20种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗的制作方法

专利名称：20种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗的制作方法
技术领域：
本发明主要围绕在人肿瘤新型诊断与治疗。以新的肿瘤诊断之特异靶基因，靶蛋 白的发现，以及其在新型肿瘤诊断中之用途和用法，与靶蛋白在肿瘤疫苗中的应用为核心。
背景技术：
癌症，为全人类共患之疾，目前，仅有放疗，化疗等非特异肿瘤治疗手段为目前临 床诊治的主要方法，其5年成活率在大部分肿瘤患者中还很低.而特异的肿瘤诊断还局限 于甲胎球蛋白在肝癌中或PSA在前列腺癌等少数肿瘤类型之应用.新的肿瘤靶抗原的发现 在过去的20年中进展缓慢，因为目前常用技术在新的肿瘤诊断的特异靶基因，靶蛋白的发 现过程中困难重重.而癌症的临床诊治应用中有急需新技术，新的肿瘤诊断的特异靶基因 为治疗提供新手段.癌症，目前仍然为仅次于心肌梗塞致死的第二位临床疾病致死的杀手.全球每年 死于癌症的患者依然以数百万记.在中国，每年常见肿瘤的发病人数在120万以上，死于癌 症的患者也有50万人之多.各国常见肿瘤类型大体说来基本相同，以肺癌，乳腺癌，胃癌， 肝癌，前列腺，结直肠癌，脑肿瘤，胰腺癌，卵巢癌等为多发病例.死亡率也大致相当.而且 一旦肿瘤转移，其临床预后极差.目前，仅有放疗，化疗等非特异肿瘤治疗手段，而且其副 作用也很大.目前，早期的特异的肿瘤诊断还仅仅局限于肝癌中甲胎球蛋白(AFP)或PSA在前 列腺癌等少数肿瘤中.大部分临床常见肿瘤仍无好的诊断方法.现在临床中使用的X光， 直肠镜，等不但费用高，病人也极受操作的痛苦.往往是因患者有临床症状而入院时则已 为晚期.而治疗中因缺少特异的肿瘤诊断指标，往往治疗效果也有盲目性.故癌症的临床 诊治中，目前急需新技术，新的肿瘤诊断的特异靶基因的发现与发展.而本技术发明正是 为此而创新.

发明内容
本发明提供了分离的多核苷酸，其选自(1)序列为SEQ ID N0:l_20之一的多核 苷酸；(2)序列为SEQID NO 1-20之一的多核苷酸的互补序列；(3)序列为SEQ ID NO :1_20 之一的多核苷酸的反义序列。本发明提供了所述多核苷酸编码的多肽。在优选的技术方案中，多肽的氨基酸序 列为 SEQ ID NO 21-40 之一。本发明提供了携带有所述分离的多核苷酸的表达载体。本发明还提供了宿主细胞，其转化或转染了所述分离的多核苷酸或者所述表达载 体。本发明提供了特异性结合所述多肽的抗体或其片段。本发明提供了所述分离的多核苷酸或多肽在制备制剂中的用途，该制剂用于刺激 或扩增能特异性识别肿瘤蛋白的T细胞。
本发明提供了用于刺激或扩增能特异性识别肿瘤蛋白的T细胞的制剂，其含有所 述分离的多核苷酸或所述多肽；以及所述制剂制备的T细胞群。本发明还提供了所述分离的多核苷酸、所述多肽或所述T细胞群在制备预防和/ 或治疗肿瘤和/或癌症的疫苗中的用途。本发明还提供了预防和/或治疗肿瘤和/或癌症的疫苗，其含有所述分离的多核 苷酸、多肽或T细胞群以及药学上可接受的佐剂。本发明还提供了所述分离的多核苷酸、多肽或T细胞群在制备诊断肿瘤和/或癌 症的试剂盒中的用途。本发明还提供了诊断肿瘤和/或癌症的试剂盒，其含有所述分离的多核苷酸、多 肽或T细胞群。本发明的肿瘤疫苗的技术方案中，我们提供如何使用靶蛋白抗体，或靶蛋白抗原 本身结合载体蛋白的技术方法.进一步来说，在本发明的肿瘤疫苗的技术方案中，我们详 细描述有1)如何将靶蛋白表达为免疫细胞表面作为靶蛋白抗原来刺激患者免疫系统，所 描述的免疫细胞包括树突状细胞，T和B淋巴细胞等.2)作为佐剂载体蛋白发展的技术内 容.这类技术包含有融合蛋白技术，其中涵盖有如何通过融合蛋白中载体蛋白部分来刺激 患者免疫系统，如何选用高免疫原性之蛋白为载体蛋白.如何借用载体蛋白部分的特点来 简化重组蛋白的纯化工艺等.在临床肿瘤的早期诊断应用方面，本发明提供具体如何使用靶蛋白及多肽结合抗 体技术开发各类常规临床检验技术抗体酶联法，组织染色法，分析质谱多肽鉴定法等技术 应用细节.同时本发明也提供具体如何使用靶基因特异D N A序列而开展的以D N A杂 交为核心而检验的技术系统如P C R技术和GeXP技术系统等.最后，在临床应用的总体诊断，治疗方面，本发明提供具体案例1)如何从患者肿 瘤组织中检测20种靶基因的表达谱，2)依据患者肿瘤组织中检测出的20种靶基因的表达谱，利 用高表达靶基因来指导临床医生即时选用预先生产好的20种靶基因所制造出的重组蛋白 作为患者特异功能肿瘤疫苗而对病人进行治疗来杀灭肿瘤细胞和预防术后肿瘤复发的具 体治疗方案.


图1为人脑源DNA CHIP_DSL人基因启动子DNA甲基化实验结果扫描图.本图为 MA校正后的结果展示图，F532坐标为对数值.读数为2时其靶基因的荧光信号实际增强了 4倍.图2为DNALI1靶基因在GNF芯片库中mRNA在各种人体组织中特异性表达图.本 结果源于用美国Affymetrix公司U133A DNA芯片杂技61种正常人体组织，16种细胞系及 一种肿瘤样品后综合分析后结果之展示.图上方读数代表其相对基因表达之水平.DNALI1 在人睾丸生殖细胞表达水平为其他组织的3500倍.同时可见这一靶基因在其他正常体组 织中几乎不表达.图3为20种靶基因的名称与基因序列号图4为14种靶基因的多肽合成序列
图5为20种靶基因的名称与基因序列号和其对应的全长cDNA克隆号图6为在大肠杆菌中表达6种靶基因的DNA引物合成序列。为克隆6个靶基因以 便在大肠杆菌中表达而获得全长重组蛋白，靶基因特异性DNA引物合成如下5’端引物加 有EcoRI酶切点，3’端引物加有Xhol酶切点.图7为二种靶蛋白SSCPDH和GSTM3表达菌在2mM IPTG诱导后靶基因在大肠杆菌 中表达SDS-PAGE图.图8为靶蛋白特异抗体进行免疫蛋白印迹(Western Blot)筛选各种肿瘤细胞及 正常人组织裂解液结果图.图9为REALTIME-PCR鉴定用靶基因DNA引物合成序列图10为人结肠癌样本REALTIME-PCR结果分析图。分析人结肠肿瘤组织的CSNK2A2 靶基因表达表达水平是用购置于美国Origene公司，(产品目录号#H CRT 101&H CRT 501)，其包含有5例正常人结肠组织(C1至C5)和19例人结肠肿瘤组织的cDNA样品.靶 基因D NA引物经美国I D T公司合成，其序列见图9 实时PCR反应用美国Bio-Rad iQ5 探测器检测，同时使用iQ SYBR Green Supermix(产品目录号170-8884)进行样品反 应.CSNK2A2靶基因表达表达水平用倍数之质而标记而小结于图10.图11为GEXP系统鉴定用靶基因DNA引物合成序列，列出了 GeXP实验中全部靶基 因和4种House-Keying基因所用的DNA引物.图12为GeXP结果之小结。因为本组靶基因分别表达在不同种类的肿瘤中，为测试 GeXP结果，我们用数种肿瘤混合的高质量RNA起始测试.美国Stratagene公司Universal human Reference RNA(产品目录号740000)，为人腺癌，肝癌，宫颈癌，睾丸肿瘤，脑胶质瘤， 黑色素瘤，脂肪瘤，白血病细胞等组织纯化的RNA混合物.本次实验中，20种靶基因于于2 种看家基因(TBP，ATP50, ACTB和GAPDH)被清晰的在肿瘤样品中检测出.C170RF39和CPD 靶基因表达水平明显增加.图13为靶蛋白C170RF39特异性抗体在人肾肿瘤组织组织化学染色(IHC)中的应 用.蛋白A柱纯化的抗C170RF39重组蛋白特异抗体的免疫组化使用浓度为lug/ml。组织 标本固定在福尔马林溶液中先固定，然后石蜡包埋，然后切成4(ym)厚度的片子。按常规 组织片微波加热处理法对组织标本片处理后用美国Ventana公司自动免疫组织化学染色 仪进行组织化学染色.染色使用AEC检测合(美国Covance公司产品).C170RF39的免疫 组化结果见下图.在部分的人肾肿瘤组织中可见高表达的细胞核染色.而对照的正常人肾 组织中几乎检测不到任何染色信号.本发明中涉及的序列说明如下序列号为1(SEQ ID N0:1)的靶基因在N C B I基因库中为基因序列号为 NM_001464的，名称为ADAM2的基因.序列号为2(SEQ ID NO 2)的靶基因在N C B I基因库中为基因序列号为 NM_024052的，名称为C170RF39的基因.序列号为3(SEQ ID NO 3)的靶基因在N C B I基因库中为基因序列号为 NM_145036的，名称为CCDC46的基因.序列号为4(SEQ ID NO 4)的靶基因在N C B I基因库中为基因序列号为 NM_001304的，名称为C P D的基因.
序列号为5(SEQ ID NO 5)的靶基因在N CB I基因库中为基因序列号为 NM_144782的，名称为CRAT的基因.序列号为6(SEQ ID NO 6)的靶基因在N C B I基因库中为基因序列号为 NM_001896的，名称为CSNK2A的基因.序列号为7(SEQ ID NO 7)的靶基因在N C B I基因库中为基因序列号为 NM_003462的，名称为DNALIl的基因.序列号为8(SEQ ID NO 8)的靶基因在N C B I基因库中为基因序列号为 NM_033214的，名称为G K 2的基因.序列号为9(SEQ ID NO 9)的靶基因在N C B I基因库中为基因序列号为 NM_153823的，名称为GSGl的基因.序列号为10(SEQ ID N0:10)的靶基因在N C B I基因库中为基因序列号为 NM_000849的，名称为GSTM3的基因.序列号为11 (SEQ ID NO=Il)的靶基因在N C B I基因库中为基因序列号为 NM_000640的，名称为IL13RA2的基因.序列号为12(SEQ ID NO 12)的靶基因在N C B I基因库中为基因序列号为 ΝΜ_016368的，名称为ISYNAl的基因.序列号为13(SEQ ID NO 13)的靶基因在N C B I基因库中为基因序列号为 NM_004923的，名称为MTL5的基因.序列号为14(SEQ ID NO 14)的靶基因在N C B I基因库中为基因序列号为 NM_153485的，名称为NUP155的基因序列号为15(SEQ ID NO 15)的靶基因在N C B I基因库中为基因序列号为 ΝΜ_015589的，名称为SAMD4A的基因序列号为16(SEQ ID NO 16)的靶基因在N C B I基因库中为基因序列号为 ΝΜ_016002的，名称为SCCPDH的基因序列号为17(SEQ ID NO 17)的靶基因在N C B I基因库中为基因序列号为 ΝΜ_019073的，名称为SPATA6的基因序列号为18(SEQ ID NO 18)的靶基因在N C B I基因库中为基因序列号为 NM_003284的，名称为TNPl的基因序列号为19(SEQ ID NO 19)的靶基因在N C B I基因库中为基因序列号为 NM_021733的，名称为TSKS的基因序列号为20(SEQ ID NO 20)的靶基因在N C B I基因库中为基因序列号为 ΝΜ_017481的，名称为UBQLN3的基因序列号为21(SEQ ID NO 21)的靶蛋白在N C B I基因库中为蛋白序列号为 NP_001455的，名称为ADAM2的蛋白.序列号为22 (SEQ ID NO 22)的靶蛋白在N C B I基因库中为蛋白序列号为NP 076957的，名称为C170RF39的蛋白序列号为23(SEQ ID NO 23)的靶蛋白在N C B I基因库中为蛋白序列号为 NP_659473的，名称为(XDC46的蛋白.序列号为24(SEQ ID NO 24)的靶蛋白在N C B I基因库中为蛋白序列号为NP_001295的，名称为CPD的蛋白·序列号为25(SEQ ID NO 25)的靶蛋白在N C B I基因库中为蛋白序列号为NP_000746的，名称为CRAT的蛋白.序列号为26(SEQ ID NO 26)的靶蛋白在N C B I基因库中为蛋白序列号为 NP_001887的，名称为CSNK2A2的蛋白.序列号为27 (SEQ ID NO 27)的靶蛋白在N C B I基因库中为蛋白序列号为 NP_003453的，名称为DNALIl的蛋白.序列号为28(SEQ ID NO 28)的靶蛋白在N C B I基因库中为蛋白序列号为 NP_149991的，名称为G K 2的蛋白.序列号为29(SEQ ID NO 29)的靶蛋白在N C B I基因库中为蛋白序列号为 NP_722545的，名称为GSGl的蛋白·序列号为30(SEQ ID NO 30)的靶蛋白在N C B I基因库中为蛋白序列号为 NP_000840的，名称为GSTM3的蛋白序列号为31(SEQ ID NO 31)的靶蛋白在N C B I基因库中为蛋白序列号为 NP_000631的，名称为IL13RA2的蛋白序列号为32(SEQ ID NO 32)的靶蛋白在N C B I基因库中为蛋白序列号为 NP_057452的，名称为ISYNAl的蛋白序列号为33(SEQ ID NO 33)的靶蛋白在N C B I基因库中为蛋白序列号为 NP_004914的，名称为MTL5的蛋白序列号为34(SEQ ID NO 34)的靶蛋白在N C B I基因库中为蛋白序列号为 NP_004289的，名称为NUP155的蛋白序列号为35(SEQ ID NO 35)的靶蛋白在N C B I基因库中为蛋白序列号为 NP_056404的，名称为SAMD4A的蛋白序列号为36(SEQ ID NO 36)的靶蛋白在N C B I基因库中为蛋白序列号为 NP_057086的，名称为SCCPDH的蛋白序列号为37 (SEQ ID NO 37)的靶蛋白在N C B I基因库中为蛋白序列号为 NP_061946的，名称为SPATA6的蛋白序列号为38(SEQ ID NO 38)的靶蛋白在N C B I基因库中为蛋白序列号为 NP_003275的，名称为TNPl的蛋白序列号为39(SEQ ID NO 39)的靶蛋白在N C B I基因库中为蛋白序列号为 NP_068379的，名称为TSKS的蛋白序列号为40(SEQ ID NO 40)的靶蛋白在N C B I基因库中为蛋白序列号为 NP_059509的，名称为UBQLN3的蛋白
具体实施例方式任何引用的参考资料如专利，发表的文献等均作为文献应用.本发明的核心之处在于发现新的多种肿瘤中特异表达的靶基因与其编码的蛋 白.同时提供详细的技术步骤说明与实验案例来指导如何应用这组肿瘤中特异表达的靶 基因与其编码的蛋白在临床诊断和重组蛋白在肿瘤疫苗发展中的应用.
在本发明的详细说明之中，会引用出许多常见的病毒学，免疫学，微生物学，分子生物学，DNA重组技术的常规技术与学术用语，在以下书籍中可查询引用 Sambrook, et al.Molecular Cloning:A LaboratoryManual (2nd Edition,1989)； Maniatis et al. Molecular Cloning :A Laboratory Manual(1982) ；Oligonucleotide Synthesis(N. Gait, ed. ,1984) ；Nucleic Acid Hybridization(B. Hames&S. Higgins, eds. ,1985) ；Transcription and Translation(B. Hames&S. Higgins, eds. ,1984) ；Animal CelICulture(R. Freshney, ed. , 1986) ；Perbal, A Practical Guide to Molecular Cloning(1984).本发明中从人肿瘤组织中分离鉴定出来的20种特指肿瘤相关抗原(TAA)的技 术路线为新型技术思路，其首先利用商业化的技术如CHIP-DSL系统寻找出正常组织中高 度DNA甲基化的基因启动子靶基因，然后从DNA芯片的阵列中很容易的查到其对应的靶基 因而同时获得靶基因的基因序列号.以此基因序列号而很快得到靶基因序列与靶蛋白序 列.同时，依据基因序列号，从NCBI库中可上网查寻完全同源的全长cDNA克隆(IMAGE克 隆号)，次开发而从ATCC或OPEN BIOSYSTEMS公司购买靶基因全长cDNA克隆.这一技术路 线避免了传统的肿瘤组织cDNA库构建，基因杂交筛选，全长cDNA克隆，序列测定时耗时的 工艺·在获得20种特指肿瘤相关抗原(TAA)后，利用其靶基因来开发有关DNA序列依赖 性诊断，重组蛋白表达的基因克隆，多肽或重组蛋白在特异抗体中的应用，以及抗体在肿瘤 早期诊断中应用等为常规发展技术路线.最后本发明还重点描述这组肿瘤相关抗原在肿 瘤疫苗发展中的使用.本文中的蛋白多肽包括靶蛋白的任何连续在8个氨基酸以上的多肽序列.同时 也包括常见的多种修饰性氨基酸而组成的多肽序列如糖基化，乙酰化，磷酸化，甲基化 等.同时本专利因涉及临床诊断和重组蛋白在肿瘤疫苗发展中的应用，如一些常用免疫概 念蛋白多肽的免疫原性，多肽表位等更是本专利的描述重点.本文中的靶蛋白有时会被 称之为肿瘤相关抗原(TAA，Tumor associated Antigen)，因其有特异的在肿瘤组织中高表 达的特点.在发现新的多种肿瘤中特异表达的这组靶基因与其编码的蛋白后，许多常规的免 疫学方法自然而然的可据此组靶基因与其编码的蛋白而发展出各种临床诊断系统如选用 靶蛋白多肽而制备特异抗体，然后建立起ELISA，胶体金，放射免疫等传统诊断试剂合.或 用这组靶基因DNA序列而发展出以PCR，DNA芯片等诊断试剂盒.这类常规技术系统在以下 文献中有详细指导说明Harlow and Lane,Antibodies :A LaboratoryManual, Cold Spring Harbor Laboratory,1988. Fundamental Immunology,3rd ed. ,243-247(RavenPress, 1993).虽然有时蛋白多肽的免疫原性不容易掌握，本发明的详细说明中有涵盖使用 靶蛋白全长序列来制备重组蛋白之技术路线.如果是细胞中穿膜蛋白在大肠杆菌中表 达，我们还阐明有如何祛除蛋白N端的信号肽与穿膜蛋白功能域等技术路线来保障重组 蛋白之高表达.此外，现今的DNA合成技术已使用的大肠杆菌蛋白合成优先码(codon optimization)成为常规技术，更进一步确保重组蛋白在大肠杆菌中高表达的技术路线之 成功率.至于重组蛋白在其他哺乳类细胞中高表达的技术路线包括使用表达载体，用较强的启动子，载体携带筛选标记基因等都在下例案例中有具体例案。在多肽化学合成的技术路线中，利用相似氨基酸来部分置换靶蛋白序列中的部分氨基酸而生产出功能类似、但序列不同的多肽其实是非常成熟的技术如Kyte 和Doolittle，1982)即在其文献出终结出氨基酸的亲水，疏水之分而给各中氨基酸 相关值来考虑氨基酸置换ta Jsoleucine (+4. 5) ；valine (+4. 2) ； leucine (+3. 8)； phenylalanine(+2. 8) ；cysteine/cystine(+2. 5) ；methionine(+1. 9) ；alanine(+1. 8)； glycine (-0. 4) ；threonine(-0. 7) ；serine (-0. 8) ；tryptophan(-0. 9) ；tyrosine(-1. 3)； proline (-1. 6) ；histidine(-3. 2) ； glutamate(-3. 5) ； glutamine(-3. 5)； aspartate (-3. 5) ；asparagine (-3. 5) ； lysine (-3. 9) ；and arginine (-4. 5).在 U. S. Pat. No. 4，554，101专利中也有具体说明·在选用靶蛋白序列中多肽来作为肿瘤疫苗发展时，其多肽序列的独特性的选择 也是治疗成功的关键因素之一.选择序列独特的多肽，可降低与其它体内蛋白的同源性 而产生交叉反应的副作用，从而使肿瘤疫苗治疗效果更特异.利用美国NCBI蛋白库中 BLAST功能来开展这一工作很方便.具体操作方法在Altschulet al. (1977)Nucl. Acids Res. 25 :3389-3402and Altschul et al. (1990) J. Mol. Biol. 215 :403_410，文献中有详细 说明.而下面的文献对多肽序列的独特性的选择提供了多方面，、更详细的说明(DNASTAR， Inc. , Dayhoff, Μ. 0. (1978)A model of evolutionary change in proteins__Matrices for detectingdistant relationships. In Dayhoff, M. 0. (ed. )Atlas of Protein Sequence and Structure, NationalBiomedical Research Foundation, Washington D.C. Vol. 5, Suppl. 3, pp. 345-358 ；Hein J. (1990)Unified Approach to Alignment and Phylogenes pp. 626-645Methods in Enzymology vol. 183, AcademicPress, Inc.，San Diego, Calif. ；Higgins, D. G. and Sharp, P.M. (1989) CABIOS 5 :151-153 ；Myers, E. W. and Muller W. (1988)CABIOS 4 :11_17 ；Robinson, E. D. (1971)Comb. Theor 11 105 ；Santou, N. Nes, Μ. (1987)Mol. Biol. Evol. 4 :406_425 ；Sneath, P. H. A. and Sokal, R. R. (1973) Numerica1Taxonomy__the Principles and Practice of Numerical Taxonomy, Freeman Press, San Francisco, Calif. ；Wilbur, W. J. and Lipman, D. J. (1983)Proc. Natl. Acad., Sci.USA 80 :726-730.有时，当多肽序列的独特性的选择已挑选出某些区域而需要经一步细化时，下列 文献中给出许多具体指导Smith and Waterman(1981)Add. APL. Math 2 :482. Needleman and Wunsch (1970) J. Mol. Biol. 48 :443. Pearson and Lipman (1988) Proc. Natl. Acad. Sci. USA 85 :2444.利用融合蛋白技术来增加靶蛋白序列中多肽的生产，方便纯化，提高靶蛋白多肽 的免疫原性等可按部分以下文献中的方法来操作.如His-Tag，HA-Tag, GST融和载体等蛋 白可经特殊的蛋白酶切点肽段如TEV等而融合，在经亲和柱纯化后用TEV蛋白酶切除融合 载体蛋白部分来获得靶蛋白多肽纯品.(Lucast et al. Biotechniques 30 =544-550,2001) 其他常用的融合载体蛋白还包括抗体中的FC段和人白蛋白等.这在以下文献中也有详细 说明(W0/2003/035105).利用融合蛋白技术来提高靶蛋白多肽的免疫原性往往以选择那些对人T细胞、树 突状细胞等有较强刺激性的载体蛋白为首选.有些亚类的IgG Fc端因能特异结合免疫细胞表面受体而具有较强的刺激性，这在文献Stoute et al.New Engl. J. Med. ,336 =86-91, 1997中有详细描述.在融合蛋白技术中有时载体蛋白与免疫靶蛋白多肽的三维结构也至关重要，需在二者结合处放置连接域小肽，如GGGGS和(GGGGS) 3等免疫原性低，而结构灵活的连接域小 肽.这类技术在以下文献中有详细描述:Maratea et al. ,Gene 40 =39-46,1985 ；Murphy et al. ,Proc. Natl. Acad. Sci. USA 83 :8258_8262，1986 ；U. S. Pat. No. 4，935，233and U. S. Pat. No. 4，751，180.并且，融合蛋白技术中还包括新近发展出的DNA-蛋白融合技术.目前，免疫细胞 表面受体中有一类RNA，DNA结合受体如TLR3，TLR9当其被激活时能极大调动免疫反应.故 TLR9结合非甲基化的DNA可化学合成并绞连上靶蛋白免疫多肽而增强其肿瘤疫苗之疗 效.以下文献对这类DNA-蛋白融合技术有较详细的描述(Stefan et al Vaccine. 2003， vol.21, pp.990-995).多肽在血中的生物半衰期也至关重要.而在多肽化学合成的技术路线中，使用 非天然氨基酸、应用修饰性氨基酸等来增加多肽稳定性也在下述文献中有详细描述(ra Arnold. The FASEB Journal, Vol 7，744-749，1993.).至于经化学合成方法来同时稳定DNA 核昔酸链的技术则更为成熟· ^niiffiphosphorothioate or 2' 0-methyl rather than phosphodiesterase linkages,用非天然喊基如inosine, queosine and wybutosine,禾口 acetyl-, methyl-, thio—等修饰 adenine, cytidine, guanine, thymineand uridine 等技 术都能增加DNA核苷酸链在血中的生物半衰期.正如专利中U. S. Patent Application 7270821描述的一样，乙肝病毒的核心蛋白 (HBcAg)为病毒之结构蛋白之一，分子量仅18KD左右.但往往以病毒核心体形式出现.每 一个核心体以180个左右的单体聚合物组成直径为27nm，具有极强的免疫原性.同时，在过 去多年的研究发现，这种病毒核心体具有特殊的密度浮力，经简单的蔗糖密度梯度离心即 可纯化出.把免疫靶蛋白多肽融合在HBcAg蛋白的C末端，然后在大肠杆菌中高表达重组 蛋白，经破菌，离心等步骤即可获得免疫原性极高的融合重组蛋白.这一技术在下例文献 中有详细的描述(W0/9940934).因16型，18型等亚型的人乳头瘤病毒的感染能诱发妇女宫颈癌而在科研领域对 其有较深入的研究.过去的数年中，利用人乳头瘤病毒的重组膜蛋白Ll而发展出的人乳头 瘤病毒的疫苗已被美国FDA批准而在临床广泛应用.重组膜蛋白Ll有极高的免疫原性，同 时其生产、纯化工艺成熟((US patent 6908613).甚至HPV18型的Ll蛋白的多肽53之194 本身已具极高的免疫原性. 其它常用的，高免疫原性的载体蛋白还包括肺炎链球菌(Streptococcus pneumoniae)的 LYTA 蛋白·其功能是负责合成 N-acetyl—L—alanine amidase.这在 Gene 43 =265-292,1986文献中有详细的说明.LYTA蛋白对磷酸胆碱和DEAE都有较高的亲和 力·所以其重组之融合蛋白可用DEAE柱来纯化.(BiotechnologylO :795-798，1992).当 LYTA蛋白的C末端，从178个氨基酸以后，(用188-305区可能更好)片段为载体蛋白与免 疫靶蛋白多肽融合即可获得重组蛋白而用于肿瘤疫苗的发展. 在载体蛋白与免疫靶蛋白多肽融合时，有功能性的肽段如增加蛋白进入细胞溶 酶体，或增加融合肽在免疫靶细胞上结合MHC class II展示等均可考虑加入融合蛋白之中.具体的技术细节在专利中U. S. Pat. No. 5，633，234.有较详细的说明.本发明中靶蛋白多肽生产技术是极为成熟的技术之一，既有多肽合成仪可买也有商业公司提供优质的多肽合成服务.文献中如Merrifield，J. Am. Chem. Soc. 85 2149-2146，1963对多肽合成原理有详细的说明，而多肽合成仪可购买美国Perkin Elmer/ Applied BioSystems Division (Foster City，Calif.)，或 ACT 公司等产品.中国上海吉尔 生化有限公司等提供多肽合成服务.本发明中所指靶蛋白多肽是指我们从人肿瘤组织中 分离鉴定出来的20种特指肿瘤相关抗原(TAA).至于本发明中分离鉴定出来一组肿瘤诊治相关的命名为序号为1至20的靶基因， 在此可指与这组基因相关的，基因片段，DNA探针，反义基因序列，RNAi等.同时也包括其 表达载体，质粒等.当然，这组基因也包含那些经DNA合成，编码已经优化但仍然产生同类 蛋白的DNA产物.在了解这一组肿瘤诊治相关靶基因特定序列后，以此而衍生出的突变体， 同源序列等是本领域的常规技术.如DNA产物与本发明中序号为1至20的靶基因有任何 75%以上的同源序列性均视为类似基因.而DNA片段之长度达15bp之长时以上时还与靶 基因有任何75%以上的同源序列性也视为为类似基因.这类常规基因工程实验操作可在 以下这类文献中有较详细的说明.Maloy et al. , 1994 ；Segal, 1976 ；Prokop and Bajpai, 1991 ；Kuby, 1994 ；and Maniatis et al.，1982. U. S. Pat. No. 4，683，202.依据序号为1至20的靶基因而衍生出反义基因序列，RNAi在抑制靶基因的 表达中的应用是显而易见的技术，这在专利文件如U. S. Pat. No. 5，739，119和U. S. Pat. No. 5，759，829中有详细的说明.用肿瘤靶基因如的反义基因序列来抑制肿瘤细胞生长在 U. S. Pat. No. 5，747，470 ；和 U. S. Pat. No. 5，591，317 以及 U. S. Pat. No. 5，783，683 等专利中 均有技术方案之介绍.故此，在本发明中所分离鉴定出来的这一组肿瘤诊治相关靶基因的 应用中，以靶基因而衍生出其反义基因序列，RNAi来治疗肿瘤就成为治疗应用之一.更多 的相关技术文献见下Int. Pat. App 1. Publ. No. WO 93/23569 和 Int. Pat. App 1. Publ. No. WO 94/02595.至于稳定小片段DNA分子的技术可从下面参考资料中获得Int. Pat. App 1. Publ. No. WO 92/07065 ；Iht. Pat. App 1. Publ. No. WO 93/15187 ；Int. Pat. App 1. Publ. No. WO 91/03162 ；Eur. Pat. App 1. Publ. No. 92110298. 4 ；U. S. Pat. No. 5，334，711 ；and Iht. Pat. App 1. Publ. No. WO 94/13688.有关 DNA-多肽杂合技术如 PNA (p印tide nucleic acids)也 在文献Good and Nielsen, Antisense Nucleic Acid Drug Dev. 19977 (4) 431-37 中有详细 的说明.这一技术的具体应用在下面的技术综述文献中有更进一步的描述=Corey et al (Trends Biotechnol 1997 June; 15 (6) 224~9).依据本发明中序号为1至20的靶基因而衍生出的DNA依赖性临床诊断中多聚核 苷酸片段将成为重要的施加部分.其纯度鉴定，质量检验，表达水平等均为重要的技术成 分.故将简要在此进行描述其一，多聚核苷酸片段的纯度鉴定，质量检验，表达水平测定等均为常规分子生 物学技术，在 Sambrooket al. , Molecular Cloning :A Laboratory Manual, Cold Spring Harbor Laboratories, Cold SpringHarbor, N. Y.，1989，书中有详细描述.同时，目前商 用的DNA合成服务公司如美国IDT公司对其DNA引物合成有严格的质量检验.各种PCR 引物之设计，PCR操作，实施PCR的使用等均有厂家提供标准试剂，和操作说明书.如DNA 连接技术衍生的 Ligation PCR 见文献:Eur. Pat. App 1. Publ. No. 320，308 和 U. S. Pat.No. 4，883，750.独特的 Qbeta R印Iicase 酶的应用见 PCT Intl. Pat. Appl. Publ. No. PCT/ US87/00880专利.链置换型DNA扩增技术(Strand Displacement Amplification)见Great Britain Pat. Appl. No. 2202328，和PCT Intl. Pat. Appl. Publ. No. PCT/US89/01025.其它技 术如转录依赖型 DNA 扩增技术(transcription-based amplification systems (TAS)见文 献 PCT Intl. Pat. Appl. Publ. No. W088/10315 等.在文献中也见报道的还有 cyclically synthesizing single-stranded RNA( “ ssRNA “ )，ssDNA, and double-stranded DNA (dsDNA).技术，见 PCT Intl. Pat. Appl. Publ. No. WO 89/06700.在 1990 年 Frohman 报导 的"RACE"技术等.至于靶基因的基因工程中，使用各种表达载体，菌株或细胞系也为常规分子生物学技术，其表达系统可在大肠杆菌，酵母，蚕螽病毒，哺乳细胞系统，转基因动，植物等系统 中进行.常用的表达启动子包括CMV，T7等强启动子系统来获得高表达.为获得稳定的哺 乳细胞表达株，包括各种筛选基因的使用，增强子(enhancer)的应用等均有商用系统如 美国EMD公司的pET载体系统，美国Invitrogen公司的pSPORTl载体系统等.至于靶基因在大肠杆菌系统的表达，美国EMD公司的pET载体系统就可作为首选 系统.这一表达系统以噬菌体T7的启动子为核心，配备各种筛选基因如青酶素，卡那酶 素等.同时还是IPTG诱导表达故表达水平高.同时，pET载体系统还配有各种融合蛋白 如GST，HIS多肽等，使表达的重组蛋白纯化变的相对简单.如果结合TEV等蛋白酶切位 点，重组蛋白的纯品可很容易的控制在融合重组蛋白或非融合重组蛋白状态.其他表达系 统，如 pIN 载体系统，常在文献 Van Heeke, G. and S. Μ. Schuster (1989) J. Biol. Chem. 264 5503-5509)中有详细说明.美国Promega公司的pGEX载体系统也是可选系统之一.如果使用酵母系统，啤酒酵母系统中与甲醇酵母表达系统均有商用系统.美国 Invitrogen公司的甲醇酵母表达系统配件与工业放大技术完善，而啤酒酵母系统在文献 Ausubel et al. (supra)and Grant et al. (1987)Methods Enzymol. 153 :516_544 中有详 细说明.有的靶蛋白的表达量达到公斤级时，为减少长期生产成本也许选用转基因植物系 统来表达靶蛋白.这一系统中常用的启动子为CaMV的35S和19S基因启动子.同时配 备烟草花叶病毒(TMV)Omega的分泌信号肽.这一技术系统在文献Takamatsu，N. (1987) EMBO J.6 :307-311中有详细说明.其它启动子系统，如RUBISC0启动子，热休克蛋白启 动子等在文献 Coruzzi，G.et al. (1984)EMBO J. 3 1671-1680 ；Broglie, R. et al. (1984) Science 224 838-843 ；and Winter, J. et al. (1991)Results Prob1. Cell Differ. 17 85-105)中有详细说明.而具体的蛋白表达之操作规程在Hobbs，S. or Murry, L. Ε. in McGraw HillYearbook of Science and Technology(1992)McGraw Hill, New York, N. Y.； pp. 191-196文献中有详细说明.蚕螽病毒(baculovirus)蛋白表达系统也为教常用的工程菌系统之一.其表达载 体，细胞株均可从美国Invitrogen公司购置.至于哺乳细胞蛋白表达系统，其技术成熟同时有许多商用系统可选择使用.如 常用的启动子有CMV，B-ACTIN等，常用的有筛选基因有G418，ZEOSIN等，常用的载体有 SV40，腺病毒载体，逆转录病毒载体等.这类系统在重组蛋白表达中的应用在文献Dachs et al. Oncology Research, Vol. 9，pp :313_325，1997 中有详细说明·
不管在哪种系统表达，常用的基因工程重组技术中的有关技术细节是需要注意 的如靶基因的转译码ATG上游的Kozak序列有助于基因在细胞中的高表达，有些蛋白在 分泌表达时选用抗体蛋白IgG的分泌信号肽有助于获得高产量.在表达载体中使用合适的 增强子有能有效提高重组蛋白表达量.这类技术在文献Scharf，D. et al. (1994)Results Prob 1. Cell Differ. 20 125-162 中有详细说明·而表达细胞系的选择目前有许多文献有这方面的专门报导.如常用的细胞系 CHO, COS, HeLa, MDCK, HEK293，and W138，列等有成熟的商用系统，其覆盖从细胞培养液的选 择，稳定克隆之筛选.细胞发酵罐技术等都有专著.当然，由于靶蛋白的糖基化修饰往往影 响其免疫原性，在表达宿主细胞选择时，其糖基化能力为考虑因素之一.因本专利所选用 的靶蛋白为肿瘤疫苗发展之用，选用强免疫原性蛋白形式为重要的选择因素之一.稳定的高表达克隆之转染，筛选技术在下列文献中有专门报道如TK基因的使 用:ffigler, Μ. et al. (1977)Cell 11 :223_32·如 adenine phosphoribosyltransferase 基因的选用(Lowy, I.et al. (1990)Cell 22:817-23).常用的筛选基因DHFR的使用常 规=Wigler, M. et al. (1980) Proc. Natl. Acad. Sci. 77 :3567_70)；新霉素和 G-418 的选用 (CoIbere-Garapin, F. et al (1981) J. Mol. Biol. 150 1-14)等·同时使用标记基因共转染 也为常见技术之一，如 beta-glucuronidase 和 Iuciferase 等在 Rhodes, C. A. et al. (1995) Methods Mol. Biol. 55 121-131.文献中有详细说明.稳定的高表达克隆之转染技术可使 用美国INVITR0GEN公司的lipofectamin，(Cat#18324_lll)和Promega公司的磷酸钙沉淀 ^去i式齐Lli ( ProFectione Mammalian Transfection System, Cat#E1200)有关稳定的高表达克隆之鉴定技术也更为成熟.通常可用靶基因的探针来测 定高表达克隆中表达载体插入染色体的基因数目，也可直接测定细胞上清中靶蛋白质的 量.这类测定可用靶蛋白特异的抗体构建的免疫酶联法enzyme-linked immunosorbent assay (ELISA),发身寸免发radioimmunoassay (RIA),和焚光标记法fluorescence activated cell sorting (FACS)等.这类常规技术在以下文献中有详细说明.Hampton，R. et al. (1990 ；Serological Methods, a Laboratory Manual, APS Press, St Paul. Minn.)和 Maddox, D. E. et al. (1983 ;J. Exp. Med. 158 1211-1216).本发明的第三部分重点阐述依据靶蛋白而生产的特异抗体的技术路线和应 用.本发明中有的涉及专业词汇如“结合蛋白体，抗体，结合物”等专指能特异结合靶蛋白 的特异结合蛋白，或蛋白多肽片段等.这种结合均指非共价的高亲和性结合，多以“结合常 数，分离常数”等来描述(Davies et al. (1990)Ann. Rev. Biochem. 59 439-473).这类技术 也在 Davies et al. (1990) Annual Rev. Biochem. 59 :439_473 等文献中有详细说明.抗体 的抗原结合部常指抗体中Fab部分，有轻，重链之分.而我们描述的这类“结合蛋白体，抗 体，结合物”专指功能上能检测出肿瘤相关抗原的结合物，其临床的使用中至少能在某种类 型的肿瘤中检测出> 20%以上的患者，其在正常人群中的假阳性率在10%以下才具一定 的使用价值.而其待检样品可以是血液样品，尿液，肿瘤组织等.最近的文献中，RNA分子 也能发展成蛋白结合物，基本的抗体有关用法的知识在Harlow and Lane, Antibodies =A Laboratory Manual, Cold Spring HarborLaboratory, 1988中有详细描述。
至于靶蛋白特异性的抗体生产其技术路线有多种，如兔的多肽免疫产生的多克隆 抗体，鼠和兔的单克隆抗体技术，噬菌体抗体展示技术等.鼠的单克隆抗体技术
早在 1976 被 Kohler and Milstein，Eur. J. Immunol. 6 :511_519，1976 发表.兔 的单克隆抗体技术在近几年才逐步发展起来，美国Epitomic公司提供客户技术服务.鼠 的单克隆抗体技术在以下文献中有更一步的报道=Inbar et al. (1972)Proc. Nat. Acad. Sci.USA 69 2659-2662 ；Hochman et al. (1976)Biochem 15 2706-2710 ；a nd Ehrlich et al. (1980)Biochem 19 4091-4096.有关在获得特异靶蛋白的稳定的鼠单克隆抗体细胞株，或从噬菌体抗体展 示技术中克隆到抗体基因，都可用单链抗体技术制成单链抗体产品(single chain Fv ‘‘ sFv")，其连接抗体轻，重链的常用绞连多肽在文献Huston et al. (1988) Proc. Nat. Acad. Sci. USA 85(16) :5879_5883中有详细描述.而如何提高单链抗体对靶抗原的亲 和性技术路线在 U. S. Pat. Nos. 5，091，513 和 5，132，405，以及 U. S. Pat. No. 4，946，778，等专 利文献中有详细指导.在获得特异的抗体基因后，如需要在临床上使用，其可经鼠单克隆抗体人 in ^ ^ IS ^ J^ (Kettleborough1 et al. Protein Engineering vol. 4 no. 7pp. 773-783,1991).这一技术中重要的抗体蛋白氨基酸Kabat资料库在Kabat et al. , in Sequences of Proteins of Immunological Interest,4th ed. , (U.S. Dept. of Health and Human Services, U. S. Government Printing Office, 1987)文献中有详细描 述·如靶蛋白为肿瘤细胞表面的膜蛋白，其特异性高亲和抗体就有可能开发成治疗 性抗体.因此，治疗性抗体蛋白可标记上放射性同位素如9°I，123I，125I，131I，186Re, 188Re, 211At, and 212Bi,或各类毒性蛋白而达到其治疗肿瘤之疗效(U. S. Pat. No. 4, 735, 792.和 U. S. Pat. No. 4，673，562).常见的毒素蛋白亚基包括ricin，abrin, diptheria toxin, cholera toxin, gelonin, Pseudomonas exotoxin, Shigellatoxin, and pokeweed antiviral protein.小分子的抗肿瘤药物也可挂在抗体上而增家抗肿瘤抗体杀瘤之疗 效.如Rodwell得专利中有实验步骤之描述U. S. Pat. No. 4,671,958.以下参考专利更进 一步阐明新的抗体与其他分子绞联的技术=U. S. Pat. No. 4，489，710，to Spitler. U. S. Pat. No. 4, 625, 014, to Senter et al. U. S. Pat. No. 4, 638, 045, to Kohn et al. . U. S. Pat. No. 4, 671, 958, to Rodwellet al. . U. S. Pat. No. 4, 569, 789, to Blattler et al.在抗肿瘤的免疫治疗中，激活T细胞是其中的重要一个环节.这类步骤既可是 体内也可是体外的免疫激活.T细胞可从骨髓中，外周血中获得.许多商用系统如美国加 洲Nexell Therapeutics公司的Isolex. TM.系统.其它实验相关技术细节在U. S. Pat. No. 5, 240, 856 ；U. S. Pat. No. 5, 215, 926 ；W089/06280 ；WO 91/16116and WO 92/07243 等专 利中也有说明.新的iPS干细胞技术也为诱导分化获得T细胞开辟了新的技术路线(Ariff et al. Journal of Cellular Biochemistry. Jan 2009，Vol. 105，No. 6 1352—1360).本组肿瘤相关性靶抗原对免疫细胞的激活可从几类技术方案中进行1)用全长 的靶抗原或其多肽直接激活T细胞.也可用表达质粒携带靶抗原cDNA转化入抗原展示细 胞(antigen presenting cell :APC)，树突状细胞(dentritic Cells)肌细胞中来表达靶 抗原而激活T细胞.这类基因介导的免疫技术为临床试验中较为成熟的技术(Diebold et al, Human Gene Therapy. March 20，1999，10 (5) :775_786·).当然，用全长的靶抗原或其多 肽直接激活T细胞时还可使用微粒载体等来加强其免疫活性.
