcttctgggaggtgaagctgaacccctgggacgtggcggcggggtgg660ctcctcgtggaagaggcgggaggaagggtgacggacctcgaggggaacccctaccgcctg720ggcagccgctacatcctcgccaccaacgggcgggtccacgaggccctgcgccggaccctc780ctgggcctggactga795<210>28<211>756<212>dna<213>人工序列(artificialsequence)<220><223>tli的核苷酸序列<400>28atgtatgaatggaatgaaatagccctaaatctcgcaaaggatattgaaagagaagtaatg60cctctctttggaacaaagaaggctggagaattcatcgggtttagcccgagcggggacaaa120actaaactcgtagataaagttgcagaagacgttgttctggagtatctcagaccccttggc180gtaaacgtagtaagtgaggagattgggaacatagaggctggaagtgaatacactattgtt240gtcgaccccatcgatggttctttcaactttatccaaggcatccctatctttggattcagc300tttgcagttttcaaaaatgaaaaacccgtctatgctatgatttacgagttcatcacaaag360aacgtttatgaaggaattcccggtgagggagcctatctaaacggggagagaatacgagtt420aggcatttaaatgaaaagtccatctcaataagcttctacacaagaggtaggggagcgagg480cttgttgaaaaagttaagagaacgagggttttaggtgctatagcagttgagcttgcttat540ctcgctagaggttctctagacggtgtaatagacataaggaactatgtaaggccgacagac600atagcagcaggttacataatcgcaaaagaagctggagctattataaccgacgatagcgga660gaggaaataaagtttagattagatgcaagggaaaagctcaacataatagcggtaaatgat720aagagactccttaagctcatacttgaagttatttaa756<210>29<211>795<212>dna<213>人工序列(artificialsequence)<220><223>gst的核苷酸序列<400>29atggcagaaaaatgggaagaaatcgaccgatacgcccggcaatggatcgacgaagcagga60aagcgcatccgcgcctcgtttgccaaacagctgacggtggaagcgaaagagaacccgaac120gacttagtgacgaatgttgaccgcgccattgagcagtttttcgccgagcacatccgtcgc180cagttccctagccaccgcctgttgggggaagaaggatttggcgaccggatcgatgccttg240gacggcgttgtttgggtgattgacccgatcgacggcacgatgaatttcgttcatcagcgg300cgccattttgctgtatcgatcggcatttttgaagatggcatcgggcagctcggctacgtg360tatgacgtcgtgtttgacgaactgtatgcggcgcaaaaagggcggggggtgtttttgaac420ggggagccgctcggcctcctgcagcccgcgccagtggcggagtcgatcatcgccatcaac480gggacatggctcatggaaaacaagcgcctcgaccatcgcccgctcatgaggctggcgaaa540gaggcgcgcggcacgcgctcgtatggttcagcggcgcttgagctcgcgtatgttgccgcc600ggccgtttggacgcctacatttcgccgcgcctgtcgccgtgggatttcgccggcgggatg660attttgattgaagaagcgggcggaatggtgacgacccttgacgggaagccgctggatctg720cttggccgcaattcggtgcttgccgcgaaacctggggtgcacgaagaaattttgcggcgc780tatcttcacgattga795<210>30<211>780<212>dna<213>人工序列(artificialsequence)<220><223>ath的核苷酸序列<400>30atgaccacttcccttcgcgatttgctcgattttgctgtggaaacggcgtatctggcagga60cgaaccaccctggcgtattttcagacgggtgttcaggcggagttcaaagccgattcttcc120cccgtgactcttgccgaccgcgccgccgagcaattgattcgtcaacgcattgaaaaacgc180ttccctcatcatgccattgtgggtgaggaatttggtgttcagggaagcgctgatgctacc240caccgctggttcattgacccgattgacggcaccaaatccttcctgcgcggcattcccctg300tacgctgtgctgttgggcttggagattgaaggcagggtgcaggtgggagtggcttattac360cccgctatggacgagatgctgtctgccgccgatggcgagggatgctggtggaacgggcgg420cgggcgcgtgtctccacggcgagccgcctggcagaagcctgggtgaccagcaccgacccc480tacaatttccagaaaaccggcaaagatgccgcctggcagcgcattcaggcggtgtcttat540caccgcggcggctggggggatgcctacggctatctgctggttgctactgggagagccgaa600gtgatgctcgatcccatcatgaacgagtgggattgcgcgccgttcccgcccatcttccgc660gaagccggtggttttttcggagactggcagggtaacgaaaccatctatggcggcgaggcg720ctggcaacaactcaggttcttctccctgaggtgctggagtgtttacattcttccctttga780780<210>31<211>804<212>dna<213>人工序列(artificialsequence)<220><223>sac的核苷酸序列<400>31atgaaaagagaagatgtagagaaggtagctagcgaagcttctaaatatatatatgaagag60agagaaaacaaggacgtcgacagagtaattaatgtacatggtaatgacgtgactagaatt120atagataagagatctgaagactttattgtggacaggttaaaaagtctaggatataatata180ctcatagttacagaagaatcaggtgtaattgacagttatgggaaaaattacgattacatt240gcaattgtagatccactggatggaagtacaaattttgtatcaggaataccatggtcttca300gtatctattgctatatacaatagggacgaagaagatatactcagttcaaatgtaggtgct360gtaagcagtattttcactccttacacattttcctacgatgaaggaagcgcttacgtaaat420ggcgtaaagatagcagagataaaaaagcctgaaaaaatactattattagcctatttctcc480aggtcaaagctacctaatctgaaattattttttgagaagattggtcagggttataaaata540agaagtttaggaagtgcatccttagatatgatattagtgtgtacaggcagggcaacaatg600ttttttgatataaggggaaaactgagaaatgtggatattgcagcctcatctaacttctgt660tccagattaggggtaattccatacgacataggtttaagaaaaattaaaagtagtttgacg720gaagtaagtgttgtaaaagatttagttataagtcttgacgaaagtcttctaagatctttc780tctcttgctttacagacggtttaa804<210>32<211>777<212>dna<213>人工序列(artificialsequence)<220><223>tta的核苷酸序列<400>32atgctagaactgctaaaggatacagcaaaaaaagcaggacgactgctaaaaataagattc60ttggaacttttgaacggcatggaaaccgaggtaagagaaaagggaaaaagcgattttgta120acaagggtagatgttgaagtggaaaactacataaaggaactgcttgctaacacaaatata180tcggtggtgggagaagaaagctttaaaggcgagataccgggcacctgcatcttcatagac240cccatagatggcacaagaaacttcatgagaaagaacccccacttcgccattaaccttgct300tatcaggagaagggaaagcttttggctggagtgacctatgaccctatgaagaatgaaatg360ttttctgctgcatttagcaaaggtgcatttttaaacggagagcgcatctacgcttccaca420aacaaggatattggcaaagccatcatcgctataggacttccatacagaggaagagaactc480atagatattcaaaccaacctttacaggaacatcttcctaaacggcgctgctacccgccac540acaggctcagcagcccttgatctggcctacatatcctgcggaagatacgatgctgctatc600tatttttatctttccccctgggacgtagctccgggcatactgctggtagaagaggcaggc660ggagaagttgaaggcaccatgggacgagaacccatccaggggtggataatagcatctaac720aaagtcatccaccccgaagttaaagacatccttgaaggctctttaaaagcggtataa777<210>33<211>765<212>dna<213>人工序列(artificialsequence)<220><223>pfu的核苷酸序列<400>33atgaagcttaagttctggagggaagtagccattgacataatctcagatttcgaaactaca60ataatgcctttctttggaaaccctgatgggggaaagctcgtaaagataagccccagtgga120gatgagacaaagctggttgataagctcgccgaggatttaatactttcaagaatcacagag180cttggagtcaacgttgtgagcgaggaagttggtgttattgataacgagagtgaatacacc240gttatagtagacccactagatggttcatataacttcattgctggaattcctttttttgct300ttgagtctagcagtatttaaaaaagataagccaatatatgcaattatctatgaacctatg360acagagagattctttgagggaatcccaggagagggagcttttttaaatggaaaaagaata420aaggtaagaaaaaccccggacgagaagccatcaataagcttttattctcgaggtaaaggc480catgaaatcgttaaacatgtaaagagaactagaaccttaggggcaatcgctctagaattg540gcatacctagctatgggtgcattagatggagtagttgatgtgaggaaatacgtaaggcca600acggacatagctgctggaacgataattgcaaaagaggcaggagcccttattaaggactcc660gcgggaaaggatatagatatttcatttaatgcaactgataggcttgatgtgatagccgtg720aacagcgaagagttgctaaaaacaattttaagcttactggagtag765<210>34<211>759<212>dna<213>人工序列(artificialsequence)<220><223>afu的核苷酸序列<400>34atggatgaaagggatgcgctgagaatttcgagggagattgcaggagaggtcagaaaggcc60attgcgtcaatgcccttgagggagagagtaaaggacgtgggaatggggaaggatggcact120ccaaccaaggcagccgacagagttgcggaagatgctgcgcttgaaattttgagaaaggag180agggttacggtggttacagaggagtcgggagttttgggggagggagatgtctttgttgcc240ctcgaccccctcgatggaaccttcaatgctacgagagggattccagtttattcagtcagc300ctctgcttctcctattcggataaactgaaagacgccttctttggctacgtttacaacctc360gcaacaggggatgaatactacgcggactccagcggggcttacagaaacggggagaggatt420gaggtgagcgatgctgaggagctttactgcaacgccataatctactatcccgacaggaag480tttccctttaagaggatgaggatttttggaagtgctgcaacggagctttgcttctttgct540gacggctcctttgactgcttcctcgacatccgccccggaaagatgcttagaatctacgat600gccgctgcgggtgtttttattgcagaaaaggcaggaggaaaggttaccgaacttgatgga660gaaagcttggggaataaaaaatttgatatgcaggaaaggctcaatatcgtcgccgcaaat720gaaaaactccatccaaagctgctggagctgattaaatga759<210>35<211>798<212>dna<213>人工序列(artificialsequence)<220><223>aac的核苷酸序列<400>35atggacaaggcatggctcgatggcctggcggaggtcgtgcgggaggctggccgcctggtg60gaggagatcgccaggcagggcttcgatacgcaattcaagcatccggaagagaggcgagat120ccggtcacgacagccgatctcgcctgtgacgcgtttctgaaagagcggctgctcacgctg180cttccggaggcgggctggctgtccgaggagacgaaggatcggccggatcgattggagaaa240aggtgggtgtggatcgtcgatcccatcgacggcacgcgggagttcgtccgccgcattccc300gagtacgccatctcggtggcgctcgcgcgggacggcgagccggtggcgggcgcggtggtg360aatcccgcgacgggggatctgtttctcggcgccgtgggcgtcggcgcatggcggaatgga420acgcccatggtttgttcccgcatccgcggcgagcggctcacgattctcggcagccggtcg480gagatgaaccgcggcgagttcgagccgtttgccggcattctcgaggtccgggcggtgggc540tccatcgcgtataagctcgcgctcgtcgcggcgggcgaggcggatggcacgttcagcctc600ggcccgaaacacgagtgggatatcgcggccggcgtcgcgctcgtcctggcggcgggcggc660cgggtacacgatggcgccgggcgcccgtttcgcttcaaccagccgcacacgctcacccgt720ggcatcgtcgcggccacgcgcgaggcgtatggcgatctcgccctgctcatcgagcggcac780gccccacggcgcgcctag798<210>36<211>777<212>dna<213>人工序列(artificialsequence)<220><223>msi的核苷酸序列<400>36atggacttgcaacgctacttggaagccgcactcgatgctgcgtacttggctcgaggcatc60cacctctactaccgcgaaaaaggcttcgccctggagagcaaaaccagtcccaccgacttg120gtcacccaagccgaccgggagagcgaagaagctatccggagtttactgctcgagcgcttc180cccgatcatgtggtgctgggtgaagaaggtgggcaggaaggccacagcgattttcgctgg240atcgtggacccgctcgacggcacggtcaactacgcccacggcttccccttctacgcggta300agcatcgccctcgaggtggggggcgaggtcgtggtgggggcagtgctggacaccgcacgg360ggcgagctgtttaccgccctcaaaggagagggagcattccaaaatggacgccccattcgc420gtttcgagcaccgcgaccctcatcggaagcctgctggcaaccggcttcccctacgatgtg480gccaaggacgccgagaacctgacctacttccagcgggctcttgccaaaggccttaccgtg540cgccgccccggagccgccgcgctggatttggcttacgtggcggcagggcggctggagggt600ttctgggaggtcaagctcaacccctgggacgtggccgcgggctggctcctcatcaccgag660gcgggcggaaccgtgagcggcatccagggcgagccttaccggctaggaaaccgctacttg720gtagccagcaacgggaagattcatgagcagctcctcgacacgctccacgggcgatga777<210>37<211>777<212>dna<213>人工序列(artificialsequence)<220><223>mruf的核苷酸序列<400>37atggatctgcgtccctacctcgaggccgctctcgatgcggcctacctagccaaaggcatt60caccagtactaccaggaaaaaggcttcacccacagcaccaaatctacccctaccgatgtg120gtcacccaggccgaccacgaggccgaggcagccatccgggctcttatcgccgagcgcttc180cccgaccacgtggtgttaggcgaggagcagggccaggaaggggagggcgagtttcgctgg240attgtagaccccttggatggcacggtgaactacgcccacggctttcccttctatggggtg300agcatcgggctggaggtacgaggagaggtggtgttgggggtggtgctggataccgcccgg360ggcgatctcttcaccgctaccaaaggcggcggggcctacctgaatggccgcccgctgcgg420gtttcccagcgaaccactttggtgggcagtctgctggctaccggttttccctacgatgtg480gcacgagacccggagaacctcacctacttccagcgggctttggccaaggggctgaccgtg540cggcggcccggggcagcggccctcgacctggccaatgtggccgcgggccggctggacggt600ttctgggaggttaagctcaacccctgggatgtggctgcgggctggctcctcatcaccgag660gcaggcggccaggtcaccgggttccagggcgaggcctaccgcctgggtaaccgctattta720gtggcctccaatgggcggattcaccaggcgctgctcgaggcccttcagggccgatag777<210>38<211>777<212>dna<213>人工序列(artificialsequence)<220><223>mta的核苷酸序列<400>38atggacttgcgtgcctacctgcaaaccgccctcgacgccgcctatttagccaaaggcatc60caccagtactaccaggaaaaaggctttacccagagcagcaagtccacccccaccgatctg120gtcacccaggccgaccacgagtcggaggcggccatccgagaactcatcgccgcgcgccac180cccgaccacgtggtgctgggcgaggagcagggccaggacaaagaaggggccttccgctgg240attgtagaccccctggacggcaccgtaaactacgcccatggttttcctttttatgcggtg300agcattggcctggaagcccacggcgaggtggtgctgggggtggttctggataccgctcga360ggcgagctgtttaccgctaccaaagggggtggggcttacctgaacggtcgtcccatccgg420gtttccactcgttcaaccctggtgggcagcctcctggccaccggatttccctacgacgta480agcaaggataccgaaaacctgacctacttccagcgggcattgaccaaagggctcatggta540cggcgccccggcgcggcggccctcgacctggcctatgtggccgccgggcgcctggacggc600ttctgggaggtgaagctgaacccctgggacgtggccgcgggctggctcatcgtcagtgag660gcgggggggcgggtaagcggccttcagggcgaggattaccggctgggcaaccgctacctg720gtggcctccaacggcctaattcatcagcccttgctggacaccctccacgggcgatag777<210>39<211>777<212>dna<213>人工序列(artificialsequence)<220><223>mch的核苷酸序列<400>39atggacctgcgccccttcctcgagaccgcgctcgccgcggcttacgtggctcgaggcatc60caccagctctaccaggacaaaggcttcacccagagcaccaagtccacccccaccgacgtc120gtgacccaagccgacaaggaggccgaggccgccatccgcgccctcatcgagcagcggcac180cccggccacgtggtgctcggcgaggaggagggccagcagggcgagggcgagtaccgctgg240atcgtggacccgctcgacggcaccgtgaattacgcccacggcttccccttctactgcgtg300agcatcgggctcgaggtgcgcggcgaggtgatggtgggcgtggtgctcgacaccgcccgc360ggcgacctcttcaccgcgacccgcggcggcggggcctacttcaacgggcggcccatgcgg420gtctcgcggtcgcccaagctgctgggcagcctcatcgccaccggcttcccctacgacgtc480gcccgcgaccgcgaaaacctgacctacctcgagcgcgtcctcttcaagggcatcaccgtg540cgccgccccggcgccgccgccctcgacctcgcctacgtggcctgcgggcggctcgacggc600ttctgggaggtcaagctcaacccctgggacgtggcggcgggctggctgctggtcgaggag660gcgggcggcaggatcaccggcatccacggcgagccctaccgcatgggcaaccgctacctc720atggcctccaacggccacatccacgaggaattgctggcgacgatccacggccggtag777<210>40<211>777<212>dna<213>人工序列(artificialsequence)<220><223>mce的核苷酸序列<400>40atggacttgcgtgtatacctgcagaccgccctcgatgcagcctatctggccaaaggcatt60caccagtactaccaggagaagggttttaccctaagcagcaagtccacccccacagacctg120gttactcaggccgaccacgagtcggaagcggccatccgagaactcattgccgcgcgccac180cccgaccacgtggtgctgggagaggagcagggccaggacagagaaggggcttttcgctgg240attgtagaccccctggatggcaccgtaaactacgcccacggttttcccttttatgcggtg300agtatcggcctcgaggcccacggcgaggtggtgctgggagtgattctcgataccgcccgg360ggcgacctattcaccgctaccaaggggggcggggccttcatgaatggccgccccatccgg420gtttcccctcgagccaccctggtgggaagcctgctggccaccggcttcccctatgacgtt480agcaaagacaccgaaaacctggtctacttccagcgggccctgaccagagggcttatggtg540cggcgtcccggcgctgccgccctcgacctggcctacgtggccgccgggcgtctggatggt600ttctgggaggtgaagctcaacccctgggatgtggccgcaggctggctcatcatccgcgag660gccgggggtacggttacgggcatggagggggaggcctaccggctgggcaaccgctacctg720gtagcctccaacggcctgattcaccagcctctgctggacaccatacacgggcgatag777<210>41<211>26<212>dna<213>人工序列(artificialsequence)<220><223>rma的正向引物dna序列<400>41ctccgccatatgatggatgcctacac26<210>42<211>25<212>dna<213>人工序列(artificialsequence)<220><223>rma的反向引物dna序列<400>42gccacctcgagggcgtagacgtggt25<210>43<211>33<212>dna<213>人工序列(artificialsequence)<220><223>tle的正向引物dna序列<400>43gtgcaaatcatatggacagaatggatttttcaa33<210>44<211>34<212>dna<213>人工序列(artificialsequence)<220><223>tle的反向引物dna序列<400>44gatattctcgagttttcttacttttttattcacc34<210>45<211>27<212>dna<213>人工序列(artificialsequence)<220><223>mrub的正向引物dna序列<400>45tgaccccatatggacttgcgtgcctat27<210>46<211>25<212>dna<213>人工序列(artificialsequence)<220><223>mrub的反向引物dna序列<400>46ttctgcctcgagtcgcccgtggatg25<210>47<211>34<212>dna<213>人工序列(artificialsequence)<220><223>dtu的正向引物dna序列<400>47taagctcatatgaagagaatacttgaagtagcta34<210>48<