在测定免疫细胞是否为靶抗原或其多肽直接激活时，可用许多方法来测定T细胞所分泌的细胞介素水平来判断其功能.如裂解靶细胞来测定细胞chromium release assay 或细胞增值法(proliferation assay)以对照细胞为基值来判断.此类技术在文献Chen et al.，Cancer Res. 54 :1065_1070，1994 中有详细描述.用放射标记的H3_thymidine掺入 法在靶抗原或其多肽直接激活时测定T细胞的细胞增殖也是常用的技术Coligan et al., Current Protocols in Immunology, vol. l,ffiley Interscience (1998).这类 T 细胞既可 是⑶4也可是⑶8细胞.临床应用时，免疫细胞在靶抗原或其多肽直接激活后，(往往也加入细胞介素如 IL2，GMCSF, IL12等)会有体内或体外增殖的过程，然后给肿瘤患者使用.T细胞重要的受体(TCR)大致分二类alpha和beta链，经二硫键而共价相连.其 结构在文献 Janeway, Travers, ffalport. Immunobiology. Fourth Ed. , 148-159. Elsevier ScienceLtd/Garland Publishing. 1999.中有详细描述.Alpha 和 beta 链一起与 CD3 蛋 白形成复合物来特异识别结合于MHC上的靶抗原多肽.T细胞受体在免疫分子的刺激下 经类似抗体基因一样的基因重组而形成众多分子类型而展显出对靶抗原多肽不同的亲和 性.这类基石出知识在文献 Janeway, Travers, ffalport. Immunobiology. Fourth Ed. , 98and 150. Elsevier Science Ltd/Garland Publishing. 1999 中有详细描述.利用本组肿瘤相 关抗原刺激T细胞后，可筛选，克隆而获得高亲和性的TCR基因.然后可以V-J orV-D-J 等基因重组形式而转入肿瘤患者的T细胞中.在组肿瘤相关抗原刺激后，这类T细胞会 高效结合MHC而产生很强的免疫激活性.有报道表明利用TCR的类似物也能产生相同之 效果(A Sette et al. AnnualReview of Immunology. Vol. 12 413-431. 1994).在靴基因 的细胞转入中，表达载体可采取许多形式TAA蛋白基因与特异TCR直接融合表达或二基 因之间加入 IRES (internal ribosome entry site)序列来分开表达等.Rolland，Crit. Rev. Therap. Drug Carrier Systems 15 :143-198，1998.有关表达载体系统有裸DNA 直接 注射，基因枪注射，腺病毒载体，逆转录病毒，腺病毒相关病毒载体AAV (adeno-associated virus)，牛豆病毒，禽痘病毒等衍生的表达载体.各系统之特点在以下文献中有详细说明 Ulmer et al. ,Science 259 :1745_1749，1993. (Rich et al. (1993)Human Gene Therapy4 461-476).禾口 Carter， B. J. (1992)Current Opinion in Biotechnology 3:533—539. Elroy-Stein andMoss, Proc.Natl.Acad.Sci.USA(1990)87 6743-6747 ；Fuerst et al. Proc. Natl. Acad. Sci. USA (1986) 83 :8122_8126. U. S. Pat. Nos. 5，843，723 ；6, 015，686 ； 6，008，035 和 6，015，694. U. S. Pat. Nos. 5，846，796 ；6，010，478 ；5，865，796 ；5，584，807 ；和 EP Patent No. 0500799.综上所述，用本发明中发现的肿瘤相关蛋白来发展“特异肿瘤疫苗”已在上述各小 结中予以阐述.经基因克隆，基因导入，T细胞刺激，激活细胞之扩增，TCR的使用等技术路 线的说明，已使具常规医学，生物学之人士能开发，使用本技术.有关更详细的肿瘤疫苗的 具体技术操作，基因转导在肿瘤疫苗中应用等总结文献见M. F. Powell and Μ. J. Newman, eds. , " Vaccine Design (the subunit and adjuvantapproach), " Plenum Press(NY, 1995). U. S. Pat. No. 5，219，740 ；Miller and Rosman(1989)BioTechniques 7:980-990.佐剂的使用在肿瘤疫苗发展中至关重要，也为本发明详细描述重点之一.而 免疫增强剂是泛指所有能增加免疫反应的任何试剂.许多佐剂的功能是让抗原分子缓释如氢氧化铝aluminum hydroxide和矿物油(mineral oil)，或非特异的增加免疫剌激，如磷脂A(lipid A)，鲍特杆菌蛋白(Bortadella pertussis)和结核杆菌蛋白 (Mycobacterium tuberculosis derived proteins).许多生产厂家有医用佐剂供应 如美国 Difco Laboratories 公司的 Freund’ s Incomplete Adjuvant and Complete Adjuvant (DifcoLaboratories, Detroit, Mich.)；美国 Merck 公司的 Adjuvant 65.美国 SmithKline Beecham 公司的 AS—2.其他产品如氧氧化招胶(aluminum hydroxide gel) (alum)or 憐酸招(aluminum phosphate) ； 丐盐(salts of calcium)，铁或锋(iron or zinc)；不溶的乙酉先化的酪胺酸(an insoluble suspension ofacylated tyrosine)；乙 酰化糖(acylated sugars)；阳离子化或阴离子化的多糖(cationically oranionically derivatized polysaccharides) ； M Μ M (polyphosphazenes)；生物 P拳 M t生 W f立 (biodegradable microspheres)；单憐酸类月旨 A(monophosphoryl lipid A) and quil A 也 为常用免疫增强剂.细胞介素，如GM-CSF，白介素2，白介素7，白介素12，等也在临床使用.因本发明重点在使用肿瘤相关抗原刺激患者自身免疫系统来达到预防与治疗效 果，我们选用免疫刺激时优先激活Thl细胞反应.高水平的Thl细胞反应将产生细胞介素 如干扰素gamma.，肿瘤坏死因子alpha (TNF. alpha).，白介素2，白介素12，其有增强体液介 导性免疫反应.当然，在肿瘤相关抗原刺激为肿瘤疫苗治疗的过程中，Thl和Th2细胞反应 都会增强.有关细胞介素的使用请阅读文献Mosmann and Coffman, Ann. Rev. Immunol. 7 145-173，1989.有的佐剂是优选刺激Thl细胞反应的，如佐剂加有单磷酸类脂A与aluminum salt 混用.在文献 U. S. Pat. Nos. 4，436，727 ；4，877，611 ；4，866，034 和 4，912，094 中有 详细说明.非甲基化CpG多核苷酸作为佐剂使用也是优选刺激Thl细胞反应.这在专利 文献中如 WO 96/02555, WO 99/33488and U. S. Pat. Nos. 6, 008, 200and 5，856，462 中也有 详细阐述.美国Aquila Biopharmaceuticals公司出售的Quil A衍生物QS21and QS7 也有刺激Thl细胞反应的效果.在佐剂混合物中加入各种微粒如壳聚糖，刺蒺藜总皂苷 和其它多聚体，聚阳离子型高分子材料polylactide and polylactide-co-glycolide particles, poly-N-acetyl glucosamine-based polymer matrix, particles composed of polysaccharides orchemicalIy modified polysaccharides, liposomes and lipid-based particles, particles composedof glycerol monoesters,等·刺漠薬总阜 苷(saponins)也可和脂质体如ISCOMs混合使用.还可加入Carbopol. RTM来增加佐剂的 粘连度(viscosity).故本专利的肿瘤疫苗的首选佐剂仿是单磷酸类脂A，刺蒺藜总皂苷 类的QS21和3D-MPL. RTM混合物为核心试剂.这与文献WO 94/00153中的方法大同小异.更多的佐剂试剂呈例在这可供选择法国S印pic公司Montanide ISA 720产 品.美国Chiron公司的SAF，ISCOMS, MF-59等产品，美国SmithKline Beecham公司的 SBAS-2or SBAS-4等产品.美国Corixa公司的RC-529产品.在专利文献中还有更多的有 关新型佐剂试剂的阐述U. S. patent application Ser. Nos. 08/853, 826and 09/074, 720, 和 WO 99/52549A1.本发明的特异肿瘤疫苗发展中细胞类型的选用以抗原递呈细胞(antigen presenting cells (APCs)的刺激为优先，这包括树突状细胞(dendritic cells)，巨噬细 胞，B细胞，单核细胞等.其他类型的细胞也有可能经处理而具备抗原递呈能力，如导入TCR基因，让HLA配型合适等技术都可用于抗原递呈细胞的生产.至于抗原递呈细胞的来源，即 可是自体，也可用异体材料.取材途径包括各种体液，器官组织等.在各类可选用的抗原递呈细胞中，树突状细胞是最理想的细胞类型.这在文献 (Banchereau andSteinman, Nature 392:245-251,1998)中有较为详细的阐述.而且配 合以上所选用的THl佐剂配方有很好的免疫刺激效果.(Timmerman and Levy, Ann. Rev. Med. 50 =507-529,1999). 一般来说，树突状细胞可从形态学上来鉴定，也可以其对颗粒物 的吞噬(take up)能力来分离.其功能往往以对特异的T细胞群的刺激能力来计量.利用 转基因TCR等技术来使肿瘤患者树突状细胞具备对特异肿瘤抗原的刺激反应能力也是本 专利指出的技术路线之一.这类技术的细节在Zitvogel et al. ,Nature Med. 4 594-600， 1998文章中有更为详细的阐述. 树突状细胞的来源可取之外周血，骨髓，入侵肿瘤组织的免疫细胞，肿瘤外周组织 中的免疫细胞，淋巴结，脾.皮肤组织，或脐带血等.从外周血中分离的单核细胞在体外细 胞培养体系中经细胞介素GM-CSF，IL-4，IL-13或TNF. alpha处理即可诱导分化成树突状 细胞.而从脐带血，骨髓中分离的CD34阳性细胞也可经GM-CSF，IL_3，TNF. alpha.，CD401 igand，LPS，flt3底物(Iigand)而体外处理来诱导分化成树突状细胞.一般来说，树突状细 胞被粗分成“成熟型和非成熟型”二种非成熟型往往指其细胞仍有很强的吞噬(take up) 能力，而且在细胞膜上仍有较高的Fe. gamma受体表达.而成熟型树突状细胞是指MHC I和 II型分子表达高的⑶54，⑶11和CD40，⑶80，⑶86及4-1BB受体蛋白都较高表达的细胞 群·在分离，纯化出树突状细胞后，应用常规分子生物技术将本专利中所描述的20种 肿瘤相关基因导入树突状细胞便为可行.这类技术在以下文献中有较为详细的阐述专利 WO 97/24447. Mahvi et al. , Immunology and cell Biology 75 :456_460，1997.常用的技 术是脂质体(Liposomes)转化技术.脂质体用与T细胞，肝细胞，细胞株PC 12中的具体应 用在 Renneisen et al.，J Biol. Chem. 1990S印· 25 ;265 (27) 16337-42 ；Muller et al.， DNA Cell Biol. 1990 April ；9(3) :221_9)等文章中有更为详细的阐述.至于免疫激活后的树突状细胞治疗使用途经，既可肌肉注射，静脉注射，也可 皮内，皮下，大网内等途经用药.这类技术在以下文献中有较为详细的阐述u. S. Pat. No. 5，543，158 ；U. S. Pat. No. 5，641，515and U. S. Pat. No. 5，399，363用药制剂中的部分免疫刺激微粒成份往往是生物降解类物质.这类组分在 免疫类药物中有常规制剂如 microparticles of poly (lactide-co-glycolide)， polyacrylate, latex, starch, cellulose, dextran ^ . S^Ll Wiiffl #^lJ ；KΦ 如:U. S. Pat. Nos. 4，897，268 ；5，075，109 ；5，928，647 ；5，811，128 ；5，820，883 ；5，853，763 ； 5，814，344，5，407，609和5，942，252中又细述.有时为达到缓释效果还在制剂中加入超 大分子物如绞联的多糖物，或使用不同磷脂类物质(phospholipid).以下文献对这类技术 有详细阐述 U. S. Pat. No. 5，151，254 and PCT applications WO 94/20078, W0/94/23701 and WO 96/06638).有关肿瘤疫苗制剂中的或使用不同磷脂类物质的使用也是常规知 识，这类技术系统在以下文献中有详细报道Lasic，Trends Biotechnol 1998 July ； 16(7) :307-21. Takakura, Nippon Rinsho 1998March ；56 (3) 691-5 ；Chandran et al., Indian J Exp Biol. 1997 August ；35 (8) :801_9 ；Margalit, Crit RevTher Drug CarrierSyst. 1995 ；12 (2-3) :233_61 ；U. S. Pat. No. 5，567，434 ；U. S. Pat. No. 5，552，157 ；U. S. Pat. No. 5，565，213 ；U. S. Pat. No. 5，738，868and U. S. Pat. No. 5，795，587.有关肿瘤疫苗制剂中的其它成分如各类缓冲液等，既可用生理盐水，也可用 PBS (phosphate bufferedsaline)液.常用的添加剂如葡萄糖，甘露糖，蔗糖，葡聚糖，甘 露醇，甘氨酸，抗氧化剂(antioxidants)，抑菌剂(bacteriostats)，螯合剂(chelating agents) EDTA或谷胱甘肽(glutathione)等.制剂类型既可是水溶液也可是冻干制剂.包 装大小可控制在一瓶一次注射剂量或一瓶中含多次注射剂量.肿瘤的生长往往是患者体内的免疫监视功能不足所致，而肿瘤的免疫治疗是基于调动患者体内自有的免疫功能来识别肿瘤细胞上的靶抗原而抑制其生长.这类基 础知识在 Jager, et al. , Oncology 2001 ；60(1) 1~7 ；Renner, et al. , Ann Hematol 2000December ；79 (12) :651_9等文献中有详细报道.目前已知体内至少有四类功能细胞 与人体抗肿瘤活动有关1) B淋巴细胞，分泌抗体来特异识别肿瘤细胞.2)单核细胞，产生 补体等直接杀死靶细胞.3)中性粒细胞(natural killer lymphocytes)，可利用特异抗体 结合肿瘤细胞后来攻击靶细胞或释放致死因子直接产生补体等直接杀死靶细胞(natural killing)，4) T细胞，处理肿瘤特异抗原而后递呈给其它免疫攻击细胞来杀伤瘤细胞，或结 合那些表面带有补体的瘤细胞· (Schreiber, H.，1989，in Fundamental Immunology (ed). W. Ε. Paul, pp. 923-955).虽然体液免疫和细胞介导的免疫都对肿瘤抑制至关重要，但实验 结果表明CD4T细胞的激活对诱导机体产生高特异抗肿瘤的抗体有关键的调节作用.而蛋 白类肿瘤相关抗原是极理想的肿瘤靶抗原而用于治疗.所以本专利中所描述的20种肿瘤 特异相关蛋白(TAA)为肿瘤治疗提供了具体内容.本组TAA主要使用在人的肿瘤治疗中使 用，它即提供了预防性治疗的可能，也为肿瘤切除术后作为肿瘤疫苗使用来阻止肿瘤复发 而提供手段.而TAA的使用，既可是蛋白形式作为抗原，也可以DNA形式来经基因转导而起 肿瘤疫苗之用.在治疗时，疗程往往是数周8数月如每月一针肿瘤疫苗，连续6针为一疗 程.而较为理想的治疗方案是同类肿瘤患者中有大于10%的病人有良好的抗肿瘤反应，能 达50%以上为理想系统.重组靶蛋白的免疫使用剂量可从病人每公斤体重注射25毫微克 (25ug)至5微克((5mg)蛋白不等.而制剂体积也从0. 1毫升到5毫升不等.以获得的TAA来刺激患者自身免疫系统的方法已在例五至例七部分有叙述.作为 被动免疫的技术之一如抗肿瘤独特型抗体(anti-idiotypic antibodies)的使用在专利文 献U. S. Pat. No. 4，918，164)有详细报道.用抗肿瘤特异性抗体来进行肿瘤诊断，治疗的成 功经验在文献中比比皆是，U. S. Pat. Nos. 6，090，365 ；6，015，542 ；5，843，398 ；5，595，721 ； and 4，708，930.抗肿瘤抗体或是标记有杀伤性同位素，或是融合表达有毒性蛋白部分.以 下专利文献的解释既围绕这一部分0ng etal, Cancer Immunol Immunother. Nov ；39 (5) 325-31,1994. Kreitman R. J. Immunotoxins. ExpertOpinion on Pharmacotherapy, Vol 1, Num 6，，pp.1117-1129. 2000.在肿瘤患者中查出肿瘤抗原是典型的肿瘤早期诊断方法之一，如肝癌中的AFP蛋 白和前列腺癌中的PSA等均为蛋白抗原.这类肿瘤蛋白抗原可从血液，尿液，肿瘤组织样品 中测出.故定量分析某些TAA为早期诊断肿瘤提供了技术可能性.测定靶基因的mRNA水平大部分临床试验中的TAA因其在正常与肿瘤组织中的表 达量不一样而成为所谓的肿瘤抗原，所以定量过程中以肿瘤组织中表达高于2倍于对照组织而成为“高表达”.本专利中有所发现的肿瘤抗原是依据其仅在早期胚胎组织中表达而获 得，其在正常组织中一般完全不表达，为测定靶基因的mRNA水平提供了准确性.近期文献中报导在肿瘤患者的外周血中有循环的肿瘤细胞存在.单用形态学指标往往很难发现于定量这类循环肿瘤细胞.而本组肿瘤特异性基因和蛋白的发现为 定量，定性这类循环肿瘤细胞提供了可能.(Muller et al. Breast Cancer Res. 8(5) 110. 2006.)用PCR技术对待测的靶基因的mRNA水平定量，用ELISA方法对肿瘤抗原测定 等已在本文中有描述.而本文试验案例中对PCR使用的DNA引物的序列，制备特异抗体而 使用的靶蛋白多肽序列等均有详细描述.而Sackett et al. , Clinical Epidemiology ABasic Science for Clinical Medicine, Little Brown and Co. , 1985, p. 106-107,等专 著对监测结果之分析也有阐述.实时PCR(real-time PCR)也是临床试验中常用的监测靶基因的mRNA水平的技术 系统之一，在本文的试验案例中提供有所有20种靶基因的实时PCR使用的DNA引物的序列 与在人结直肠癌肿瘤样品定量分析之结果.(图11).实时PCR技术结合抗体-磁珠抓获靶 细胞技术为许多血液肿瘤提供了方便快捷的方法.这类磁珠抓获靶细胞技术在以下几个 公司中可买到Dynal磁珠，美国Dynal Biotech公司，StemS印磁珠.(加拿大StemCell Technologies，公司)，和 RosetteS印磁珠(美国 StemCell Technologies 公司)·这类靴抗 体_磁珠系统常常挂有⑶抗体，如下例抗体都有商用产品⑶2，⑶3，⑶4，⑶5，⑶8，⑶10， CDllb, CD14, CD15, CD16, CD19, CD20, CD24, CD25, CD29, CD33, CD34, CD36, CD38, CD41，CD45， CD45RA, CD45R0, CD56, CD66B, CD66e, HLA-DR，IgE, and TCR. alpha. · beta..从靶细胞抓获， 细胞裂解，RNA快纯化，RT反应及PCR都可从QIAGEN公司购买带商用试剂盒.这类试剂也 已批准在临床试验中应用.(QIAamp DNABlood Maxi Kit 50, Cat#51194).虽然实时PCR(real-time PCR)能准确测定靶基因表达水平，但因操作是一种基因 一个PCR样品来进行，有时有样品之间的读数误差.为克服这一难题，本专利更进一步展示 出新技术GeXP定量测定技术.在这一技术系统中，20种靶基因的DNA特异引物与4种看家 (Housekeeping)基因在单一 PCR试管中进行反应，使定量测定靶基因在正常于肿瘤组织中 表达水平更准确.为指导下一步的肿瘤疫苗提供正确信息.这一 GeXP定量测定技术的具 体应用在试验实例(六)中有详细说明.本试验实例部分的叙述仅为提供部分试验手段作为应用样板，依据常规的基因工 程，细胞学，免疫学，肿瘤学等技术方法可经不同技术系统而取得同类试验结果.故本发明 中20种人肿瘤特异性相关抗原在肿瘤诊断，治疗的应用不局限于本试验的实例中.实施例1、鉴定人正常组织中基因启动子DNA甲基化之靶基因人基因组中基因启动子DNA甲基化是组织分化过程中重要的基因表达调控步骤 之一，在肿瘤发病早期，许多重要的组织分化相关的基因，因肿瘤细胞的甲基化调控失调， 而由原静默状态而成为活跃表达，人AFP(肝中甲胎球蛋白)就是很好的例子.本实验旨 在获得那些人基因组中基因启动子其在正常情况被高度甲基化，而在肿瘤细胞中有可能是 去甲基化状态.这一信息为寻找肿瘤细胞中基因去甲基化提供了基础.本实验使用美国 AVIVA SYSTEMS BIOLOGY (ASB)公司CHIP_DSL技术系统.本技术系统已在2006年PNAS中 发表(March 20，Vol. 104. 2007，PNAS pp :4852_4857)，可用购买的ASB公司商用试剂盒进 行·
本实验结合特异抗胞嘧啶甲基化单克隆抗体进行人组长的，纯化后染色体DNA抗 体免疫沉淀技术与基因启动子芯片技术相结合而进行.IA 染色体DNA抗体免疫沉淀(ChIP Assays)与DNA产物扩增准备以下实验以美国ASB公司CHIP_DSL技术系统操作说明书为基本实验步骤之指导.试剂特异抗胞嘧啶甲基化单克隆抗体(5-Methylcytosine Antibody)为Eurogentec公司产品，(目录号BI-MECY_0100) · 20K人基因启动子芯片为AVIVA产品(目 录号AK0504).正常多种人体组织基因组DNA来源于脑，肝，肾，胎盘，肺，胃，小肠，直肠结 肠，胰腺，胎儿胸腺，肌肉，单个核细胞，将这些组织均勻研磨后放入消化缓冲液(IOOmM氯 化钠，IOmM TrisCl, pH值为8. 0，25mM四乙酸，pH值8.0，0.5% SDS)和0. 1毫克/毫升蛋 白酶K中，50°C放置12-18小时，乙醇-异丙醇法提纯。提取的DNA经超声裂解后成为长度 为500-800bp的片断.10微克基因组DNA先在95°C变性10分钟，迅速在冰上冷却，然后加 入Iug单克隆抗体IgG并同时加入20微升蛋白A-磁珠和50微升PBS溶液后在37°C保温 60分钟.最后用1毫升PBS溶液经离心法洗4次.重悬磁珠样品于50微升TE溶液中.经 抗体沉淀的样品甲基化DNA样品在95°C变性3分钟，经离心去除磁珠，保留上清中DNA样
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BFI ·沉淀的DNA样品(约200-300ng DNA)按CHIP_DSL试剂盒中引物指导的DNA连接 反应完成DSL反应，最后一步用荧光标记的T7引物(Cy3和Cy5)标记扩增，经QIAquick PCR 扩增纯化试剂盒(Qiagen公司产品)纯化出的DNA样品溶解在50 μ L的水中。经95°C变性 5分钟后加入至20K人启动子芯片(见详细的制备说明书)于50°C保温过夜。杂交后的芯 片按制备说明书经离心洗片后再经离心干燥，用GenePix 4000B扫描仪(Axon设备)扫描。 用Axon提取软件从寡核苷酸阵列扫描图象中得到荧光强度数值，对于每个位置上的数组， Cy5标记试验样品的log2比率和Cy3标记的参考样品进行比较，然后，log2比率减去各点， 而进行每个通道正常化矫正。Cy5标记的芯片样品为抗体沉淀的样品甲基化DNA，而Cy3标 记是来源于未经抗体处理的对照基因组DNA.IB 数据分析使用GenePix 5000扫描仪与GenePixPro 5. 1软件分析Cy5和Cy3荧光强度。分 析中去掉标记的异常点(例如，那些污染散斑，碎片或杂交中产生的杂带)本实验使用常用的芯片结果分析软件包R. (http//www. R-proiect. org)为基础 以 Erroe 模型进行分析.本方法发表在(Nature, 2005. Aur 11:436(7052) 876~80. Epub 2005Tun 29.) .Erroe模型分析可简述如下对所有的扫描数据。根据P值的检测结果，先 将结果正常化.(以阴性对照的计算得出的结果为参量)。检测P值计算公式为1的R/n， 其中R是指基因信号与阴性对照的相对值，N是阴性对照的数量。检测P值可以解释为在 没有特异性目标探针杂交时检测到的一定的信号水平。低表达的信号(LEGs)我们设定了 P 值的阈值有几挡0. 05(0. 05彡？值< 1)，中等表达(MEGs)我们设定P值阈值是大于0，但 “小于0. 05(0 < P值< 0. 05)高表达(HEGs)的P值等于0。图1为人脑源DNA CHIP_DSL 实验结果扫描图.AVIVA人20K的启动子芯片阵列中使用了每一个基因启动子区以单一 DNA引物 来代表的阵列设计.其包含在美国NCBI RefSeq数据库中的全部18560特异性基因，其中 2912个基因因其具备双启动子区而选定设计有2个启动子DNA引物。每个芯片点阵的确切的设计详见ASB芯片制备说明书。芯片结果分析数据按点回归至每一基因的启动子区，而 得知其甲基化程度.本实验一共获得常见的在多种正常组织中其基因的启动子区高甲基 化的靶基因约721个.实施例2、鉴定肿瘤特异性表达基因之实验。因本发明志在获得肿瘤特异性靶基因，故实验的第二步集中在寻找在正常组织中 基本不表达而在肿瘤组织中高表达的基因.据实验方案一中介绍，在许多肿瘤组织细胞中因其甲基化调控有失调的现象，许多早期胚胎阶段特异基因却在成人肿瘤组织细胞中异常高表达.这类基因往往在正常组 织中关闭良好而一点也不表达.据此理论，实验的第二步集中于胚胎早期特异表达基因的 鉴定.因人体中生殖细胞最接近胚胎早期状态，故使用多种人组织与人生殖细胞的mRNA的 特异性表达的芯片测定库结果开展对比来分析研究.本实验方法是以721个在实验方案一中所发现的基因启动子区高甲基化的靶 基因为起点，把每一基因放入GNF symAtlas多组织mRNA的特异性表达的芯片测定库中 (http://symatlas. gnf. org/SymAtlas/),寻求R在人睾丸牛殖细胞表汰之基因.以此获 的172个目标基因.此组基因仅在人睾丸生殖细胞表达，在正常组织中其mRNA表达几乎测 不出.为证明是这172个靶基因可能是肿瘤相关抗原，我们进一步通过symAtlas数据库继 续鉴定NCI 60种肿瘤细胞中这172个目标基因之表达情况.经此步筛选，这172个目标基 因在多种肿瘤细胞中均有不同程度的表达.图2为DNALIl靶基因在GNF芯片库中mRNA的 特异性表达代表性图.实施例3、肿瘤特异性表达基因蛋白靶抗体之生产为进一步证明肿瘤细胞中这172个目标基因确实在不同的肿瘤细胞中有相关的 蛋白之表达，我们进一步选出20种目标基因蛋白进行兔多克隆抗体生产.(图3中列有20 种目标基因名称和NCBI基因库基因序列号).其中14个靶蛋白以多肽合成获得其特异多 肽抗原，每一靶蛋白设计有二个特异多肽抗原.另6个靶蛋白以全长基因重组获得蛋白抗 原而进行靶蛋白特异抗体的生产.3A:靶蛋白的多肽合成14个靶蛋白的多肽合成在ACT公司Apex多肽合仪中ACT公司提供的操之指导手 册常规进行以Wang树脂为固相载体，改进Fmoc (芴甲氧羰基)/tBu法进行.Fmoc经HPTU 激活。特定多肽序列按序列以Wang树脂为固相载体开始反应，整个反应在一个固定的表面 进行。从固定的支持物上剪切的全长多肽经三氟乙酸无水乙醇茴香硫醚水苯酚 (40 1 2 2 3)。裂解后2小时后，最后多肽沉淀在冷的甲基叔丁基醚中。肽颗粒 溶解含0. 三氟乙酸的水中，用C18柱反相高效液相色谱法纯化并冻干。14个靶蛋白多 肽氨基酸序列见图4(每一靶蛋白设计有二个特异多肽抗原).3B =KLH载体蛋白之绞连匙孔戚血藍蛋白(KLH)为美国EMD公司产品(产品目录号#374805)，suIfo-SMCC 为美国SOLTEVentures公司产品(产品目录号#C L 207).本项目合成的多肽均在其C末端加有cys将提供出S H基团，在水溶性S MCC 的作用在与K L H蛋白表面的N H 2基团经maleimide反应而产生共价绞连.具体反应操 作如下取等量(3mg)K L H和待绞连的靶基因蛋白多肽(3mg)，加入50毫升烧杯中，然后缓慢加入经水溶解好的sulfo-SMCC约(1.8mg).室温搅拌30分钟而完成绞连反应.绞连 蛋白经Si^phadex G25柱分离收集.测定蛋白浓度以备动物免疫之用.3C:抗血清生产过程对于图4这14个TAA蛋白，从每一靶抗原独特的区域中选定了两个多肽进行合成，在连接到KLH后，冻存，直到免疫接种需要时使用。多克隆抗血清生产过程用400微克靶抗原多肽-KLH结合物(0. 5毫生)，加上等 量的完全弗氏佐剂(CFA)的混合，皮下注射((S. C.)成兔。4周后，400微克抗原多肽-KLH 结合物与等量的不完全弗氏佐剂(IFA)的混合皮下注射兔2次，每次间隔4周。最后一次 兔静脉注射100微克纯抗原KLH结合物。最后一次免疫7天后取兔血，将血放置4度孵育 12-24小时，离心后得到血清。经蛋白A柱纯化而获得兔IgG抗体.特异抗体效价之测定在96孔板中以50ul多肽包被4度过夜.多肽浓度通常是 lug/50ul.封闭非特异性反映是在每孔加入250ul BSA室温下孵育2小时.多克隆抗血清 50ul (通常是2ug/ul)经倍比稀释之1 20万(用PBS/0. 1 % Tween/0. 1 %牛血清白蛋白 稀释兔抗体)，加入孔中与抗原特异性结合，4度条件下用在96孔板孵育2小时。板洗按以 下条件进行含0. 吐温的PBS洗涤6次。然后每孔加入50ul 1 10，000稀释的山羊抗 兔辣根过氧化物酶(HRP)室温孵育30分钟而显色。加入IOOul的IN的H. sub. 2S0. sub. 4 终止反应，并立即在450nm左右波长下读数。各抗体的效价多在> 1 10000以上.3D 靶基因全长cDNA克隆制备在CHIP_DSL芯片结果分析后，数据按芯片上的点阵回归至每一基因的启动子区 而获得靶基因的基因库代号(accession^.然后依据靶基因的基因库代号而经美国NCBI 国立生物技术信息中心数据库数据库进行搜索而获得靶基因全长cDNA序列.依此靶基因 全长cDNA序列，经一步搜索美国M G C数据库数据库而获得靶基因的IMAGE克隆，以100% 的同源性为准.然后从美国Open Biosystems公司购买目标特异性IMAGE克隆，根据不同 的载体，用不同的D N A引物在ABI的D N A测序仪对每个克隆在纯化进一步验证其D N A 序列之正确性.对应的靶基因IMAGE克隆小结在图5中.3E:重组靶蛋白在大肠杆菌中的表达本实验选用6 个靶蛋白:C170RF39, CSNK2A2, DNALI1，GSGl，GSTM3，IL13RA2.进行
其重组蛋白的表达和纯化，用于抗体生产6个靶蛋白基因经P C R按常规技术进行全长编码基因人的扩增，具体D N A引物 见图6.靶蛋白基因的IMAGE克隆均从美国Open Biosystems公司购买经质粒载体DNA纯化 后为PCR反应之底物DNA，PC R片段经胶纯化后用内切酶NdeI-XhoI双酶切后插入pET41a 改造的表达载体中，在PET41载体重组表达系统T7启动子下端有MASMTGGQQMGRHHHHHH融 合在靶蛋白质的N-末端.在大肠杆菌高表达用BL21star细菌(Invitrogen公司产品)转化表达载体，力口 入LB液并以kanamycin抗生素为筛选在37度摇床过夜生长。第二天早上，10毫升过夜培 养的细菌液加入到500ml LB培养液中，kanamycin抗生素。摇菌至培养的光密度在560纳 米达到0.4-0. 6时，加入IPTG(ImM)诱导四个小时，最后离心收获重组菌。菌体经PBS液 离心洗涤细胞。丢弃上清。加入IOml裂解液并震荡重悬(含溶菌酶)。大肠杆菌细胞在 16，OOOpsi压力下裂解，重新离心细胞，离心管底为大肠杆菌重组包涵体蛋白.
将包涵体蛋白重悬在IOmM Tris pH 8. 0，1 % CHAPS液中重新离心洗涤包涵体。重 复这个过程2次。然后在样品蛋白液中慢慢加入终浓度8M固体尿素，经搅拌至包涵体完全 溶解。可溶性蛋白样品经5毫升镍螯合树脂(Qiagen公司His Tag column)中孵育45分 钟至1小时，室温连续搅拌。将树脂和蛋白质混合物通过一次性浇柱收集。用12-20倍体 积的缓冲溶液洗涤柱子(2M尿素P B S液).经IOmM的Tris pH值8. 0和300mM咪唑收集 3毫升组分。跑SDS-PAGE凝胶，以确定纯化蛋白之纯度。如蛋白不够纯化，最后的纯化步 骤，用阴离子交换树脂，Hi-Pr印Q(Biorad),与缓冲液和混合液一起装入柱子中。利用增长 的盐浓度洗脱每种抗原。收集各组分，跑SDS-PAGE凝胶，以确定各组分而获得纯化重组蛋 白。重组蛋白组分用IOmM Tris pH 8.0重新透析。最后用0. 22微米的过滤器过滤蛋白装 入安剖瓶中免疫接种或直接把抗原冻存备用。实施例4、用免疫印迹(Western blot)检测癌症样本。试剂各肿瘤细胞株均购置于美国ATCC，并按其细胞株培养条件进行细胞培养.人肝癌细胞株H印G2，(产品目录号HB-8065)，人T细胞肿瘤株Jurkat，(产品目录号 TIB-152).人乳腺癌细胞株MCF7 :(产品目录号HTB_22)人B细胞株721B，为美国THOMAS JEFFERSON医学院Feitelson教授之赠品.人畸胎瘤细胞NTERA2，(产品目录号CRL-1973)， 人宫颈癌细胞株Hela，(产品目录号CCL-2).人的正常组织如脑，心，肺，肝，横纹肌与胎 盘组织均提取于22周水囊引产的胎儿样品.蛋白样品制备5X IO7培养细胞或0. 4克的组织勻浆重悬与1毫升新鲜配置的 裂解液中(150mMNaCl，50mM Tris-HCl, pH7. 4, ImM EDTA, 1% NP_40，0. 5%脱氧胆酸，0. 1% SDS, ImM PMSF, lug/ml Leupeptin, Aprotinin, P印statin).混勻后加入等体积 2X 的 S D S-P A G E上样液(2XSDS Loading Buffer 0. 2M DTT,4% SDS,0. 2% Bromopheno 1 blue, 20% Sucrose or glycerol, 0. IM Tris_HCl，pH6. 8).进一步混勻后放至 95C 水浴中处理 5 分钟.经离心后取30ul样品上样进行蛋白电泳.蛋白电泳在8%聚丙烯酰胺电泳中按常规进行，分离蛋白后用Bio-RAd公司电转 移系统将蛋白转移到PVDF杂交膜上，经干奶粉/P B S液4°C封闭过夜后，然后通过一 抗(1 4000稀释(终浓度为lug/ml)，保温37°C，2小时.经0. 1 % NP-40/P B S液洗膜5 次，然后加入1 50，00稀释的H R P标记的羊抗兔二抗抗体，保温37°C，1小时，经0. NP-40/P B S液洗膜5次后将膜经E C L溶液(PIERCE公司产品，产品目录号#32160)处理 后夹入X光片中经显影，定影完成免疫印迹的检测.图8为靶蛋白UBQLN3和DNALIl在肿 瘤及正常组织中表达的情况.实施例5、实时PCR法分析特异性靶基因在多种肿瘤中的表达。实时PCR(Real-time PCR)分析肿瘤样本用实时PCR检测前列腺癌和乳腺肿瘤样 本在各种肿瘤和正常组织中的表达技术为一般常规技术。简言之，用PCR扩增几个周期后 的DNA高于背景的对数值来定量。利用连续荧光监测，每个PCR反应很容易确定循环次数下 的DNA扩增倍数的阈值。目前，一些商业荧光检测系统可供选择。一个是基于双链DNA的特 异性结合荧光染料染色。另外使用含有荧光定量探针染料标记在5’端(FAM)同时Quencher 染料在标记3’端(TAMRA) (Perkin Elmer/应用生物系统公司，福斯特城，加利福尼亚州)。 目标的PCR扩增产物从含有的核酸探针AmpliTaq Gold. TM. (Perkin Elmer/应用生物系统 公司，福斯特城，加利福尼亚州)的Quencher染料裂解和释放。因此，从释放的标记染料比例确定PCR产物产生的荧光信号。这两种检测方法，发现了相同的结果。要比较多种组织 样品中基因表达的相对水平，采用组织的RNA经逆转录生成cDNA，然后用靶基因特异性引 物对每个相同的cDNA量进行实时PCR。每个基因样本通常是在重复进行。最后每个基因的 实时PCR结果一般是平均拷贝数的基因值。实时PCR反应的定量结果可在用SYBR Green I 染色的GeneAmp 5700检测或用TaqMan探针(Perkin Elmer//应用生物系统公司司，福斯 特城，加利福尼亚州)的ABI PRISM公司7700探测器检测。试剂人结肠肿瘤组织的cDNA样品均购置于美国Origene公司，( 产品目录号#H CRT 101&HC R T 501，其包含有5例正常人结肠组织和19例人结肠肿瘤组织的cDNA样 品.靶基因D N A引物经美国I D T公司合成，其序列见图9 实时PCR反应用美国Bio-Rad iQ5探测器检测，同时使用iQ SYBRGreen Supermix(产品目录号170-8884)进行样品反 应·实时PCR分析人结肠肿瘤组织的CSNK2A2靶基因表达水平在0. 2ml离心管中按下 列操作程序进行0. 2ml离心管中加入12. 5ul iQ SYBR Green Supermix液，2ul人结肠肿 瘤组织cDNA样品，2ul(20pM/ul)有义引物和2ul (20pM/ul)反义引物，6. 5ul去离子水.振 荡样品管，然后短暂离心。将管放到预热的Bio-Rad iQ5探测器检测上开始P C R反应.按 下列程序开始循环预变性94°C 4分钟1次，变性94°C 1分钟，退火55°C 1分钟，延伸72°C 1 分钟，循环38次.本反应使用看家基因(house-ke印ing geneas control)GAPDH(甘油酸 脱氢酶)转录水平来校正待测靶基因表达水平.CSNK2A靶基因在实时PCR获得在各人结肠 肿瘤组织中的相对表达水平，结果总结于图10。实施例6、使用GeXP系统检测人类癌症组织。GeXP是一种新的，改良的多种靶D N A样品在同一试管中进行RT-PCR法(逆转录 聚合酶链式反应)的反应与定量分析.它可以用通用引物转换多种RT-PCR的两个引物，最 终产物能够通过现有的BeckmanCoulter公司的GenomeLab GeXP遗传分析系统进行检测。 这一检测方法消除连单个靶基因在单管中进行实时PCR法时的样品人为操作误差.为临床 检测多种靶基因表达水平的定量测定提供了可能.本实验意在证明使用GeXP技术结合20 种新型肿瘤靶基因之监测效果.试剂人多肿瘤组织的mRNA样品分别均购置于美国strategene公司，(产品目 录号740000，其包含有数种肿瘤细胞提取的mRNA样品.靶基因D N A引物经美国Allele Bioscience公司合成，其序列见图11 实时PCR反应用美国ABI公司Verit 96孔板P C R 机进行.反应样品反应用Beckman Coulter公司的GenomeLabGeXP遗传分析系统进行检 测.(http//www, beckman. com/products/instrument/geneticanalysis/Rexp instdcr. asp)根据GenomeLab GeXP分析系统使用说明书，20种靶基因的引物与4种看家基因为 参照基因而结合设计.如T B P (TATA盒结合蛋白)，ATP50(ATP合成酶，H+的转运，线粒体 的Fl复合体，0亚基)，ACTB (beta Actin)，GAPD (甘油三磷酸脱氢酶)。基因的引物设计 原理在GenomeLab GeXP分析系统使用说明书中有详细介绍，同时Beckman Coulter公司还 为客户提供基因的引物设计服务.GeXP样品制备与检测均按GenomeLab GeXP分析系统使用说明书进行简言之，纯 化的肿瘤组织mRNA(美国stratagene公司产品)，第一轮将mRNA反转录成cDNA的特异性靶基因采用反向引物见表X，其中逆转录酶及试剂盒购自Invitrogen公司。根据使用说明书，前3个循环采用通用型/基因特异的引物进行PCR反应，混合后加入F和R PCR引物扩 增30个循环。5，端(The forward)引物标记了 D4染料，其来源于GenomeLab GeXP制备 试剂盒中。GenomeLab GeXP系统借助于合成的靶基因因每一基因P C R产品长度之间有 2-5bp之差别用毛细吸管凝胶电涌读取P C R产物的荧光X度而定性定量.根据GeXP系统 分析获得一定数量的前列腺癌和乳腺肿瘤样本的结果，见图12。实施例7、靶基因C170RF39的免疫组织化学对人类癌症组织分析靶基因C170RF39为全新尚无任何文献报道之基因.依据前期实时PCR (Real-time PCR)分析肿瘤样本时发现C170RF39在部分人肿瘤样品中高表达现象，本实验在制备其特 异抗体后对临床获得的肿瘤样品开展初级免疫组织化学筛选.经实验方法三中步骤对包涵体蛋白纯化，复性后获得纯化重组蛋白和纯化抗体。 为了确定C170RF39蛋白在各种正常和各种癌组织的表达，用亲和纯化的C170RF多克隆抗 体进行免疫组织化学分析。具体地说，组织标本固定在福尔马林溶液中12-24小时，石蜡包 埋，然后切成4 (ym)厚度的片子。按常规组织片微波加热处理法对组织标本片处理后用美 国Ventana公司自动免疫组织化学染色仪(型号Ventana320-202)进行组织化学染色.染 色使用AEC检测盒(美国Covance公司产品)，C170RF多克隆抗体使用浓度为(lug/ml). C170RF39的免疫组化结果见图13.在部分的人肾肿瘤组织中可见高表达的细胞核染色.而 对照的正常人肾组织中几乎检测不到任何染色信号.
0188]SEQUENCE LISTING1
0189]<110>北京思安创新生物技术有限公司
0190]段，陵浔
0191]<120>20种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗
0192]<130>NM 001464
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<160>1
<170>PatentIn version 3. 5
<210>1
<211>2657
<212>DNA
<213>Homo sapiens <220> <221>exon <222>(1).. (130) <220>
<221>exon
<222>(131).
<220>
<221>exon
<222>(208).
<220>
(207)
(263)<221>exon <222>(264). . (342) <220> <221>exon <222>(343). . (419) <220> <221>exon <222>(420).. (588) <220> <221>exon <222>(589). . (645) <220> <221>exon <222>(646).. (717) <220> <221>exon <222>(718). . (884) <220> <221>exon <222>(885). . (966) <220> <221>exon <222>(967).. (1103) <220> <221>exon
<222>(1104).. (1287)
<220>
<221>exon
<222>(1288).. (1386) <220> <221>exon
<222>(1387).. (1582)
<220>
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<222>(1583). . (1688)
<220>
<221>exon
<222>(1689).. (1872) <220>
0
2 3
15
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17
18
19
20
CNgatgtggcatttttacttgt 1t tac aga gaa aag tea aat tat gtt ggt912
AspValAlaPheLeuLeuValTyr Arg Glu Lys Ser Asn Tyr Val Gly
290300
gcaacctttcaagggaagatgtgtgat gca aac tat gca gga ggt gtt960
AlaThrPheGinGlyLysMetCysAsp Ala Asn Tyr Ala Gly Gly Val
305310315 320
gttctgcaccccagaaccataagtctg gaa tea ctt gca gtt att tta1008
ValLeuHisProArgThrlieSerLeu Glu Ser Leu Ala Val lie Leu
325330 335
getcaattattgagecttagtatgggg ate act tat gat gac att aac1056
AlaGinLeuLeuSerLeuSerMetGly lie Thr Tyr Asp Asp lie Asn
340345 350
tgccagtgcteaggagetgtctgc att atg aat cca gaa gca at t1104
LysCysGinCysSerGlyAlaValCys lie Met Asn Pro Glu Ala lie
355360365
catttcagtggtgtgaagatetttagt aac tgc age ttc gaa gac ttt1152
HisPheSerGlyValLysliePheSer Asn Cys Ser Phe Glu Asp Phe
370375380
gcacattttattteaaagcagaagtcc cag tgt ctt cac aat cag cct1200
AlaHisPhelieSerLysGinLysSer Gin Cys Leu His Asn Gin Pro
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cgcttagatccttttttccagcaa gca gtg tgt ggt aat gca aag1248
ArgLeuAspProPhePheLysGinGin Ala Val Cys Gly Asn Ala Lys
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ctggaagcaggagaggagtgtgactgt ggg act gaa cag gat tgt gcc1296
LeuGluAlaGlyGluGluCysAspCys Gly Thr Glu Gin Asp Cys Ala
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cttattggagaaacatgctgtgatatt gcc aca tgt aga ttt aaa gcc1344
LeulieGlyGluThrCysCysAsplie Ala Thr Cys Arg Phe Lys Ala
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ggtteaaactgtgetgaaggaccatgc tgc gaa aac tgt eta ttt atg1392
GlySerAsnCysAlaGluGlyProCys Cys Glu Asn Cys Leu Phe Met
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teagaaagaatgtgtaggccttcc ttt gaa gaa tgc gac ctc cct1440
SerLysGluArgMetCysArgProSer Phe Glu Glu Cys Asp Leu Pro
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gaatattgcaatggateatctgcatea tgc cca gaa aac cac tat gtt1488
GluTyrCysAsnGlySerSerAlaSer Cys Pro Glu Asn His Tyr Val
485490 495
taccat tcc 3-3-3- CC3. atg aga tgg cca ttt ttc tta ttc att cct ttc 2160
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段，陵浔<120>20种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗<130>NM_024052<160>1<170>PatentIn version 3. 5<210>1<211>4232<212>DNA<213>Homo sapiens<220><221>exon<222>(1). . (606)<220><221>exon<222>(607). . (666)<220><221>exon<222> (667). . (774)<220><221>exon<222>(775). . (876)<220><221>exon<222>(877).. (1007)<220><221>exon<222>(1008)·· (4232)<220><221>polyA_signal<222>(1094)·· (1099)<220><221>polyA_site<222>(1117)·· (1117)<220><221>polyA_site<222> (1151)·· (1151)<220><221>STS
<222>(3822)·· (3950)
<400>1gcc get cgc gca ccg cgc cac gtg ttt tct cct ccc gcc aat ggg egg 48Ala Ala Arg Ala Pro Arg His Val Phe Ser Pro Pro Ala Asn Gly Arg
151015aag cgc cgc ctg age ctg ctc ccg ccc ccc agg ctc gcc tga gcc aat 96Lys Arg Arg Leu Ser Leu Leu Pro Pro Pro Arg Leu Ala Ala Asn202530egg aag ggg egg ggt gtg tgt gtg tct gtg tgt gtt tgt gtg ttg tgt 144Arg Lys Gly Arg Gly Val Cys Val Ser Val Cys Val Cys Val Leu Cys354045gtc tgt gag tgt ctg tgt gtg tgt atg tgt gcg cga ggg caa gtc ggg 192Val Cys Glu Cys Leu Cys Val Cys Met Cys Ala Arg Gly Gln Val Gly505560agg ggg acc cag ctc agg act ggg agg ccc tgc tcg cag gtc cct ggg 240Arg Gly Thr Gln Leu Arg Thr Gly Arg Pro Cys Ser Gln Val Pro Gly657075tcc egg tgg egg ccg gag cgc ttg ctc cgc agg cag egg gcg ggt ggt 288Ser Arg Trp Arg Pro Glu Arg Leu Leu Arg Arg Gln Arg Ala Gly Gly80859095cgc ccc tcc cgc ccc cac ccc gcg cgt gcg cgc ccc ggc ctc tcc ctc 336Arg Pro Ser Arg Pro His Pro Ala Arg Ala Arg Pro Gly Leu Ser Leu100105110ccc gcc acc ctc ctc ggc tcc cgc gcg gcg gcg gcg gtt cct ctc cca 384Pro Ala Thr Leu Leu Gly Ser Arg Ala Ala Ala Ala Val Pro Leu Pro115120125ctc ccc cca gcc ctg get ccg ggg gac ccc gcg atg ccg gtc cgc acc 432Leu Pro Pro Ala Leu Ala Pro Gly Asp Pro Ala Met Pro Val Arg Thr130135140gag tgt ccc ccg ccg gcc ggt gcc tcc get gcc tcc gcg gcc tea ctc 480Glu Cys Pro Pro Pro Ala Gly Ala Ser Ala Ala Ser Ala Ala Ser Leu145150155ate ccg ccg ccg ccc ate aac acc cag cag ccc ggc gtg gcc acc age 528lie Pro Pro Pro Pro lie Asn Thr Gln Gln Pro Gly Val Ala Thr Ser160165170175ctg ctc tac age ggc tcc aag ttc cgc ggc cac cag aag age aag ggg 576Leu Leu Tyr Ser Gly Ser Lys Phe Arg Gly His Gln Lys Ser Lys Gly180185190aac tcg tac gac gta gag gtg gtg ctg cag cac gtg gac acg ggg aac 624Asn Ser Tyr Asp Val Glu Val Val Leu Gln His Val Asp Thr Gly Asn
195200205
tct tac ctttgt ggg tac ttg aag att aaa ggc ctt act gag gag tat 672Ser Tyr LeuCys Gly Tyr Leu Lys lie Lys Gly Leu Thr Glu Glu Tyr210215220cca acc cttaca acc ttc ttc gaa gga gaa ata ate age aaa aaa cac 720Pro Thr LeuThr Thr Phe Phe Glu Gly Glu lie lie Ser Lys Lys His225230235cct ttc ttaact cgc aag tgg gat gca gat gaa gat gtt gat egg aaa 768Pro Phe LeuThr Arg Lys Trp Asp Ala Asp Glu Asp Val Asp Arg Lys240245250255cac tgg ggcaag ttt ctg get ttt tat cag tat gca aaa tea ttt aac 816His Trp GlyLys Phe Leu Ala Phe Tyr Gln Tyr Ala Lys Ser Phe Asn260265270tea gat gacttt gat tat gaa gag ctg aag aat gga gac tac gtc ttc 864Ser Asp AspPhe Asp Tyr Glu Glu Leu Lys Asn Gly Asp Tyr Val Phe275280285atg agg tggaag gaa cag ttt ctg gtc cca gat cac acg ate aaa gac 912Met Arg TrpLys Glu Gln Phe Leu Val Pro Asp His Thr lie Lys Asp290295300ate agt ggtget tct ttt gcc ggg ttc tac tac ate tgc ttt cag aag 960lie Ser GlyAla Ser Phe Ala Gly Phe Tyr Tyr lie Cys Phe Gln Lys305310315tea gca gcctcc ata gag ggc tac tac tac cat agg agt tea gaa tg g 1008Ser Ala Ala Ser lie Glu Gly Tyr Tyr Tyr His Arg Ser Ser Glu Trp320325330tat cag tccctc aat cta acc cat gtt cct gaa cac agt gca ccc ate 1056Tyr Gln SerLeu Asn Leu Thr His Val Pro Glu His Ser Ala Pro lie340345350tat gaa ttcegg tga caa egg ttc aga aca gca acc aaa taa aac tga 1104Tyr Glu PheArg Gln Arg Phe Arg Thr Ala Thr Lys Asn355360act tgg caaaaa aga act ttg ccg aga aaa ttg tgt acc tgc cag aac 1152Thr Trp GlnLys Arg Thr Leu Pro Arg Lys Leu Cys Thr Cys Gln Asn365370375380cag gag aagtgt gtt cct gtt tct tea cga gca gac tcg cat cac aaa 1200Gln Glu LysCys Val Pro Val Ser Ser Arg Ala Asp Ser His His Lys385390395gca tga atgtta acc cac aga ate caa gga gca tgg ctg gcc cgt ggg 1248Ala MetLeu Thr His Arg lie Gln Gly Ala Trp Leu Ala Arg Gly
400405410
gca ggt gga ggg age agt ctt cgt tct tcc tcc cct cag tgg cag ttt 1296Ala Gly Gly Gly Ser Ser Leu Arg Ser Ser Ser Pro Gln Trp Gln Phe415420425ggt ctt cac ctg ttt tta age tac ctt aaa cgc act ttt cct tcc tgc 1344Gly Leu His Leu Phe Leu Ser Tyr Leu Lys Arg Thr Phe Pro Ser Cys430435440aca get aac ttc tac ate act gaa atg ccc att cct tcc tcc gtc cca 1392Thr Ala Asn Phe Tyr lie Thr Glu Met Pro lie Pro Ser Ser Val Pro445450455cct cca gcc gaa tag aag gtc tgc tcc egg ate acc ctc age ctt ggt 1440Pro Pro Ala Glu Lys Val Cys Ser Arg lie Thr Leu Ser Leu Gly460465470get cag tgg tcc cga ggc cct aga ccc cca ccc ccc gcc agt tgc ttt 1488Ala Gln Trp Ser Arg Gly Pro Arg Pro Pro Pro Pro Ala Ser Cys Phe475480485490gtc tgg tag ctc aag aga agg cag age ccc age acc tct gtg ccc ccc 1536Val Trp Leu Lys Arg Arg Gln Ser Pro Ser Thr Ser Val Pro Pro495500505aga get ctg tgc agg gag ttg gcc age tgc cgc ate ate ggc cac caa 1584Arg Ala Leu Cys Arg Glu Leu Ala Ser Cys Arg lie lie Gly His Gln510515520ggg cac aag agg egg agg ctc cag tcc ctg ctg ggc tgc ctc agt ctt 1632Gly His Lys Arg Arg Arg Leu Gln Ser Leu Leu Gly Cys Leu Ser Leu525530535cag tgc tga ttg tgt cac ggg tea gcg tgc get get gag ccc tgt act 1680Gln Cys Leu Cys His Gly Ser Ala Cys Ala Ala Glu Pro Cys Thr540545550gtt aac agt gca aag cta gtg agt age tgt cag gtt ccc tgg gcc tgc 1728Val Asn Ser Ala Lys Leu Val Ser Ser Cys Gln Val Pro Trp Ala Cys555560565cat cag gga tea cta aat tta agg ctt cca gat ccc tgg cag gaa cag 1776His Gln Gly Ser Leu Asn Leu Arg Leu Pro Asp Pro Trp Gln Glu Gln570575580att cca gtc ctg ctt act cag tac att tgc tcc aaa ctt ttc aac ttg 1824lie Pro Val Leu Leu Thr Gln Tyr lie Cys Ser Lys Leu Phe Asn Leu585590595600agg gca ata cta aac tta aaa ata aga gtt ttt tta tta caa aat tat 1872Arg Ala lie Leu Asn Leu Lys lie Arg Val Phe Leu Leu Gln Asn Tyr
605610615ttt tat ggt ccc ttt aac gag gac cct atg gca aat gca caa tta ttc 1920Phe Tyr Gly Pro Phe Asn Glu Asp Pro Met Ala Asn Ala Gln Leu Phe
620625630aga att aca ttt tat cgt ttt cac att gaa aac age aaa tgt tga ctt 1968Arg lie Thr Phe Tyr Arg Phe His lie Glu Asn Ser Lys Cys Leu635640645agt aat ttt tat ate gat ate tat atg tat atg tat att taa tea acc 2016Ser Asn Phe Tyr lie Asp lie Tyr Met Tyr Met Tyr lie Ser Thr650655660age agt ttt gaa act agt cat cct ggt aca aaa gtt ttg cag cat tgc 2064Ser Ser Phe Glu Thr Ser His Pro Gly Thr Lys Val Leu Gln His Cys665670675cta aat tat agt get caa cac aag aac tgt ttt tgg ggc cag ttt age 2112Leu Asn Tyr Ser Ala Gln His Lys Asn Cys Phe Trp Gly Gln Phe Ser680685690att tgt gcc gcc tcc ttt tgc tac tea aaa cag caa tgc ttg gcg gca 2160lie Cys Ala Ala Ser Phe Cys Tyr Ser Lys Gln Gln Cys Leu Ala Ala695700705710gcc ttc cat gag gca gaa ggg gtc gtc gtt ctc tga aaa cag tga ctc 2208Ala Phe His Glu Ala Glu Gly Val Val Val Leu Lys Gln Leu715720tga aat gtt ggg aca ggg gaa ggg gtg gga aac atg aac atg ctc aaa 2256Asn Val Gly Thr Gly Glu Gly Val Gly Asn Met Asn Met Leu Lys725730735taa ctc gag get cac gtg cca aag tgt gtg tgt gtg tgt gtg tgt gtg 2304Leu Glu Ala His Val Pro Lys Cys Val Cys Val Cys Val Cys Val740745750tgt gtg tgt atg atg ttt tgt ttt ttt aaa tgt ttt aaa age tta ata 2352Cys Val Cys Met Met Phe Cys Phe Phe Lys Cys Phe Lys Ser Leu lie755760765770ggt tgg cat cag ttg tag ccc aca aac atg get agg get ttg ggg att 2400Gly Trp His Gln Leu Pro Thr Asn Met Ala Arg Ala Leu Gly lie775780785tgg cat ttt tct ggt ggt tta caa gac tta ctc cga ata caa gag gaa 2448Trp His Phe Ser Gly Gly Leu Gln Asp Leu Leu Arg lie Gln Glu Glu790795800aag ctt taa aaa caa gat tgc ate ata acc ccc agg age aaa gca acc 2496Lys Leu Lys Gln Asp Cys lie lie Thr Pro Arg Ser Lys Ala Thr
805810815tgg agg ctg cat ate cat ggg egg gca gca taagag aat gtg aag ccc 2544Trp Arg Leu His lie His Gly Arg Ala Ala Glu Asn Val Lys Pro820825830ttc agt gag acg aga cga gca atg gga aac ctt ttc tgt tct taa gaa 2592Phe Ser Glu Thr Arg Arg Ala Met Gly Asn Leu Phe Cys Ser Glu835840845gtg act tta att tta ttg aca tat gta ctg tat gtt act gag att gaa 2640Val Thr Leu lie Leu Leu Thr Tyr Val Leu Tyr Val Thr Glu lie Glu850855860tgt tag ggg aag cct taa ggt aga ggt att tgg gaa agt age cag tgg 2688Cys Gly Lys Pro Gly Arg Gly lie Trp Glu Ser Ser Gln Trp865870875gca ctt gtg ata tct aaa ate tgt tat ccc agg act gtg tac aga ggg 2736Ala Leu Val lie Ser Lys lie Cys Tyr Pro Arg Thr Val Tyr Arg Gly880885890gca acc tac cca ttc agg aag gtc agg get ttg gaa ccc taa aaa gtt 2784Ala Thr Tyr Pro Phe Arg Lys Val Arg Ala Leu Glu Pro Lys Val895900905ggc ccg atg act taa agg gaa aaa taa ttc att ccc aga gat gag tea 2832Gly Pro Met Thr Arg Glu Lys Phe lie Pro Arg Asp Glu Ser910915920gaa cag tct cct caa tcc tga aat tea aca agg cat cag aag ggc tgg 2880Glu Gln Ser Pro Gln Ser Asn Ser Thr Arg His Gln Lys Gly Trp925930935ctg tgg tea age cca get get gtc atg tga gga gat get cac tgt ggt 2928Leu Trp Ser Ser Pro Ala Ala Val Met Gly Asp Ala His Cys Gly940945950ctt gtt gag ctg atg gcc ttg gtt gag ctg atg gac aag tga agg agg 2976Leu Val Glu Leu Met Ala Leu Val Glu Leu Met Asp Lys Arg Arg955960965cca tgg ggc tgt get gtc ctt cct gcc gta cgt gcc att cca ctc tct 3024Pro Trp Gly Cys Ala Val Leu Pro Ala Val Arg Ala lie Pro Leu Ser970975980tea get ctc ccc tea aca gca tgc gag ccc ata cct tct gca ttt ttc 3072Ser Ala Leu Pro Ser Thr Ala Cys Glu Pro lie Pro Ser Ala Phe Phe985990995cag gcc tgt gag gga tat agg cct ccc ctt gga gca ctg agt ccg 3117
Gln Ala Cys Glu Gly Tyr Arg Pro Pro Leu Gly Ala Leu Ser Pro
100010051010
gag gtc atecct gag ctc ate cac ggc tea tgc tgt ggc tcc gag 3162Glu Val liePro Glu Leu lie His Gly Ser Cys Cys Gly Ser Glu101510201025tag tgg tttgag tgg gca gga agg gcc att tgc aaa cac tgc tgt 3207Trp PheGlu Trp Ala Gly Arg Ala lie Cys Lys His Cys Cys103010351040gtt cta gacaaa caa ccc aag tcc tat ggc agg att tct tcc ttc 3252Val Leu AspLys Gln Pro Lys Ser Tyr Gly Arg lie Ser Ser Phe104510501055ctt cct ttttta ccc agg tga tga agt gca ccc att gta ctg gga 3297Leu Pro PheLeu Pro ArgSer Ala Pro lie Val Leu Gly106010651070aga atg aag agg tga tac ctt tac tag ate ctt cag aca cat cta tga 3345Arg Met Lys Arg Tyr Leu Tyr lie Leu Gln Thr His Leu10751080gaa gat ttgttc att taa aag tct gcc cac tga gga tag gga aag 3390Glu Asp LeuPhe lie Lys Ser Ala His Gly Gly Lys108510901095gat taa gga ttt ttc cac ctc ctc tta gta act cct gaa tta cca 3435Asp Gly Phe Phe His Leu Leu Leu Val Thr Pro Glu Leu Pro11001105aca tea acttct ttc tct ccg ttc ctg aag gaa ctt tgg gga ate 3480Thr Ser ThrSer Phe Ser Pro Phe Leu Lys Glu Leu Trp Gly lie111011151120ate ttc atecgt agt tac get ttc ctg aac ctt ctc agt ggt tta 3525lie Phe lieArg Ser Tyr Ala Phe Leu Asn Leu Leu Ser Gly Leu112511301135cat gcc tctgaa act atg tgc aat att ttt ggt tga cac ttg tat 3570His Ala SerGlu Thr Met Cys Asn lie Phe GlyHis Leu Tyr114011451150cca tcc ttaaga aat tag tgc aga ttg cag atg ttc tgt ctt cca 3615Pro Ser LeuArg AsnCys Arg Leu Gln Met Phe Cys Leu Pro115511601165tcc caa acaage ctg cca tga ggt agg ate cta ggg gtt tgg ctg 3660Ser Gln ThrSer Leu Pro Gly Arg lie Leu Gly Val Trp Leu117011751180ttc tta ctc cac tgc tea gaa age ttc cat tct get gtc ctc age 3705Phe Leu Leu His Cys Ser Glu Ser Phe His Ser Ala Val Leu Ser
118511901195ctc gac cct ctt ctg tgc tgg act tcc acc cac cca ccc ttt tea 3750Leu Asp Pro Leu Leu Cys Trp Thr Ser Thr His Pro Pro Phe Ser120012051210
agt tta gat agt gtt tea get get gtc cca aag taa caa aac aat 3795Ser Leu Asp Ser Val Ser Ala Ala Val Pro Lys Gln Asn Asn121512201225acc tac tgt taa gta gga agt caa tct gtg tgc tgg ttt tgt tgg 3840Thr Tyr Cys Val Gly Ser Gln Ser Val Cys Trp Phe Cys Trp12301235tta tgt gga aat cac aaa ctc cag agg age aaa ggg gtt ttt caa 3885Leu Cys Gly Asn His Lys Leu Gln Arg Ser Lys Gly Val Phe Gln124012451250tgt ctt ttg ctt tea gaa aca gag aat ata ata ttc tea cta agg 3930Cys Leu Leu Leu Ser Glu Thr Glu Asn lie lie Phe Ser Leu Arg125512601265cct tga att gat ttt ttt ctc ata aaa tag tct gat aag cta att 3975Prolie Asp Phe Phe Leu lie Lys Ser Asp Lys Leu lie127012751280ttttgc cat taa ctg tgt tgt ate gtg gtt taa aat4020Phe Lys Lys Lys Cys His Leu Cys Cys lie Val Val Asn128512901295tac taa caa gtt gtg gga aag aaa aat aat att tga ttt tga ate tta 4068Tyr Gln Val Val Gly Lys Lys Asn Asn lie Phe lie Leu13001305aat gtt ttt aaa aat tag act tga atg ggc aca aag tat aaa tat4113Asn Val Phe Lys Asn Thr Met Gly Thr Lys Tyr Lys Tyr131013151320ttt gtt tct tta tgg agg aca tgt gga agg agt ttg agg gtt tgg4158Phe Val Ser Leu Trp Arg Thr Cys Gly Arg Ser Leu Arg Val Trp132513301335gat ggg aga agt att ttg cag age tat caa tgt cca aat aat tta4203Asp Gly Arg Ser lie Leu Gln Ser Tyr Gln Cys Pro Asn Asn Leu134013451350aaa aaa aaa ata aag gta ttt aag cag tg4232Lys Lys Lys lie Lys Val Phe Lys Gln13551360SEQUENCE LISTING3<110>北京思安创新生物技术有限公司
段，陵浔
<120>20种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗<130>NM_145036<160>1<170>PatentIn version 3. 5<210>1<211>3525<212>DNA<213>Homo sapiens<220><221>exon<222>(1). . (211)<220><221>exon<222>(212). . (325)<220><221>exon<222>(326). . (516)<220><221>exon<222>(517). . (689)<220><221>exon<222>(690). . (783)<220><221>exon<222>(784). . (861)<220><221>exon<222>(862). . (909)<220><221>exon<222>(910). . (987)<220><221>exon<222> (988).. (1074)<220><221>exon<222>(1075)·· (1174)
<220><221>exon<222>(1175)·· (1293)<220><221>exon<222> (1294)·· (1437)<220><221>exon<222>(1438)·· (1592)<220><221>exon<222>(1593)·· (1722)<220><221>exon<222>(1723)·· (1815)<220><221>exon<222> (1816)·· (1875)<220><221>exon<222> (1876) ·· (1955)<220><221>exon<222>(1956).. (2091)<220><221>exon<222>(2092).. (2199)<220><221>exon<222>(2200).. (2382)<220><221>exon<222>(2383).. (2613)<220><221>exon<222>(2614).. (2676)<220><221>exon
<222>(2677).. (2826)
<220><221>exon<222>(2827).. (2916)
<220><221>exon<222> (2917) ·· (3018)<220><221>exon<222> (3019).. (3083)<220><221>STS<222>(3024)·· (3150)<220><221>exon<222>(3084).. (3518)<400>1ggg ccc gtt ctc egg gcc tgg ggg gcg cgc gcc cct ggg gtc ccg gcg 48Gly Pro Val Leu Arg Ala Trp Gly Ala Arg Ala Pro Gly Val Pro Ala151015ctg ccc get gtg gtg tcc ccg gca ggc aga gcg cga tcc agg ccg gga 96Leu Pro Ala Val Val Ser Pro Ala Gly Arg Ala Arg Ser Arg Pro Gly202530cga ctt ggg aaa gtt cta agt aaa gtt tga tea gcc ggt ttg ate ggt 144Arg Leu Gly Lys Val Leu Ser Lys Val Ser Ala Gly Leu lie Gly354045ctg acg tgg tcc aag agt ctg acc caa ggg ttt gcg act ggg aga gga 192Leu Thr Trp Ser Lys Ser Leu Thr Gln Gly Phe Ala Thr Gly Arg Gly505560aat ctg tgt cct gga caa g at tgg att atg gaa gtg gga agt gaa gaa 240Asn Leu Cys Pro Gly Gln Asp Trp lie Met Glu Val Gly Ser Glu Glu6575gaa aaa tgg gag aag ctg gat gca gaa ttt gat cac ttt gtg gtt gat 288Glu Lys Trp Glu Lys Leu Asp Ala Glu Phe Asp His Phe Val Val Asp80859095atg aag ccc ttt gtt cta aaa tta cct cat agg aca g aa egg cag agg 336Met Lys Pro Phe Val Leu Lys Leu Pro His Arg Thr Glu Arg Gln Arg100105110tgt get ctt tgg att aga aag ctg tgc gaa cct tea gga aca ggt gca 384Cys Ala Leu Trp lie Arg Lys Leu Cys Glu Pro Ser Gly Thr Gly Ala
115120125gga ata atgggg agg aag aat egg aac ctg tat gca aaa ttg tta ttg 432Gly lie MetGly Arg Lys Asn Arg Asn Leu Tyr Ala Lys Leu Leu Leu
130135140cat atg cttaaa cga ggt gcg ctt gaa ggc cct ttt aca cac cga cct 480His Met LeuLys Arg Gly Ala Leu Glu Gly Pro Phe Thr His Arg Pro145150155gaa ccc gggaca cta aaa att cta cct tea tac atg tcc ate tat ttt 528Glu Pro GlyThr Leu Lys lie Leu Pro Ser Tyr Met Ser lie Tyr Phe160165170175gat gaa ccaaat cca gca cga gca aaa ggt tea age cca gag gga tta 576Asp Glu ProAsn Pro Ala Arg Ala Lys Gly Ser Ser Pro Glu Gly Leu180185190cca gcc tgggta ctg ggt gag ctg gag aca agt gaa cac aaa tta aat 624Pro Ala TrpVal Leu Gly Glu Leu Glu Thr Ser Glu His Lys Leu Asn195200205gaa tea tggaaa ctc tct tct gga gaa gat aac act tta gta cag tcg 672Glu Ser TrpLys Leu Ser Ser Gly Glu Asp Asn Thr Leu Val Gln Ser210215220cca act gatgtc tac ag c agg gaa cag tac act ggg aag ctc cga gtg 720Pro Thr AspVal Tyr Ser Arg Glu Gln Tyr Thr Gly Lys Leu Arg Val225230235aga tea cactcc ttg agt cca act cac aga gaa gat gga caa aat att 768Arg Ser HisSer Leu Ser Pro Thr His Arg Glu Asp Gly Gln Asn lie240245250255acc cca aagatt tgt gaa gtt tat tcg aaa aaa tct cct gtt tct ttg 816Thr Pro Lyslie Cys Glu Val Tyr Ser Lys Lys Ser Pro Val Ser Leu260265270gat gat agtgac att gaa get cgc ctt aat agt tgg aat ctt ggg ata 864Asp Asp SerAsp lie Glu Ala Arg Leu Asn Ser Trp Asn Leu Gly lie275280285gaa aat cctcga tac ctg aga cag aag cct ate cca gtt tct ctg atg 912Glu Asn ProArg Tyr Leu Arg Gln Lys Pro lie Pro Val Ser Leu Met290295300aca ccg aaattc age ctg aga aaa tcc age agt ttc cat gat gat cat 960Thr Pro LysPhe Ser Leu Arg Lys Ser Ser Ser Phe His Asp Asp His305310315ttt ctc tctcga ata cgt gag aaa gag ctg gac atg aaa aca aaa atg 1008Phe Leu SerArg lie Arg Glu Lys Glu Leu Asp Met Lys Thr Lys Met
320325330335atg gaa get aaa ttt cat gaa gaa aag ctt aaa ctg caa cag aaa cat 1056Met Glu Ala Lys Phe His Glu Glu Lys Leu Lys Leu Gln Gln Lys His340345350gat get gat gtt cag aag att tta gaa aga aag aat aat gaa ata gaa 1104Asp Ala Asp Val Gln Lys lie Leu Glu Arg Lys Asn Asn Glu lie Glu355360365gaa ctg aaa act tta tac agg agt aaa caa cat gaa act gaa gag act 1152Glu Leu Lys Thr Leu Tyr Arg Ser Lys Gln His Glu Thr Glu Glu Thr370375380att aga aag ctt gaa aag aaa g tt cag aca ctg ata cgt gac tgt caa 1200lie Arg Lys Leu Glu Lys Lys Val Gln Thr Leu lie Arg Asp Cys Gln385390395gtt ate aga gag act aaa gaa gac cag att gca gag ctg aaa aag ata 1248Val lie Arg Glu Thr Lys Glu Asp Gln lie Ala Glu Leu Lys Lys lie400405410415tgt gaa caa agt acg gaa tct cta aat aat gac tgg gaa aaa aag ctt 1296Cys Glu Gln Ser Thr Glu Ser Leu Asn Asn Asp Trp Glu Lys Lys Leu420425430cac aat get gta gca gaa atg gaa cag gaa aag ttt gat ctt caa aag 1344His Asn Ala Val Ala Glu Met Glu Gln Glu Lys Phe Asp Leu Gln Lys435440445caa cac act gaa aac att caa gaa ttg ctt gag gat aca aat gtg cgt 1392Gln His Thr Glu Asn lie Gln Glu Leu Leu Glu Asp Thr Asn Val Arg450455460ctg aat aaa atg gag agt gaa tac atg gcg caa aca cag tcc aca aac 1440Leu Asn Lys Met Glu Ser Glu Tyr Met Ala Gln Thr Gln Ser Thr Asn465470475cac atg ate aaa gaa ctg gag gcc cgt gtc cag cag ctg act gga gaa 1488His Met lie Lys Glu Leu Glu Ala Arg Val Gln Gln Leu Thr Gly Glu480485490495gca gag aac agt aat tta cag agg cag aaa tta att caa gaa aaa gca 1536Ala Glu Asn Ser Asn Leu Gln Arg Gln Lys Leu lie Gln Glu Lys Ala500505510gaa ctt gaa aga tgt tac cag ata acg tgt agt gaa tta caa gaa gta 1584Glu Leu Glu Arg Cys Tyr Gln lie Thr Cys Ser Glu Leu Gln Glu Val515520525aag gca ag g cgt aac aca ctg cat aaa gag aag gac cat ctt gta aat 1632
Lys Ala Arg Arg Asn Thr Leu His Lys Glu Lys Asp His Leu Val Asn
535540gat tat gag caa aac atg aaa ctg tta caa acc aaa tat gat get gat 1680Asp Tyr Glu Gln Asn Met Lys Leu Leu Gln Thr Lys Tyr Asp Ala Asp
545550555ata aac ctt cta aaa caa gaa cat get ctt tea get tct aag gca tct 1728lie Asn Leu Leu Lys Gln Glu His Ala Leu Ser Ala Ser Lys Ala Ser560565570575agt atg att gaa gaa tta gag cag aat gtc tgt caa tta aaa cag cag 1776Ser Met lie Glu Glu Leu Glu Gln Asn Val Cys Gln Leu Lys Gln Gln580585590tta cag gaa tea gaa ctt caa aga aag caa caa cta agg gat caa gaa 1824Leu Gln Glu Ser Glu Leu Gln Arg Lys Gln Gln Leu Arg Asp Gln Glu595600605aat aag ttt cag atg gag aaa agt cat tta aaa cac ate tat gaa aaa 1872Asn Lys Phe Gln Met Glu Lys Ser His Leu Lys His lie Tyr Glu Lys610615620aag get cat gac ttg cag agt gaa ctt gat aaa gga aaa gaa gat act 1920Lys Ala His Asp Leu Gln Ser Glu Leu Asp Lys Gly Lys Glu Asp Thr625630635caa aag aaa att cat aaa ttt gag gaa get ttg aa g gaa aaa gag gag 1968Gln Lys Lys lie His Lys Phe Glu Glu Ala Leu Lys Glu Lys Glu Glu640645650655cag cta act cgt gtg act gaa gtt cag agg ttg cag gcc cag cag gca 2016Gln Leu Thr Arg Val Thr Glu Val Gln Arg Leu Gln Ala Gln Gln Ala660665670gat gcc get ctg gaa gag ttt aag egg cag gtg gaa ctg aac tea gag 2064Asp Ala Ala Leu Glu Glu Phe Lys Arg Gln Val Glu Leu Asn Ser Glu675680685aaa gtc tat get gaa atg aaa gag cag atg gaa aaa gtg gag gca gat 2112Lys Val Tyr Ala Glu Met Lys Glu Gln Met Glu Lys Val Glu Ala Asp690695700cta act aga tcc aaa tct ctt cgt gag aaa caa tea aag gag ttt tta 2160Leu Thr Arg Ser Lys Ser Leu Arg Glu Lys Gln Ser Lys Glu Phe Leu705710715tgg caa ctg gag gac ate aga cag egg tat gaa caa cag ata gta gag 2208Trp Gln Leu Glu Asp lie Arg Gln Arg Tyr Glu Gln Gln lie Val Glu720725730735ctg aag ctg gag cat gaa cag gag aag acg cac cta tta cag cag cat 2256Leu Lys Leu Glu His Glu Gln Glu Lys Thr His Leu Leu Gln Gln His