211>34<212>dna<213>人工序列(artificialsequence)<220><223>dtu的反向引物dna序列<400>48acaaaagactcgagaaatttttcatttctaatga34<210>49<211>34<212>dna<213>人工序列(artificialsequence)<220><223>msi的正向引物dna序列<400>49atatacatatggacttgcaacgctacttggaagc34<210>50<211>31<212>dna<213>人工序列(artificialsequence)<220><223>msi的反向引物dna序列<400>50tggtggtcgactcgcccgtggagcgtgtcga31<210>51<211>34<212>dna<213>人工序列(artificialsequence)<220><223>mruf的正向引物dna序列<400>51atatacatatggatctgcgtccctacctcgaggc34<210>52<211>30<212>dna<213>人工序列(artificialsequence)<220><223>mruf的反向引物dna序列<400>52tggtggtcgactcggccctgaagggcctcg30<210>53<211>34<212>dna<213>人工序列(artificialsequence)<220><223>mta的正向引物dna序列<400>53atatacatatggacttgcgtgcctacctgcaaac34<210>54<211>31<212>dna<213>人工序列(artificialsequence)<220><223>mta的反向引物dna序列<400>54tggtggtcgactcgcccgtggagggtgtcca31<210>55<211>34<212>dna<213>人工序列(artificialsequence)<220><223>mch的正向引物dna序列<400>55atatacatatggacctgcgccccttcctcgagac34<210>56<211>31<212>dna<213>人工序列(artificialsequence)<220><223>mch的反向引物dna序列<400>56tggtggtcgacccggccgtggatcgtcgcca31<210>57<211>34<212>dna<213>人工序列(artificialsequence)<220><223>mce的正向引物dna序列<400>57atatacatatggacttgcgtgtatacctgcagac34<210>58<211>31<212>dna<213>人工序列(artificialsequence)<220><223>mce的反向引物dna序列<400>58tggtggtcgactcgcccgtgtatggtgtcca31<210>59<211>823<212>prt<213>人工序列(artificialsequence)<220><223>ct1的氨基酸序列<400>59metleulyslysleuprogluasnleugluhisleuglugluleuala151015tyrasnleutrptrpsertrpserargproalaglnargleutrparg202530lysileaspprogluglytrpglugluhisargasnprovallysile354045leulysgluvalseraspgluargleuglugluleuserlysaspasp505560asppheileserleutyrgluleuthrilegluargphelysasptyr65707580metglulysgluaspthrtrppheasnvalasntyrproglutrpasp859095glulysilevaltyrmetcysmetglutyrglyleuthrlysalaleu100105110proiletyrserglyglyleuglyileleualaglyasphisleulys115120125seralaseraspleuglyleuproleuilealaileglyleuleutyr130135140lyshisglytyrphethrglnglnileaspargaspglylysglnile145150155160gluilepheproasptyrasnprogluaspleuprometlysproleu165170175lysaspglulysglyasnglnvalilevalgluvalproleuaspser180185190thrvalvallysalaargvalphegluvallysvalglyargvalser195200205leutyrleuleuaspproaspileglugluasnglugluargtyrarg210215220lysilecysasntyrleutyrasnprogluproaspvalargvalser225230235240glngluileleuleuglyileglyglymetlysleuleuargalaleu245250255asnleulysproglyvalilehisleuasngluglyhisproalaphe260265270serserleugluargilelysasntyrmetglugluglytyrserphe275280285thrglualaleugluilevalargglnthrservalphethrthrhis290295300thrprovalproalaglyhisaspargphepropheaspleuvalglu305310315320lyslysleuserlysphephegluglypheglulysargasnleuleu325330335metaspleuglylysaspgluthrglyserpheasnmetthrtyrleu340345350alaleuargthrserserpheileasnglyvalserlysleuhisala355360365gluvalserargargmetphelysasnvaltrpglnglyvalproval370375380glugluileproilegluglyilethrasnglyvalhismetglythr385390395400trpileasnargglumetarglysleutyraspargtyrleuglyarg405410415valtrpargasphisthraspleugluglyiletrptyrglyvalasp420425430argileproaspglugluleutrpglnalahisleuargalalyslys435440445argpheileglutyrilelysgluservalargargargasngluarg450455460leuglyileaspgluaspvalproasnileaspgluasnserleuile465470475480ileglyphealaargargphealathrtyrlysargalavalleuleu485490495leuseraspleugluargleulyslysileleuasnaspprogluarg500505510provaltyrvalvaltyralaglylysalahisproargaspaspala515520525glylysglupheleulysargiletyrgluvalserglnmetproglu530535540phelysasnargileilevalleugluasntyraspileglymetala545550555560argleumetvalserglyvalaspvaltrpleuasnasnproargarg565570575prometglualaserglythrserglymetlysalaalaalaasngly580585590valleuasnalaservaltyraspglytrptrpvalgluglytyrasn595600605glyargasnglytrpvalileglyaspgluservalleuprogluthr610615620gluvalaspaspproargaspalaglualaleutyraspleuleuglu625630635640asngluileileprothrtyrtyrgluasnlysglulystrpilephe645650655metmetlysgluserilelysservalalaproargpheserthrthr660665670argmetleulysglutyrthrglulysphetyrilelysglyleuval675680685asnlysglutrpleugluarglysgluasnalagluargpheglyala690695700trplysgluargileleuargasntrpserservalserilegluarg705710715720ilevalleugluaspthrargservalgluvalthrvallysleugly725730735aspleuserproaspaspvalleuvalgluleuleuileglyarggly740745750glusermetgluaspleugluiletrplysvalileglnilearglys755760765hisargarggluglyaspleupheiletyrsertyrvalasnglyala770775780leuglyhisleuglyserproglytrpphetyralavalargvalleu785790795800protyrhisprolysleuprothrargpheleuprogluileproval805810815valtrplyslysvalleugly820<210>60<211>2472<212>dna<213>人工序列(artificialsequence)<220><223>ct1的核苷酸序列<400>60atgctgaagaaactcccggagaatctggagcatctggaagaactcgcctacaacctctgg60tggagctggtctaggcccgctcagagactctggagaaagatagatccggaaggctgggag120gaacacagaaaccccgttaaaatactgaaagaagtttctgatgaaaggctcgaagaactt180tcaaaagatgatgatttcatatccctctacgaactcaccattgaaaggttcaaggattac240atggagaaagaagacacctggttcaacgtgaactaccccgaatgggacgagaagatcgtc300tacatgtgtatggagtacggtttgaccaaagcccttccgatctactccggtggtcttgga360atcctcgcgggagaccatctcaaatccgcaagcgatcttggacttcctctcatagcgatc420ggacttctctacaaacatggatatttcacccagcagatcgacagagatggaaaacagata480gagattttccctgattacaacccagaggacttacccatgaagcccctgaaggatgaaaag540ggaaaccaggtgatcgtggaggttcctctcgacagtaccgtggtgaaggcacgtgttttt600gaagtgaaggtaggaagggtgagtctgtacctgctcgatccggacatcgaggaaaacgag660gaacgatacagaaagatctgcaactacctttacaacccggaacccgatgtgagggtctcc720caggagatactcctcggaattgggggaatgaagcttctcagggctctgaacctgaaacca780ggagtcatccatctgaacgaaggacatccggcgttctcttccctcgaaaggataaagaac840tacatggaagaaggatattccttcacagaggcccttgagatcgtgagacagacgagtgtg900tttacaacccacacacccgttcccgctggacacgacagatttccctttgacctcgtggaa960aagaaactttcgaaattcttcgaaggattcgaaaagagaaatcttctcatggatcttggg1020aaagatgaaacaggcagtttcaacatgacgtatcttgccctgagaacgtcctctttcata1080aacggcgtgagcaaactgcatgcggaagtttccagaaggatgttcaaaaacgtgtggcag1140ggtgttcccgtggaggaaataccgatcgaagggataacgaacggcgttcacatgggaacc1200tggatcaaccgtgagatgagaaaactgtacgacagatatctcggaagggtatggagagat1260cacaccgaccttgagggtatctggtacggtgttgacaggattccagatgaagaactctgg1320caggctcacctgagggcaaagaagagattcatcgagtacataaaagaatcggtaagaaga1380agaaacgagagactgggaatcgacgaagatgtgccgaacatcgatgaaaattcgctcatc1440ataggttttgcaagaaggtttgccacttacaagagggcagttctcctgctcagcgatctg1500gagagactcaagaagatcctcaacgatccagaaagacccgtttacgtggtctatgcgggg1560aaggcccatccaagggacgatgcggggaaggaatttttgaaacgcatctacgaagtctcg1620cagatgcctgagttcaaaaacaggatcatcgtactggaaaactacgacattggaatggca1680cggctcatggtgtcgggagtggatgtgtggctgaacaacccgagaagacccatggaagca1740agtggaacaagcggaatgaaggcagcagccaacggagttcttaacgcgagtgtttacgat1800ggatggtgggttgaagggtacaacggcagaaacggctgggtcataggcgatgaaagcgtt1860cttccagagacggaagtggacgatcccagggacgcagaagcactctacgatctcctcgaa1920aacgaaatcatcccaacctactacgaaaacaaagaaaagtggatcttcatgatgaaagag1980agcataaagagtgttgctccaagattcagcaccaccagaatgctcaaagaatacacggag2040aagttctacataaagggacttgtgaacaaagaatggcttgaaagaaaagaaaacgccgaa2100aggtttggtgcatggaaggaaaggatcctcagaaactggagcagcgtttccatagaaaga2160atcgtccttgaggacacaaggagtgttgaggtgacggtgaaactgggagacctttcacct2220gatgatgtactggttgaacttttgattggaagaggagaaagcatggaagatctggagatc2280tggaaggtgatacagataagaaagcacagaagggaaggggatctgttcatctacagttat2340gtcaacggtgccctcggtcatcttggctctccgggatggttctacgcggtgagggtgcta2400ccttatcatccgaaacttcccaccagattcttgccggagatacctgtggtgtggaaaaag2460gttctcgggtga2472<210>61<211>441<212>prt<213>人工序列(artificialsequence)<220><223>ct2的氨基酸序列<400>61metileglytyrglniletyrvalargserpheargaspglyasnphe151015aspglyvalglyaspphelysglyleulysglyalailesertyrleu202530lysgluleuglyvalaspphevaltrpleumetprovalpheserser354045ileserphehisglytyraspvalvalaspphetyrserphelysala505560glutyrglyaspglulysasppheargglumetileglualaphehis65707580aspasnglyilelysvalvalleuaspleuproilehishisthrgly859095pheleuhisvaltrppheglnlysalaleulysglyaspprohistyr100105110argasptyrtyrvaltrpalaserglulysthraspleuaspgluarg115120125argglutrpaspasngluargiletrphisproleugluaspglyarg130135140phetyrargglyleupheglyproleuserproaspleuasntyrasp145150155160asnproglnvalphegluglumetlyslysvalvaltyrhisleuleu165170175glumetglyvalaspglypheargpheaspalaalalyshismetarg180185190aspthrleugluglnasnvalargphetrpargtyrpheleuserasp195200205ilegluglyilepheleualagluiletrpalagluserlysvalval210215220aspgluhisglyargilepheglytyrmetleuasnpheaspthrser225230235240hiscysilelysglualavaltrplysgluasnphelysvalleuile245250255gluserilegluargalaleuvalglylysasptyrleuprovalasn260265270phethrserasnhisaspmetserargleualaserphegluglygly275280285leusergluglulysvallysleuserleuserileleuphethrleu290295300proglyvalproleuilephetyrglyaspgluleuglymetlysgly305310315320iletyrarglysproasnthrgluvalvalleuasppropheprotrp325330335sergluasnmetcysvalgluglyglnthrphetrplystrpproala340345350tyrasnasppropheserglyvalservalglutyrglnargargasn355360365argaspserileleuserhisthrmetargtrpalaglyphearggly370