740745750aac gca gag aag gat age cta gtc cga gac cat gaa egg gaa att gaa 2304Asn Ala Glu Lys Asp Ser Leu Val Arg Asp His Glu Arg Glu lie Glu 755760765aac ctg gaa aaa cag ctt cgc get gcc aat atg gag cac gaa aat caa 2352Asn Leu Glu Lys Gln Leu Arg Ala Ala Asn Met Glu His Glu Asn Gln770775780att cag gag ttc aag aaa cga gat gca cag gtt att gcc gac atg gag 2400lie Gln Glu Phe Lys Lys Arg Asp Ala Gln Val lie Ala Asp Met Glu785790795gcc cag gtt cac aag ttg aga gaa gaa ttg ate aat gtg aac tea cag 2448Ala Gln Val His Lys Leu Arg Glu Glu Leu lie Asn Val Asn Ser Gln800805810815egg aaa cag cag ctg gta gag ctt ggt ctt ctt cgt gaa gag gaa aag 2496Arg Lys Gln Gln Leu Val Glu Leu Gly Leu Leu Arg Glu Glu Glu Lys820825830caa agg get aca agg gaa cat gag att gtc gtc aat aaa ctg aag get 2544Gln Arg Ala Thr Arg Glu His Glu lie Val Val Asn Lys Leu Lys Ala835840845gaa tea gaa aag atg aaa ata gag ctg aaa aag act cat get get gag 2592Glu Ser Glu Lys Met Lys lie Glu Leu Lys Lys Thr His Ala Ala Glu850855860aca gag atg aca ctg gaa aag gcc aac age aag ctg aag cag att gag 2640Thr Glu Met Thr Leu Glu Lys Ala Asn Ser Lys Leu Lys Gln lie Glu865870875aag gaa tac act cag aag ctt gcc aaa tct tea cag ate ata gca gaa 2688Lys Glu Tyr Thr Gln Lys Leu Ala Lys Ser Ser Gln lie lie Ala Glu880885890895ctt cag aca acc att tcc tct ctg aaa gaa gag aac age cag cag cag 2736Leu Gln Thr Thr lie Ser Ser Leu Lys Glu Glu Asn Ser Gln Gln Gln900905910ctt get gca gaa agg egg ctc cag gat gtt aga caa aag ttt gaa gat 2784Leu Ala Ala Glu Arg Arg Leu Gln Asp Val Arg Gln Lys Phe Glu Asp915920925gag aag aag cag ctg att aga gat aat gac caa gca ate aag gtt tta 2832Glu Lys Lys Gln Leu lie Arg Asp Asn Asp Gln Ala lie Lys Val Leu930935940caa gat gaa tta gaa aac cgt tct aat cag gtg cga tgt gca gag aaa 2880Gln Asp Glu Leu Glu Asn Arg Ser Asn Gln Val Arg Cys Ala Glu Lys[1027]945950955[1028]aaa tta caa cac aaa gaa ttg gag tea cag gaa cag ata act tac ata 2928Lys Leu Gln His Lys Glu Leu Glu Ser Gln Glu Gln lie Thr Tyr lie960965970975cga caa gaa tat gaa aca aaa ttg aaa gga ttg atg cca gca tcc cta 2976Arg Gln Glu Tyr Glu Thr Lys Leu Lys Gly Leu Met Pro Ala Ser Leu980985990aga caa gaa ctt gaa gac acc att tcc tcc cta aaa tea cag gtt aat 3024Arg Gln Glu Leu Glu Asp Thr lie Ser Ser Leu Lys Ser Gln Val Asn99510001005ttt ctg caa aag aga get tcc ate ctt cag gaa gaa ctg act aca 3069Phe Leu Gln Lys Arg Ala Ser lie Leu Gln Glu Glu Leu Thr Thr101010151020tat caa ggc aga ag g taa ctg cac gag aga atg caa egg atg caa 3114Tyr Gln Gly Arg ArgLeu His Glu Arg Met Gln Arg Met Gln102510301035ttt cca ggc tgt get gtg gac ttc ttc cag cag gtt tga aga ttt3159Phe Pro Gly Cys Ala Val Asp Phe Phe Gln Gln Val Arg Phe104010451050gga tat tgt aaa ctg tga gat cag tgg cat ttt tta aat cct taa atg 3207Gly Tyr Cys Lys Leu Asp Gln Trp His Phe Leu Asn Pro Met10551060taa tta aga tea taa aaa cat gca tea ttc ata cac tag tgg ggt 3252Leu Arg Ser Lys His Ala Ser Phe lie His Trp Gly106510701075ggt ttt att tgt gtc tgc cta agt tat ttt ttt aac att cct tta 3297Gly Phe lie Cys Val Cys Leu Ser Tyr Phe Phe Asn lie Pro Leu108010851090ttt ccg att ttc atg ttt gtg tgc att tga gta tgt get gag aac 3342Phe Pro lie Phe Met Phe Val Cys lie Val Cys Ala Glu Asn109511001105tgc tta tat tgg gca aag tga ttt cct atg ata tgc ctt gtt aat 3387Cys Leu Tyr Trp Ala Lys Phe Pro Met lie Cys Leu Val Asn11101115[1061]cct ttt gca tag aat ttt acc agt tgc gta cga tea aaa tea cgt 3432Pro Phe Ala Asn Phe Thr Ser Cys Val Arg Ser Lys Ser Arg112011251130 ttg tag tat cat ate aaa aat tct aac ctg ttt aca ttg ttt tea 3477Leu Tyr His lie Lys Asn Ser Asn Leu Phe Thr Leu Phe Ser113511401145tgt tea tgt tcc tat gtt att aaa ata tta ttt tgt act ta 3525Cys Ser Cys Ser Tyr Val lie Lys lie Leu Phe Cys Thr115011551160 SEQUENCE LISTING4<110>北京思安创新生物技术有限公司段，陵浔<120>20种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗<130>NM_001304<160>1<170>PatentIn version 3. 5<210>1<211>8025<212>DNA<213>Homo sapiens<220><221>exon<222>(1). . (761)<220〉<221>sig一peptide<222>(16). . (105)<220><221>misc_feature<222> (106). . (1395)<220><221>exon<222> (762).. (1009)<220><221>exon<222>(1010).· (1152)<220>[1096]<221>exon<222> (1153) ·· (1322)<220><221>exon<222> (1323) ·· (1672)<220><221>misc_feature<222> (1396) ·· (1497)<220><221>misc_feature<222> (1498).. (2616)<220><221>exon<222> (1673). . (1864)<220><221>exon<222> (1865). . (2032)<220><221>exon<222> (2033). . (2142)<220><221>exon<222> (2143). . (2245)<220><221>exon<222> (2246). . (2388)<220><221>exon<222> (2389) ·· (2558)<220><221>exon<222> (2559) ·· (2888)<220><221>misc_feature<222> (2617). . (2703)<220><221>misc_feature[1133]<222> (2704). . (3912)<220>[1135]<221>exon<222> (2889). . (3084) <220><221>exon<222> (3085). . (3220)<220><221>exon<222> (3221) ·· (3352)<220><221>exon<222> (3353) ·· (3448)<220><221>exon<222> (3449). . (3513)<220><221>exon<222> (3524) ·· (3645)<220><221>exon<222> (3646). . (3831)<220><221>exon<222> (3832) ·· (3931)<220><221>misc—feature<222> (3904) ·· (3981)<220><221>misc_feature<222> (3904). . (3981)<220><221>exon<222> (3932). . (8025)<220><221>STS<222>(4493). . (4622)<220><221>STS<222> (5585). . (5734)<220>[1174]<221>polyA_site[1175]<222> (5764). . (5764)<220><221>STS<222> (6399). . (6505)<220><221>STS<222> (6820). . (6939)<220><221>STS<222> (6830). . (6978)<220><221>polyA_signal<222> (7014) ·· (7019)<220><221>polyA_signal<222> (7018). . (7023)<220><221>polyA_site<222> (7042) ·· (7042)<220><221>STS<222> (7838). . (7967)<220><221>polyA_site<222> (8025). . (8025)<400>1egg cgc tgc tgg aag atg gcg age ggc egg gac gag egg ccg cct tgg 48Arg Arg Cys Trp Lys Met Ala Ser Gly Arg Asp Glu Arg Pro Pro Trp151015egg cta ggg egg ctc ctg ttg ctc atg tgc ctg ctg ctg ctg ggg age 96Arg Leu Gly Arg Leu Leu Leu Leu Met Cys Leu Leu Leu Leu Gly Ser202530tcg gcc egg gcg get cac ate aag aag gcg gag gcg act acc aca act 144Ser Ala Arg Ala Ala His lie Lys Lys Ala Glu Ala Thr Thr Thr Thr354045acg age gcg ggc gcc gag gcg gcc gag ggc cag ttc gac cgc tac tac 192Thr Ser Ala Gly Ala Glu Ala Ala Glu Gly Gln Phe Asp Arg Tyr Tyr505560[1213]cac gaa gag gagttg gag tcg gcg ctg agg gag gcg gcg gcc gcg ggc 240[1214]His Glu Glu GluLeu Glu Ser Ala Leu Arg Glu Ala Ala Ala Ala Gly65707580ctc ccc ggc ctggcc cgc ctc ttt age ate ggc cgc tcg gtg gaa ggc 288Leu Pro Gly LeuAla Arg Leu Phe Ser lie Gly Arg Ser Val Glu Gly859095egg ccg ctg tgggtg ctt cgc ctc acc gcc ggc ctg ggg tcg cta ate 336Arg Pro Leu TrpVal Leu Arg Leu Thr Ala Gly Leu Gly Ser Leu lie100105110cct gag ggc gacgcg ggg cct gac get gcc ggg ccc gac get gcg ggg 384Pro Glu Gly AspAla Gly Pro Asp Ala Ala Gly Pro Asp Ala Ala Gly115120125ccg ctg ctg cccggc egg ccc cag gtg aag ctg gtg ggc aac atg cat 432Pro Leu Leu ProGly Arg Pro Gln Val Lys Leu Val Gly Asn Met His130135140ggc gac gag accgtg tcg cgc cag gtg ttg ate tac ttg gcc cgc gag 480Gly Asp Glu ThrVal Ser Arg Gln Val Leu lie Tyr Leu Ala Arg Glu145150155160ctg gcg gcc ggctac cgc cgc ggg gac ccg cgc ctg gtc cgc ctg ctc 528Leu Ala Ala GlyTyr Arg Arg Gly Asp Pro Arg Leu Val Arg Leu Leu165170175aac acc acc gacgtg tac ctg ctg ccc age ctc aac ccc gat ggc ttc 576Asn Thr Thr AspVal Tyr Leu Leu Pro Ser Leu Asn Pro Asp Gly Phe180185190gag cgt gcc cgcgag ggc gac tgt ggc ttc ggc gac ggc ggc ccg tcc 624Glu Arg Ala ArgGlu Gly Asp Cys Gly Phe Gly Asp Gly Gly Pro Ser195200205ggg gcc age ggccgc gac aat agt cgc ggc cgc gac ctc aac cga age 672Gly Ala Ser GlyArg Asp Asn Ser Arg Gly Arg Asp Leu Asn Arg Ser210215220ttt ccc gac cagttt age acc ggc gaa ccc ccc gcc ctg gac gag gtg 720Phe Pro Asp GlnPhe Ser Thr Gly Glu Pro Pro Ala Leu Asp Glu Val225230235240ccc gag gtg cgcgcc ctc ate gag tgg ate cgc agg aac aa g ttt gtg 768ProGlu Val Arg Ala Leu lie Glu Trp lie Arg Arg Asn Lys Phe Val245250255ctt tct gga aatctg cat ggt ggc tea gtg gta gca age tat cct ttt 816Leu Ser Gly AsnLeu His Gly Gly Ser Val Val Ala Ser Tyr Pro Phe260265270[1252]gat gat tct cca gaa cat aag gcc act gga ate tat age aaa acc tea 864Asp Asp Ser Pro Glu His Lys Ala Thr Gly lie Tyr Ser Lys Thr Ser275280285gat gat gaa gta ttt aaa tac ttg gca aaa get tat get tea aac cac 912Asp Asp Glu Val Phe Lys Tyr Leu Ala Lys Ala Tyr Ala Ser Asn His290295300ccc ata atg aaa act ggt gag cct cat tgt cca gga gat gaa gac gag 960Pro lie Met Lys Thr Gly Glu Pro His Cys Pro Gly Asp Glu Asp Glu305310315320act ttc aaa gat gga ate aca aac ggc gca cat tgg tat gat gtg gaa g 1009Thr Phe Lys Asp Gly lie Thr Asn Gly Ala His Trp Tyr Asp Val Glu325330335gt ggt atg caa gat tac aat tat gtg tgg gcc aac tgt ttt gag ate 1056Gly Gly Met Gln Asp Tyr Asn Tyr Val Trp Ala Asn Cys Phe Glu lie340345350aca tta gaa ctg tct tgt tgc aag tac cca cct get tea cag ctt cga 1104Thr Leu Glu Leu Ser Cys Cys Lys Tyr Pro Pro Ala Ser Gln Leu Arg355360365cag gaa tgg gag aac aat cgt gag tct ttg ate aca ttg att gaa aag 1152Gln Glu Trp Glu Asn Asn Arg Glu Ser Leu lie Thr Leu lie Glu Lys370375380gtt cac att gga gtg aaa gga ttt gtt aaa gat tcc ata aca gga tct 1200Val His lie Gly Val Lys Gly Phe Val Lys Asp Ser lie Thr Gly Ser385390395400ggg tta gag aat gca acc ate tea gtg get ggt att aat cat aat ate 1248Gly Leu Glu Asn Ala Thr lie Ser Val Ala Gly lie Asn His Asn lie405410415aca aca ggc aga ttt ggt gat ttc tac cga tta ctt gtt cct gga act 1296Thr Thr Gly Arg Phe Gly Asp Phe Tyr Arg Leu Leu Val Pro Gly Thr420425430tac aac ctt aca gta gtt tta act gg g tat atg cca ttg act gtt act 1344Tyr Asn Leu Thr Val Val Leu Thr Gly Tyr Met Pro Leu Thr Val Thr435440445aat gta gtg gtg aaa gaa gga cca gcc aca gag gtg gat ttt tct ctt 1392Asn Val Val Val Lys Glu Gly Pro Ala Thr Glu Val Asp Phe Ser Leu450455460agg cca act gta act tea gta ate cct gac acg aca gag get gta tea 1440Arg Pro Thr Val Thr Ser Val lie Pro Asp Thr Thr Glu Ala Val Ser[1290]465470475480[1291]act get age aca gtt get ata cct aat att ctt tct gga aca tea tcc 1488Thr Ala Ser Thr Val Ala lie Pro Asn lie Leu Ser Gly Thr Ser Ser485490495 tcc tac cag cca att cag cca aag gac ttt cac cac cac cat ttc cct 1536Ser Tyr Gln Pro lie Gln Pro Lys Asp Phe His His His His Phe Pro500505510gat atg gaa ate ttc ttg aga agg ttt gcc aat gaa tat cct aac att 1584Asp Met Glu lie Phe Leu Arg Arg Phe Ala Asn Glu Tyr Pro Asn lie515520525acc egg ctt tat tcc ttg gga aaa tea gta gag tea aga gaa ctt tat 1632Thr Arg Leu Tyr Ser Leu Gly Lys Ser Val Glu Ser Arg Glu Leu Tyr530535540gtg atg gag ata tct gat aat ccg ggt gtc cat gaa cca g gt gaa cca 1680Val Met Glu lie Ser Asp Asn Pro Gly Val His Glu Pro Gly Glu Pro545550555560gaa ttt aag tac att gga aat atg cat gga aat gaa gtg gtt gga aga 1728Glu Phe Lys Tyr lie Gly Asn Met His Gly Asn Glu Val Val Gly Arg565570575gaa ctg ctg ttg aac ctc ata gaa tac ctt tgt aag aac ttt gga aca 1776Glu Leu Leu Leu Asn Leu lie Glu Tyr Leu Cys Lys Asn Phe Gly Thr580585590gac cct gaa gtc aca gat ttg gtt cat aac act aga att cac ctt atg 1824Asp Pro Glu Val Thr Asp Leu Val His Asn Thr Arg lie His Leu Met595600605cca tcc atg aat cct gat ggg tat gaa aag tcc cag gaa g ga gat tea 1872Pro Ser Met Asn Pro Asp Gly Tyr Glu Lys Ser Gln Glu Gly Asp Ser610615620ata agt gta att ggc aga aac aac age aac aac ttt gac ctg aac cga 1920lie Ser Val lie Gly Arg Asn Asn Ser Asn Asn Phe Asp Leu Asn Arg625630635640aat ttc cca gac cag ttt gtt cag ate aca gat cct acg caa cca gaa 1968Asn Phe Pro Asp Gln Phe Val Gln lie Thr Asp Pro Thr Gln Pro Glu645650655act att get gta atg age tgg atg aag tcc tat cca ttt gta ctt tea 2016Thr lie Ala Val Met Ser Trp Met Lys Ser Tyr Pro Phe Val Leu Ser660665670gca aac ctg cat gga g gt tct ttg gtg gtt aac tac cct ttt gat gat 2064Ala Asn Leu His Gly Gly Ser Leu Val Val Asn Tyr Pro Phe Asp Asp675680685[1330]gat gaa caa gga ctt gcc aca tat agt aaa tea cca gat gat get gtg 2112[1331]Asp Glu Gln Gly Leu Ala Thr Tyr Ser Lys Ser Pro Asp Asp Ala Val690695700ttc caa caa ata gca ctt tct tat tcc aag gaa aat tcc cag atg ttt 2160Phe Gln Gln lie Ala Leu Ser Tyr Ser Lys Glu Asn Ser Gln Met Phe705710715720caa ggt aga cct tgc aag aat atg tat cct aat gaa tat ttt cct cat 2208Gln Gly Arg Pro Cys Lys Asn Met Tyr Pro Asn Glu Tyr Phe Pro His725730735gga ata aca aat gga get agt tgg tat aat gtg cca g ga gga atg cag 2256Gly lie Thr Asn Gly Ala Ser Trp Tyr Asn Val Pro Gly Gly Met Gln740745750gac tgg aac tat tta caa aca aat tgc ttt gaa gtg act att gaa cta 2304Asp Trp Asn Tyr Leu Gln Thr Asn Cys Phe Glu Val Thr lie Glu Leu755760765ggt tgt gtg aaa tat cca ctt gag aaa gag ctg cca aac ttt tgg gaa 2352Gly Cys Val Lys Tyr Pro Leu Glu Lys Glu Leu Pro Asn Phe Trp Glu770775780cag aat cga aga tea cta ate cag ttt atg aaa cag gtt cat cag ggc 2400Gln Asn Arg Arg Ser Leu lie Gln Phe Met Lys Gln Val His Gln Gly785790795800gtc aga gga ttt gtt cta gat gcc aca gat ggc agg ggt ata tta aat 2448Val Arg Gly Phe Val Leu Asp Ala Thr Asp Gly Arg Gly lie Leu Asn805810815gcc acc att agt gtt get gag att aat cac cca gtg act act tac aaa 2496Ala Thr lie Ser Val Ala Glu lie Asn His Pro Val Thr Thr Tyr Lys820825830act gga gat tac tgg cgt ctc ttg gtt cca gga act tat aaa ate aca 2544Thr Gly Asp Tyr Trp Arg Leu Leu Val Pro Gly Thr Tyr Lys lie Thr835840845gca tct get cga gg g tat aat cca gtt acc aag aat gtg act gtc aag 2592Ala Ser Ala Arg Gly Tyr Asn Pro Val Thr Lys Asn Val Thr Val Lys850855860agt gaa ggc get att cag gtc aac ttc aca ctt gtt cga tcc tea aca 2640Ser Glu Gly Ala lie Gln Val Asn Phe Thr Leu Val Arg Ser Ser Thr865870875880gat tea aac aat gaa tea aag aaa gga aaa ggg get age age age acc 2688Asp Ser Asn Asn Glu Ser Lys Lys Gly Lys Gly Ala Ser Ser Ser Thr885890895[1369]aat gat gcc agt gat cca act act aaa gag ttt gaa act tta att aaa 2736Asn Asp Ala Ser Asp Pro Thr Thr Lys Glu Phe Glu Thr Leu lie Lys900905910gac ctt tea gcg gag aat ggt ttg gaa age ctc atg tta cgc tcc tcc 2784Asp Leu Ser Ala Glu Asn Gly Leu Glu Ser Leu Met Leu Arg Ser Ser915920925tea aat ctg get ctg get ctt tat cga tac cat tcc tac aaa gac tta 2832Ser Asn Leu Ala Leu Ala Leu Tyr Arg Tyr His Ser Tyr Lys Asp Leu930935940tea gag ttt ctg aga gga ctt gta atg aac tat cca cat att aca aat 2880Ser Glu Phe Leu Arg Gly Leu Val Met Asn Tyr Pro His lie Thr Asn945950955960ctt acc aa t ttg gga cag age act gaa tat cgt cac att tgg tcc ctt 2928Leu Thr Asn Leu Gly Gln Ser Thr Glu Tyr Arg His lie Trp Ser Leu965970975gaa ate tcc aat aag ccc aat gta tct gag cct gaa gaa cca aag att 2976Glu lie Ser Asn Lys Pro Asn Val Ser Glu Pro Glu Glu Pro Lys lie980985990cgt ttt gtt get ggt ate cat gga aat gcg cca gtt gga act gaa ctg 3024Arg Phe Val Ala Gly lie His Gly Asn Ala Pro Val Gly Thr Glu Leu99510001005ctt ttg get ctg gca gaa ttt ctc tgc ctg aac tac aaa aag aac 3069Leu Leu Ala Leu Ala Glu Phe Leu Cys Leu Asn Tyr Lys Lys Asn101010151020cca get gtt acc caa ttg gtt gac agg act agg att gtg att gtc 3114Pro Ala Val Thr Gln Leu Val Asp Arg Thr Arg lie Val lie Val102510301035cct tct cta aat cca gat ggg cga gag aga get caa gag aaa gac 3159Pro Ser Leu Asn Pro Asp Gly Arg Glu Arg Ala Gln Glu Lys Asp104010451050tgt act tea aaa ata gga caa aca aat get cgt ggc aaa gat ttg 3204Cys Thr Ser Lys lie Gly Gln Thr Asn Ala Arg Gly Lys Asp Leu105510601065gat aca gac ttc aca a at aat gcc tcc caa cct gag acc aaa gcc 3249Asp Thr Asp Phe Thr Asn Asn Ala Ser Gln Pro Glu Thr Lys Ala107010751080ate att gaa aat ttg att caa aaa cag gac ttt agt ctt tct gtt 3294[1406]lie lie Glu Asn Leu lie Gln Lys Gln Asp Phe Ser Leu Ser Val108510901095[1408]gcc tta gat ggt ggt tcc atg ctg gtc aca tat cct tat gac aag 3339Ala Leu Asp Gly Gly Ser Met Leu Val Thr Tyr Pro Tyr Asp Lys110011051110 cca gta cag aca g tg gaa aat aaa gag act ctg aag cat ttg gca 3384Pro Val Gln Thr Val Glu Asn Lys Glu Thr Leu Lys His Leu Ala111511201125tct ctt tat gca aat aat cat cca tcc atg cac atg ggt cag ccc 3429Ser Leu Tyr Ala Asn Asn His Pro Ser Met His Met Gly Gln Pro113011351140agt tgc cca aat aaa tea g at gag aat att cca gga gga gta atg 3474Ser Cys Pro Asn Lys Ser Asp Glu Asn lie Pro Gly Gly Val Met11451155cgt gga gca gaa tgg cat agt cac ctg ggc age atg aag gattatagtg 3523Arg Gly Ala Glu Trp His Ser His Leu Gly Ser Met Lys116011651170tea cct atg gcc att gtc egg aaa tea cag tat aca caa get get 3568Ser Pro Met Ala lie Val Arg Lys Ser Gln Tyr Thr Gln Ala Ala117511801185gtt act ttc cta gtg ctg cac gac tcc ctt cct tgt ggg cag aca 3613Val Thr Phe Leu Val Leu His Asp Ser Leu Pro Cys Gly Gln Thr119011951200ata aga gat ctc ttc tta gta tgt tag tgg ag g ttc aca agg gag 3658lie Arg Asp Leu Phe Leu Val Cys Trp Arg Phe Thr Arg Glu120512101215ttc atg gat ttg tta aag ata aga ctg gaa age caa tct cta aag 3703Phe Met Asp Leu Leu Lys lie Arg Leu Glu Ser Gln Ser Leu Lys122012251230cag tea ttg tac tta atg aag gaa taa agg tac aaa caa aag agg 3748Gln Ser Leu Tyr Leu Met Lys Glu Arg Tyr Lys Gln Lys Arg12351240gag gtt att tcc atg tac tct tag cgc cag gtg tcc ata aca tta 3793Glu Val lie Ser Met Tyr Ser Arg Gln Val Ser lie Thr Leu124512501255ttg cca tcg ctg atg ggt acc age aac aac att cac ag g tct ttg 3838Leu Pro Ser Leu Met Gly Thr Ser Asn Asn lie His Arg Ser Leu126012651270tgc ate atg atg cag cta gtt ctg tgg tga tag tct ttg aca cag 3883Cys lie Met Met Gln Leu Val Leu TrpSer Leu Thr Gln127512801285[1447]ata acc gga tat ttg gtt tgc caa ggg age ttg tgg taa ctg tat 3928[1448]lie Thr Gly Tyr Leu Val Cys Gln Gly Ser Leu Trp Leu Tyr129012951300cag gtg cta cta tgt egg cat tga tcc taa cag ctt gca tta ttt ggt 3976Gln Val Leu Leu Cys Arg His Ser Gln Leu Ala Leu Phe Gly13051310gca tct get caa tea agt cta ata gac aca agg atg get ttc ate 4021Ala Ser Ala Gln Ser Ser Leu lie Asp Thr Arg Met Ala Phe lie131513201325ggc tea ggc age ate atg atg agt atg aag atg aaa ttc gca tga 4066Gly Ser Gly Ser lie Met Met Ser Met Lys Met Lys Phe Ala133013351340tgt cta ccg get cca aga agt ccc tcc taa gcc atg agt tcc agg 4111Cys Leu Pro Ala Pro Arg Ser Pro Ser Ala Met Ser Ser Arg134513501355atg aaa cag aca ctg aag agg aaa cat tat att cta gca aac att 4156Met Lys Gln Thr Leu Lys Arg Lys His Tyr lie Leu Ala Asn lie136013651370gaa aaa cac att ttg cat ate tcc cag cat aag tac caa gca aaa 4201Glu Lys His lie Leu His lie Ser Gln His Lys Tyr Gln Ala Lys137513801385ttacag ttc ctc ttg gga gaa cac tgc att aag aag aga gac tct 4246Leu Gln Phe Leu Leu Gly Glu His Cys lie Lys Lys Arg Asp Ser139013951400ctt get tct tea aag age ttt ggg aaa tta aat tgc taa att tgt 4291Leu Ala Ser Ser Lys Ser Phe Gly Lys Leu Asn Cys lie Cys140514101415att ctc tgt gaa ttt cac tgg cag ttt tga act tcc ctt cct taa 4336lie Leu Cys Glu Phe His Trp Gln PheThr Ser Leu Pro14201425agt act cta aac ctt taa aaa aaa ate tga ttt atg cag cag aga 4381Ser Thr Leu Asn Leu Lys LysIle Phe Met Gln Gln Arg143014351440tgg gac age cac ttt ttc ttt tta att taa gat gag cta ttt gga 4426Trp Asp Ser His Phe Phe Phe Leu lie Asp Glu Leu Phe Gly144514501455get tat gta ata atg gca taa age caa cta gag gat gtt gta ttt 4471Ala Tyr Val lie Met Ala Ser Gln Leu Glu Asp Val Val Phe146014651470[1486]tgc aca tea gat gtt tac tag tgg ctt tag tat ttt tct ttg ttt taa 4519Cys Thr Ser Asp Val Tyr Trp Leu Tyr Phe Ser Leu Phe 14751480atg gcc aaa aga ate cag aaa cat taa ggc agg gac age agt cag 4564Met Ala Lys Arg lie Gln Lys His Gly Arg Asp Ser Ser Gln148514901495aat cga cat aaa get tta aaa act caa ggt ttt ttc aac cta ctg 4609Asn Arg His Lys Ala Leu Lys Thr Gln Gly Phe Phe Asn Leu Leu150015051510agg agt act ttt ctc tag ttg tta aat age tgg agt ttt tct tat 4654Arg Ser Thr Phe Leu Leu Leu Asn Ser Trp Ser Phe Ser Tyr151515201525tea ggt tta atg gag gtt gaa ttg att ttt aaa cac ata taa cag 4699Ser Gly Leu Met Glu Val Glu Leu lie Phe Lys His lie Gln153015351540tag gaa atg aat aaa tgg get tct gca ttt ggc ttt cta cct gtt 4744Glu Met Asn Lys Trp Ala Ser Ala Phe Gly Phe Leu Pro Val15451550cca agg cta gat egg aac tgg tag act acg ctg taa gca gga ttt 4789Pro Arg Leu Asp Arg Asn Trp Thr Thr Leu Ala Gly Phe155515601565cac tac ctc tct taa ggt tta gca aac ttc taa ata gcc cat ttt 4834His Tyr Leu Ser Gly Leu Ala Asn Phe lie Ala His Phe157015751580aag gga gaa ctt act aac ttt att gtg aaa ggt cta aat gcc cac 4879Lys Gly Glu Leu Thr Asn Phe lie Val Lys Gly Leu Asn Ala His158515901595ttg aat gaa get gag aga gag ate tag caa aag cta aaa ctc atg 4924Leu Asn Glu Ala Glu Arg Glu lie Gln Lys Leu Lys Leu Met16001605ttg tct ate ttt gaa ctt ggt aaa aac cca cag gtg ctg ctg ctt 4969Leu Ser lie Phe Glu Leu Gly Lys Asn Pro Gln Val Leu Leu Leu161016151620ata tct gtg aag cac tag ctt att cta gga atg cct gat tct tta 5014lie Ser Val Lys His Leu lie Leu Gly Met Pro Asp Ser Leu162516301635ata ttg cct aaa tcg gaa cct ttt tct atg ttg CSC Q-CQ- tgg ttt 5059lie Leu Pro Lys Ser Glu Pro Phe Ser Met Leu His Thr Trp Phe164016451650[1525]tea gat gac cca gcc ate tac aag ate tga att cta ctg aaa ata 5104[1526]Ser Asp Asp Pro Ala lie Tyr Lys lie lie Leu Leu Lys lie165516601665tct aga aat gtg gaa gag acc tac ttg cac att ctt aac ctg tat 5149Ser Arg Asn Val Glu Glu Thr Tyr Leu His lie Leu Asn Leu Tyr167016751680ttg aac aca aaa tat cta tac ttc atg ctc cag ccc aag cct ata 5194Leu Asn Thr Lys Tyr Leu Tyr Phe Met Leu Gln Pro Lys Pro lie168516901695ccc tgt aat age ata cta tta ttg aaa tcg ctt gac egg tct tgt 5239Pro Cys Asn Ser lie Leu Leu Leu Lys Ser Leu Asp Arg Ser Cys170017051710tea cat agg cct ctg gga gtg att tgg ttc ttt gcc cta atg ttt 5284Ser His Arg Pro Leu Gly Val lie Trp Phe Phe Ala Leu Met Phe171517201725cat ttg acg gtc tct ttt tga tea acc aat ttt tct aaa agt tea 5329His Leu Thr Val Ser Phe Ser Thr Asn Phe Ser Lys Ser Ser173017351740gtc gaa age ttt taa gta tag ctt cct ccc ttg aaa aaa aat gta 5374Val Glu Ser PheVal Leu Pro Pro Leu Lys Lys Asn Val17451750aac tat gac tgc tga gtg ata aaa cac tgt ggt gtg aaa gtg tea 5419Asn Tyr Asp Cys Val lie Lys His Cys Gly Val Lys Val Ser175517601765tct tea ctg cca ate agg caa aga ccg gaa aga ttt gca ttt tat 5464Ser Ser Leu Pro lie Arg Gln Arg Pro Glu Arg Phe Ala Phe Tyr177017751780tat gtc tgt ctt ate atg caa tgg aaa tga tgc ttt ttg taa gta 5509Tyr Val Cys Leu lie Met Gln Trp Lys Cys Phe Leu Val178517901795tgc ate tta cca atg atg taa egg ttt aat acc ttt gaa tgt ttt5554Cys lie Leu Pro Met Met Arg Phe Asn Thr Phe Glu Cys Phe180018051810aat aac caa gtt get get gaa ctt ata cta aat cag ggg acc aaa5599Asn Asn Gln Val Ala Ala Glu Leu lie Leu Asn Gln Gly Thr Lys181518201825aaa ctt get ctt ate ttc tea aat tgt att cta tat cca tta atg5644Lys Leu Ala Leu lie Phe Ser Asn Cys lie Leu Tyr Pro Leu Met183018351840[1564]tat cag tta tcc caa age ctt cag gtg gag ggg ttt acc acc ttc5689Tyr Gln Leu Ser Gln Ser Leu Gln Val Glu Gly Phe Thr Thr Phe184518501855cta ggt cgt tea acc agg ttt tgt gag gaa tgc att caa agt ggc5734Leu Gly Arg Ser Thr Arg Phe Cys Glu Glu Cys lie Gln Ser Gly186018651870ttt ata aaa gaa gat ttt ctt tag caa gaa taa tga ggt cat gtc att 5782Phe lie Lys Glu Asp Phe Leu Gln GluGly His Val lie18751880tgt taa taa gta tct gtg ata aat ccg tgg ttc aag gtt aag cca 5827CysVal Ser Val lie Asn Pro Trp Phe Lys Val Lys Pro188518901895ttc tgg tat tct ggt att age aac tgt aaa ttc tgc cac ctc ata 5872Phe Trp Tyr Ser Gly lie Ser Asn Cys Lys Phe Cys His Leu lie190019051910cat gga aca gag ctt gtg gga tgc taa tag tta gtg aag tat aca tga 5920His Gly Thr Glu Leu Val Gly CysLeu Val Lys Tyr Thr19151920ttt aat ttc taa taa tct tta tgt ttt ctt taa gga tgg tgg tgt 5965Phe Asn Phe Ser Leu Cys Phe LeuGly Trp Trp Cys192519301935att get ctt ttt cag ctt tat ttt taa gag tac agt cag gaa acc6010lie Ala Leu Phe Gln Leu Tyr Phe Glu Tyr Ser Gln Glu Thr194019451950aac aag ggg cct aag agt ggc tgc ccc tgc ttg gga cat tac age6055Asn Lys Gly Pro Lys Ser Gly Cys Pro Cys Leu Gly His Tyr Ser195519601965aag tga aac aaa gtt aat gtg aca age ttt get ttg tta tea ttg 6100Lys Asn Lys Val Asn Val Thr Ser Phe Ala Leu Leu Ser Leu19701975gtc ttc act aga gga tac ctt tta cat gta ctt ctc tct tgg ate 6145Val Phe Thr Arg Gly Tyr Leu Leu His Val Leu Leu Ser Trp lie198019851990aaa tat gtc ttt aac tgt aca tct cag tgg ctg gag gcc atg cct 6190Lys Tyr Val Phe Asn Cys Thr Ser Gln Trp Leu Glu Ala Met Pro[1599]199520002005ttt aag cat gtg taa aat ttt taa aga aat gaa cat aca cat agt 6235Phe Lys His Val Asn Phe Arg Asn Glu His Thr His Ser201020152020[1603]tat ttt agt aat att tcc tga aag aaa aac caa att ctg cta taa 6280Tyr Phe Ser Asn lie Ser Lys Lys Asn Gln lie Leu Leu202520302035 gtc ttg ate ttc aat gaa ctt tta aat aat gca ttt age tgg aaa 6325Val Leu lie Phe Asn Glu Leu Leu Asn Asn Ala Phe Ser Trp Lys204020452050aca aga ctt tcc cag ctt gta tta cct aga age gtg aat gta tag 6370Thr Arg Leu Ser Gln Leu Val Leu Pro Arg Ser Val Asn Val20552060gat acc tga cta cta aga cta tat tct cag ccc tgc cct gtc ttt 6415Asp Thr Leu Leu Arg Leu Tyr Ser Gln Pro Cys Pro Val Phe206520702075tat ttg egg gtc taa tct aat att aga 3- 3- tat taa ccg ctt aag 6460Tyr Leu Arg Val Ser Asn lie Arg lie Tyr Pro Leu Lys208020852090gca ttg aag cca tat ggg atg ggg aat gca ttt ctt cag tgt ttc 6505Ala Leu Lys Pro Tyr Gly Met Gly Asn Ala Phe Leu Gln Cys Phe209521002105tcc gag aga ctt tcc att tcc ttg gag tta tgg egg caa gta agt 6550Ser Glu Arg Leu Ser lie Ser Leu Glu Leu Trp Arg Gln Val Ser211021152120ate ata gta tta aga aat ttg cct aaa tct gag ttg tgc ctt tct 6595lie lie Val Leu Arg Asn Leu Pro Lys Ser Glu Leu Cys Leu Ser212521302135tta ctc aca agg cat ggg ctt tgt cct ggt gat cag ttt gta age 6640Leu Leu Thr Arg His Gly Leu Cys Pro Gly Asp Gln Phe Val Ser214021452150ctt ctt cct tcc cag ctc ctt aat aaa age aaa gtg att gag tag 6685Leu Leu Pro Ser Gln Leu Leu Asn Lys Ser Lys Val lie Glu215521602165gta atg ttc aaa gtg tct gcc tgt gta cat gta ctt gta ttg att 6730Val Met Phe Lys Val Ser Ala Cys Val His Val Leu Val Leu lie217021752180atg tag ttc agt aag atg tgc cca agt cat ttc aga aag aaa gac 6775Met Phe Ser Lys Met Cys Pro Ser His Phe Arg Lys Lys Asp21852190cct tea gtt ttg atg cat ttt get gaa cac ttg ggt agt gag tgg 6820Pro Ser Val Leu Met His Phe Ala Glu His Leu Gly Ser Glu Trp219522002205[1642]gat cct ate cag ttg agg aat get tgc aat get cat tga agg gat 6865[1643]Asp Pro lie Gln Leu Arg Asn Ala Cys Asn Ala His Arg Asp221022152220ttg ctt tgg gac ttt gtc ate ttc cag aaa gga aac ata ttg tat 6910Leu Leu Trp Asp Phe Val lie Phe Gln Lys Gly Asn lie Leu Tyr222522302235att tgg ccc agt gtg att gat tgc ttt ate ttt ggt aac ttt tac 6955lie Trp Pro Ser Val lie Asp Cys Phe lie Phe Gly Asn Phe Tyr224022452250ttg aat ggg att tgc tga att aat gac tat tga att taa aac taa 7000Leu Asn Gly lie Cys lie Asn Asp Tyr lie Asn22552260tta tga gttggt agt gtt at gtc tga get 7045LeuVal Asp Lys lie Lys Gly Ser Val Tyr ValAla226522702275tat tgt gtt tga get aac acc agg tta ctc agt aac cat gac ctg 7090Tyr Cys Val Ala Asn Thr Arg Leu Leu Ser Asn His Asp Leu228022852290ctc ctc cat ttc cat tta ttc tea aca tta aat agt ttt ate ttg 7135Leu Leu His Phe His Leu Phe Ser Thr Leu Asn Ser Phe lie Leu229523002305ttg ttg cca gaa atg cac ttg tgc cag gta ttg tcc ctg ctg tat 7180Leu Leu Pro Glu Met His Leu Cys Gln Val Leu Ser Leu Leu Tyr231023152320gaa aag ctt ctt ggc aat gaa ttc tgt aat agt gcc cta cat tat 7225Glu Lys Leu Leu Gly Asn Glu Phe Cys Asn Ser Ala Leu His Tyr232523302335ggt ttt ctg gtg gaa ttg ttt taa cag tga caa ccc agg att tcc aat 7273Gly Phe Leu Val Glu Leu Phe Gln Gln Pro Arg lie Ser Asn23402345ata ttt ttg ttt tat tgt tat tac