375380gluasnhistrpleuaspargalaasnileglupheleucyslysglu385390395400glulysleuleuvaltyrargleuvalaspgluglyargserleulys405410415valilehisasnleuserasnglyglumetvalphegluglyvalarg420425430valglnprotyrserthrgluvalval435440<210>62<211>1326<212>dna<213>人工序列(artificialsequence)<220><223>ct2的核苷酸序列<400>62atgataggctaccagatctacgtgagatcattcagggatggaaacttcgatggtgtgggg60gatttcaaaggattgaaaggtgcgatttcctacctgaaagaactgggtgttgattttgtc120tggctcatgcccgtcttttcctccatttccttccacgggtatgacgtggtggatttttat180tctttcaaagccgagtacggagacgagaaagactttagagagatgatcgaggcgttccac240gacaacggtataaaagtcgttctcgatcttcccatccatcatactggtttcctccatgtg300tggtttcagaaagccctgaaaggagatccacactacagggattattacgtatgggcgagt360gaaaaaacggatctggacgaaagaagagagtgggacaacgaaaggatctggcatcctctg420gaggacggaaggttctacagaggacttttcggtcccctctcacccgatctgaactacgat480aacccgcaggtttttgaagagatgaagaaggtggtttatcaccttcttgaaatgggagtg540gacggattcagattcgacgcagcaaagcacatgagagatactctggaacagaacgttcgc600ttttggaggtatttcctctccgatattgagggaatattccttgcggaaatctgggcagaa660tccaaagttgtggatgaacacggcaggatattcggctacatgctaaatttcgatacctca720cactgtattaaggaagcggtgtggaaggaaaacttcaaagtgttgatcgagtcgatcgaa780agggccctggttggaaaagattatctgccggtgaacttcacatcgaaccatgatatgtca840aggcttgcgagtttcgaaggagggttgagtgaagagaaggtgaaactctcactttccatt900ctgttcacgcttcccggggttcctctcatattctacggagacgaactgggaatgaaagga960atctatcgaaaaccgaacacggaagtcgtgctggatccgttcccctggagcgaaaacatg1020tgtgttgaaggccagacattttggaaatggcccgcgtataacgatccattctccggtgtt1080tctgttgagtatcagaggagaaatcgtgattcgattctctcacacacgatgaggtgggca1140ggattcagaggggaaaatcactggctggacagggcaaacatcgaatttctgtgcaaagaa1200gaaaaactgctcgtgtacagactggtcgatgaagggcgttctctgaaagtgatacacaac1260ctgtcgaatggtgaaatggtgtttgagggagtgcgcgtacaaccctacagcacggaggtg1320gtttga1326<210>63<211>451<212>prt<213>人工序列(artificialsequence)<220><223>tn1的氨基酸序列<400>63metlyslysmetalaleulyspheasppheserasnleupheglupro151015asnileserglyglyleuargglugluaspleugluserthrlysglu202530lysvalileglualailelysasnphethrgluasnthrproaspphe354045alaargleuasparglystrpileaspservallysgluleugluglu505560trpvalvalasnpheaspthrvalvalvalleuglyileglyglyser65707580glyleuglyasnleualaleuhistyrserleuargproleuasntrp859095asnglumetserargglugluargasnglytyralaargvalpheval100105110valaspasnvalaspproaspleumetalaservalleuaspargile115120125aspleulysthrthrleupheasnvalileserlysserglyserthr130135140alagluvalmetalaasntyrserilealaargglyileleugluala145150155160asnglyleuaspprolysgluhisileleuilethrthraspproglu165170175lysglypheleuarglysvalvallysglugluglypheargserleu180185190gluvalproproglyvalglyglyargpheservalleuthrproval195200205glyleupheseralametalagluglyileaspileglugluleuhis210215220aspglyalaargaspalaphegluargcyslyslysgluaspleuphe225230235240gluasnproalaalametilealaleuthrhistyrleutyrleulys245250255argglylysserileservalmetmetalatyrserasnargmetthr260265270tyrleuvalasptrptyrargglnleutrpalagluserleuglylys275280285argtyrasnleulysglyglugluvalphethrglyglnthrproval290295300lysalaileglyalathraspglnhisserglnileglnleutyrasn305310315320gluglyproasnasplysvalilethrpheleuargleugluasnphe325330335asparggluileileileproaspthrglyargglugluleulystyr340345350leualaarglysargleusergluleuleuleualagluglnthrgly355360365thrgluglualaleuarglysasnaspargproasnmetlysvalile370375380pheaspargleuthrsertyrasnvalglyglnphephealatyrtyr385390395400glualaalathralaphemetglytyrleuleugluileasnprophe405410415aspglnproglyvalgluleuglylyslysilethrphealaleumet420425430glyarggluglytyrglutyrgluilelysaspargthrlyslysval435440445ileileglu450<210>64<211>1356<212>dna<213>人工序列(artificialsequence)<220><223>tn1的核苷酸序列<400>64atgaaaaagatggctttgaaatttgatttttcaaatctttttgaaccgaacatctccggt60ggactgagagaggaagatctggaaagcacaaaagaaaaggtgatagaggcgataaagaat120ttcactgagaacacaccggattttgccagactggacagaaaatggatcgattcggtgaag180gaactcgaggagtgggtggtgaacttcgacacggtggtcgttctgggaattgggggatcc240ggtcttggaaaccttgcccttcattattcgttgagaccactgaactggaacgagatgtcg300agagaggaaagaaacggttatgcgagagtcttcgtggtggacaacgtagatcccgatctc360atggcctccgtccttgataggatagatctgaagacaacgctgttcaacgtgatctcaaaa420tctggatccacggctgaggttatggcgaattactcgatcgcaaggggaatcctggaggct480aatggtctggacccgaaagaacacatcctcatcacaacagatccagagaagggctttttg540agaaaagtagtgaaagaagagggcttcagaagtcttgaggtccctcccggcgttggagga600aggttcagcgtgctgacgcccgttggcctcttctctgccatggcggagggtatcgacata660gaagaactccacgacggtgcccgggatgcgttcgagagatgcaagaaggaagacctgttc720