ttc cac tat gat tga 7318lie Phe Leu Phe Tyr Cys Tyr Tyr Gln Lys Phe His Tyr Asp235023552360tgt tea gtg att ttc tat age aac ttt ttt ggt aac tct ttg ggt 7363Cys Ser Val lie Phe Tyr Ser Asn Phe Phe Gly Asn Ser Leu Gly236523702375ttc tga ttt gtt tta get aaa att ttg ggg ata tga ttt ggg tct 7408Phe Phe Val Leu Ala Lys lie Leu Gly lie Phe Gly Ser238023852390[1681]ttg att aat gtc age tga act tgg att tct agt tea tga aga aat7453Leu lie Asn Val Ser Thr Trp lie Ser Ser Ser Arg Asn23952400ctc tcc caa tac cca ttt ate cta ttt tta gca ata att cgt taa7498Leu Ser Gln Tyr Pro Phe lie Leu Phe Leu Ala lie lie Arg240524102415tga ttc cac ttg att ttc aga ata ttg tcc tgg ttg att ttg att 7543Phe His Leu lie Phe Arg lie Leu Ser Trp Leu lie Leu lie242024252430tga cag cat aca tta tga aat ttg aaa gta ggt tac cat ttt gag7588Gln His Thr Leu Asn Leu Lys Val Gly Tyr His Phe Glu243524402445gca gtt gga tat aaa tta tgt aaa tat gta tga tta tga ttt tta taa 7636Ala Val Gly Tyr Lys Leu Cys Lys Tyr ValLeu Phe Leu24502455atg gca taa cat gag tgt act aac tac ctt cta tgc tgg cca tgc 7681Met AlaHis Glu Cys Thr Asn Tyr Leu Leu Cys Trp Pro Cys246024652470tac aga ttt tct gga ggt atg aca ata gta ttt ttt tat get cag 7726Tyr Arg Phe Ser Gly Gly Met Thr lie Val Phe Phe Tyr Ala Gln247524802485att aaa aat cag ctt ttc acc tct cca gtt ttt cca agt gat act 7771lie Lys Asn Gln Leu Phe Thr Ser Pro Val Phe Pro Ser Asp Thr249024952500ccc agt tct aga gca ate tac age tgt tta tgt gag gtg ccc aac 7816Pro Ser Ser Arg Ala lie Tyr Ser Cys Leu Cys Glu Val Pro Asn250525102515acc cat tea tct caa gtg ctt cag tct ttg gtt tat ttc atg cac7861Thr His Ser Ser Gln Val Leu Gln Ser Leu Val Tyr Phe Met His252025252530tgt gcc ttc aaa atg aaa ttt tta aaa ggg act tta aat gaa gtt 7906Cys Ala Phe Lys Met Lys Phe Leu Lys Gly Thr Leu Asn Glu Val253525402545gaa tag tag ttt tta aaa gtc aat ttg taa ttt atg tga aat cta act 7954GluPhe Leu Lys Val Asn Leu Phe Met Asn Leu Thr25502555[1717]gta atg agg tcc ttt ctg ttt ttt ata tgt aaa cag ate tac taa 7999Val Met Arg Ser Phe Leu Phe Phe lie Cys Lys Gln lie Tyr256025652570[1720]tcc tgt ata aaa gtt att tta cga tg 8025Ser Cys lie Lys Val lie Leu Arg25752580 SEQUENCE LISTING5<110>北京思安创新生物技术有限公司段，陵浔<120>20种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗<130>NM_144782<160>1<170>PatentIn version 3.5<210>1<211>2558<212>DNA<213>Homo sapiens<220><221>STS<222>(80). . (312)<220><221>transit_peptide<222>(296). · (328)<220><221>misc_feature<222> (1139) ·· (1140)<220><221>STS<222> (1688). . (2398)<220><221>STS<222> (1921) ·· (2176)<220><221>STS<222> (2156) ·· (2430)<220><221>STS<222> (2348) ·· (2493)<220><221>polyA一signal<222> (2509) ·· (2514)<220>[1759]<221>polyA_site[1760]<222> (2533). . (2533)<400>1atcgatgccc cgcccccgcc ccacagccaa cctcccgggc accgttgccc gcccggggcc 60cgctaccggc ccaggccccg cctcgcgagt cctcctcccc gggtgcctgc ccgcagcccg 120ctcggcccag agggtgggcg cggggctgcc tctaccggct ggcggctgta actcagcgac 180cttggcccga aggctctagc aaggacccac cgaccccagc cgcggcggcg gcggcgcgga 240ctttgcccgg tgtgtggggc ggagcggact gcgtgtccgc ggacgggcag cgaagatgtt 300agccttcgct gccaggaccg tggtgaagcc tctgggcttc ctgaagccct tctccttgat 360gaaggcttcc agccgcttca aggcacacca ggatgcactg ccacggctgc ccgtgccccc 420tctccagcag tccctggacc actacctgaa ggcgctgcag cccatcgtga gtgaggagga 480gtgggcccac accaagcagc tggtggatga gtttcaggcc tcaggaggtg taggggagcg 540cctgcagaag gggctggagc gtcgggccag gaagacggag aactggctgt ctgagtggtg 600gctcaagacc gcctacctcc agtaccgcca gcctgtggtc atctactcga gcccaggcgt 660gatgctaccc aagcaggact tcgtggacct gcagggtcag ctccgatttg ctgccaaact 720cattgagggt gtgttggatt tcaaggtcat gattgacaac gagaccctgc ccgtggagta 780cctggggggg aagccactgt gcatgaacca gtactatcag atcttgtcct cctgccgagt 840gccgggcccc aagcaggaca cagtcagcaa cttcagcaag accaagaagc ctcccacgca 900catcaccgtg gtacacaact accagttttt tgagctggat gtgtaccaca gtgacgggac 960acccctcact gcggatcaga tctttgtgca gctggagaag atctggaact catccctaca 1020gaccaacaag gagcctgtgg gcatcctcac ctccaaccac cgcaactcct gggccaaggc 1080[1780]atacaacacc ctcatcaaag acaaggtgaa ccgggattcc gtgcgctcca tccagaagaa 1140acccgagctt gtgcggtctc ccatggtgcc cctgcccatg cccaagaagc tgcggttcaa 1200 [1782]catcaccccc gagatcaaga gcgacatcga gaaggccaag cagaacctca gcatcatgat 1260ccaggacctg gatatcaccg tgatggtgtt ccaccatttt ggaaaagact tccccaagtc 1320ggagaagcta agcccagatg ccttcatcca gatggctttg cagctggcct actacaggat 1380ctacggacag gcatgtgcca cctatgaaag tgcctccctg cgcatgtttc acctgggccg 1440caccgacacc atccgctcgg cttccatgga ctcactcacc tttgtcaagg ccatggatga 1500ctccagcgtc acggagcacc agaaggtgga gctgctgcgg aaggccgtgc aggcccaccg 1560aggctacacc gaccgggcca tccgcgggga ggcctttgat cgacacctgc tgggcctgaa 1620gctgcaggcc atcgaggacc tggtgagcat gcccgacatc ttcatggaca cctcctacgc 1680catcgccatg cacttccacc tctccaccag ccaggtccct gccaagacag actgtgtcat 1740gttcttcggg cccgtggtcc ccgacggcta cggtgtctgc tataacccca tggaggccca 1800catcaacttc tccctgtcgg cctacaacag ctgcgcggag accaacgccg cccgcctggc 1860gcattacctg gagaaggcgc tcctggacat gcgtgccctg ctgcagagcc acccccgggc 1920caagctctga gcccctagga ctcaggcctg ccaatgccac agccaagccc accctgggat 1980gggccaccca ccagggctca gctccttggt tccctcttcc ttggttccct cttccctggt 2040ccccccaaat ctactgagcc acggaccgca tcctccaggg ggcctgcagg ccccagccaa 2100gtgccttccg tgggtcatcc cagcacctgc cagggcccga cctggggctg agtgcagagg 2160ctgagcagga cgttaggccc gggcccctgg cacgtcctcc accggtgcct cctctgggga 2220agggaaccag ccctccagag caggagactg gcaagagctc tttgtctcac cagctcagcc2280ccggccactc cctgccaact ccatgaccag gccaccatct gtaccctgct tcccaaactc 2340ccaggacctg gagacaggat tgtctggggc cgaggggcca gggtgtgagg tttcacctcc 2400gttgcggctg tgctcctgtg gataacattg ctagcgagcc gcctctggtt ccactcagct 2460tggttcctgc ccccgccctg ctgtatgata ttaatgtgga aggtcatcaa taaaggggaa 2520ttgtgttggt gaaaaaaaaa aaaaaaaaaa aaaaaaaa 2558SEQUENCE LISTING6<110>北京思安创新生物技术有限公司 [1807]段，陵浔<120>20种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗<130>NM_001896<160>1<170>PatentIn version 3. 5<210>1<211>1674<212>DNA<213>Homo sapiens<220><221>exon<222>(1). . (246)<220><221>exon<222>(247). . (358)<220><221>exon<222>(359). . (460)<220><221>exon<222>(461). . (511)<220><221>exon<222>(512). . (571)<220><221>exon[1833]<222>(572). . (655)<220><221>exon<222>(656). . (766) <220><221>exon<222>(767). . (868)<220><221>STS<222>(783). . (987)<220><221>exon<222>(869). . (969)<220><221>exon<222> (970). . (1118)<220><221>exon<222>(1119). . (1212)<220><221>exon<222>(1213)·· (1659)<220><221>STS<222> (1293) ·· (1569)<220><221>STS<222>(1376)·· (1558)<400>1gcg gcc gcc cgc cgc cgc get cct cct cct cct cct cca gcg ccc ggc 48Ala Ala Ala Arg Arg Arg Ala Pro Pro Pro Pro Pro Pro Ala Pro Gly151015ggc ccg ctg cct cct ccg ccc gac gcc ccg cgt ccc ccg ccg cgc cgc 96Gly Pro Leu Pro Pro Pro Pro Asp Ala Pro Arg Pro Pro Pro Arg Arg202530cgc cgc cac cct ctg cgc ccc gcg ccg ccc ccc ggt ccc gcc cgc cat 144Arg Arg His Pro Leu Arg Pro Ala Pro Pro Pro Gly Pro Ala Arg His354045gcc egg ccc ggc cgc ggg cag cag ggc ccg ggt cta cgc cga ggt gaa 192[1872]Ala Arg ProGly Arg Gly Gln Gln Gly Pro Gly Leu Arg Arg Gly Glu[1873]505560cag tct gaggag ccg cga gta ctg gga cta cga ggc tea cgt ccc gag 240Gln Ser GluGlu Pro Arg Val Leu Gly Leu Arg Gly Ser Arg Pro Glu65707580ctg βββ taatea aga tga tta cca act ggt tcg aaa act tgg tcg ggg 288Leu GlySer Arg Leu Pro Thr Gly Ser Lys Thr Trp Ser Gly8590aaa ata tagtga agt att tga ggc cat taa tat cac caa caa tga gag 336Lys lieSer lie Gly HisTyr His Gln Gln Glu95100105agt ggt tgtaaa aat cct gaa g cc agt gaa gaa aaa gaa gat aaa acg 384Ser Gly CysLys Asn Pro Glu Ala Ser Glu Glu Lys Glu Asp Lys Thr110115120aga ggt taagat tct gga gaa cct tcg tgg tgg aac aaa tat cat taa 432Arg GlyAsp Ser Gly Glu Pro Ser Trp Trp Asn Lys Tyr His125130135get gat tgacac tgt aaa gga ccc cgt g tc aaa gac acc age ttt ggt 480Ala AspHis Cys Lys Gly Pro Arg Val Lys Asp Thr Ser Phe Gly140145150att tga atatat CQ-Q- 3-3- tac aga ttt taa g ca act cta cca gat cct 528lie lieTyr Gln Tyr Arg PheAla Thr Leu Pro Asp Pro155160gac aga ctttga tat ccg gtt tta tat gta tga act act taa a gc tct 576Asp Arg Leu Tyr Pro Val Leu Tyr Val Thr ThrSer Ser165170gga tta ctgcca cag caa ggg aat cat gca cag gga tgt gaa acc tea 624Gly Leu LeuPro Gln Gln Gly Asn His Ala Gln Gly Cys Glu Thr Ser180185190caa tgt catgat aga tea cca aca gaa aaa g ct gcg act gat aga ttg 672Gln Cys HisAsp Arg Ser Pro Thr Glu Lys Ala Ala Thr Asp Arg Leu195200205ggg tct ggcaga att cta tea tcc tgc tea gga gta caa tgt tcg tgt 720Gly Ser GlyArg lie Leu Ser Ser Cys Ser Gly Val Gln Cys Ser Cys210215220age ctc aaggta ctt caa ggg acc aga get cct cgt gga cta tea g at 768Ser Leu LysVal Leu Gln Gly Thr Arg Ala Pro Arg Gly Leu Ser Asp225230235gta tga ttatag ctt gga cat gtg gag ttt ggg ctg tat gtt age aag 816[1911]Val Leu Leu Gly His Val Glu Phe Gly Leu Tyr Val Ser Lys245250cat gat ctt tcg aag gga acc att ctt cca tgg aca gga caa cta tga 864His Asp Leu Ser Lys Gly Thr lie Leu Pro Trp Thr Gly Gln Leu255260265cca g ct tgt tcg cat tgc caa ggt tct ggg tac aga aga act gta tgg 912Pro Ala Cys Ser His Cys Gln Gly Ser Gly Tyr Arg Arg Thr Val Trp270275280285gta tct gaa gaa gta tea cat aga cct aga tcc aca ctt caa cga tat 960Val Ser Glu Glu Val Ser His Arg Pro Arg Ser Thr Leu Gln Arg Tyr290295300cct ggg aca aca ttc acg gaa acg ctg gga aaa ctt tat cca tag tga 1008Pro Gly Thr Thr Phe Thr Glu Thr Leu Gly Lys Leu Tyr Pro305310315gaa cag aca cct tgt cag ccc tga ggc cct aga tct tct gga caa act 1056Glu Gln Thr Pro Cys Gln Pro Gly Pro Arg Ser Ser Gly Gln Thr320325330tct gcg ata cga cca tea aca gag act gac tgc caa aga ggc cat gga 1104Ser Ala lie Arg Pro Ser Thr Glu Thr Asp Cys Gln Arg Gly His Gly335340345gca ccc ata ctt ct a ccc tgt ggt gaa gga gca gtc cca gcc ttg tgc 1152Ala Pro lie Leu Leu Pro Cys Gly Glu Gly Ala Val Pro Ala Leu Cys350355360aga caa tgc tgt get ttc cag tgg tct cac ggc age acg atg aag act 1200Arg Gln Cys Cys Ala Phe Gln Trp Ser His Gly Ser Thr Met Lys Thr365370375gga aag cga egg gtc tgt tgc ggt tct ccc act ttt cca taa gca gaa 1248Gly Lys Arg Arg Val Cys Cys Gly Ser Pro Thr Phe Pro Ala Glu380385390caa gaa cca aat caa acg tct taa cgc gta tag aga gat cac gtt ccg 1296Glu Glu Pro Asn Gln Thr Ser Arg Val Arg Asp His Val Pro395400405tga gca gac aca aaa egg tgg cag gtt tgg cga gca cga act aga cca 1344Ala Asp Thr Lys Arg Trp Gln Val Trp Arg Ala Arg Thr Arg Pro410415420[1946]age gaa ggg cag ccc acc acc gta tat caa acc tea ctt ccg aat gta 1392Ser Glu Gly Gln Pro Thr Thr Val Tyr Gln Thr Ser Leu Pro Asn Val425430435aaa ggc tea ctt gcc ttt ggc ttc ctg ttg act tct tcc cga ccc aga 1440[1950]Lys Gly Ser Leu Ala Phe Gly Phe Leu Leu Thr Ser Ser Arg Pro Arg440445450 [1952]aag cat ggg gaa tgt gaa ggg tat gca gaa tgt tgt tgg tta ctg ttg 1488Lys His Gly Glu Cys Glu Gly Tyr Ala Glu Cys Cys Trp Leu Leu Leu455460465470ctc ccc gag ccc ctc aac tcg tcc cgt ggc cgc ctg ttt ttc cag caa 1536Leu Pro Glu Pro Leu Asn Ser Ser Arg Gly Arg Leu Phe Phe Gln Gln475480485acc acg cta act age tga cca cag act cca cag tgg ggg gac ggg cgc 1584Thr Thr Leu Thr Ser Pro Gln Thr Pro Gln Trp Gly Asp Gly Arg490495500agt atg tgg cat ggc ggc agt tac ata tta tta ttt taa aag tat ata 1632Ser Met Trp His Gly Gly Ser Tyr lie Leu Leu Phe Lys Tyr lie505510515tta ttg aat aaa agg ttt taa aag aaa aaaaaaaaaaa aaaaa1674Leu Leu Asn Lys Arg Phe Lys Lys520SEQUENCE LISTING7<110>北京思安创新生物技术有限公司段，陵浔<120>20种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗<130>NM_003462<160>1<170>PatentIn version 3. 5<210>1<211>2663<212>DNA<213>Homo sapiens<220><221>exon<222>(1). . (157)<220><221>exon<222>(158). . (303)<220><221>exon<222>(304). . (473)<220><221>exon[1989]<222>(474). . (652)[1990]<220><221>exon<222>(653). . (817)<220><221>STS<222>(653). . (817)<220><221>exon<222> (818) ·· (2649)<220><221>STS<222>(1639)·· (1769)<220><221>STS<222> (2489)·· (2621)<220><221>polyA_signal<222>(2628)·· (2633)<220><221>polyA_site<222>(2649). . (2649)<400>1ggc ggt ttc cat ggt gac ggc aaa caa ggc cca cac tgg aca ggg cag 48Gly Gly Phe His Gly Asp Gly Lys Gln Gly Pro His Trp Thr Gly Gln151015ctg ctg ggt tgc tac tct cgc ctc cgc cat gat tcc gcc cgc aga ctc 96Leu Leu Gly Cys Tyr Ser Arg Leu Arg His Asp Ser Ala Arg Arg Leu202530ttt get caa gta cga cac ccc agt get ggt gag ccg gaa cac gga gaa 144Phe Ala Gln Val Arg His Pro Ser Ala Gly Glu Pro Glu His Gly Glu354045acg gag ccc caa g gc tcg get act gaa agt cag ccc cca gca gcc tgg 192Thr Glu Pro Gln Gly Ser Ala Thr Glu Ser Gln Pro Pro Ala Ala Trp505560acc ttc agg ttc age ccc aca gcc acc caa gac caa get ccc ctc aac 240Thr Phe Arg Phe Ser Pro Thr Ala Thr Gln Asp Gln Ala Pro Leu Asn65707580tcc ctg tgt ccc aga tcc tac aaa gca ggc aga aga aat ctt gaa tgc 288[2028]Ser Leu CysPro Arg Ser Tyr Lys Ala Gly Arg Arg Asn Leu Glu Cys859095cat act accccc aag gga gtg ggt gga aga cac gca get atg gat cca 336His Thr ThrPro Lys Gly Val Gly Gly Arg His Ala Ala Met Asp Pro100105110gca ggt gtccag cac ccc tag cac cag gat gga cgt ggt gca cct cca 384Ala Gly ValGln His Pro His Gln Asp Gly Arg Gly Ala Pro Pro115120125 gga gca gttaga ctt aaa get gca gca gcg gca ggc cag gga aac agg 432Gly Ala ValArg Leu Lys Ala Ala Ala Ala Ala Gly Gln Gly Asn Arg130135140cat ctg ccctgt ccg cag gga act cta ctc aca gtg ttt tg a tga gtt 480His Leu ProCys Pro Gln Gly Thr Leu Leu Thr Val PheVal145150155gat ccg ggaggt cac cat caa ctg tgc gga gag ggg get get get get 528Asp Pro GlyGly His His Gln Leu Cys Gly Glu Gly Ala Ala Ala Ala160165170gcg agt ccggga cga gat ccg cat gac cat cgc tgc cta cca gac cct 576Ala Ser ProGly Arg Asp Pro His Asp His Arg Cys Leu Pro Asp Pro175180185190gta cga gagcag cgt ggc gtt tgg cat gag gaa ggc act gca ggc tga 624Val Arg GluGln Arg Gly Val Trp His Glu Glu Gly Thr Ala Gly195200205gca ggg gaagtc aga cat gga gag gaa a at cgc aga att gga gac gga 672Ala Gly GluVal Arg His Gly Glu Glu Asn Arg Arg lie Gly Asp Gly210220aaa gag agacct gga gag gca agt gaa cga gca gaa ggc aaa atg tga 720Lys Glu ArgPro Gly Glu Ala Ser Glu Arg Ala Glu Gly Lys Met225230235age cac tgagaa gcg gga gag cga gag gcg gca ggt gga gga gaa gaa 768Ser HisGlu Ala Gly Glu Arg Glu Ala Ala Gly Gly Gly Glu Glu240245250gca caa tgagga gat tea gtt cct gaa gcg aac aaa tea gca get gaa g 817Ala GlnGly Asp Ser Val Pro Glu Ala Asn Lys Ser Ala Ala Glu255260265gc cca actgga agg cat tat tgc acc aaa gaa gtg ata att tcc aca 864Gly Pro ThrGly Arg His Tyr Cys Thr Lys Glu Val lie lie Ser Thr270275280tga tta atttcc aac aag aca ctt ggg agt tat tta ctg tgt tcc tct 912[2067]Leu lie Ser Asn Lys Thr Leu Gly Ser Tyr Leu Leu Cys Ser Ser285290295ggc age caa taa aat cat cat aag ccc ttt gta ata aaa age tag ttt 960 [2070]Gly Ser Gln Asn His His Lys Pro Phe Val lie Lys Ser Phe300305310cct gag tga aca age cat aac ctc ccc taa aca cca cct agg tat ttg 1008Pro Glu Thr Ser His Asn Leu Pro Thr Pro Pro Arg Tyr Leu315320325tta gaa gtc aca cta tta ctc caa tgt cat cag aca cct aag gtc tgc 1056Leu Glu Val Thr Leu Leu Leu Gln Cys His Gln Thr Pro Lys Val Cys330335340cag cca ggc tcc tgg ctg ggc aat gga aga tgg tgt ggc cct gtt agt 1104Gln Pro Gly Ser Trp Leu Gly Asn Gly Arg Trp Cys Gly Pro Val Ser345350355ctc cgt gtg tgg ctt act age cag cct tgg gaa ctg cca act caa att 1152Leu Arg Val Trp Leu Thr Ser Gln Pro Trp Glu Leu Pro Thr Gln lie360365370cta aga aag cca ctg ctt tct cat cat cac tct ata cca ata ctt att 1200Leu Arg Lys Pro Leu Leu Ser His His His Ser lie Pro lie Leu lie375380385tct ggc caa atg aat ctg ctt ctc tgc ccc tea aac ttt tag ttc aca 1248Ser Gly Gln Met Asn Leu Leu Leu Cys Pro Ser Asn Phe Phe Thr390395400att cat ctt cta cct taa ctt ggg ctt ctt ggg cct ctg gcc ttc ctt 1296lie His Leu Leu Pro Leu Gly Leu Leu Gly Pro Leu Ala Phe Leu405410415act taa tgt ctt ctt ttc cct act cta atg cat ttc taa ctc act ttg 1344Thr Cys Leu Leu Phe Pro Thr Leu Met His Phe Leu Thr Leu420425430gag ctt tgg ttt tct aat gta tta tcc cca ctt gcc agt caa ctg gac 1392Glu Leu Trp Phe Ser Asn Val Leu Ser Pro Leu Ala Ser Gln Leu Asp435440445ccc ttc ctc ctc ggt ttc aga ctg cct aca tta gga aac aat ggc agt 1440Pro Phe Leu Leu Gly Phe Arg Leu Pro Thr Leu Gly Asn Asn Gly Ser450455460465caa acc cat ggc ttt gga gaa agt aaa tgt ttg cca gaa agg aat act 1488Gln Thr His Gly Phe Gly Glu Ser Lys Cys Leu Pro Glu Arg Asn Thr470475480agt cac agt ggc ctt tgt gag ttg tct gca act cag ctc ttc ccc cag 1536[2106]Ser His Ser Gly Leu Cys Glu Leu Ser Ala Thr Gln Leu Phe Pro Gln485490495cac aga tct gtt ccc ctt ate ctg cag aaa ate aag ccc tga ctc tgc 1584 [2109]His Arg Ser Val Pro Leu lie Leu Gln Lys lie Lys Pro Leu Cys500505510act ccc cga agt agt gat gtt aat taa caa ctg aag agg taa cta aat 1632Thr Pro Arg Ser Ser Asp Val Asn Gln Leu Lys Arg Leu Asn515520525ctc aca tgc agg tct aat gac taa taa ttg gag tac ggc tgc tag aca 1680Leu Thr Cys Arg Ser Asn AspLeu Glu Tyr Gly Cys Thr530535act gca ttt tag tat ttc tct tcc att ctc ctg gtt ttg tag acc cag 1728Thr Ala Phe Tyr Phe Ser Ser lie Leu Leu Val Leu Thr Gln540545550aag att gaa tga gtg aca taa ate ttt agt tcg ggg caa gcc agg gtg 1776Lys lie Glu Val Thr lie Phe Ser Ser Gly Gln Ala Arg Val555560565ggc tag ggt ggt aag ctg gag gac ttc ate ctt cag tta ggc tgc aca 1824Gly Gly Gly Lys Leu Glu Asp Phe lie Leu Gln Leu Gly Cys Thr570575580agt aac att acc taa aag gca cta aca tgc tea ggt tcc cca gaa aga 1872Ser Asn lie Thr Lys Ala Leu Thr Cys Ser Gly Ser Pro Glu Arg585590595ggc gta aga agg gcc tct cct tag cag age ttc cac ctg cca tcc gtc 1920Gly Val Arg Arg Ala Ser Pro Gln Ser Phe His Leu Pro Ser Val600605610ttg ggt tea gtg age ttc aag get cac aat gga age act gtc att tcc 1968Leu Gly Ser Val Ser Phe Lys Ala His Asn Gly Ser Thr Val lie Ser615620625cca gaa aag ctg tgt tcc cta tgc tga aca cac cat aca cat tct cat 2016Pro Glu Lys Leu Cys Ser Leu Cys Thr His His Thr His Ser His630635640ctg gaa tct aag gag cag ctt tta ccc tga tcc agt ate ctg agg aat 2064Leu Glu Ser Lys Glu Gln Leu Leu Pro Ser Ser lie Leu Arg Asn645650655ttt aag cct cca ctc aaa tga cct gcc tgt gtt gtc att tcc atg gga 2112Phe Lys Pro Pro Leu Lys Pro Ala Cys Val Val lie Ser Met Gly660665670aag aac tct ttc cac gag ate tgc tag ttc cag gcc tct aag aca gga 2160[2145]LysAsn Ser Phe His Glu lie CysPhe Gln Ala Ser Lys Thr Gly675680685acg tat gtg cca taa gtg ggt cta ctt cac aga ctc aat gag gca gaa 2208Thr Tyr Val ProVal Gly Leu Leu His Arg Leu Asn Glu Ala Glu690695700attatt gta gtt ttc tcc tat ttc ttc tgc 3-CC C3.3. Ctt tct CCt tgt 2256lielie Val Val Phe Ser Tyr Phe Phe Cys Thr Gln Leu Ser Pro Cys705710715 [2153]atttea aag gcc agg cca tgt aca cta acg tcc ttg aaa ttt gca gtt 2304lieSer Lys Ala Arg Pro Cys Thr Leu Thr Ser Leu Lys Phe Ala Val720725730735ctg tat get tct att cca aat cat tea tta cca ata aaa acg aaa tac 2352Leu Tyr Ala Ser lie Pro Asn His Ser Leu Pro lie Lys Thr Lys Tyr740745750caccct ttc cat ttt ata gac ctc ate ccc tat ttc tgt cag aca gtt 2400HisPro Phe His Phe lie Asp Leu lie Pro Tyr Phe Cys Gln Thr Val755760765ata tga cag ggt gac tgt gga acc tct tag ttc ate caa agt cta cct 2448lieGln Gly Asp Cys Gly Thr SerPhe lie Gln Ser Leu Pro770775780gaa gtg cta gac ttt cag act ctt ate act gaa ate ctt aag gtt gag 2496Glu Val Leu Asp Phe Gln Thr Leu lie Thr Glu lie Leu Lys Val Glu785790795gag get tta ttt ccc tag cac tgg tga agg get tea act gtc aaa cct 2544Glu Ala Leu Phe ProHis TrpArg Ala Ser Thr Val Lys Pro800805810cag aac aaa tgc att agg gcc tta gaa atg tea atg ggg cag gaa gaa 2592Gln Asn Lys Cys lie Arg Ala Leu Glu Met Ser Met Gly Gln Glu Glu815820825aacaca att tct aac tgc ctg ttt ttg tat aat tta ata aaa acc ttt 2640Asn Thr lie Ser Asn Cys Leu Phe Leu Tyr Asn Leu lie Lys Thr Phe830835840 2663Thr Leu845SEQUENCE LISTING8<110>北京思安创新生物技术有限公司段，陵浔[2183]<120>20种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗<130>NM_033214<160>1<170>PatentIn version 3.5 <210>1<211>1899<212>DNA<213>Homo sapiens<220><221>exon<222>(1). . (1866)<220><221>CDS<222>(19). . (1680)<220><221>STS<222> (943). . (1214)<220><221>STS<222>(1362)·· (1690)<220><221>STS<222>(1686)·· (1872)<400>1age cga cct act ggt gtc atg gca gcc cca aag aca gca get gtg ggg 48Ser Arg Pro Thr Gly Val Met Ala Ala Pro Lys Thr Ala Ala Val Gly151015ccg ttg gtg gga gcg gtg gtc cag ggc acc aac tcc act cgc ttt ctg 96Pro Leu Val Gly Ala Val Val Gln Gly Thr Asn Ser Thr Arg Phe Leu202530gtt ttc aat tea aaa aca gcg gaa cta ctt agt cat cac aaa gtg gaa 144Val Phe Asn Ser Lys Thr Ala Glu Leu Leu Ser His His Lys Val Glu354045tta aca caa gag ttc cca aaa gaa gga tgg gtg gaa caa gac cct aaa 192Leu Thr Gln Glu Phe Pro Lys Glu Gly Trp Val Glu Gln Asp Pro Lys505560gaa att ctt cag tct gtc tac gag tgt ata gcg aga acg tgt gag aaa 240Glu lie Leu Gln Ser Val Tyr Glu Cys lie Ala Arg Thr Cys Glu Lys65707580[2222]ctt gac gaactg aat att gat ata tcc aac ata aaa get gtt ggt gtc 288Leu Asp GluLeu Asn lie Asp lie Ser Asn lie Lys Ala Val Gly Val859095age aat cagagg gaa acc act gta ate tgg gac aag tta aca gga gag 336Ser Asn GlnArg Glu Thr Thr Val lie Trp Asp Lys Leu Thr Gly Glu100105110cct ctc tacaat get gtg gtg tgg ctt gat cta aga acc cag act act 384 [2229]Pro Leu TyrAsn Ala Val Val Trp Leu Asp Leu Arg Thr Gln Thr Thr115120125gtt gag gatctt agt aaa aaa att cca gga aat agt aac ttc gtc aag 432Val Glu AspLeu Ser Lys Lys lie Pro Gly Asn Ser Asn Phe Val Lys130135140tct aag acaggc ctt cca ctc age act tac ttc agt gca gta aaa ctt 480Ser Lys ThrGly Leu Pro Leu Ser Thr Tyr Phe Ser Ala Val Lys Leu145150155160cgt tgg atgctt gac aat gtg aga aac gtc caa aag get gtt gaa gaa 528Arg Trp MetLeu Asp Asn Val Arg Asn Val Gln Lys Ala Val Glu Glu165170175ggt aga getctt ttt ggt acc att gat tea tgg ctt ate tgg agt ttg 576Gly Arg Ala Leu Phe Gly Thr lie Asp Ser Trp Leu lie Trp Ser Leu180185190aca gga ggagtt aat gga ggc gtg cat tgt aca gat gta aca aat gca 624Thr Gly GlyVal Asn Gly Gly Val His Cys Thr Asp Val Thr Asn Ala195200205agt agg acaatg ctt ttt aat ate cat tct ttg gaa tgg gat aaa gag 672Ser Arg ThrMet Leu Phe Asn lie His Ser Leu Glu Trp Asp Lys Glu210215220ctc tgt gacttt ttt gaa att cca atg gac ctt ctt cca aat gtc ttc 720Leu Cys AspPhe Phe Glu lie Pro Met Asp Leu Leu Pro Asn Val Phe225230235240agt tct tctgag ate tat ggc cta att aaa act gga gcc ctg gaa ggt 768Ser Ser SerGlu lie Tyr Gly Leu lie Lys Thr Gly Ala Leu Glu Gly245250255gtg cca atatct ggg tgt ttg ggg gac caa tgt get gca tta gta gga 816Val Pro lieSer Gly Cys Leu Gly Asp Gln Cys Ala Ala Leu Val Gly260265270caa atg tgcttc cag gag gga caa gcc aaa aac acc tat gga aca ggt 864Gln Met CysPhe Gln Glu Gly Gln Ala Lys Asn Thr Tyr Gly Thr Gly275280285[2261]tgc ttc tta ctg tgt aat acg ggt cgt aaa tgt gtg ttt tct gaa cat 912[2262]Cys Phe Leu Leu Cys Asn Thr Gly Arg Lys Cys Val Phe Ser Glu His290295300ggc ctt ttg acc aca gta get tac aaa cta ggc aga gag aag cca gca 960Gly Leu Leu Thr Thr Val Ala Tyr Lys Leu Gly Arg Glu Lys Pro Ala305310315320tat tat gca ctg gaa ggt tct gtt get ata gca ggt get gtt att cgt 1008Tyr Tyr Ala Leu Glu Gly Ser Val Ala lie Ala Gly Ala Val lie Arg325330335tgg cta aga gac aat ctt gga att ata gag acc tea gga gac att gaa 1056Trp Leu Arg Asp Asn Leu Gly lie lie Glu Thr Ser Gly Asp lie Glu340345350aga ctt get aaa gaa gta gga act tct tat ggc tgt tac ttt gtc cca 1104Arg Leu Ala Lys Glu Val Gly Thr Ser Tyr Gly Cys Tyr Phe Val Pro355360365gcc ttt tea ggg tta tat gca cct tat tgg gag ccc agt gca aga ggg 1152Ala Phe Ser Gly Leu Tyr Ala Pro Tyr Trp Glu Pro Ser Ala Arg Gly370375380ata ctc tgt ggc ctc act cag ttt acc aat aaa tgt cat att get ttt 1200lie Leu Cys Gly Leu Thr Gln Phe Thr Asn Lys Cys His lie Ala Phe385390395400get gca tta gaa get gtt tgt ttc caa acc cga gag att ttg gaa gcc 1248Ala Ala Leu Glu Ala Val Cys Phe Gln Thr Arg Glu lie Leu Glu Ala405410415atg aac cgt gac tgt gga att cca ctt cgt cat ttg cag gta gat gga 1296Met Asn Arg Asp Cys Gly lie Pro Leu Arg His Leu Gln Val Asp Gly420425430gga atg acc aac aac aaa gtt ctt atg cag cta caa gca gat att ctt 1344Gly Met Thr Asn Asn Lys Val Leu Met Gln Leu Gln Ala Asp lie Leu435440445cat att cca gta ata aaa ccc ttt atg cct gaa aca act gca cta gga 1392His lie Pro Val lie Lys Pro Phe Met Pro Glu Thr Thr Ala Leu Gly450455460get gcc atg gca gca ggg get gca gag gga gta age gtt tgg age ctt 1440Ala Ala Met Ala Ala Gly Ala Ala Glu Gly Val Ser Val Trp Ser Leu465470475480gaa ccc cag get ttg tea gtt ctc agg atg gaa cga ttt gaa cca cag 1488Glu Pro Gln Ala Leu Ser Val Leu Arg Met Glu Arg Phe Glu Pro Gln485490495[2300]ate cag gcc aca gaa agt gaa att cgt tat gcc aca tgg aag aaa gcc 1536lie Gln Ala Thr Glu Ser Glu lie Arg Tyr Ala Thr Trp Lys Lys Ala500505510gta atg aag tea atg ggt tgg gtt acc agt cag tct cct gaa ggt ggt 1584Val Met Lys Ser Met Gly Trp Val Thr Ser Gln Ser Pro Glu Gly Gly515520525gat cct tct ate ttc tct agt ctg cct ttg gga ttt ttt ata gtg agt 1632Asp Pro Ser lie Phe Ser Ser Leu Pro Leu Gly Phe Phe lie Val Ser530535540age atg gta atg cta att gga gca aga tat ate tcg ggt gtg cca taa 1680Ser Met Val Met Leu lie Gly Ala Arg Tyr lie Ser Gly Val Pro545550555taa tac cag cta tgg agt ccc aag atg taa aca ttt tac ata atg aaa 1728Tyr Gln Leu Trp Ser Pro Lys Met Thr Phe Tyr lie Met Lys560565570gca tat age agt tct gcc tct taa tat aat gac act att ata gac tct 1776Ala Tyr Ser Ser Ser Ala Ser Tyr Asn Asp Thr lie lie Asp Ser575580585gat ttt gtt tat aaa cca ctt get aca tga ccc atg aaa acc cta gac 1824Asp Phe Val Tyr Lys Pro Leu Ala Thr Pro Met Lys Thr Leu Asp590595600ttg tga cct gaa ata aag aaa ata cgt tag aag age act gta1866Leu Pro Glu lie Lys Lys lie Arg Lys Ser Thr Val605610615 1899SEQUENCE LISTING9<110>北京思安创新生物技术有限公司段，陵浔<120>20种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗<130>NM_153823<160>1<170>PatentIn version 3. 5<210>1<211>1942<212>DNA<213>Homo sapiens<220>[2337]<221>exon[2338]<222>(1). · (177)<220><221>exon<222>(178). . (493)<220><221>exon<222>(494). . (541) [2345]<220><221>exon<222>(542). . (694)<220><221>exon<222>(695). . (806)<220><221>exon<222> (807) ·· (1942)<220><221>STS<222> (884)·· (1736)<220><221>polyA_site<222> (1225)·· (1225)<400>1age att taa tea tga ggc act cca aag aat atg aga tag aag ggg agg 48Ser lie Ser Gly Thr Pro Lys Asn Met Arg Lys Gly Arg1510ggt agg agg aga ggg agg agg gta gga aga cag ttt gca ttc ttg caa 96Gly Arg Arg Arg Gly Arg Arg Val Gly Arg Gln Phe Ala Phe Leu Gln152025cat taa acc aaa ggg act tgg agt gca gat ggc ate ctt egg ttc ttc 144His Thr Lys Gly Thr Trp Ser Ala Asp Gly lie Leu Arg Phe Phe303540cag aca age tgc aag acg ctg acc atg gcc aag atg gag ctc tcg aag 192Gln Thr Ser Cys Lys Thr Leu Thr Met Ala Lys Met Glu Leu Ser Lys45505560gcc ttc tct ggc cag egg aca ctc cta tct gcc ate ctc age atg cta 240Ala Phe Ser Gly Gln Arg Thr Leu Leu Ser Ala lie Leu Ser Met Leu657075tea ctc age ttc tcc aca aca tcc ctg ctc age aac tac tgg ttt gtg 288[2377]Ser Leu SerPhe Ser Thr Thr Ser Leu Leu Ser Asn Tyr Trp Phe Val808590ggc aca cagaag gtg ccc aag ccc ctg tgc gag aaa ggt ctg gca gcc 336Gly Thr GlnLys Val Pro Lys Pro Leu Cys Glu Lys Gly Leu Ala Ala 95100105aag tgc tttgac atg cca gtg tcc ctg gat gga gat acc aac aca tcc 384Lys Cys PheAsp Met Pro Val Ser Leu Asp Gly Asp Thr Asn Thr Ser110115120acc cag gaggtg gta caa tac aac tgg gag act ggg gat gac egg ttc 432Thr Gln GluVal Val Gln Tyr Asn Trp Glu Thr Gly Asp Asp Arg Phe125130135140tcc ttc eggage ttc egg agt ggc atg tgg cta tcc tgt gag gaa act 480Ser Phe ArgSer Phe Arg Ser Gly Met Trp Leu Ser Cys Glu Glu Thr145150155gtg gaa gaacca g gg gag agg tgc cga agt ttc att gaa ctt aca cca 528Val Glu GluPro Gly Glu Arg Cys Arg Ser Phe lie Glu Leu Thr Pro160165170cca gcc aagaga g aa ate cta tgg tta tcc ctg gga acg cag ate acc 576Pro Ala LysArg Glu lie Leu Trp Leu Ser Leu Gly Thr Gln lie Thr175180185tac ate ggactt caa ttc ate age ttc ctc ctg cta cta aca gac ttg 624Tyr lie GlyLeu Gln Phe lie Ser Phe Leu Leu Leu Leu Thr Asp Leu190195200cta ctc actggg aac cct gcc tgt ggg ctc aaa ctg age gcc ttt get 672Leu Leu ThrGly Asn Pro Ala Cys Gly Leu Lys Leu Ser Ala Phe Ala205210215220get gtt tcctct gtc ctg tea g gt ctc ctg ggg atg gtg gcc cac atg 720Ala Val SerSer Val Leu Ser Gly Leu Leu Gly Met Val Ala His Met225230235atg tat teacaa gtc ttc caa gcg act gtc aac ttg ggt cca gaa gac 768Met Tyr SerGln Val Phe Gln Ala Thr Val Asn Leu Gly Pro Glu Asp240245250tgg aga ccacat gtt tgg aat tat ggc tgg gcc ttc ta c atg gcc tgg 816Trp Arg ProHis Val Trp Asn Tyr Gly Trp Ala Phe Tyr Met Ala Trp255260ctc tcc ttcacc tgc tgc atg gcg tcg get gtc acc acc ttc aac acg 864Leu Ser PheThr Cys Cys Met Ala Ser Ala Val Thr Thr Phe Asn Thr270275280tac acc aggatg gtg ctg gag ttc aag tgc aag cat agt aag age ttc 912[2416]Tyr Thr Arg Met Val Leu Glu Phe Lys Cys Lys His Ser Lys Ser Phe285290295300aag gaa aac ccg aac tgc cta cca cat cac cat cag tgt ttc cct egg 960Lys Glu Asn Pro Asn Cys Leu Pro His His His Gln Cys Phe Pro Arg305310315egg ctg tea agt gca gcc ccc accgtg ggt cct ttg acc age tac cac 1008Arg Leu Ser Ser Ala Ala Pro Thr Val Gly Pro Leu Thr Ser Tyr His [2423]320325330cag tat cat aat cag ccc ate cac tct gtc tct gag gga gtc gac ttc 1056Gln Tyr His Asn Gln Pro lie His Ser Val Ser Glu Gly Val Asp Phe335340345tac tcc gag ctg egg aac aag gga ttt caa aga ggg gcc age cag gag 1104Tyr Ser Glu Leu Arg Asn Lys Gly Phe Gln Arg Gly Ala Ser Gln Glu350355360ctg aaa gaa gca gtt agg tea tct gta gag gaa gag cag tgt tag gag 1152Leu Lys Glu Ala Val Arg Ser Ser Val Glu Glu Glu Gln Cys Glu365370375tta age ggg ttt ggg gag tag get tga gcc cta cct tac acg tct get 1200Leu Ser Gly Phe Gly Glu Ala Ala Leu Pro Tyr Thr Ser Ala380385390gat tat caa cat gtg ctt aag cca aca tcc gtc tct tga gca tgg ttt 1248Asp Tyr Gln His Val Leu Lys Pro Thr Ser Val Ser Ala Trp Phe395400405tta gag get acg aat aag get atg aat aag ggt tat ctt taa gtc cta 1296Leu Glu Ala Thr Asn Lys Ala Met Asn Lys Gly Tyr Leu Val Leu410415420agg gat tcc tgg gtg cca ctg ctc tct ttt cct cta cag ctc cat ctt 1344Arg Asp Ser Trp Val Pro Leu Leu Ser Phe Pro Leu Gln Leu His Leu425430435gtt tea ccc acc cca cat ctc aca cat cca gaa ttc cct tct tta ctg 1392Val Ser Pro Thr Pro His Leu Thr His Pro Glu Phe Pro Ser Leu Leu440445450455ata gtt tct gtg cca ggt tct ggg cta aac cat gga gat aaa aag aag 1440lie Val Ser Val Pro Gly Ser Gly Leu Asn His Gly Asp Lys Lys Lys460465470agt aaa ata cac ttc ccg acc tta agg ate tga tac ate tag tta ttg 1488Ser Lys lie His Phe Pro Thr Leu Arg lie Tyr lie Leu Leu475480485gtc tgg aaa ttg gag aaa tgg agg tga gca aat taa egg tag tgt gac 1536[2455]Val Trp Lys Leu Glu Lys Trp Arg Ala Asn Arg Cys Asp490495tgt ggc tea act ttc cat ggt acg cgc gaa ttg ttc tgg aag tea cag 1584 [2458]Cys Gly Ser Thr Phe His Gly Thr Arg Glu Leu Phe Trp Lys Ser Gln500505510atg gga aga cag aag cgc ccc tga gtg ctt tgt cat ata gca cag aag 1632Met Gly Arg Gln Lys Arg Pro Val Leu Cys His lie Ala Gln Lys515520525tcc tcc aca ctg cct ggg aca gtg tcc tgc ate cac aca gta ggt acc 1680Ser Ser Thr Leu Pro Gly Thr Val Ser Cys lie His Thr Val Gly Thr530535540545cag taa aag tgt tat taa ata ttg tcc cag gag ccc tta gga gag tgg 1728Gln Lys Cys Tyrlie Leu Ser Gln Glu Pro Leu Gly Glu Trp550555ttt aac atg aga agg cgc caa cag cac aca ggg tea ggg tea ctt cta 1776Phe Asn Met Arg Arg Arg Gln Gln His Thr Gly Ser Gly Ser Leu Leu560565570575gaa ata gcc cac caa ggc tat tea gat tgg act gtg gtt ccc ttc tgg 1824Glu lie Ala His Gln Gly Tyr Ser Asp Trp Thr Val Val Pro Phe Trp580585590aca gtc aca cca ggg act tcc cag tga aca agg ggg cat agg ctc ctc 1872Thr Val Thr Pro Gly Thr Ser Gln Thr Arg Gly His Arg Leu Leu595600605tct gga tcc cac ctt ttc ate cct tct gag aaa tgt acc cta tta ttt 1920Ser Gly Ser His Leu Phe lie Pro Ser Glu Lys Cys Thr Leu Leu Phe610615620cct tea taa aga atg ccaccc a 1942Pro Ser Arg Met Pro Pro625SEQUENCE LISTING10<110>北京思安创新生物技术有限公司段，陵浔<120>20种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗<130>NM_000849<160>1<170>PatentIn version 3. 5<210>1<211>4144[2493]<212>DNA<213>Homo sapiens<220><221>exon<222>(1). . (86)<220><221>exon<222>(87). . (358) [2501]<220><221>exon<222>(359). . (434)<220><221>exon<222>(435). . (499)<220><221>exon<222>(500). . (581)<220><221>exon<222>(582). . (682)<220><221>exon<222>(683). . (778)<220><221>exon<222>(779). . (889)<220><221>STS<222>(826).. (1064)<220〉<221>exon<222>(890).. (4127)<220><221>polyA_site<222>(1084)·. (1084)<220><221>STS<222>(1092)·. (1277)<220>[2532]<221>polyA_site<222> (1133)·· (1133)<220><221>polyA_signal<222>(1532)·· (1537)<220><221>polyA_site<222> (1557)·· (1557)<220><221>polyA_signal<222>(4098). . (4103)<220><221>polyA_site<222>(4127). . (4127)<400>1ggt tgg ttc tga gaa ggc ttc aag gaa tag gca gac att tea gca agg 48Gly Trp Phe Glu Gly Phe Lys Glu Ala Asp lie Ser Ala Arg1510ctg ctg agg aag gat agg ctg tgg aaa tta gga tgc ag c act cct gcc 96Leu Leu Arg Lys Asp Arg Leu Trp Lys Leu Gly Cys Ser Thr Pro Ala15202530egg gtc ccg cct egg ggt cgc cag gcc ctg aac ccc aac gcc ggc att 144Arg Val Pro Pro Arg Gly Arg Gln Ala Leu Asn Pro Asn Ala Gly lie354045agt cgc gcc tgc gca egg ccc tgt gga gcc gcg gag gca agg gac gga 192Ser Arg Ala Cys Ala Arg Pro Cys Gly Ala Ala Glu Ala Arg Asp Gly505560gaa egg ggc gga ggc gga gtc agg gcg ccc gcg cgt ggg ccc cgc ccc 240Glu Arg Gly Gly Gly Gly Val Arg Ala Pro Ala Arg Gly Pro Arg Pro657075ctt atg tcg ggt ata aag ccc ctc ccg ctc aca gtt tcc cta gtc ctc 288Leu Met Ser Gly lie Lys Pro Leu Pro Leu Thr Val Ser Leu Val Leu808590gaa ggc tcg gaa gcc cgt cac cat gtc gtg cga gtc gtc tat ggt tct336Glu Gly Ser Glu Ala Arg His His Val Val Arg Val Val Tyr Gly Ser95100105110egg gta ctg gga tat tcg tgg g ct ggc gca cgc cat ccg cct get cct 384Arg Val Leu Gly Tyr Ser Trp Ala Gly Ala Arg His Pro Pro Ala Pro[2570]115120125[2571]gga gtt cac gga tac ctc tta tga gga gaa acg gta cac gtg egg gga432[2572]Gly Val His Gly Tyr Leu Leu Gly Glu Thr Val His Val Arg Gly130135140ag c tcc tga cta tga tcg aag cca atg get gga tgt gaa att caa get 480Ser Ser Leu Ser Lys Pro Met Ala Gly Cys Glu lie Gln Ala145150155aga cct gga ctt tcc taa t ct gcc cta cct cct gga tgg gaa gaa caa 528Arg Pro Gly Leu SerSer Ala Leu Pro Pro Gly Trp Glu Glu Gln160165170gat cac cca gag caa tgc cat ctt gcg cta cat cgc tcg caa gca caa576Asp His Pro Glu Gln Cys His Leu Ala Leu His Arg Ser Gln Ala Gln175180185cat gt g tgg tga gac tga aga aga aaa gat tcg agt gga cat cat aga 624His Val Trp Asp Arg Arg Lys Asp Ser Ser Gly His His Arg190195200gaa cca agt aat gga ttt ccg cac aca act gat aag get ctg tta cag672Glu Pro Ser Asn Gly Phe Pro His Thr Thr Asp Lys Ala Leu Leu Gln205210215ctc tga cca c ga aaa act gaa gcc tea gta ctt gga aga get acc tgg 720Leu Pro Arg Lys Thr Glu Ala Ser Val Leu Gly Arg Ala Thr Trp220225230aca act gaa aca att ctc cat gtt tct ggg gaa att ctc atg gtt tgc768Thr Thr Glu Thr lie Leu His Val Ser Gly Glu lie Leu Met Val Cys235240245egg gga aaa g ct cac ctt tgt gga ttt tct cac cta tga tat ctt gga 816Arg Gly Lys Ala His Leu Cys Gly Phe Ser His Leu Tyr Leu Gly250255260tea gaa ccg tat att tga ccc caa gtg cct gga tga gtt ccc aaa cct864Ser Glu Pro TyrIlePro Gln Val Pro Gly Val Pro Lys Pro265270275gaa ggc ttt cat gtg ccg ttt tga g gc ttt gga gaa aat cgc tgc cta 912Glu Gly Phe His Val Pro PheGly Phe Gly Glu Asn Arg Cys Leu280285290ctt aca gtc tga tea gtt ctg caa gat gcc cat caa caa caa gat ggc 960Leu Thr Val Ser Val Leu Gln Asp Ala His Gln Gln Gln Asp Gly295300305cca gtg ggg caa caa gcc tgt atg ctg age agg agg cag act tgc aga 1008Pro Val Gly Gln Gln Ala Cys Met Leu Ser Arg Arg Gln Thr Cys Arg310315320[2610]get tgt ttt gtt tea tcc tgt ccg taa ggg gtc age get ctt get ttg 1056Ala Cys Phe Val Ser Ser Cys Pro Gly Val Ser Ala Leu Ala Leu325330335ctc ttt tea atg aat age act tat gtt act ggt gtc cag ctg agt ttc 1104Leu Phe Ser Met Asn Ser Thr Tyr Val Thr Gly Val Gln Leu Ser Phe340345350tct tgg gta taa agg cta aaa ggg aaa aag gat atg tgg aga ate ate 1152Ser Trp Val Arg Leu Lys Gly Lys Lys Asp Met Trp Arg lie lie 355360365aag ata tga att gaa tcg ctg cga tac tgg cat ttc cct act ccc caa 1200Lys lie lie Glu Ser Leu Arg Tyr Trp His Phe Pro Thr Pro Gln370375380ctg agt tea agg get gta ggt tea tgc cca age cct gag agt ggg tac 1248Leu Ser Ser Arg Ala Val Gly Ser Cys Pro Ser Pro Glu Ser Gly Tyr385390395tag aaa aaa cga gat tgc aca gtt gga gag age agg tgt gtt aaa tgg 1296Lys Lys Arg Asp Cys Thr Val Gly Glu Ser Arg Cys Val Lys Trp400405410gac tgg agt ccc tgt gaa gac tgg gtg agg ata aca caa gta aaa ctg 1344Asp Trp Ser Pro Cys Glu Asp Trp Val Arg lie Thr Gln Val Lys Leu415420425430tgg tac tga tgg act taa ccg gag ttc gga aac cgt cct gtg tac aca 1392Trp Tyr Trp Thr Pro Glu Phe Gly Asn Arg Pro Val Tyr Thr435440tgg gag ttt agt gtg ata aag gca gta ttt cag act ggt ggg cta gcc 1440Trp Glu Phe Ser Val lie Lys Ala Val Phe Gln Thr Gly Gly Leu Ala445450455460aat aga gtt ggg aca att get tac tea tta aaa ata ata gag ccc cac 1488Asn Arg Val Gly Thr lie Ala Tyr Ser Leu Lys lie lie Glu Pro His465470475ttg aca cta ttc act aaa att aat ctg gaa ttt aag gcc caa cat taa 1536Leu Thr Leu Phe Thr Lys lie Asn Leu Glu Phe Lys Ala Gln His480485490aca caa age tgt tga aat att gat gaa aat gta aga att ttt gtg acc 1584Thr Gln Ser Cys Asn lie Asp Glu Asn Val Arg lie Phe Val Thr495500505acg ggg tag gag aag ttt ctt taa aga cgt aaa aag aaa aaa acc ata 1632Thr Gly Glu Lys Phe Leu Arg Arg Lys Lys Lys Lys Thr lie510515520[2649]ggc tgc tct gcc tgt gga gta gcc att ctt tat tct ttt act ttt taa 1680Gly Cys Ser Ala Cys Gly Val Ala lie Leu Tyr Ser Phe Thr Phe525530535 [2652]aat aat aac aat cac aaa gga aag att tgc caa ttg aat agg ate aaa 1728Asn Asn Asn Asn His Lys Gly Lys lie Cys Gln Leu Asn Arg lie Lys540545550ttc tac aac ttt tcc taa ggc acc cac aaa taa atg aca agt gag aca 1776Phe Tyr Asn Phe Ser Gly Thr His Lys Met Thr Ser Glu Thr555560565aaa age act agt ata aag att ata taa aga cct ata aat cat taa gac 1824Lys Ser Thr Ser lie Lys lie lie Arg Pro lie Asn His Asp570575aac tga gtt ttt aaa aat gga caa aat cca tga aca att tat aaa ata 1872Asn Val Phe Lys Asn Gly Gln Asn Pro Thr lie Tyr Lys lie580585590gaa ata tag cca aaa atg tct aaa aga tgt tta tga tgt aag gaa tta 1920Glu lie Pro Lys Met Ser Lys Arg Cys Leu Cys Lys Glu Leu595600605tea att aca aca gtg aga aac cat att tcc ttc tat ccg atg agt aaa 1968Ser lie Thr Thr Val Arg Asn His lie Ser Phe Tyr Pro Met Ser Lys610615620aag ttt cta aag tgt tgg caa ggc tta gga aag tga tat gca ttt cct 2016Lys Phe Leu Lys Cys Trp Gln Gly Leu Gly Lys Tyr Ala Phe Pro625630635get ggt caa gtt gta aat tag tat age tac ttt gga gga caa ttg taa 2064Ala Gly Gln Val Val Asn Tyr Ser Tyr Phe Gly Gly Gln Leu640645650aat tta aat cat ate cta tga ccc age aat tcc atg cta taa aaa ctc 2112Asn Leu Asn His lie Leu Pro Ser Asn Ser Met Leu Lys Leu655660665ate cat ggc egg get tgg tgg ctc acg cct gta ate cca gca ctt tgg 2160lie His Gly Arg Ala Trp Trp Leu Thr Pro Val lie Pro Ala Leu Trp670675680gag get gag gtg ggc aga tea cga ggt caa gag ate gag acc ate ctg 2208Glu Ala Glu Val Gly Arg Ser Arg Gly Gln Glu lie Glu Thr lie Leu685690695gcc aac atg atg aaa ccc cgt ttc tac taa aaa tac aaa aat tag ctg 2256Ala Asn Met Met Lys Pro Arg Phe Tyr Lys Tyr Lys Asn Leu700705710[2688]ggc atg gtg gca tgt gcc tat agt ccc age tac tea gga ggc tga ggc 2304Gly Met Val Ala Cys Ala Tyr Ser Pro Ser Tyr Ser Gly Gly Gly715720725 agg aga att get tga acc tgg gag gca gag gtt gca gtg age cga gat 2352Arg Arg lie Ala Thr Trp Glu Ala Glu Val Ala Val Ser Arg Asp730735740cac gcc act gca ctc cag cct ggc tac agg gcg aga ctc tgt ctc aaa 2400His Ala Thr Ala Leu Gln Pro Gly Tyr Arg Ala Arg Leu Cys Leu Lys745750755aaa aaa aaa aaa aaa aaa aaa aaa aca ttc aaa gag aca tat act aag 2448Lys Lys Lys Lys Lys Lys Lys Lys Thr Phe Lys Glu Thr Tyr Thr Lys760765770atg ttc act gtg gca ttg tct gta atg aca aat aag tgg aaa ccg tgt 2496Met Phe Thr Val Ala Leu Ser Val Met Thr Asn Lys Trp Lys Pro Cys775780785790aaa tac ctg tea ata cta tgg agt acc atg tgg caa egg aag aat gag 2544Lys Tyr Leu Ser lie Leu Trp Ser Thr Met Trp Gln Arg Lys Asn Glu795800805act gaa ctg tgt gaa cta aca tgc aaa gat ccc caa aac agg cca ggt 2592Thr Glu Leu Cys Glu Leu Thr Cys Lys Asp Pro Gln Asn Arg Pro Gly810815820gtg gtt get cac age tgt aat aac aac acc ttc aga ggc tga ggt gag 2640Val Val Ala His Ser Cys Asn Asn Asn Thr Phe Arg Gly Gly Glu825830835agg ate agt tga ggc cag gag ttt aag acc age ctg ggc aac ata gtg 2688ArgIle SerGly Gln Glu Phe Lys Thr Ser Leu Gly Asn lie Val840845850aga ccc ctg tct ccc aaa aat ttt ttt taa tta get gtg cgc aat tgc 2736Arg Pro Leu Ser Pro Lys Asn Phe Phe Leu Ala Val Arg Asn Cys855860865tea tgc ata gtc cca get acc cag gag get gag gtg gga gga tea ctt 2784Ser Cys lie Val Pro Ala Thr Gln Glu Ala Glu Val Gly Gly Ser Leu870875880gag ccc agg aat ttg aag ctg cag tga get gtg ttc ttg cca ctg cac 2832Glu Pro Arg Asn Leu Lys Leu Gln Ala Val Phe Leu Pro Leu His885890895tcc aat ctg ggt gac tga gca aga ccc tgt ctc tta aaa aaa taa aaa 2880Ser Asn Leu Gly Asp Ala Arg Pro Cys Leu Leu Lys Lys Lys900905910[2727]aga tct cca age ata gag aag agt ctg gag gga aac acc aaa ctc ata 2928Arg Ser Pro Ser lie Glu Lys Ser Leu Glu Gly Asn Thr Lys Leu lie915920925aca gtc tta ctg cag gca agt ggg ata aag gcc cag act cca tgg tgg 2976Thr Val Leu Leu Gln Ala Ser Gly lie Lys Ala Gln Thr Pro Trp Trp930935940aag tta aag ggc att tcc aag tta agg cta aga ctt get ttt cta act 3024Lys Leu Lys Gly lie Ser Lys Leu Arg Leu Arg Leu Ala Phe Leu Thr945950955960aag aga atg tgc tea tgc att get tgt gta gta gaa act agt ttt tag 3072Lys Arg Met Cys Ser Cys lie Ala Cys Val Val Glu Thr Ser Phe965970975aaa aga aag caa act taa gaa aca ctg act cct gtg gag atg act tgg 3120Lys Arg Lys Gln Thr Glu Thr Leu Thr Pro Val Glu Met Thr Trp980985990cac cac tct cct ttc aca gag cag agt ctg aat agt ctt cag aga tag 3168His His Ser Pro Phe Thr Glu Gln Ser Leu Asn Ser Leu Gln Arg99510001005gcc tgt ggg cca gat tgc cat ccc cta tgg acc aga age caa gga3213Ala Cys Gly Pro Asp Cys His Pro Leu Trp Thr Arg Ser Gln Gly101010151020tct ctc tag tga tgg tea gag ggc cca aat ggc agg gat acc cag tga 3261Ser LeuTrp Ser Glu Gly Pro Asn Gly Arg Asp Thr Gln10251030tgt cag gag gaa tag tac aga cag aag gtg cta age aga caa ttc 3306Cys Gln Glu Glu Tyr Arg Gln Lys Val Leu Ser Arg Gln Phe103510401045aac tgc cat gtt ttg cca ccc cct gtg age agg gat tag gtg ttc 3351Asn Cys His Val Leu Pro Pro Pro Val Ser Arg Asp Val Phe105010551060agg cca gta tct tgg gca tgg ggg age ctt tgg cca gaa gag gta 3396Arg Pro Val Ser Trp Ala Trp Gly Ser Leu Trp Pro Glu Glu Val106510701075taa age tea gaa gtt ttt cag tct gat aac tat tga tat aat ttc 3441Ser Ser Glu Val Phe Gln Ser Asp Asn Tyr Tyr Asn Phe10801085[2763]cat agt gtg agg gag egg tat get cta ccc ttg tgt att taa ggc 3486His Ser Val Arg Glu Arg Tyr Ala Leu Pro Leu Cys lie Gly109010951100[2766]agg aca gag aaa ttg agg atg ccc ctg ggg cta gat cga tga tat 3531[2767]Arg Thr Glu Lys Leu Arg Met Pro Leu Gly Leu Asp Arg Tyr110511101115gac cag aaa tea aaa agg gaa tgc att att tat tgc tgt gta aca 3576Asp Gln Lys Ser Lys Arg Glu Cys lie lie Tyr Cys Cys Val Thr112011251130ctg tta aga gag gag gta gtt aag gga tgg ata ggc aca tag aga 3621Leu Leu Arg Glu Glu Val Val Lys Gly Trp lie Gly ThrArg113511401145ggg aag gtc tea gga gaa ggg ata gaa agg aac tat gtt cac aag 3666Gly Lys Val Ser Gly Glu Gly lie Glu Arg Asn Tyr Val His Lys115011551160aac aac ggc aag aca gtg cct ata ctg cct cta cag tta ata gga 3711Asn Asn Gly Lys Thr Val Pro lie Leu Pro Leu Gln Leu lie Gly116511701175aaa acg gaa gaa gtt acc ctt aag cta gga tgt tgc tea gaa gtg 3756Lys Thr Glu Glu Val Thr Leu Lys Leu Gly Cys Cys Ser Glu Val118011851190acc ate cca act ttg gtg egg gca caa taa ate age cta aat gtc 3801Thr lie Pro Thr Leu Val Arg Ala Glnlie Ser Leu Asn Val119512001205cct aat tta acc cag ctc att ata atg tea tta aac atg aca tta 3846Pro Asn Leu Thr Gln Leu lie lie Met Ser Leu Asn Met Thr Leu121012151220gca ttg tgg ttt tag cac ccc cat ggg ttt tgc tta ggc act cat 3891Ala Leu Trp Phe His Pro His Gly Phe Cys Leu Gly Thr His12251230ggg taa taa cca aga tgg agt ccc ttt ggc aaa act tag gca tgc 3936GlyPro Arg Trp Ser Pro Phe Gly Lys Thr Ala Cys123512401245aca get gta gta ccc caa gaa gaa aat gtt act tct ctc ate tgg 3981Thr Ala Val Val Pro Gln Glu Glu Asn Val Thr Ser Leu lie Trp125012551260gca aaa ccc aca gaa gac ttc cca get tct gcc aca taa aag aca 4026Ala Lys Pro Thr Glu Asp Phe Pro Ala Ser Ala Thr Lys Thr126512701275cag aac aca gac gcc tta ctg gca acc tgc ttt caa gac ccc tgt 4071Gln Asn Thr Asp Ala Leu Leu Ala Thr Cys Phe Gln Asp Pro Cys128012851290[2805]ctt tgc tga gag ctt tcc ttt ttc cca ata aat tct act ctg ccc 4116Leu Cys Glu Leu Ser Phe Phe Pro lie Asn Ser Thr Leu Pro [2807]12951300tac tea ctc tc aaaaaaaaaa aaaaaaa4144Tyr Ser Leu1305SEQUENCE LISTING11<110>北京思安创新生物技术有限公司段，陵浔<120>20种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗<130>NM_000640<160>1<170>PatentIn version 3. 5<210>1<211>1376<212>DNA<213>Homo sapiens<220><221>exon<222>(1). . (92)<220><221>exon<222>(93). . (219)<220><221>sig一peptide<222>(126). . (203)<220><221>exon<222>(220). . (371)<220><221>exon<222>(372). . (525)<220><221>exon<222>(526)·· (646)<220><221>exon<222>(647). . (831)<220>[2844]<221>exon[2845]<222>(832). . (977)<220><221>exon<222> (978)·· (1122)<220><221>exon<222>(1123)·· (1241)<220><221>exon<222>(1242)·· (1373)<220><221>polyA_signal<222>(1343)·· (1348)<220><221>polyA_site<222〉(1366)·· (1366)<220><221>polyA_site<222>(1373)·· (1373)<400>1gta aga aca ctc tcg tga gtc taa egg tct tcc gga tga agg cta ttt48Val Arg Thr Leu Ser Val Arg Ser Ser Gly Arg Leu Phe1510gaa gtc gcc ata acc tgg tea gaa gtg tgc ctg tcg gcg ggg ag a gag 96Glu Val Ala lie Thr Trp Ser Glu Val Cys Leu Ser Ala Gly Arg Glu152025gca ata tea agg ttt taa ate tcg gag aaa tgg ctt tcg ttt get tgg 144Ala lie Ser Arg Phe lie Ser Glu Lys Trp Leu Ser Phe Ala Trp303540cta tcg gat get tat ata cct ttc tga taa gca caa cat ttg get gta 192Leu Ser Asp Ala Tyr lie Pro PheAla Gln His Leu Ala Val455055ctt cat ctt cag aca ccg aga taa aag tta acc ctc ctc agg att ttg 240Leu His Leu Gln Thr Pro Arg Lys Leu Thr Leu Leu Arg lie Leu606570aga tag tgg ate ccg gat act tag gtt ate tct att tgc aat ggc aac 288Arg Trp lie Pro Asp Thr Val lie Ser lie Cys Asn Gly Asn758085[2883]ccc cac tgtctc tgg ate att tta agg aat gca cag tgg aat atg aac 336Pro His CysLeu Trp lie lie Leu Arg Asn Ala Gln Trp Asn Met Asn9095100taa aat accgaa aca ttg gta gtg aaa cat gga ag a cca tea tta cta 384Asn ThrGlu Thr Leu Val Val Lys His Gly Arg Pro Ser Leu Leu [2888]105110115aga ate tacatt aca aag atg ggt ttg ate tta aca agg gca ttg aag 432Arg lie Tyrlie Thr Lys Met Gly Leu lie Leu Thr Arg Ala Leu Lys120125130cga aga tacaca cgc ttt tac cat ggc aat gca caa atg gat cag aag 480Arg Arg TyrThr Arg Phe Tyr His Gly Asn Ala Gln Met Asp Gln Lys135140145150ttc aaa gttcct ggg cag aaa cta ctt att gga tat cac cac aag gaa 528Phe Lys ValPro Gly Gln Lys Leu Leu lie Gly Tyr His His Lys Glu155160165ttc cag aaacta aag ttc agg ata tgg att gcg tat att aca att ggc 576Phe Gln LysLeu Lys Phe Arg lie Trp lie Ala Tyr lie Thr lie Gly170175180aat att tactct gtt ctt gga aac ctg gca tag gtg tac ttc ttg ata 624Asn lie TyrSer Val Leu Gly Asn Leu Ala Val Tyr Phe Leu lie185190195cca att acaact tgt ttt act g gt atg agg get tgg ate atg cat tac 672Pro lie ThrThr Cys Phe Thr Gly Met Arg Ala Trp lie Met His Tyr200210agt gtg ttgatt aca tea agg ctg atg gac aaa ata tag gat gca gat 720Ser Val Leulie Thr Ser Arg Leu Met Asp Lys lieAsp Ala Asp215220225ttc cct atttgg agg cat cag act 3. 3. aag att tct ata ttt gtg tta 768Phe Pro lieTrp Arg His Gln Thr lie Lys lie Ser lie Phe Val Leu230235240atg gat catcag aga aca age cta tea gat cca gtt att tea ctt ttc 816Met Asp HisGln Arg Thr Ser Leu Ser Asp Pro Val lie Ser Leu Phe245250255260age ttc aaaata tag tta aac ctt tgc cgc cag tct ate tta ctt tta 864Ser Phe Lyslie Leu Asn Leu Cys Arg Gln Ser lie Leu Leu Leu265270275ctc ggg agagtt cat gtg aaa tta age tga aat gga gca tac ctt tgg 912Leu Gly ArgVal His Val Lys Leu Ser Asn Gly Ala Tyr Leu Trp280285290[2922]gacetattccagcaaggtgttttg att atg aaa ttg aga tea gag aag960[2923]AspLeuPheGinGinGly ValLeu lie Met Lys Leu Arg Ser Glu Lys[2924]295300 305[2925]atgataetaccttggtg a-ctg；eta cag ttg aaa atg aaa cat aca cct1008[2926]MetlieLeuProTrpLeu Leu Gin Leu Lys Met Lys His Thr Pro[2927]310315 320[2928]tgacaacaaatgcccgac aat tat get ttg tag taa gaa gca1056[2929]LysGinGinMetLys Pro Asp Asn Tyr Ala Leu Glu Ala[2930]325330 335[2931]aagtgaatatttattgetcagatg acg gaa ttt gga gtg agt gga gtg1104[2932]LysliePhelieAlaGinMet Thr Glu Phe Gly Val Ser Gly Val[2933]340345 350[2934]ataaacaatgetgggaaggtgaag acc tat cga aga aaa ctt tgc tac1152[2935]lieAsnAsnAlaGlyLysValLys Thr Tyr Arg Arg Lys Leu Cys Tyr[2936]355360 365[2937]gtttctggctaccatttggtttea tct taa tat tag tta tat ttg taa1200[2938]ValSerGlyTyrHisLeuValSer Ser Tyr Leu Tyr Leu[2939]370375[2940]ccggtctgcttttgcgtaagecaa aca cct acc caa aaa tg a ttc cag1248[2941]ProValCysPheCysValSerGin Thr Pro Thr Gin Lys Phe Gin[2942]380385390 395[2943]aatttttctgtgatacatgaagac ttt cca tat caa gag aca tgg tat1296[2944]AsnPheSerVallieHisGluAsp Phe Pro Tyr Gin Glu Thr Trp Tyr[2945]400405 410[2946]tgactcaacagtttccagteatgg cca aat gtt caa tat gag tct caa1344[2947]LeuAsnSerPheGinSerTrp Pro Asn Val Gin Tyr Glu Ser Gin[2948]415420 425[2949]taaactgaatttttcttgcgaatg ttg aa aaa1376[2950]ThrGluPhePheLeuArgMet Leu[2951]430[2952]SEQUENCE LISTING12[2953]<110>北京思安创新生物技术有限公司[2954]段，陵浔[2955]<120>20 种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗[2956]<130>NM__016368[2957]<160>1[2958]<170>PatentIn version 3.,5[2959]<210>1[2960]<211>1852CN 101845438 A_说 明 书_96/189 页<212>DNA<213>Homo sapiens<220><221>exon<222>(1). . (41)<220><221>exon [2967]<222>(42).<220><221>exon<222>(171)<220><221>exon<222>(333)<220><221>exon<222>(466)<220><221>exon<222>(660)<220><221>STS<222>(715)<220><221>exon<222>(810)<220><221>exon<222>(1026).. (1190)<220><221>exon<222>(1191) (1304)<220><221>exon<222>(1305).. (1522)<220><221>exon<222>(1523) (1825)<220><221>STS
.(170)
. (332)
. (465)
(659)
. (809)
(970)
(1025)
98[3000<222>(1559).(1791)[3001<400>1[3002ccgcgctgtccgccgccg ctg cct gag teg act ctg cgc eg c ccg ccg48[3003ProArgCysProProPro Leu Pro Glu Ser Thr Leu Arg Arg Pro Pro[30041510 15[3005cgatggaggccgccgccc agt tct teg teg aga gcc egg acg tgg tct96[3006ArgTrpArgProProPro Ser Ser Ser Ser Arg Ala Arg Thr Trp Ser[30072025 30[3008acggccccgaggccateg agg cgc aat acg agt acc gga cga cgc gcg144[3009ThrAlaProArgProSer Arg Arg Asn Thr Ser Thr Gly Arg Arg Ala[30103540 45[3011teagccgcgagggtggcg ttc tea ag g tgc acc cca cgt cca cgc get192[3012SerAlaAlaArgValAla Phe Ser Arg Cys Thr Pro Arg Pro Arg Ala[30135055 60[3014teaccttccggaccgccc ggc agg tgc ccc ggc teg ggg tea tgc ttg240[3015SerProSerGlyProPro Gly Arg Cys Pro Gly Ser Gly Ser Cys Leu[30166570 75 80[3017teggetggggcgggaaca acg get cca cac tea ccg ccg egg tgc tgg288[3018SerAlaGlyAlaGlyThr Thr Ala Pro His Ser Pro Pro Arg Cys Trp[30198590 95[3020ccaategactgcgtttgt cct ggc cca cgc gca gcg gcc gca ag g agg336[3021ProlieAspCysValCys Pro Gly Pro Arg Ala Ala Ala Ala Arg Arg[3022100105 110[3023ccaactactacggetcgc tga ctc agg egg gca ccg tga gcc tgg gcc384[3024ProThrThrThrAlaArg Leu Arg Arg Ala Pro Ala Trp Ala[3025115120 125[3026tggacgccgagggccagg agg tgt teg tac cct tea gcg egg tgc tgc432[3027TrpThrProArgAlaArg Arg Cys Ser Tyr Pro Ser Ala Arg Cys Cys[3028130135 140[3029ccatggtggcgcccaacg acc teg tgt teg atg get ggg aca tct cgt480[3030ProTrpTrpArgProThr Thr Ser Cys Ser Met Ala Gly Thr Ser Arg[3031145150 155[3032cgctgaacctggccgagg cga tgc ggc gcg cga agg tgc tgg act ggg528[3033ArgThrTrpProArg Arg Cys Gly Ala Arg Arg Cys Trp Thr Gly[3034160165 170[3035ggctgcaggageaactgt ggc cgc aca tgg agg ccc tgc ggc ccc ggc576[3036GlyCysArgSerAsnCys Gly Arg Thr Trp Arg Pro Cys Gly Pro Gly[3037175180 185[3038cttctgtttacatccccg aat tea teg egg cca acc aga gcg cgc gcg624[3039]LeuLeuPheThrSerProAsnSerSerArgProThrArg Ala Arg Ala[3040]190195200205 [3041]eggacaaccteatcccaggetcgcgtgcgcageagc tgg age aga tcc672[3042]ArgThrThrSerSerGinAlaArgValArgSerSerTrp Ser Arg Ser[3043]210215220[3044]gcagggacatccgagacttccggtetagcgeggggctggacaaag tea720[3045]AlaGlyThrSerGluThrSerGlyLeuAlaArgGlyTrp Thr Lys Ser[3046]225230235[3047]tagtgctgtggaeggcgaacaeggagegettct:gtg aggtga,ttc cag768[3048]CysCysGlyArgArgThrArgSerAlaSerValArgPhe Gin[3049]240245250[3050]gccteaacgacacagccgagaacctgctgcgcaccattgagc teg gtc816[3051]AlaSerThrThrGinProArgThrCysCysAlaProLeuSerSer Val[3052]255260[3053]tggaggtgtcgccctccacgctcttegccgtggccagcatcctgg agg864[3054]TrpArgCysArgProProArgSerSerProTrpProAlaSerTrp Arg[3055]270275280[3056]getgtgccttccteaatgggtctccgcagaacaccctggtgcccg gag912[3057]AlaValProSerSerMetGlyLeuArgArgThrProTrpCysPro Glu[3058]285290295[3059]ctcttgagetegcgtggcageaccgggtttttgtgggcggagatg act960[3060]LeuLeuSerSerArgGlySerThrGlyPheLeuTrpAlaGluMet Thr[3061]300305310315[3062]teaagtcaggccagaccaaagtea agtccgtgc -ttg tgg act tcc tea1008[3063]SerSerGinAla Arg Pro Lys Ser Ser Pro CysLeu Trp Thr Ser Ser[3064]320325330[3065]ttggetccggccteaaga cca tgt cca tcgtgagttacaaccacc tgg1056[3066]LeuAlaProAla Ser ArgPro Cys Pro SerVal Thr Thr Thr Trp[3067]335340345[3068]gcaacaacgatggggagaacctateggcgc丨cat “tgc agt tcc get eta1104[3069]AlaThrThrMet Gly Arg Thr Tyr Arg ArgHisCysSerSerAla Leu[3070]350355360[3071]aggaggtgt cca aga gca acg tggtggacgacatgg tgc aga gcaacc1152[3072]ArgArg Cys Pro Arg Ala Thr Trp Trp ThrThrTrp Cys Arg Ala Thr[3073]365370375[3074]cagtgctctata cgc ccg gcg aag age ctg acc act ；gcg tgg teatea1200[3075]GinCysSerlie Arg Pro Ala Lys Ser Leu Thr Thr AlaTrp Ser SerCN 101845438A 呪明书99/189 页[3076]380 385390[3077]agtatg tgc cgt acg tgg gtg aca gcaagegcgcgc tgg atg agtata1248[3078]SerMet Cys Arg Thr Trp Val Thr Ala SerAlaArgTrp Met Ser lie[3079]395400405410[3080]cctegg age tga tgc tgg gcg gaa ccaacacactgg tgc tgc acaaca1296[3081]ProArg Ser Cys Trp Ala Glu Pro ThrHisTrpCys Cys Thr Thr[3082]415420425[3083]cgtgtg ag g act cgc tgc tgg ccg caccca tea tgc tgg acc tagcgc1344[3084]ArgVal Arg Thr Arg Cys Trp Pro His Pro Ser Cys Trp Thr Arg [3085]430 435440[3086]tgctga ccg age tgt gcc age gcg tgagettctgca ctg aca tggacc1392[3087]CysPro Ser Cys Ala Ser Ala AlaSerAlaLeu Thr Trp Thr[3088]445450[3089]ccgage cgc aga cct tcc acc ccg tgctgtccctgc tea get tcctct1440[3090]ProSer Arg Arg Pro Ser Thr Pro Cys CysProCysSer Ala Ser Ser[3091]455460465470[3092]teaagg cgc cac tag tgc cgc ccg gcagcceggtgg tea atg cgcttt1488[3093]SerArg Arg His Cys Arg Pro Ala AlaArgTrpSer Met Arg Phe[3094]475480485[3095]tccgcc age gca get gca teg aga aca tcc tea ggg cct gcg tggggc1536[3096]SerAla Ser Ala Ala Ala Ser Arg Thr SerSerGly Pro Ala Trp Gly[3097]490 495500[3098]tcccgc cac aga acc aca tgc tcc tggaacacaaaa tgg age gcccag1584[3099]SerArg His Arg Thr Thr Cys Ser Trp AsnThrLysTrp Ser Ala Gin[3100]505 510515[3101]ggccca gcc tea age gag ttg gac ccgtggctgcca cct acc etatgt1632[3102]GlyPro Ala Ser Ser Glu Leu Asp Pro TrpLeuProPro Thr Leu Cys[3103]520 525530[3104]tgaaca aga aag gac egg tac ccg ctgccaccaatg get gca ccggtg1680
1013105]Thr Arg Lys Asp Arg Tyr Pro Leu Pro ProMet Ala Ala Pro Val3106]535 5405453107]atg cca atg ggc ate tgc aag agg age ccccaa tgc cca cca cct gag7283108]Met Pro Met Gly lie Cys Lys Arg Ser Pro GinCys Pro Pro Pro Glu3109]550 5555603110]gcc ccg gtc aca cag ttt ctc ggc tct tcctcc ccg ctg ccc ccc acg7763111]Ala Pro Val Thr Gin Phe Leu Gly Ser Ser SerPro Leu Pro Pro Thr3112]565 570 5755803113]acc eta cct tga agg ccc cca caa ata aag gcg ctg cca ctc age cct c8253114]Thr Leu Pro Arg Pro Pro Gin lie Lys AlaLeu Pro Leu Ser Pro3115]585 5905953116]8523117]SEQUENCE LISTING133118]<110>北京思安创新生物技术有限公司3119]段，陵浔3120]<120>20种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗3121]<130>NM_0049233122]<160>13123]<170>PatentIn version 3. 53124]<210>13125]<211>25353126]<212>DNA3127]<213>Homo sapiens3128]<220>3129]<221>exon3130]<222>(1). . (101)3131]<220>3132]<221>exon3133]<222>(102). . (611)3134]<220>3135]<221>exon3136]<222>(612). . (770)3137]<220>3138]<221>exon3139]<222>(771). . (891)3140]<220>
3141]<221>exon
3142]<222>(892). . (968)
3143]<220>
3144]<221>exon
3145]<222> (969)(1057)
3146]<220>
3147]<221>exon
3148]<222> (1058) (1160)
3149]<220>
3150]<221>exon
3151]<222>(1161) (1298)
3152]<220>
3153]<221>exon
3154]<222> (1299) (1474)
3155]<220>
3156]<221>exon
3157]<222> (1475)(2535)
3158]<220>
3159]<221>STS
3160]<222>(2123) (2304)
3161]<220>
3162]<221>polyA_signal
3163]<222> (2512)(2517)
3164]<220>
3165]<221>polyA_site
3166]<222> (2535)(2535)
3167]<400>1
3174]
3175]
3176]
3177]
3178]
CCC
Pro
agg Arg
ag Arg
agg Arg
ccc 48 Pro
ggc 96 Gly
g tgc gcg gcc ccc gcc ate ccg ccc cgc cgc cct gcg cca tgg 144 Cys Ala Ala Pro Ala lie Pro Pro Arg Arg Pro Ala Pro Trp 354045
gcc ctc tgc egg gcg ggc tgc cca gcc ccg agg atg cga tgg 192 Ala Leu Cys Arg Ala Gly Cys Pro Ala Pro Arg Met Arg Trp
3168]acccttcccccttctccaaccgccaacggccgcctgcgcctgagg3169]ThrLeuProProSerProThrAlaAsnGlyArgLeuArgLeuArg3170]1510153171]ggcccacgccccateccgctcgetccacgctgcggcagecccggc3172]GlyProArgProlieProLeuAlaProArgCysGlySerProGly3173]202530
103[3179505560[3180tgaeggagetcttaagccccgagggtccgttegctteggagaacateg240[3181ArgSerSerAlaProArgValArgSerLeuArgArgThrSer[3182657075[3183gcctgaaggcccccgtgaagtacgaggaggacgagttccacgtcttea288[3184AlaArgProProSerThrArgArgThrSerSerThrSerSer[3185808590[3186aagaagcgtacctgggcceggeggaccccaaggaacccgtcctgcacg336[3187LysLysArgThrTrpAlaArgArgThrProArgAsnProSerCysThr[318895100105[3189cgtteaaccccgcgctgggcgccgactgcaagggccaggteaaggcga384[3190ArgSerThrProArgTrpAlaProThrAlaArgAlaArgSerArgArg[3191110115120[3192agetegeggggggcgacagcgacggcggggagetccteggggagtacc432[3193SerSerArgGlyAlaThrAlaThrAlaGlySerSerSerGlySerThr[3194125130135140[3195ccgggatcccagageteagcgcgctggaggacgtegcgctcctgcagg480[3196ProGlySerGinSerSerAlaArgTrpArgThrSerArgSerCysArg[3197145150155[3198ccccgcagecgcccgcctgcaacgtgcacttcctgtcctcgctgctac528[3199ProArgSerArgProProAlaThrCysThrSerCysProArgCysTyr[3200160165170[3201ccgcgcaccgcagcceggeggtgttgcccctgggcgcctgggtcctgg576[3202ProArgThrAlaAlaArgArgCysCysProTrpAlaProGlySerTrp[3203175180185[3204aaggagcctcccaccegggcgtccgcatgatcccag ttg aaa tea agg624[3205LysGluProProThrArgAlaSerAlaSerGinLeu Lys Ser Arg[3206190195200[3207aagcaggtggtaetaetacaagtaataateeggaagaagcaactttgc672[3208LysGinValValLeuLeuGinVallielieArgLysLysGinLeuCys[3209205210215[3210agaatettcttgctcaggaatcctgttgcaagttcccatcgtcccagg720[3211ArgliePheLeuLeuArgAsnProValAlaSerSerHisArgProArg[3212220225230235[3213aactagaggatgcctcctgetgttctcttaagaaagattccaacccaa768[3214AsnArgMetProProAlaValLeuLeuArgLyslieProThrGin[3215240245250[3216tgg tga tat gcc aat tga aag ggg gca cac aaa tgc tat gta tag aca816[3217TrpTyr Ala AsnLys Gly Ala His Lys Cys Tyr ValThr[3218]255260[3219]atteta gaacaa gag aac taa aag cac tccatttgg ttc ctc agt ate864[3220]lieLeu GluGin Glu Asn Lys His Serlie Trp Phe Leu Ser lie[3221]265270275[3222]aagate 3-3-3-ata att ate tac agt cag atgtcceta aac caa tga ctg912[3223]LysIle Lyslie Ilelle Tyr Ser Gin Met Ser Leu Asn Gin Leu[3224]280285290[3225]ctttag taggga gat ttt tgc cag cat caacaaaat taa ate tea tta960[3226]LeuGly Asp Phe Cys Gin His Gin Gin Asnlie Ser Leu[3227]295300305[3228]cacc ttg agg gag cct tac categg tag tea acg ggt ctgctt1008[3229]HisAsn AsnLeu Arg Glu Pro Tyr His ArgSer Thr Gly Leu Leu[3230]310 315320[3231]tcccct egggat caa ctc ttc cag gac cac caa aaa taa ctt tgg ctg g1057[3232]SerPro Arg Asp Gin Leu Phe Gin Asp HisGinLys Leu Trp Leu[3233]325 330335[3234]gtact gtg act get ttg cca gtg gggactttt gca aca act gcaatt1104[3235]GlyThr ValThr Ala Leu Pro Val Gly ThrPheAla Thr Thr Ala lie[3236]340 345350[3237]gtaata attgtt gca aca act tgc ateatgata ttg aac ggt ttaaag1152[3238]Vallie lieVal Ala Thr Thr Cys lie MetlieLeu Asn Gly Leu Lys[3239]355360365[3240]ccatta agg cat gtc ttg gta gaa atecag aag ctt tcc age caa1200[3241]ProLeu ArgHis Val Leu Val Glu lie Gin Lys Leu Ser Ser Gin Lys[3242]370375380[3243]ttggga agg■ gcc aat tgg gca atg teaageccc age aca aca aagggt1248[3244]LeuGly Arg Ala Asn Trp Ala Met Ser SerProSer Thr Thr Lys Gly[3245]385390395400[3246]gcaact gcagga ggt cag get gcc tgaagaatt act gcg agt getatg1296[3247]AlaThr Ala Gly Gly Gin Ala Ala ArglieThr Ala Ser Ala Met[3248]405 410415[3249]agg ccc aaa tta tgt gtt ctt eta tttgca aat gca ttg gtt gca