gaaaatccagcggcgatgatcgccctcacacactatctctatctgaagagaggaaagagc780atctccgtcatgatggcctactccaacaggatgacctacctcgtggactggtacagacag840ctgtgggcagaaagtctgggaaagagatacaacctgaaaggagaggaggtcttcacgggt900cagaccccggtgaaggcaataggagccaccgatcagcactctcagatacagctttacaac960gagggcccaaacgacaaagtgataacgtttttgcggttggaaaacttcgatagagagatc1020ataataccggacaccggaagagaagagctcaaataccttgcaagaaaaagactctctgaa1080cttctccttgcagaacagacaggaacagaggaagccctaaggaaaaacgacagaccgaac1140atgaaggtgatcttcgacagactcacctcttacaatgtgggccagttcttcgcttattat1200gaagccgcaactgctttcatggggtatctcctcgagatcaacccgtttgatcagccgggt1260gtggaacttggaaagaagatcacgtttgccctcatgggaagggaaggttacgaatacgaa1320ataaaagatcgcaccaagaaggtgatcatagaatga1356<210>65<211>221<212>prt<213>人工序列(artificialsequence)<220><223>fp3e的氨基酸序列<400>65metmetvallysilealaalaserileleualacysaspleualaarg151015leualaaspgluvallysargvalglugluhisileaspmetvalhis202530pheaspvalmetaspglyhisphevalproasnileserpheglyleu354045provalleulysalaleuarglysgluthrserleuproileserval505560hisleumetilethrasnprogluasptyrvalaspargphevalglu65707580gluglyalaaspmetvalalavalhistyrgluthrthrprohisleu859095hisargilevalhisargilelysaspleuglyalalysalapheval100105110alaleuasnprohisthrprovalpheleuleusergluileilethr115120125aspvalaspglyvalleuvalmetservalasnproglyphesergly130135140glnargpheilealaargserleuglulysileargserleulyslys145150155160metileargaspleuglyleugluthrgluilemetvalaspglygly165170175valasnglugluasnalaserileleuilelysasnglyalathrile180185190leuvalmetglytyrglyilephelysasngluasntyrvalgluleu195200205valargserilelysglngluargglygluseralagly210215220<210>66<211>666<212>dna<213>人工序列(artificialsequence)<220><223>fp3e的核苷酸序列<400>66atgatggtaaagatcgccgcttcaatccttgcgtgtgatcttgcaagactcgccgatgag60gtaaaaagggtggaagaacacatagacatggttcacttcgatgtcatggatggacacttc120gttccgaacatctcgttcggattgcccgttctcaaagccctgagaaaagaaaccagcctt180cctataagtgttcatctgatgatcacaaatccagaggactatgtggaccgtttcgtggaa240gagggagcggacatggtggcggtccactacgagacaacgccgcaccttcacaggatagtg300cacaggataaaggatctcggggcgaaggcgttcgtcgccctcaacccacacacaccggtt360tttctcctgtctgagatcataacggatgtggatggcgtactcgtgatgagtgtgaacccg420ggcttttctggtcagagattcattgcaaggagtctggaaaaaataaggagtctgaagaag480atgataagggatctgggactcgaaacggagatcatggtcgatggtggtgtcaacgaagaa540aacgcttctatcttaataaagaacggtgcgacgatccttgtaatggggtacggtatcttc600aaaaacgaaaactatgtggaactggtgagatccatcaagcaggaaagaggggaatctgct660ggctga666<210>67<211>441<212>prt<213>人工序列(artificialsequence)<220><223>tn2的氨基酸序列<400>67metileglytyrglniletyrvalargserpheargaspglyasnphe151015aspglyvalglyaspphelysglyleulysglyalailesertyrleu202530lysgluleuglyvalaspphevaltrpleumetprovalpheserser354045ileserphehisglytyraspvalvalaspphetyrserphelysala505560glutyrglyaspglulysasppheargglumetileglualaphehis65707580aspasnglyilelysvalvalleuaspleuproilehishisthrgly859095pheleuhisvaltrppheglnlysalaleulysglyaspprohistyr100105110argasptyrtyrvaltrpalaserglulysthraspleuaspgluarg115120125argglutrpaspasngluargiletrphisproleugluaspglyarg130135140phetyrargglyleupheglyproleuserproaspleuasntyrasp145150155160asnproglnvalphegluglumetlyslysvalvaltyrhisleuleu165170175glumetglyvalaspglypheargpheaspalaalalyshismetarg180185190aspthrleugluglnasnvalargphetrpargtyrpheleuserasp195200205ilegluglyilepheleualagluiletrpalagluserlysvalval210215220aspgluhisglyargilepheglytyrmetleuasnpheaspthrser225230235240hiscysilelysglualavaltrplysgluasnphelysvalleuile245250255gluserilegluargalaleuvalglylysasptyrleuprovalasn260265270phethrserasnhisaspmetserargleualaserphegluglygly275280285leusergluglulysvallysleuserleuserileleuphethrleu290295300proglyvalproleuilephetyrglyaspgluleuglymetlysgly305310315320iletyrarglysproasnthrgluvalvalleuasppropheprotrp325330335sergluasnmetcysvalgluglyglnthrphetrplystrpproala340345350tyrasnasppropheserglyvalservalglutyrglnargargasn355360365argaspserileleuserhisthrmetargtrpalaglyphearggly370375380gluasnhistrpleuaspargalaasnileglupheleucyslysglu385390395400glulysleuleuvaltyrargleuvalaspgluglyargserleulys405410415valilehisasnleuserasnglyglumetvalphegluglyvalarg420425430valglnprotyrserthrgluvalval435440<210>68<211>1326<212>dna<213>人工序列(artificialsequence)<220><223>tn2的核苷酸序列<400>68atgataggctaccagatctacgtgagatcattcagggatggaaacttcgatggtgtgggg60gatttcaaaggattgaaaggtgcgatttcctacctgaaagaactgggtgttgattttgtc120tggctcatgcccgtcttttcctccatttccttccacgggtatgacgtggtggatttttat180tctttcaaagccgagtacggagacgagaaagactttagagagatgatcgaggcgttccac240gacaacggtataaaagtcgttctcgatcttcccatccatcatactggtttcctccatgtg300tggtttcagaaagccctgaaaggagatccacactacagggattattacgtatgggcgagt360gaaaaaacggatctggacgaaagaagagagtgggacaacgaaaggatctggcatcctctg420gaggacggaaggttctacagaggacttttcggtcccctctcacccgatctgaactacgat480aacccgcaggtttttgaagagatgaagaaggtggtttatcaccttcttgaaatgggagtg540gacggattcagattcgacgcagcaaagcacatgagagatactctggaacagaacgttcgc600ttttggaggtatttcctctccgatattgagggaatattccttgcggaaatctgggcagaa660tccaaagttgtggatgaacacggcaggatattcggctacatgctaaatttcgatacctca720cactgtattaaggaagcggtgtggaaggaaaacttcaaagtgttgatcgagtcgatcgaa780agggccctggttggaaaagattatctgccggtgaacttcacatcgaaccatgatatgtca840aggcttgcgagtttcgaaggagggttgagtgaagagaaggtgaaactctcactttccatt900ctgttcacgcttcccggggttcctctcatattctacggagacgaactgggaatgaaagga960atctatcgaaaaccgaacacggaagtcgtgctggatccgttcccctggagcgaaaacatg1020tgtgttgaaggccagacattttggaaatggcccgcgtataacgatccattctccggtgtt1080tctgttgagtatcagaggagaaatcgtgattcgattctctcacacacgatgaggtgggca1140ggattcagaggggaaaatcactggctggacagggcaaacatcgaatttctgtgcaaagaa1200gaaaaactgctcgtgtacagactggtcgatgaagggcgttctctgaaagtgatacacaac1260ctgtcgaatggtgaaatggtgtttgagggagtgcgcgtacaaccctacagcacggaggtg1320gtttga1326<210>69<211>34<212>dna<213>人工序列(artificialsequence)<220><223>ct1的正向引物dna序列<400>69aggagaaactcatatgctgaagaaactcccggag34<210>70<211>21<212>dna<213>人工序列(artificialsequence)<220><223>ct1的反向引物dna序列<400>70agccccctcgagcccgagaac21<210>71<211>26<212>dna<213>人工序列(artificialsequence)<220><223>ct2的正向引物dna序列<400>71aaagggcatatgatcctgtttggaac26<210>72<211>27<212>dna<213>人工序列(artificialsequence)<220><223>ct2的反向引物dna序列<400>72ataccagtctcgagcagtttcaggatc27<210>73<211>34<212>dna<213>人工序列(artificialsequence)<220><223>tn1的正向引物dna序列<400>73ttactgagggcatatgaaaaagatggctttgaaa34<210>74<211>30<212>dna<213>人工序列(artificialsequence)<220><223>tn1的反向引物dna序列<400>74aagacgcgtcgacttctatgatcaccttct30<210>75<211>34<212>dna<213>人工序列(artificialsequence)<220><223>fp3e的正向引物dna序列<400>75ggaacatatgatggtaaagatcgccgcttcaatc34<210>76<211>27<212>dna<213>人工序列(artificialsequence)<220><223>fp3e的反向引物dna序列<400>76catactcgagcttcccctctcctatct27<210>77<211>270<212>prt<213>人工序列(artificialsequence)<220><223>来源于嗜热异常球菌(deinococcusgeothermalis)的多聚磷酸依赖型葡萄糖激酶<400>77metleualaalaseraspserserglnhisglyglylysalavalthr151015leuserprometservalileleuglyileaspileglyglysergly202530ilelysglyalaprovalaspthralathrglylysleuvalalaglu354045arghisargileprothrprogluglyalahisproaspalavallys505560aspvalvalvalgluleuvalarghispheglyhisalaglyproval65707580glyilethrpheproglyilevalglnhisglyhisthrleuserala859095alaasnvalasplysalatrpileglyleuaspalaaspthrleuphe100105110thrglualathrglyargaspvalthrvalileasnaspalaaspala115120125alaglyleualaglualaargpheglyalaglyalaglyvalprogly130135140gluvalleuleuleuthrpheglythrglyileglyseralaleuile145150155160tyrasnglyvalleuvalproasnthrglupheglyhisleutyrleu165170175lysglyasplyshisalagluthrtrpalaseraspargalaargglu180185190glnglyaspleuasntrplysglntrpalalysargvalserargtyr195200205leuglntyrleugluglyleupheserproaspleupheileilegly210215220glyglyvalserlyslysalaasplystrpglnprohisvalalathr225230235240thrargthrargleuvalproalaalaleuglnasnglualaglyile245250255valglyalaalametvalalaalaglnargserglnglyasp260265270<210>78<211>253<212>prt<213>人工序列(artificialsequence)<220><223>来源于嗜热厌氧绳菌(anaerolineathermophila)的多聚磷酸依赖型葡萄糖激酶<400>78metglyargglnglymetgluileleuglyileaspileglyglyser151015glyilelysglyalaprovalaspvalgluthrglyglnleuthrala202530gluargtyrargleuprothrprogluasnalaleuproglugluval354045alaleuvalvalalaglnilevalgluhispheglntrplysglyarg505560valglyalaglypheproalaalailelyshisglyvalalaglnthr65707580alaalaasnilehisprothrtrpileglyleuhisalaglyasnleu859095pheserglulyscysglycysprovalservalleuasnaspalaasp100105110alaalaglyleualaglumetilepheglyalaglylysglyglnlys115120125glyvalvalleumetilethrileglythrglyileglythralaleu130135140phethraspglyileleuvalproasnthrgluleuglyhisileglu145150155160ileargglylysaspalagluglnargserserglualaalaarggln165170175arglysasptrpthrtrpglnglntrpalalysargleuasngluhis180185190leugluargleuglualaleuphetrpproaspleupheileleugly195200205glyglyalavallysasnhisglulysphepheprotyrleulysleu210215220argthrprophevalalaalalysleuglyasnleualaglyileval225230235240glyalaalatrptyralahisthrglngluthrglnala245250当前第1页12