105[3250]ArgPro Lys Leu Cys Val Leu Leu Phe Ala Asn Ala Leu Val Ala Lys[3251]420425430[3252]attatgaag aaa gcc cag aac gaa agacactaa tga gca tgc caaact1392[3253]lieMetLys Lys Ala Gin Asn Glu Arg HisAla Cys Gin Thr[3254]435440445[3255]acatgcaga ctg gag gtt tgg aag gcagccatt acc tgc cac caacga1440[3256]ThrCysArg Leu Glu ValTrp Lys Ala AlalieThr Cys His Gin Arg[3257]450455460[3258]aattttcaggac ttc caa gat tea gtc acg ata g gc ggc ctt cctcat1488[3259]AsnPheGin Asp Phe Gin Asp Ser Val ThrlieGly Gly Leu Pro His[3260]465470475[3261]gcatctcct ggg agg tgg tgg agg ccacatgcg cct gcc tgc ttgctc1536[3262]AlaSerPro Gly Arg Trp Trp Arg Pro HisAlaPro Ala Cys Leu Leu[3263]480485490[3264]agggagaag agg ccg aga aag aac actgetcca agt gcc tgg cagage1584[3265]ArgGluLysArgProArgLysAsn Thr AlaProSer Ala Trp Gin Ser[3266]495500505[3267]agatgatcctggaggaatttggaa ggt gettatcac aga ttc tcc aca1632[3268]ArgSerTrpArgAsnLeuGlu Gly Ala Tyr His Arg Phe Ser Thr[3269]510515520[3270]ctgagtttaaatetaagggattga aaa tgg agtaga gta taa agt gtg1680[3271]LeuSerLeuAsnLeuArgAspLys Trp Ser Arg Val Ser Val[3272]525530535[3273]aatgcatgttgattttgtcttagt eta gaaatetct agt tta gaa agg1728[3274]AsnAlaCysPheCysLeuSer Leu Glulie Ser Ser Leu Glu Arg[3275]540545550[3276]atgtttaggggaacatgaggctgg ctc tgcageaac aac cag get ccc1776[3277]MetPheArgGlyThrGlyTrp Leu CysSerAsn Asn Gin Ala Pro[3278]555560565[3279]ctgcatccctgggcccagggagtt tac teagagctc tct gaa gat gtg1824[3280]LeuHisProTrpAlaGinGlyVal Tyr Ser Glu Leu Ser Glu Asp Val[3281]570575580[3282]gcaacccatgcccccttttctgag gag gtgcatggc ctg age att gtt1872[3283AlaThrHisAlaProPheSerGluGlu Val His Gly Leu Ser lie Val[3284585590595 600[3285tgtctggcccagaggagagagcttggg ttc cca tag tcc tgg gag agt1920[3286CysLeuAlaGinArgArgGluLeuGly Phe Pro Ser Trp Glu Ser[3287605610 615[3288gtctgcagggcggcggagggcagagca ggg cag ggg agg gca cag cag1968[3289ValCysArgAlaAlaGluGlyArgAla Gly Gin Gly Arg Ala Gin Gin[3290620625 630[3291gccctgcgaagggcagageggggcagg gga ggg cag age agg ccc tgc2016[3292AlaLeuArgArgAlaGluArgGlyArg Gly Gly Gin Ser Arg Pro Cys[3293635640 645[3294ggagagctcactctggtcgactcttcc tct cag aga atg ttg ctc tgg2064[3295GlyGluLeuThrLeuValAspSerSer Ser Gin Arg Met Leu Leu Trp[3296650655660[3297aggctgctctgcatgaccetaatg gtt tct tgt ttg ttt ttc aaa2112[3298ArgLeuLeuCysMetLysThrLeuMet Val Ser Cys Leu Phe Phe Lys[3299665670675[3300ttatttagaaataagttctccggatgg get gtt gtg ata cca ctt aaa2160[3301LeuPheArgAsnLysPheSerGlyTrp Ala Val Val lie Pro Leu Lys[3302680685690 695[3303atetctagagaaetactgaacacctaa aga ttt tct gta gcg tag ata2208[3304lieSerArgGluLeuLeuAsnThrArg Phe Ser Val Ala lie[3305700705[3306tttccccagaggcacgcgaactgtcag tct ttc eta agg ccc ccg gga2256[3307PheProGinArgHisAlaAsnCysGin Ser Phe Leu Arg Pro Pro Gly[3308710715720 725[3309gacgcaggcaatggggcctegcaggcc agg ctt gca cca gca tgt ctt2304[3310AspAlaGlyAshGlyAlaSerGinAla Arg Leu Ala Pro Ala Cys Leu[3311730735 740[3312gagttagaggacttattatccagt ttc ttc tgt gtt tct act tga2352[3313GluLeuGluAspLeuLysLeuSerSer Phe Phe Cys Val Ser Thr[3314745750 755[3315attgtggaaaagctctattatccaatt aac ttc tcc ata att att gtt2400[3316lieValGluLysLeuTyrTyrProlie Asn Phe Ser lie lie lie Val[3317760765 770[3318gtaatattattattgtttgtaclclclcat ggt tea cat aac tag ctt gtg2448[3319VallieLeuLeuLeuPheValLysHis Gly Ser His Asn Leu Val[3320775780785[3321gaaaccageaggtaaaatgaattctta agt tga cgc ttt tgg ttc tgt2496[3322]Glu Thr Ser Arg Asn Glu Phe Leu Ser Arg Phe Trp Phe Cys790795800tgt aaa gca aag atg aat aaa aat ttc caa tgt ctt caa 2535Cys Lys Ala Lys Met Asn Lys Asn Phe Gin Cys Leu Gin805810SEQUENCE LISTING14<110>北京思安创新生物技术有限公司段，陵浔<120>20种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗<130>NM_153485<160>1<170>PatentIn vetsion 3. 5<210>1<211>4355<212>DNA<213>Homo sapiens<220><221>exon<222>(1). . (275)<220><221>exon<222>(276). . (413)<220><221>exon<222>(414). . (510)<220><221>exon<222>(511). . (581)<220><221>exon<222>(582). . (674)<220><221>exon<222>(675). . (841)<220><221>exon<222>(842). . (947)<220><221>exon
108[3361<222>(948)..(1021)[3362<220>[3363<221>exon[3364<222>(1022)..(1113)[3365<220>[3366<221>exon[3367<222>(1114)..(1211)[3368<220>[3369<221>exon[3370<222>(1212). (1364)[3371<220>[3372<221>exon[3373<222>(1365). (1465)[3374<220>[3375<221>exon[3376<222>(1466)..(1636)[3377<220>[3378<221>exon[3379<222>(1637)..(1747)[3380<220>[3381<221>exon[3382<222>(1748). (1842)[3383<220>[3384<221>exon[3385<222>(1843)..(1931)[3386<220>[3387<221>exon[3388<222>(1932). (1994)[3389<220>[3390<221>exon[3391<222>(1995)..(2142)[3392<220>[3393<221>exon[3394<222>(2143). (2209)[3395<220>[3396<221>exon[3397<222>(2210)..(2325)[3398<220>[3399<221>exonCN 10845438 A[3400<222>(2326).(2423)[3401<220>[3402<221>exon[3403<222>(2424).(2554)[3404<220>[3405<221>exon[3406<222>(2555).(2746)[3407<220>[3408<221>exon [3409<222>(2747). (2885)[3410<220>[3411<221>exon[3412<222>(2886). (3021)[3413<220>[3414<221>exon[3415<222>(3022). (3175)[3416<220>[3417<221>exon[3418<222>(3176). (3280)[3419<220>[3420<221>exon[3421<222>(3281). (3435)[3422<220>[3423<221>exon[3424<222>(3436). (3565)[3425<220>[3426<221>STS[3427<222>(3492). (3653)[3428<220>[3429<221>exon[3430<222>(3566). (3679)[3431<220>[3432<221>exon[3433<222>(3680). (3800)[3434<220>[3435<221>exon[3436<222>(3801). (3911)[3437<220>[3438<221>exon
说明书
108/189 页
110[3439<222>(3912). (4048)[3440<220>[3441<221>exon[3442<222>(4049). (4155)[3443<220>[3444<221>exon[3445<222>(4156). (4355)[3446<220>[3447<221>polyA_signal[3448<222>(4333). (4338)[3449<220>[3450<221>polyA_site[3451<222>(4355). (4355)[3452<400>1[3453aaggcgctcgtttggcgcgcgcgccctaggeggeggate taa get aac48[3454LysAlaLeuValTrpArgAlaArgProArgArgArglie Ala Asn[3455151015[3456ttcttggatcttttcttgtttctccttggtttttgactt ttt ctg gac96[3457PheLeuAspLeuPheLeuPheLeuLeuGlyPheLeu Phe Leu Asp[3458202530[3459cctggtgtctacgatttccgagatgccgtcttctttgtt ggg cgc ggc144[3460ProGlyValTyrAspPheArgAspAlaValPhePheVal Gly Arg Gly[3461354045[3462gatgccggcctctacatetgccgcagecctgcaggaage tct gga aaa192[3463AspAlaGlyLeuTyrlieCysArgSerProAlaGlySer Ser Gly Lys[3464505560[3465tgctggacggetcatcgaccgteagttgcaagaggaccg cat gta ccc240[3466CysTrpThrAlaHisArgProSerValAlaArgGlyPro His Val Pro[3467657075[3468ggacctttccgagetgettatggtgtctgccccaaa taa tcc cac cgt288[3469GlyProPheArgAlaAlaTyrGlyValCysProLysSer His Arg[34708085[3471ttctggcatgtcagatatggattatcctttacaaggacc tgg ttt get336[3472PheTrpHisValArgTyrGlyLeuSerPheThrArgThr Trp Phe Ala[347395100105[3474gtccgtacccaaccttccagagatcagttccatccgaag agt tcc tct384[3475ValArgThrGinProSerArgAspGinPheHisProLys Ser Ser Ser[3476110115120125[3477cccacctgaacttgttgagcagtttggaca tat gca gtg taa ttg cat432[3478]ProThrThr CysAla Val Trp ThrTyr Ala Val Leu His[3479]130135[3480]gatgggtgtgtt ccctcctat cag cag agettg get cac aat tga cag480[3481]AspGlyCysVal ProSerTyr Gin Gin SerLeu Ala His Asn Gin[3482]140145150[3483]tgatatattcat gtggaaeta tga gga tggagg aga cct tgc eta ttt528[3484]TyrlieHis ValGluLeu Gly TrpArg Arg Pro Cys Leu Phe[3485]155160165[3486]tgatggacttag tgagactat tct tgc tgtggg get tgt gaa gcc aaa576[3487]TrpThrAspTyr Ser Cys CysGly Ala Cys Glu Ala Lys[3488]170175 180[3489]ageagg cat ctt tea acc tea tgt gcg aca cct cct ggt ttt ggc gac624[3490]SerArgHis Leu Ser Thr Ser Cys Ala Thr Pro Pro Gly Phe Gly Asp[3491]185190 195[3492]ccctgtagacat agtaattct tgg act cageta tgc taa ttt gca aac672[3493]ProCysArg His Ser Asn Ser Trp Thr Gin Leu Cys Phe Ala Asn[3494]200205210[3495]agg ttc tgg agt tct taa tga tag ttt gtc tgg tgg aat gca gtt get720[3496]ArgPhe Trp Ser SerPhe Val Trp Trp Asn Ala Val Ala[3497]215220[3498]tccagatccttt atattctct tcc tac tgataa tac tta cct ttt aac768[3499]SerArgSerPhe liePheSer Ser TyrTyr Leu Pro Phe Asn[3500]225230235[3501]aataacttccac tgataatgg cag aat ttt ctt ggc tgg aaa gga tgg816[3502]AsnAsnPheHisTrp Gin Asn Phe Leu Gly Trp Lys Gly Trp[3503]240245250[3504]ctgtttatatga agtageeta cca g gc tga age agg gtg gtt tag cca864[3505]LeuPhelieSerSerLeu Pro GlySer Arg Val Val Pro[3506]255265[3507]aagatgtaggaa aatclclclcca ctc aaa gag ctc act ttc ttt cct tgt912[3508]LysMetGlu AsnLysPro Leu Lys Glu Leu Thr Phe Phe Pro Cys[3509]270275 280[3510]tccttccttget acaattcac gtt ctc aga aga tg a tcc tat tct tea960[3511]SerPheLeuAla ThrlieHis Val Leu ArgArg Ser Tyr Ser Ser[3512]285290295[3513]aattgcaat tga taa ttc tag aaa tatttt ata tac acg ate tgagaa1008[3514]AsnCys AsnPheLys Tyr Phelie Tyr Thr lie Glu[3515]300305[3516]aggagtaat aca g gt gta tga ttt gggaca aga tgg aca agg aatgag1056[3517]ArgSerAsn Thr Gly Val Phe Gly Thr Arg Trp Thr Arg Asn Glu[3518]310315320[3519]cagagttgc ctc tgt gtc aca gaa tgccat tgt ctc tgc tgc tggtaa1104[3520]GinSerCys Leu Cys Val Thr Glu Cys HisCys Leu Cys Cys Trp[3521]325330335[3522]cattgctag gac cat cga teg ttc tgtttt taa acc aat tgt ccaaat1152[3523]HisCysAsp His Arg Ser Phe Cys PheThr Asn Cys Pro Asn[3524]340345350[3525]ageagtgat tga aaa ttc tga ate actgga ctg tea gtt att ggctgt1200[3526]SerSerAsp Lys Phe lie Thr GlyLeu Ser Val lie Gly Cys[3527]355 360365[3528]cacacatgc ag g tgt tag gtt ata ttttag cac ttg tcc att cagaca1248[3529]HisThrCys Arg Cys Val lie PheHis Leu Ser lie Gin Thr[3530]375 380[3531]gccattage acg gcc taa tac act gacget ggt tea tgt ccg cttacc1296[3532]AlalieSer Thr Ala Tyr Thr Asp AlaGly Ser Cys Pro Leu Thr[3533]385390 395[3534]tcctggatt ctc age ate ttc aac cgtgga aaa gcc ttc aaa agtaca1344[3535]SerTrplie Leu Ser lie Phe Asn Arg GlyLys Ala Phe Lys Ser Thr[3536]400 405410[3537]tagagetct tta tag taa ag g tat tctatt gat ggc age ctc aga1392[3538]SerSer Leu Arg Tyr Ser lie Asp Gly Ser Leu Arg Lys[3539]420[3540]tgaggataa tga tat ttt atg gtg tgtcaa cca tga tac ttt tccttt1440[3541]GlyTyr Phe Met Val Cys Gin Pro Tyr Phe Ser Phe[3542]425430435[3543]ccagcc aat gat gga aac cca g atgac age tgg tgt tga tggtea1488[3544]ProLysAla Asn Asp Gly Asn Pro Asp Asp Ser Trp Cys Trp SerCN 101845438A 呪明书112/189页[3545]440450[3546]ttcctg ggc tct ttc tgc gat aga tgaatt gaa agt aga taa aataat1536[3547]PheLeu Gly Ser Phe Cys Asp Arg lieGlu Ser Arg Asn Asn[3548]455 460465[3549]tacacc ttt aaa caa gga tea tat tccaat aac tga ttc acc agttgt1584[3550]TyrThr Phe Lys Gin Gly Ser Tyr Ser Asn Asn Phe Thr Ser Cys[3551]470 475480 [3552]tgtaca gca gca cat gtt acc tcc gaa gaa att tgt tct cct ctc agc1632[3553]CysThr Ala Ala His Val Thr Ser Glu Glulie Cys Ser Pro Leu Ser[3554]485 490495[3555]acag gg gag cct tat gtt tea taa acttag acc tgt aga tea actgag1680[3556]ThrGly Glu Pro Tyr Val Ser ThrThr Cys Arg Ser Thr Glu[3557]500505 510[3558]gcatct act tgt gag taa tgt ggg aggaga tgg aga aga gat tgaaag1728[3559]AlaSer Thr Cys Glu Cys Gly Arg Arg Trp Arg Arg Asp Lys[3560]515520[3561]attctt taa att aca tea g ga aga ccaggc ttg tgc aac ttg ccttat1776[3562]lieLeu lie Thr Ser Gly Arg Pro Gly Leu Cys Asn Leu Pro Tyr[3563]525535[3564]tcttgc ttg ctc cac tgc tgc ctg tgatag aga agt ate tgc ctgggc1824[3565]SerCys Leu Leu His Cys Cys LeuArg Ser lie Cys Leu Gly[3566]540545550[3567]tacteg ggc ttt ctt tag gta tgg tggtga age aca gat gag atttcc1872[3568]TyrSer Gly Phe Leu Val Trp TrpSer Thr Asp Glu lie Ser[3569]555 560565[3570]aaccac tct tcc gcc tcc aag taa tgttgg tcc cat ctt ggg gtctcc1920[3571]AsnHis Ser Ser Ala Ser Lys Cys Trp Ser His Leu Gly Val Ser[3572]570 575580[3573]tgteta ttc ta g ttc tcc tgt tcc tagtgg tag tcc eta tcc aaatcc1968
114[3574]CysLeuPhePhe Ser Cys Ser Trp Ser Leu Ser Lys Ser[3575]585590 595[3576]atectttttggg aac acc gtc tea tg g tat aca gcc tcc tgc catgtc2016[3577]lieLeuPheGlyAsnThrValSer Trp Tyr Thr Ala Ser Cys His Val[3578]600610[3579]aactccagtgtgtgctctgggaaaccc age aac tea ggc cac aaa tat2064[3580]AsnSerSerValCysSerGlyLys Pro Ser Asn Ser Gly His Lys Tyr[3581]615620 625[3582]gagttgtgtgactggaccagagat tgt gta ctc tgg aaa aca caa tgg2112[3583]GluLeuCysAspTrpThrArgAsp Cys Val Leu Trp Lys Thr Gin Trp[3584]630635640[3585]tatttgcatttacttttcteggat cat ggg aaa cat ttg gga tgc aag2160[3586]TyrLeuHisLeuLeuPheSerAsp His Gly Lys His Leu Gly Cys Lys[3587]645650655 660[3588]cttagttgtggagagaatattcaa gag tgg caa cag aga gat cac tgc a2209[3589]LeuSerCysGlyGluAsnlieGin Glu Trp Gin Gin Arg Asp His Cys[3590]665670 675[3591]attgaaagtagtgttccctgcca act get aga gtc agt get aca aga2256[3592]AsnLysCysSerLeuPro Thr Ala Arg Val Ser Ala Thr Arg[3593]680685 690[3594]actaaa ggg ttt:gcal gga,atttct aga cag aaa ctc cca gtt tgc agg2304[3595]ThrLys Gly Phe Ala Gly lie Ser Arg Gin Lys Leu Pro Val Cys Arg[3596]695700 705[3597]aggaccattaggtccclclcltac tac tgc taa agt gca gca gag get2352[3598]ArgThrlieArgLysSerLysTyr Tyr Cys Ser Ala Ala Glu Ala[3599]710715 720[3600]gataggattcatgcgtcctgaaaa egg aaa tcc cca gca aat gca aca2400[3601]AspArglieHisAlaSerLys Arg Lys Ser Pro Ala Asn Ala Thr[3602]725730 735[3603]ggaactgcagaggaagttteatg a ggc tea act aag tga aaa gat ttc2448[3604]GlyThrAlaGluGluValSerGly Ser Thr Lys Lys Asp Phe[3605]740745 750[3606]actteaggcaatteagcagttggt teg aaa ate ata tea ggc tct ggc2496[3607]ThrSerGlyAsnSerAlaValGly Ser Lys lie lie Ser Gly Ser Gly[3608]755760 765[3609]tttatggaaacttctttgtgaaca tea att cac tat cat tgt ggc aga2544[3610]PheMetGluThrSerLeuThr Ser lie His Tyr His Cys Gly Arg[3611]770775 780CN 10845438A祝 明 书 114/189页[3612actteagaag ga act tea aga gcagct gaa gat cac cac ctt taa aga2592[3613ThrSerGluGly Thr Ser Arg Ala Ala Glu Asp His His Leu Arg[3614785790 795[3615tcttgtaatcaggga caa aga act cac agg ggc att aat tgc ttc tct2640[3616SerCysAsnGinGly Gin Arg Thr His Arg Gly lie Asn Cys Phe Ser[3617800805 810[3618tatcaactgetacat cag aga taa tgc cgc tgt tga tgg cat tag ttt2688[3619TyrGinLeuLeuHis Gin Arg Cys Arg Cys Trp His Phe[3620815820 825[3621acatttacaggatat ctg ccc act tct ata tag cac tga tga tgc aat2736[3622ThrPheThrGlyTyr Leu Pro Thr Ser lie His Cys Asn[3623830835[3624ttgttctaag gcaaa tga get tct cca gcg ttc ccg aca agt tea aaa2784[3625LeuPheGly Lys Ala Ser Pro Ala Phe Pro Thr Ser Ser Lys [3626840845 850[3627taagactgaclclclaga aag aat gtt aag gga ate att aaa gga ata tea2832[3628AspLys Arg Lys Asn Val Lys Gly lie lie Lys Gly lie Ser[3629855860 865[3630clclclaattagcaatea agt gga cct ttc caa tgt ttg tgc tea gta tag2880[3631LysAsnGin Ser Ser Gly Pro Phe Gin Cys Leu Cys Ser Val[3632870 875 880[3633acaagt gag att tta tga ggg tgt ggt gga act ttc tct tac ggc tgc2928[3634ThrSerGlu lie Leu Gly Cys Gly Gly Thr Phe Ser Tyr Gly Cys[3635885 890 895[3636agagaaclclclagatcc tea agg tct tgg get tea ttt eta taa aca tgg2976[3637ArgGluLysArgSer Ser Arg Ser Trp Ala Ser Phe Leu Thr Trp[3638900905 910[3639agaaccagaagaaga cat agt tgg act tea ggc ctt cca aga aag att3024[3640ArgThrArgArgArg His Ser Trp Thr Ser Gly Leu Pro Arg Lys lie[3641915920 925[3642cagttacaaatg cat tac aga cac act tea aga act ggt aaa tea3072[3643LysGinLeuGinMet His Tyr Arg His Thr Ser Arg Thr Gly Lys Ser[3644930935 940[3645aagtaaggccgctcc tea gtc tcc cag tgt acc caa aaa acc tgg tcc3120[3646LysGlyArgSer Ser Val Ser Gin Cys Thr Gin Lys Thr Trp Ser[3647945950 955[3648tccagtgttgtcate tga tcc aaa tat get gag taa tga aga age agg3168[3649SerSerValVallie Ser Lys Tyr Ala Glu Arg Ser Arg[3650960965 970
116[3651acateat tt tga aca aat get taa att gtc aca gcg ate caa gga tga3216[3652ThrSerPhe Thr Asn Ala lie Val Thr Ala lie Gin Gly[3653975 980[3654getctttag tat tgc cct tta taa ttg get aat aca agt cga cct tgc3264[3655AlaLeuTyr Cys Pro Leu Leu Ala Asn Thr Ser Arg Pro Cys[3656985990 995[3657agataaget get aca g gt tgc ttc tcc att tct gga gcc aca tct3309[3658ArgAla Ala Thr Gly Cys Phe Ser lie Ser Gly Ala Thr Ser[36591000 1005 1010[3660agtccgaat ggc caa agt tga tea aaa cag agt teg tta tat gga3354[3661SerProAsn Gly Gin Ser Ser Lys Gin Ser Ser Leu Tyr Gly[36621015 1020 1025[3663tttactctg gcg gta tta cga gaa gaa cag aag ttt cag taa tgc3399[3664PheThrLeu Ala Val Leu Arg Glu Glu Gin Lys Phe Gin Cys[36651030 1035 1040[3666tgctegtgt act gtc cag act ggc tga cat gca tag cac aga aat ttc3447[3667CysSerCys Thr Val Gin Thr Gly His Ala His Arg Asn Phe[36681045 1050[3669acttea gca gcg act aga gta cat tgc teg age cat tct tag tgc3492[3670ThrSer Ala Ala Thr Arg Val His Cys Ser Ser His Ser Cys[367110551060 1065[3672caaaagttc cac tgc cat ttc ate aat age tgc cga tgg tga att3537[3673GinLysPhe His Cys His Phe lie Asn Ser Cys Arg Trp lie[36741070 1075 1080[3675tctteatga att aga aga aaa aat gga g gt tgc tag gat cca act3582[3676SerSerlie Arg Arg Lys Asn Gly Gly Cys Asp Pro Thr[36771085 1090 1095[3678teagataca gga gac act aca aag gca gta ttc cca tea ttc tt3627[3679SerAspThr Gly Asp Thr Thr Lys Ala Val Phe Pro Ser Phe Phe[36801100 1105 1110[3681tgtacagga tgc agt ttc tea get gga ttc tga get gat gga cat3672[3682CysThrGly Cys Ser Phe Ser Ala Gly Phe Ala Asp Gly His[36831115 1120[3684aactaa g ct tta tgg gga att tgc tga ccc att taa act tgc aga3717[3685AsnAla Leu Trp Gly lie Cys Pro lie Thr Cys Arg[368611251130 1135[3687gtgcaaact tgc aat aat tea ttg tgc egg tta ttc aga ccc tat3762[3688ValGinThr Cys Asn Asn Ser Leu Cys Arg Leu Phe Arg Pro Tyr[36891140 1145 11503690]attggtgca gac act ttg gca aga tat cat aga gaa ag a att gag38073691]lieGly Ala Asp Thr Leu Ala Arg Tyr His Arg Glu Arg lie Glu3692]1155 1160 11653693]tgacagtgt gac att gag ctc ctc gga tag aat gca tgc tct tag tct38553694]GinCys Asp lie Glu Leu Leu Gly Asn Ala Cys Ser Ser3695]1170 11753696]caagattgt tct cct tgg caa aat tta tgc tgg cac acc acg ctt39003697]GinAspCys Ser Pro Trp Gin Asn Leu Cys Trp His Thr Thr Leu3698]11801185 11903699]ctttccttt ag a ttt tat tgt aca gtt ttt aga aca gca ggt ttg39453700]LeuSerPhe Arg Phe Tyr Cys Thr Val Phe Arg Thr Ala Gly Leu3701]11951200 12053702]tactttgaa ctg gga tgt ggg ctt cgt aat aca gac aat gaa tga39903703]TyrPheGlu Leu Gly Cys Gly Leu Arg Asn Thr Asp Asn Glu3704]12101215 12203705]aattggagt acc att acc tag act act aga agt tta tga tea gtt40353706]AsnTrpSer Thr lie Thr Thr Thr Arg Ser Leu Ser Val3707]12251230 12353708]gttcaaate acg g ga tcc att ctg gaa cag aat gaa gaa gcc act40803709]ValGinlie Thr Gly Ser lie Leu Glu Gin Asn Glu Glu Ala Thr3710]1240 1245 12503711]gcacctttt gga ttg tat aca tgt att att gat aag ata tgt tga41253712]AlaProPhe Gly Leu Tyr Thr Cys lie lie Asp Lys lie Cys3713]1255 1260 12653714]gaatcccag cca agt ttt aaa ttg tga aag gag aag att tac aaa41703715]GluSerGin Pro Ser Phe Lys Leu Lys Glu Lys lie Tyr Lys3716]1270 12753717]tctctgcct gga tgc tgt ttg tgg tta cct tgt tga get cca gtc42153718]SerLeuPro Gly Cys Cys Leu Trp Leu Pro Cys Ala Pro Val3719]12801285 12903720]tatgagctc gtc agt age agt aca age cat cac tgg gaa ttt taa42603721]TyrGluLeu Val Ser Ser Ser Thr Ser His His Trp Glu Phe3722]12951300 13053723]atetcttea age taa att aga acg get tea tta att act gtg ata43053724]lieSerSer Ser lie Arg Thr Ala Ser Leu lie Thr Val lie3725]1310 1315 13203726]tactttcat ccg ttt tat gat ctg taa aat aaa aac tea get ggg43503727]TyrPheHis Pro Phe Tyr Asp Leu Asn Lys Asn Ser Ala Gly3728]1325 1330 1335[3729]tccag
4355SerSEQUENCE LISTING15<110>北京思安创新生物技术有限公司 段，陵浔<120>20种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗<130>NM_015589<160>1<170>PatentIn version 3. 5<210>1<211>2356<212>DNA<213>Homo sapiens<220><221>exon<222>(1). . (194)<220><221>exon<222>(195).<220><221>exon<222>(714).<220><221>exon<222>(978).<220><221>exon<222>(1088)<220><221>exon<222>(1175)<220><221>exon<222>(1509)<220><221>exon<222>(1595)<220>
.(713)
(977)
(1087)
..(1174)(1508)
(1594)
. (1713)
119[3767]<221>exon<222> (1714)·· (1915) [3769]<220><221>exon<222> (1916)·· (2042)<220><221>exon<222>(2043).. (2126)<220><221>exon<222>(2127)·· (2338)<400>1gat gtt tcg cga cca ggt egg ggt get ggc ggg ctg gtt taa ggg ctg 48Asp Val Ser Arg Pro Gly Arg Gly Ala Gly Gly Leu Val Gly Leu151015gaa cga gtg cga gca gac tgt tgc get get gtc get get caa gcg cgt 96Glu Arg Val Arg Ala Asp Cys Cys Ala Ala Val Ala Ala Gln Ala Arg202530gag cca gac cca ggc ccg ctt cct cca get ctg cct gga gca ctc get 144Glu Pro Asp Pro Gly Pro Leu Pro Pro Ala Leu Pro Gly Ala Leu Ala354045ggc cga ctg cgc cga get gca cgt cct cga acg cga ggc caa cag ccc 192Gly Arg Leu Arg Arg Ala Ala Arg Pro Arg Thr Arg Gly Gln Gln Pro505560eg g aat cat taa cca atg gca aca gga ate caa gga taa agt gat ttc 240Arg Asn His Pro Met Ala Thr Gly lie Gln Gly Ser Asp Phe657075cct cct gtt aac tea tct gcc ttt get gaa gcc agg aaa cct cga cgc 288Pro Pro Val Asn Ser Ser Ala Phe Ala Glu Ala Arg Lys Pro Arg Arg808590gaa agt aga ata tat gaa act get gcc caa aat cct ggc tea ctc tat 336Glu Ser Arg lie Tyr Glu Thr Ala Ala Gln Asn Pro Gly Ser Leu Tyr95100105tga aca caa cca gca cat tga gga gag cag gca get get gtc cta tgc 384Thr Gln Pro Ala His Gly Glu Gln Ala Ala Ala Val Leu Cys110115120ttt gat aca tcc age cac ttc gtt aaa aga ccg tag tgc ttt age cat 432Phe Asp Thr Ser Ser His Phe Val Lys Arg Pro Cys Phe Ser His125130135[3806]gtg get gaa tea ctt gga gga ccg cac gtc gac cag ctt tgg tgg cca 480Val Ala Glu Ser Leu Gly Gly Pro His Val Asp Gln Leu Trp Trp Pro140145150gaa ccg agg ccg ctc aga ctc tgt gga tta tgg aca gac aca cta cta 528Glu Pro Arg Pro Leu Arg Leu Cys Gly Leu Trp Thr Asp Thr Leu Leu155160165170tea cca aag aca gaa ctc tga tga caa get caa tgg gtg gca gaa ctc 576 [3813]Ser Pro Lys Thr Glu LeuGln Ala Gln Trp Val Ala Glu Leu175180tcg gga ttc tgg gat ttg cat caa tgc ctc caa ctg gca gga caa aag 624Ser Gly Phe Trp Asp Leu His Gln Cys Leu Gln Leu Ala Gly Gln Lys185190195200cat ggg gtg tga gaa tgg cca tgt gcc cct cta ctc ctc ctc ate tgt 672His Gly Val Glu Trp Pro Cys Ala Pro Leu Leu Leu Leu lie Cys205210215ccc cac cac aat caa tac gat tgg aac cag cac aag tac aa t tct ctc 720Pro His His Asn Gln Tyr Asp Trp Asn Gln His Lys Tyr Asn Ser Leu220225230agg cca ggc aca cca cag ccc ttt gaa acg ate tgt gtc cct tac ccc 768Arg Pro Gly Thr Pro Gln Pro Phe Glu Thr lie Cys Val Pro Tyr Pro235240245acc cat gaa tgt gcc aaa cca gcc tct agg aca tgg atg gat gtc tea 816Thr His Glu Cys Ala Lys Pro Ala Ser Arg Thr Trp Met Asp Val Ser250255260tga gga ctt acg age tag agg acc cca gtg cct ccc ate cga tea tgc 864Gly Leu Thr Ser Arg Thr Pro Val Pro Pro lie Arg Ser Cys265270275ccc cct gtc tcc aca aag cag tgt age ctc ttc agg aag tgg tgg gag 912Pro Pro Val Ser Thr Lys Gln Cys Ser Leu Phe Arg Lys Trp Trp Glu280285290tga aca ctt aga aga tea gac cac tgc tcg taa cac att cca aga aga 960Thr Leu Arg Arg Ser Asp His Cys Ser His lie Pro Arg Arg295300305agg tag tgg gat gaa ag a tgt tcc age ctg get gaa aag cct ccg cct 1008Arg Trp Asp Glu Arg Cys Ser Ser Leu Ala Glu Lys Pro Pro Pro310315320gca caa ata tgc cgc get ttt ctc cca gat gac cta tga gga gat gat 1056Ala Gln lie Cys Arg Ala Phe Leu Pro Asp Asp Leu Gly Asp Asp325330335[3845]ggc cct cac cga gtg cca get gga ggc gca g aa tgt tac caa agg tgc 1104Gly Pro His Arg Val Pro Ala Gly Gly Ala Glu Cys Tyr Gln Arg Cys340345350 aag aca caa aat tgt cat cag tat tea gaa get caa aga aag aca aaa 1152Lys Thr Gln Asn Cys His Gln Tyr Ser Glu Ala Gln Arg Lys Thr Lys355360365tct cct gaa gtc ttt gga aag g ga cat cat cga ggg ggg cag cct gcg 1200Ser Pro Glu Val Phe Gly Lys Gly His His Arg Gly Gly Gln Pro Ala370375380385cat ccc get cca gga act gca cca gat gat cct gac tcc gat caa ggc 1248His Pro Ala Pro Gly Thr Ala Pro Asp Asp Pro Asp Ser Asp Gln Gly390395400cta cag ctc ccc gag cac cac ccc cga ggc tcg ccg ccg gga gcc cca 1296Leu Gln Leu Pro Glu His His Pro Arg Gly Ser Pro Pro Gly Ala Pro405410415ggc ccc gcg tea gcc ctc act gat ggg ccc cga gag cca gag ccc cga 1344Gly Pro Ala Ser Ala Leu Thr Asp Gly Pro Arg Glu Pro Glu Pro Arg420425430ctg caa aga tgg ggc cgc age cac tgg cgc cac ggc cac ccc ctc ggc 1392Leu Gln Arg Trp Gly Arg Ser His Trp Arg His Gly His Pro Leu Gly435440445egg ggc cag egg ggg get cca gcc gca cca get gag cag ctg cga tgg 1440Arg Gly Gln Arg Gly Ala Pro Ala Ala Pro Ala Glu Gln Leu Arg Trp450455460465gga get ggc cgt cgc ccc cct gcc aga ggg gga cct ccc egg gca gtt 1488Gly Ala Gly Arg Arg Pro Pro Ala Arg Gly Gly Pro Pro Arg Ala Val470475480cac acg cgt cat ggg gaa ag t gtg cac aca get ctt ggt ctc cag acc 1536His Thr Arg His Gly Glu Ser Val His Thr Ala Leu Gly Leu Gln Thr485490495tga tga gga aaa tat aag ttc cta ttt aca get cat aga caa gtg tct 1584Gly Lys Tyr Lys Phe Leu Phe Thr Ala His Arg Gln Val Ser500505510aat tea tga g gc att tac aga gac aca gaa aaa aag att gtt gtc atg 1632Asn SerGly lie Tyr Arg Asp Thr Glu Lys Lys lie Val Val Met515520525gaa aca gca ggt gca gaa get ctt tcg gtc ttt ccc tcg gaa aac cct 1680Glu Thr Ala Gly Ala Glu Ala Leu Ser Val Phe Pro Ser Glu Asn Pro530535540[3884]tct aga cat ate agg ata tcg aca gca aag aaa tcg agg ctt tgg gca 1728Ser Arg His lie Arg lie Ser Thr Ala Lys Lys Ser Arg Leu Trp Ala545550555ate caa ctc cct ccc gac ggc tgg ctc tgt ggg egg tgg cat ggg cag 1776lie Gln Leu Pro Pro Asp Gly Trp Leu Cys Gly Arg Trp His Gly Gln560565570acg gaa ccc gcg cca gta cca gat ccc ctc tcg gaa cgt ccc ttc cgc 1824Thr Glu Pro Ala Pro Val Pro Asp Pro Leu Ser Glu Arg Pro Phe Arg575580585590ccg cct ggg cct ctt ggg cac cag tgg att cgt cag ctc caa cca gcg 1872Pro Pro Gly Pro Leu Gly His Gln Trp lie Arg Gln Leu Gln Pro Ala595600605caa cac cac age tac ccc cac cat cat gaa aca agg aag aca g aa cct 1920Gln His His Ser Tyr Pro His His His Glu Thr Arg Lys Thr Glu Pro610615620gtg gtt tgc caa ccc egg ggg cag caa tag cat gcc aag ccg cac cca 1968Val Val Cys Gln Pro Arg Gly Gln Gln His Ala Lys Pro His Pro625630635cag ctc agt cca gag gac ccg ctc get gcc cgt gca cac ttc ccc aca 2016Gln Leu Ser Pro Glu Asp Pro Leu Ala Ala Arg Ala His Phe Pro Thr640645650gaa cat get gat gtt cca gca gcc ag a att cca get tcc cgt gac cga 2064Glu His Ala Asp Val Pro Ala Ala Arg lie Pro Ala Ser Arg Asp Arg655660665acc tga cat caa caa cag get gga gtc gtt gtg cct cag tat gac cga 2112Thr His Gln Gln Gln Ala Gly Val Val Val Pro Gln Tyr Asp Arg670675680aca cgc cct ggg ag a egg ggt tga ccg gac ctc cac cat cta gaa get 2160Thr Arg Pro Gly Arg Arg Gly Pro Asp Leu His His Leu Glu Ala685690695gaa gat gag agt gac cgc get ggc cgt gaa ate gac tgc tgc ggg tcc 2208Glu Asp Glu Ser Asp Arg Ala Gly Arg Glu lie Asp Cys Cys Gly Ser700705710715agt gtc cgc cat ctt cag ggt tgc aca gaa tcc tcc aag ata ctt tgc 2256Ser Val Arg His Leu Gln Gly Cys Thr Glu Ser Ser Lys lie Leu Cys[3919]720725730age ctt ttt tcc ccc tgg tcc ctc tcc cgt ttt gat ttt gtg aga gcg 2304Ser Leu Phe Ser Pro Trp Ser Leu Ser Arg Phe Asp Phe Val Arg Ala735740745[3923]tag gtc ate ctc gta aac ata tea gta gac ctg g aaaaaaaaaa aaaaaaaa 2356Val lie Leu Val Asn lie Ser Val Asp Leu750755SEQUENCE LISTING16<110>北京思安创新生物技术有限公司段，陵浔<120>20种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗<130>NM_016002 <160>1<170>PatentIn version 3. 5<210>1<211>2482<212>DNA<213>Homo sapiens<220><221>exon<222>(1). . (537)<220><221>exon<222>(538). . (650)<220><221>exon<222>(651). . (731)<220><221>exon<222>(732). . (861)<220><221>exon<222>(862). . (911)<220><221>exon<222>(912). . (1042)<220><221>exon<222>(1043)·· (1160)<220><221>exon<222> (1161).. (1280)<220>[3962]<221>exon[3963]<222> (1281) ·· (1337)<220><221>exon<222>(1338)·· (1449)<220><221>exon<222>(1450).. (1531)<220><221>exon<222> (1532). . (2474)<220><221>STS<222>(1577).. (1739)<220><221>STS<222> (1901). . (2027)<220><221>STS<222>(2019)·· (2168)<220><221>STS<222>(2020)·· (2144)<220><221>STS<222>(2020)·· (2144)<220><221>STS<222>(2184).. (2428)<220><221>STS<222>(2247)·· (2421)<400>1tgc get gcc egg ggc ggc cag age cct gca gaa caa acc gcc gca egg 48Cys Ala Ala Arg Gly Gly Gln Ser Pro Ala Glu Gln Thr Ala Ala Arg151015aat tag ggg ccg ccg tcc gcg cct gcc agg ctt gga age gga ctc gcc 96Asn Gly Pro Pro Ser Ala Pro Ala Arg Leu Gly Ser Gly Leu Ala202530[4001]ccc agg gcc accgcc cta gga aac ctc ccg tcg gaa gcc tcc caa gca 144Pro Arg Ala ThrAla Leu Gly Asn Leu Pro Ser Glu Ala Ser Gln Ala354045age cgc tea cccgcg get ccc gcg cga ggc ggc gtg ggc gtg ggc gcg 192Ser Arg Ser ProAla Ala Pro Ala Arg Gly Gly Val Gly Val Gly Ala505560gcc ctg gcg eggegg cgc gcg egg ggt gaa ggg age ccg gcg cgc tgg 240Ala Leu Ala ArgArg Arg Ala Arg Gly Glu Gly Ser Pro Ala Arg Trp [4009]657075ctg egg cga eggegg ccg ttg ctg cgc egg gta ctg ggg tcg ctg cct 288Leu Arg Arg ArgArg Pro Leu Leu Arg Arg Val Leu Gly Ser Leu Pro80859095gag gcg cag gcgccg tgg act cca ccc gcc ccg ggg cct ggg ctc get 336Glu Ala Gln AlaPro Trp Thr Pro Pro Ala Pro Gly Pro Gly Leu Ala100105110gtg gac tcg teatgg cga ccg age aga ggc ctt tcc acc tgg tgg tgt 384Val Asp Ser SerTrp Arg Pro Ser Arg Gly Leu Ser Thr Trp Trp Cys115120125tcg gcg cgt ctgget tea ccg gcc agt tcg tga ccg agg agg tgg ccc 432Ser Ala Arg LeuAla Ser Pro Ala Ser Ser Pro Arg Arg Trp Pro130135140ggg age agg tggacc egg age gga get ccc gcc tgc cct ggg ccg tgg 480Gly Ser Arg TrpThr Arg Ser Gly Ala Pro Ala Cys Pro Gly Pro Trp145150155egg gcc get cccggg aga age tgc age ggg tgc tgg aga agg egg ccc 528Arg Ala Ala ProGly Arg Ser Cys Ser Gly Cys Trp Arg Arg Arg Pro160165170tga age tgg gaagac caa cac tgt cat ctg aag ttg gaa tea tea tct 576Ser Trp GluAsp Gln His Cys His Leu Lys Leu Glu Ser Ser Ser175180185gtg ata ttg ctaate cag cct cgc ttg atg aaa tgg cta aac agg caa 624Val lie Leu Leulie Gln Pro Arg Leu Met Lys Trp Leu Asn Arg Gln190195200205cag ttg tcc teaatt gcg tag gac ca t ate ggt ttt atg gag aac ctg 672Gln Leu Ser Serlie Ala Asp His lie Gly Phe Met Glu Asn Leu210215220taa taa aag catgta ttg aaa atg gag cca gtt gta tcg aca tea gtg 720Lys HisVal Leu Lys Met Glu Pro Val Val Ser Thr Ser Val225230[4040]gag aac ctc ag t ttc tgg aac taa tgc aac tga agt ate atg aga aag 768Glu Asn Leu Ser Phe Trp Asn Cys Asn Ser lie Met Arg Lys235240245 ctg cag aca aag ggg ttt ata tea ttg gaa gca gcg get ttg act cca 816Leu Gln Thr Lys Gly Phe lie Ser Leu Glu Ala Ala Ala Leu Thr Pro250255260ttc cag cag ate tgg gag taa tat ata cca gaa ata aaa tga atg gta 864Phe Gln Gln lie Trp Glu Tyr lie Pro Glu lie Lys Met Val265270275ctt tga ctg ctg tgg aaa gtt tcc tga cta tac att cag gac ctg ag g 912Leu Leu Leu Trp Lys Val SerLeu Tyr lie Gln Asp Leu Arg280285290ggt tga gca ttc atg atg gta cct gga agt cag caa ttt atg gtt ttg 960Gly Ala Phe Met Met Val Pro Gly Ser Gln Gln Phe Met Val Leu295300305gag ate aga gta att tga gaa aac taa gaa atg tat caa ate tga aac 1008Glu lie Arg Val lie Glu Asn Glu Met Tyr Gln lie Asn310315320ctg tcc cgc tea ttg gtc caa aat tga aga gaa g gt ggc caa ttt ctt 1056Leu Ser Arg Ser Leu Val Gln Asn Arg Glu Gly Gly Gln Phe Leu325330335att gtc ggg aac tea aag gtt att cca ttc ctt tta tgg gat ctg atg 1104lie Val Gly Asn Ser Lys Val lie Pro Phe Leu Leu Trp Asp Leu Met340345350tgt ctg ttg taa gga gga ctc aac gtt act tgt atg aaa att tag agg 1152Cys Leu Leu Gly Gly Leu Asn Val Thr Cys Met Lys lie Arg355360365aat cac ca g ttc agt atg ctg cgt atg taa ctg tgg gag gca tea cct 1200Asn His Gln Phe Ser Met Leu Arg Met Leu Trp Glu Ala Ser Pro370375380ctg tta tta age tga tgt ttg cag gac ttt tct ttt tgt tct ttg tga 1248Leu Leu Leu Ser Cys Leu Gln Asp Phe Ser Phe Cys Ser Leu385390ggt ttg gaa ttg gaa ggc aac ttc tea taa aa t tcc cat ggt tct tct 1296Gly Leu Glu Leu Glu Gly Asn Phe Ser Asn Ser His Gly Ser Ser395400405cct ttg get att ttt caa aac aag gcc caa cac aaa aac ag a ttg atg 1344Pro Leu Ala lie Phe Gln Asn Lys Ala Gln His Lys Asn Arg Leu Met410415420425[4079]ctg cct cat tea cgc tga cat tct ttg gtc aag gat aca gcc aag gca 1392Leu Pro His Ser Arg His Ser Leu Val Lys Asp Thr Ala Lys Ala430435440 [4082]ctg gta cag ata aga aca aac caa ata tea aaa ttt gta ctc agg tga 1440Leu Val Gln lie Arg Thr Asn Gln lie Ser Lys Phe Val Leu Arg445450455aag gac cag agg ctg get atg tgg cta ccc cca tag cta tgg ttc agg 1488Lys Asp Gln Arg Leu Ala Met Trp Leu Pro Pro Leu Trp Phe Arg460465470cag cca tga ctc ttc taa gtg atg ctt ctc ate tgc cta agg c gg gcg 1536Gln Pro Leu Phe Val Met Leu Leu lie Cys Leu Arg Arg Ala475480ggg tct tea cac ctg gag cag ctt ttt cca aaa caa agt tga ttg aca 1584Gly Ser Ser His Leu Glu Gln Leu Phe Pro Lys Gln Ser Leu Thr485490495gac tea aca aac acg gta ttg agt tta gtg tta tta gca get ctg aag 1632Asp Ser Thr Asn Thr Val Leu Ser Leu Val Leu Leu Ala Ala Leu Lys500505510515tct aaa cac tgg aag aat taa ctg aag tea taa cgt gcg tga att aac 1680Ser Lys His Trp Lys Asn Leu Lys Ser Arg Ala lie Asn520525age ttc tct att tga tat ttg aaa ttc ttc tgt aag cct gtc tga gtg 1728Ser Phe Ser lie Tyr Leu Lys Phe Phe Cys Lys Pro Val Val530535540tat gtg gaa acg att gtc aaa tct aaa ata tct ata tat taa aaa gta 1776Tyr Val Glu Thr lie Val Lys Ser Lys lie Ser lie Tyr Lys Val545550555gga aat tgt cct age tta ccc taa att tea aat ctg agt tga ttt tgt 1824Gly Asn Cys Pro Ser Leu Pro lie Ser Asn Leu Ser Phe Cys560565570gat ttt att get tat aac aga gaa ctc ata ttt gac ata ttt ttt tea 1872Asp Phe lie Ala Tyr Asn Arg Glu Leu lie Phe Asp lie Phe Phe Ser575580585ttg atg tgt tcc tgg tag att ttc acg aat gag ctg gca ggt cta atg 1920Leu Met Cys Ser Trp lie Phe Thr Asn Glu Leu Ala Gly Leu Met590595600ggg gag gcg gcg tcc cag tct gtg ttg cag cag cat tct cat egg ggg 1968Gly Glu Ala Ala Ser Gln Ser Val Leu Gln Gln His Ser His Arg Gly605610615[4118]tgc gca cac cat cgt tac tgt egg gca gta act gcc get tgc ctt gcc 2016Cys Ala His His Arg Tyr Cys Arg Ala Val Thr Ala Ala Cys Leu Ala620625630gca gta gga ggg aaa tct cac ctt cct tcc aca tac tgt ctt gag cct 2064Ala Val Gly Gly Lys Ser His Leu Pro Ser Thr Tyr Cys Leu Glu Pro635640645650ttg cta aat taa act gca ctt ttt get att ttt gcc tag ttt ttc gcc 2112 [4125]Leu Leu Asn Thr Ala Leu Phe Ala lie Phe Ala Phe Phe Ala655660aat cta cac tga ttt tgg act gtt acc taa gtt gaa aaa taa aag gtt 2160Asn Leu His Phe Trp Thr Val Thr Val Glu Lys Lys Val665670675gt c aat cga atg gtg gtt taa tgt ttg gac ctg ccg atg tat ttg tat 2208Val Asn Arg Met Val ValCys Leu Asp Leu Pro Met Tyr Leu Tyr680685690agt ggt aga aac atg ctg ctt aag tgg cct aac ctg ttt ctt gcc aat 2256Ser Gly Arg Asn Met Leu Leu Lys Trp Pro Asn Leu Phe Leu Ala Asn695700705aag tag get tat cat ttt ate ttt acg taa ttc tat ate tgt gac tag 2304Lys Ala Tyr His Phe lie Phe ThrPhe Tyr lie Cys Asp710715720gtt ttt aag gat aca get tat aag ttg cta tea att ttc act acc taa 2352Val Phe Lys Asp Thr Ala Tyr Lys Leu Leu Ser lie Phe Thr Thr725730735gca gaa ttt ttc tct aat tta ctt ttt gta ttt taa cta ggt ttt aca 2400Ala Glu Phe Phe Ser Asn Leu Leu Phe Val Phe Leu Gly Phe Thr740745750tgg aag ccc taa aat aag gca aaa gac ttt ttc ttt tgt aat aag cat 2448Trp Lys Pro Asn Lys Ala Lys Asp Phe Phe Phe Cys Asn Lys His755760765ata ata aac acg tat ata cat age aa aaaaaaaa2482lie lie Asn Thr Tyr lie His Ser770SEQUENCE LISTING 17<110>北京思安创新生物技术有限公司段，陵浔<120>20种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗<130>NM_019073<160>1[4157]<170>PatentIn version 3.5<210>1<211>1739 <212>DNA<213>Homo sapiens<220><221>exon<222>(1). . (216)<220><221>exon<222>(217). . (354)<220><221>exon<222>(355). . (403)<220><221>exon<222> (404) · · (445)<220><221>exon<222>(446). . (570)<220><221>exon<222>(571). . (651)<220><221>exon<222>(652). . (945)<220><221>exon<222> (946) ·. (1033)<220><221>exon<222>(1034). . (1074)<220><221>exon<222> (1075)·· (1259)<220><221>exon<222> (1260)·· (1359)<220>[4196<221>exon[4197<222>(1360)..(1451)[4198<220>[4199<221>exon[4200<222>(1452)..(1739)[4201<400>1[4202gettctccggcggaacccaggctggaccgcgggccceggcctgggggc48[4203AlaSerProAlaGluProArgLeuAspArgGlyProArgProGlyGly[4204151015[4205cactgctgccaccgccgccgccacctcgtctcctccccgtccccgccc96[4206HisCysCysHisArgArgArgHisLeuValSerSerProSerProPro[4207202530[4208agecccaggtctccccgcctcactegggcccgtggceggggtcactcc144[4209SerProArgSerProArgLeuThrArgAlaArgGlyArgGlyHisSer[4210354045[4211ccgccgcccctccccgcaeggatgccgaaggtgaaggcgctgcagtgc192[4212ProProProLeuProAlaArgMetProLysValLysAlaLeuGinCys[4213505560[4214gccctggcgctggagateageteagtaacttgcccaggagtcgtgctt240[4215AlaLeuAlaLeuGlulieSerSerValThrCysProGlyValValLeu[421665707580[4217gacgaggacatetatcttageatetgtgtgtttggccaatac288[4218LysAspLysGluAsplieTyrLeuSerlieCysValPheGlyGinTyr[4219859095[4220aagacacaatgtgtcccagccacttttcccctggtcttcaatgcc336[4221LysLysThrGinCysValProAlaThrPheProLeuValPheAsnAla[4222100105110[4223agaatggtgtttgaaaaggtgttcccggacgcagtagatcctggagat384[4224ArgMetValPheGluLysValPheProAspAlaValAspProGlyAsp[4225115120125[4226gtggttacacagcttgaat atgat aca gca gtg ttc gag ttg ata cag432[4227ValValThrGinLeuGluTyr Asp Thr Ala Val Phe Glu Leu lie Gin[4228130140[4229etagttccaccag tg ggt gaa aca ctg tct acg tat gac gaa aat aca480[4230LeuValProProVal Gly Glu Thr Leu Ser Thr Tyr Asp Glu Asn Thr[4231145150155160[4232cgagatttcatgtttccgggtccaaaccaaatgtctggacaccatgat528[4233ArgAspPheMetPhePro Gly Pro AsnGin Met Ser Gly His His Asp[4234165170175[4235]tea aac cgc cag gtt acc atg agg agg att tct ggc ctt cga gga aat 576[4236]Ser Asn Arg Gln Val Thr Met Arg Arg lie Ser Gly Leu Arg Gly Asn180185190get cca agg ctg gaa ttt tct acg act tea gtg att act gaa tgt ctg 624Ala Pro Arg Leu Glu PheSer Thr Thr Ser Val lie Thr Glu Cys Leu195200205ata agt tea agg aaa tgc cac act cag gat aaa ttt att tac cat ttg 672lie Ser Ser Arg Lys Cys His Thr Gln Asp Lys Phe lie Tyr His Leu210215220get cca gtt gaa aaa tea cat ggc aga ctg caa aac aga aca tea aga 720Ala Pro Val Glu Lys Ser His Gly Arg Leu Gln Asn Arg Thr Ser Arg225230235240tea caa aag aaa aaa tcc aag tea cct gag aga agt aaa tac tgt ata 768Ser Gln Lys Lys Lys Ser Lys Ser Pro Glu Arg Ser Lys Tyr Cys lie245250255aat gca aaa aac tac gaa cag cct aca att tct tea aaa tea cac tct 816Asn Ala Lys Asn Tyr Glu Gln Pro Thr lie Ser Ser Lys Ser His Ser260265270cca tct ccc tac aca aaa aga cgc atg tgt gag cta tct gaa gac acc 864Pro Ser Pro Tyr Thr Lys Arg Arg Met Cys Glu Leu Ser Glu Asp Thr275280285agg egg egg ctg gcc cat tta aat ctg gga ccc tat gag ttc aaa aaa 912Arg Arg Arg Leu Ala His Leu Asn Leu Gly Pro Tyr Glu Phe Lys Lys290295300gaa aca gat aaa cct cca ttt gtg att aga cat gtt gat ccc cca agt 960Glu Thr Asp Lys Pro Pro Phe Val lie Arg His Val Asp Pro Pro Ser305310315320ccc agg get gat act tta ttg gga tct tct gga aga gac tgt gaa aga 1008Pro Arg Ala Asp Thr Leu Leu Gly Ser Ser Gly Arg Asp Cys Glu Arg325330335gat gga tgg tea agg gtg cac aat g at cat tct cat ctt ggc tgc tgc 1056AspGly Trp Ser Arg Val His Asn Asp His Ser His Leu Gly Cys Cys340350cga ccc aag gat tat aag gtt ate agg aca ccc cat ggg aga gac ttc 1104Arg Pro Lys Asp Tyr Lys Val lie Arg Thr Pro His Gly Arg Asp Phe355360365gat gac tct tta gaa aaa tgt gaa gag tat ttg age cca agg tcg tgt 1152Asp Asp Ser Leu Glu Lys Cys Glu Glu Tyr Leu Ser Pro Arg Ser Cys370375380CN 10845438A说明 书 131/189页[4274agtaagccceggcatteagcg agg acc ttg eta gtc cat tea gca ccc1200[4275SerLysProArg HisSer Ala Arg Thr Leu Leu Val His Ser Ala Pro[4276385390395 400[4277teaacaatgccaaagcattct cca age cct gtg tta aat aga get tct1248[4278SerThrMetProLysHisSer Pro Ser Pro Val Leu Asn Arg Ala Ser[4279405410 415[4280ctcagggaaag a tttcattct gat tgg tgt tea cct tea aac tgc gat1296[4281LeuArgGluArgPhe His Ser Asp Trp Cys Ser Pro Ser Asn Cys Asp[4282425 430[4283gagatecatgacegggtaaaa aat gtc ttg aaa tea cat cag get cat1344[4284GlulieHisAspArgValLys Asn Val Leu Lys Ser His Gin Ala His[4285435440 445 [4286caaagacatttatatgatgag aga gac eta gag aaa gat gat gaa ctg1392[4287GinArgHisLeuTyrAspGlu Arg Asp Leu Glu Lys Asp Asp Glu Leu[4288450455 460[4289gaactgagaagtctttta tgt aga gac tct gcc tat gac agt gac1440[4290GluLeuLysArgSerLeuLeu Cys Arg Asp Ser Ala Tyr Asp Ser Asp[4291465470475 480[4292cccgagtatag cteatgtcag cag cca cgt ggc act ttc cat ttg gat1488[4293ProGluTyrSerSer Cys Gin Gin Pro Arg Gly Thr Phe His Leu Asp[4294485490 495[4295gacggtgaatactggtccaac agg gca gcc tct tat aag gga aaa tcc1536[4296AspGlyGluTyrTrpSerAsn Arg Ala Ala Ser Tyr Lys Gly Lys Ser[4297500505 510[4298caccgacccatetttgagaac age atg gac aag atg tac agg aac tta1584[4299HisArgProliePheGluAsn Ser Met Asp Lys Met Tyr Arg Asn Leu[4300515520 525[4301tacclclclaaggcctgtagttct get tea cat aca cag gaa age ttc tga1632[4302TyrLysLysAlaCysSerSer Ala Ser His Thr Gin Glu Ser Phe[4303530535 540[4304gaccatccatgataaacctea tta gtg tcc gtg tea att tct caa tga1680[4305AspHisProThrSer Leu Val Ser Val Ser lie Ser Gin[4306545550 555[4307tgtttgatgtgateaata tct gta ttc ctt ttt tta aaa aaa atg1728[4308LysCysLeuMetSerlie Ser Val Phe Leu Phe Leu Lys Lys Met[4309560565 570[4310ggatt a taa ct1739[4311]GlyLeu
133[4312]SEQUENCE LISTING18<110>北京思安创新生物技术有限公司段，陵浔 <120>20种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗<130>NM_003284<160>1<170>PatentIn version 3. 5<210>1<211>415<212>DNA<213>Homo sapiens<220><221>exon<222>(1). . (164)<220><221>exon<222>(165). . (402)<220><221>STS<222>(213). . (393)<400>1gcc cct cat ttt ggc aga act tac cat gtc gac cag ccg caa att aaa 48Ala Pro His Phe Gly Arg Thr Tyr His Val Asp Gln Pro Gln lie Lys151015gag tea tgg cat gag gag gag caa gag ccg ate tcc tea caa ggg agt 96Glu Ser Trp His Glu Glu Glu Gln Glu Pro lie Ser Ser Gln Gly Ser202530caa gag agg tgg cag caa aag aaa ata ccg taa ggg caa cct gaa aag 144Gln Glu Arg Trp Gln Gln Lys Lys lie Pro Gly Gln Pro Glu Lys354045tag gaa acg ggg cga tga eg c caa tcg caa tta ccg ctc cca ctt gtg 192Glu Thr Gly Arg Arg Gln Ser Gln Leu Pro Leu Pro Leu Val505560age ccc cag egg get ctg ccc tgg tgc get tea cac age acc aag cag 240Ser Pro Gln Arg Ala Leu Pro Trp Cys Ala Ser His Ser Thr Lys Gln657075caa caa gaa cag cag aag ggg aac tgc caa gga gac ctg atg tta gat 288Gln Gln Glu Gln Gln Lys Gly Asn Cys Gln Gly Asp Leu Met Leu Asp808590caa age cag aga gga gcc tat gga atg tgg ate aaa tgc cag ttg tga 336 Gin Ser Gin Arg Gly Ala Tyr Gly Met Trp lie Lys Cys Gin Leu
95100105
cga aat gag gaa tgt ata tgt tgg ctg ttt ttc ccc aac ate tea ata 384 Arg Asn Glu Glu Cys lie Cys Trp Leu Phe Phe Pro Asn lie Ser lie
110115120
aaa ctt tga aag cag aaa aaaaaaaaaa aaa415
Lys Leu Lys Gin Lys 125
SEQUENCE LISTING19 <110>北京思安创新生物技术有限公司 段，陵浔
<120>20种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗
<130>NM_021733
<160>1
<170>PatentIn version 3. 5 <210>1 <211>1812 <212>DNA
<213>Homo sapiens <220> <221>exon <222>(1). . (181) <220> <221>exon <222>(182).. (410) <220> <221>exon <222>(411).. (506) <220> <221>exon <222>(507).. (590) <220> <221>exon <222>(591). . (674) <220> <221>exon <222>(675).. (1003) <220><221>exon<222> (1004)·· (1198)<220><221>exon<222>(1199)·· (1372)<220><221>exon [4397]<222>(1373)·· (1508)<220><221>exon<222>(1509)·· (1633)<220><221>exon<222> (1634)·· (1812)<220><221>polyA_signal<222>(1786)·· (1791)<220><221>polyA_site<222>(1812)·· (1812)<400>1acc ccc aca cca tgg cga gcg tgg tgg tga aga cga tct ggc agt cca 48Thr Pro Thr Pro Trp Arg Ala Trp Trp Arg Arg Ser Gly Ser Pro151015aag aga tcc atg agg ccg ggg aca ccc cca egg ggg tgg aga get get 96Lys Arg Ser Met Arg Pro Gly Thr Pro Pro Arg Gly Trp Arg Ala Ala202530ccc age tag tcc cag agg ctc ccc gga ggg tga cca gcc ggg cca agg 144Pro Ser Ser Gln Arg Leu Pro Gly Gly Pro Ala Gly Pro Arg354045gga tcc cga aga aaa aga agg ccg tgt cgt tcc acg g gg tgg age ccc 192Gly Ser Arg Arg Lys Arg Arg Pro Cys Arg Ser Thr Gly Trp Ser Pro505560aga tgt ccc ate age cca tgc act ggt gcc tga acc tea aac ggt cct 240Arg Cys Pro lie Ser Pro Cys Thr Gly Ala Thr Ser Asn Gly Pro657075egg cct gca cca acg tgt cac tgc tea acc tgg ccg cca tgg age cca 288Arg Pro Ala Pro Thr Cys His Cys Ser Thr Trp Pro Pro Trp Ser Pro808590[4429]ctg act cca egg gga cag act cca cag tgg aag acc tea gcg gcc aac 336Leu Thr Pro Arg Gly Gln Thr Pro Gln Trp Lys Thr Ser Ala Ala Asn95100105 [4432]tea cac tgg ctg ggc ccc ctg cct ccc cta ccc tac cct ggg ate egg 384Ser His Trp Leu Gly Pro Leu Pro Pro Leu Pro Tyr Pro Gly lie Arg110115120atg acg cag aca tea egg aaa tcc tg a gtg ggg tea aca gtg gat tgg 432Met Thr Gln Thr Ser Arg Lys SerVal Gly Ser Thr Val Asp Trp125130135140tcc gcg cca aag act cca tea cca get tga agg aaa aga cca acc ggg480Ser Ala Pro Lys Thr Pro Ser Pro Ala Arg Lys Arg Pro Thr Gly145150155tta acc age acg tgc agt ctc tgc ag a gcg agt gtt ctg tgc tga gcg 528Leu Thr Ser Thr Cys Ser Leu Cys Arg Ala Ser Val Leu Cys Ala160165170aga ate tgg aga gaa ggc ggc aag agg cag aag agt tgg agg ggt act576Arg lie Trp Arg Glu Gly Gly Lys Arg Gln Lys Ser Trp Arg Gly Thr175180185gca ttc aac tea ag g aga act get gga agg tga ccc ggt ctg tgg aag 624Ala Phe Asn Ser Arg Arg Thr Ala Gly Arg Pro Gly Leu Trp Lys190195200atg ctg aaa tea aaa cca acg tct tga age aga att ctg ccc tgc tgg672Met Leu Lys Ser Lys Pro Thr Ser Ser Arg lie Leu Pro Cys Trp205210215ag g aga age tgc get acc tcc age age age tgc agg atg aga cgc cgc 720Arg Arg Ser Cys Ala Thr Ser Ser Ser Ser Cys Arg Met Arg Arg Arg220225230gac ggc agg agg ccg age tgc agg age egg agg aga age agg age egg768Asp Gly Arg Arg Pro Ser Cys Arg Ser Arg Arg Arg Ser Arg Ser Arg235240245agg aga age agg age egg agg aga age aga age egg agg ctg gcc tct816Arg Arg Ser Arg Ser Arg Arg Arg Ser Arg Ser Arg Arg Leu Ala Ser250255260cct gga aca gcc tgg gcc ccg ccg cca cgt ccc agg get gcc ccg gcc864Pro Gly Thr Ala Trp Ala Pro Pro Pro Arg Pro Arg Ala Ala Pro Ala265270275280cgc cag gga gtc ccg aca aac cct cgc ggc cac acg gcc tgg tcc ccg912Arg Gln Gly Val Pro Thr Asn Pro Arg Gly His Thr Ala Trp Ser Pro285290295[4468]cag get ggg gaa tgg ggc ctc ggg ctg gcg agg gcc cct acg tga gcg960Gln Ala Gly Glu Trp Gly Leu Gly Leu Ala Arg Ala Pro Thr Ala300305310age agg aat tgc aga age tgt tea ccg gca tcg aag age tga g ga gag 1008Ser Arg Asn Cys Arg Ser Cys Ser Pro Ala Ser Lys Ser Gly Glu315320 [4474]agg tgt cct cac tga ccg ccc ggt ggc ate agg agg agg ggg egg tgc 1056Arg Cys Pro His Pro Pro Gly Gly lie Arg Arg Arg Gly Arg Cys330335340agg aag ccc tgc ggc tgc tcg ggg gcc tgg gcg gca ggg tcg acg get 1104Arg Lys Pro Cys Gly Cys Ser Gly Ala Trp Ala Ala Gly Ser Thr Ala345350355tcc tag gcc agt ggg age ggg cac age gcg aac agg cac aga egg cgc 1152Ser Ala Ser Gly Ser Gly His Ser Ala Asn Arg His Arg Arg Arg360365370ggg act tgc agg age tgc gag gtc ggg egg atg age tgt gca cca t gg 1200Gly Thr Cys Arg Ser Cys Glu Val Gly Arg Met Ser Cys Ala Pro Trp375380385tgg age ggt cag cag tgt ctg tgg ctt cac tga gga gcg aac tgg agg 1248Trp Ser Gly Gln Gln Cys Leu Trp Leu His Gly Ala Asn Trp Arg390395400ggc tgg gcc cac tga aac cca ttc tgg agg agt tcg ggc ggc aat ttc 1296Gly Trp Ala His Asn Pro Phe Trp Arg Ser Ser Gly Gly Asn Phe405410415aga act ctc gaa gag ggc ctg acc tct cca tga acc tgg ate ggt ccc 1344Arg Thr Leu Glu Glu Gly Leu Thr Ser Pro Thr Trp lie Gly Pro420425430acc aag gca act gtg ccc get gtg cca g cc agg ggt cgc agt tgt cta 1392Thr Lys Ala Thr Val Pro Ala Val Pro Ala Arg Gly Arg Ser Cys Leu435440445egg agt ccc tgc age age tgc tgg acc gag cac tga cct cac tag tgg 1440Arg Ser Pro Cys Ser Ser Cys Trp Thr Glu HisPro His Trp450455460acg agg tga age aga ggg gcc tga ctc ctg cct gtc cca get gtc aga 1488Thr Arg Ser Arg Gly Ala Leu Leu Pro Val Pro Ala Val Arg465470475ggc tac aca aga aga ttc tg g age tgg age gcc agg cct tag cca aac 1536Gly Tyr Thr Arg Arg Phe Trp Ser Trp Ser Ala Arg Pro Pro Asn[4506]480485490acg tea ggg cag agg ccc tga get cca ccc ttc ggc tgg ccc aag acg 1584Thr Ser Gly Gln Arg Pro Ala Pro Pro Phe Gly Trp Pro Lys Thr 495500505agg ccc tgc ggg cca aga acc tac tgc tga cag aca aga tga age cag a 1633Arg Pro Cys Gly Pro Arg Thr Tyr Cys Gln Thr Arg Ser Gln510515520gg aga aga tgg cca ctc tgg acc ate tac act tga aga tgt get ccc 1680Arg Arg Arg Trp Pro Leu Trp Thr lie Tyr Thr Arg Cys Ala Pro525530535tcc acg ate ate tea gca acc tgc cac ttg agg ggt cca egg gaa caa 1728Ser Thr lie lie Ser Ala Thr Cys His Leu Arg Gly Pro Arg Glu Gln540545550tgg ggg gag gca gca gtg cag gaa ccc ccc caa aac agg ggg get cag 1776Trp Gly Glu Ala Ala Val Gln Glu Pro Pro Gln Asn Arg Gly Ala Gln555560565ccc ctg aac aat aaa tgg cct ctc atg cta gca tga1812Pro Leu Asn Asn Lys Trp Pro Leu Met Leu Ala570575SEQUENCE LISTING20<110>北京思安创新生物技术有限公司段，陵浔<120>20种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗<130>NM_017481<160>1<170>PatentIn version 3. 5<210>1<211>2347<212>DNA<213>Homo sapiens<220><221>exon<222>(1). . (50)<220><221>exon<222>(51). . (2345)<220><221>CDS<222>(87). . (2054)[4545]<220>[4546]<221>STS<222>(197). . (443)<220><221>STS<222>(257)·· (1042)<220><221>STS<222>(422). . (665)<220><221>STS<222>(642). . (889)<220><221>STS<222>(783).. (1984)<220><221>STS<222>(860).. (1110)<220><221>STS<222>(1056)·· (1303)<220><221>STS<222>(1131)·· (1949)<220><221>STS<222>(1282)·· (1549)<220><221>STS<222>(1520)·· (1748)<220><221>STS<222>(1707)·· (1947)<220><221>STS<222> (1919)·· (2178)<400>1ggg agg ttt gga gcc ctg cat aaa gag aag gac ggg acc aca get gac 48Gly Arg Phe Gly Ala Leu His Lys Glu Lys Asp Gly Thr Thr Ala Asp[4584]151015tg c tgt gtc ccc aca gat ctg ggc ctc ctg ctg cca cca tgg cca aag 96Cys Cys Val Pro Thr Asp Leu Gly Leu Leu Leu Pro Pro Trp Pro Lys202530gtg gag aag ccc tgc cac agg gca gcc cag cac cag tcc agg ate ccc 144Val Glu Lys Pro Cys His Arg Ala Ala Gln His Gln Ser Arg lie Pro354045 [4591]acc tea tea agg tga cag tga aga cgc cca aag aca agg agg att tct 192Thr Ser Ser Arg Gln Arg Arg Pro Lys Thr Arg Arg lie Ser505560cag tta cag aca cat gca cta tcc age age tga agg aag aga tat ctc 240Gln Leu Gln Thr His Ala Leu Ser Ser Ser Arg Lys Arg Tyr Leu657075age get tta agg ccc acc ccg ate age ttg ttc taa tct ttg ctg gca 288Ser Ala Leu Arg Pro Thr Pro lie Ser Leu Phe Ser Leu Leu Ala808590aaa tcc tea agg ate ctg act cac tgg cac agt gtg gag tgc gag atg 336Lys Ser Ser Arg lie Leu Thr His Trp His Ser Val Glu Cys Glu Met95100105gcc tea ctg tcc acc tgg tea tea aga ggc age acc gtg cca tgg gca 384Ala Ser Leu Ser Thr Trp Ser Ser Arg Gly Ser Thr Val Pro Trp Ala110115120atg agt gcc cag ctg cct ctg tcc cta ccc agg gcc caa gtc ctg gat 432Met Ser Ala Gln Leu Pro Leu Ser Leu Pro Arg Ala Gln Val Leu Asp125130135140cac tcc ctc age caa get cca ttt acc cag cag atg ggc ccc ctg cct 480His Ser Leu Ser Gln Ala Pro Phe Thr Gln Gln Met Gly Pro Leu Pro145150155tta get tag gtc tcc tea cag gcc tea gta ggc tgg get tgg cct ate 528Leu Ala Val Ser Ser Gln Ala Ser Val Gly Trp Ala Trp Pro lie160165170gtg get tcc ctg acc age caa get ccc tga tgc ggc age atg tgt ctg 576Val Ala Ser Leu Thr Ser Gln Ala Pro Cys Gly Ser Met Cys Leu175180185tgc ctg agt ttg tga ctc age tea ttg atg acc cct tea tcc egg gtc 624Cys Leu Ser Leu Leu Ser Ser Leu Met Thr Pro Ser Ser Arg Val190195200tgc tgt cca aca cag gcc tag tac gcc age tgg ttc ttg aca acc ccc 672Cys Cys Pro Thr Gln Ala Tyr Ala Ser Trp Phe Leu Thr Thr Pro[4623]205210215ata tgc age age tga tcc age aca acc ctg aga ttg ggc ata ttc tta 720lie Cys Ser Ser Ser Ser Thr Thr Leu Arg Leu Gly lie Phe Leu [4626]220225230aca acc egg aaa tta tgc ggc aga cac tgg agt ttt tac gta acc ctg 768Thr Thr Arg Lys Leu Cys Gly Arg His Trp Ser Phe Tyr Val Thr Leu235240245cca tga tgc agg aga tga tac gta gcc agg acc ggg tgc tea gta act 816ProCys Arg Arg Tyr Val Ala Arg Thr Gly Cys Ser Val Thr250255260tgg aga gca ttc ctg gtg get aca atg tgc ttt gca cta tgt aca cag 864Trp Arg Ala Phe Leu Val Ala Thr Met Cys Phe Ala Leu Cys Thr Gln265270275ata tta tgg acc caa tgc tta acg cag tcc agg age agt ttg gcg gca 912lie Leu Trp Thr Gln Cys Leu Thr Gln Ser Arg Ser Ser Leu Ala Ala280285290ate cct ttg cca ctg cca cta ctg ata atg cca cca cca cca cca gcc 960lie Pro Leu Pro Leu Pro Leu Leu lie Met Pro Pro Pro Pro Pro Ala295300305aac ctt caa gga tgg aga att gtg acc ctc tcc cca acc cct gga ctt 1008Asn Leu Gln Gly Trp Arg lie Val Thr Leu Ser Pro Thr Pro Gly Leu310315320325cca cac atg gag get cag gta gca ggc aag gaa ggc agg atg ggg ate 1056Pro His Met Glu Ala Gln Val Ala Gly Lys Glu Gly Arg Met Gly lie330335340agg atg cac ctg aca tta gaa ata ggt ttc caa act ttc tgg gta tta 1104Arg Met His Leu Thr Leu Glu lie Gly Phe Gln Thr Phe Trp Val Leu345350355taa ggc tct atg act ate tcc age aat tac acg aga acc ccc agt ccc 1152Gly Ser Met Thr lie Ser Ser Asn Tyr Thr Arg Thr Pro Ser Pro360365370tag gaa ctt ate tac agg gga ctg cat ctg ccc tea gcc aaa gcc agg 1200Glu Leu lie Tyr Arg Gly Leu His Leu Pro Ser Ala Lys Ala Arg375380385aac cac cac cat cag taa aca gag ttc ccc cat cgt cac cct cat ctc 1248Asn His His His Gln Thr Glu Phe Pro His Arg His Pro His Leu390395400agg age ctg ggt cag gcc age ctc tcc ccg agg agt cag tag caa tea 1296Arg Ser Leu Gly Gln Ala Ser Leu Ser Pro Arg Ser Gln Gln Ser[4662]405410415agg gaa ggt cct cct gcc cag ctt tcc tga gat acc cca cag aga aca 1344Arg Glu Gly Pro Pro Ala Gln Leu Ser Asp Thr Pro Gln Arg Thr [4665]420425430gta ctg gac aag gtg gag acc aag atg gtg cag gga aaa get cta ctg 1392Val Leu Asp Lys Val Glu Thr Lys Met Val Gln Gly Lys Ala Leu Leu435440445gac ata gca caa act tgc ctg ate ttg tct egg ggc tgg gag att ctg 1440Asp lie Ala Gln Thr Cys Leu lie Leu Ser Arg Gly Trp Glu lie Leu450455460cca aca ggg ttc cat ttg ctc cct tat ctt ttt ccc cca egg cag cca 1488Pro Thr Gly Phe His Leu Leu Pro Tyr Leu Phe Pro Pro Arg Gln Pro465470475480ttc ctg gaa tcc ctg age ctc cct ggc tgc cat ccc egg ctt ate caa 1536Phe Leu Glu Ser Leu Ser Leu Pro Gly Cys His Pro Arg Leu lie Gln485490495gat ctc tga ggc cag atg gca tga ate cag ctc cac agt tac agg atg 1584AspLeuGly Gln Met Ala lie Gln Leu His Ser Tyr Arg Met500505510aga tac aac cac age tgc cac tgc tga tgc acc ttc agg cag cca tgg 1632Arg Tyr Asn His Ser Cys His Cys Cys Thr Phe Arg Gln Pro Trp515520525caa acc ccc gtg ccc tgc aag ccc tgc ggc aga ttg age agg gtc tac 1680Gln Thr Pro Val Pro Cys Lys Pro Cys Gly Arg Leu Ser Arg Val Tyr530535540agg tcc tag cta ctg aag cac ctc gcc tcc tac tct ggt tea tgc ctt 1728Arg Ser Leu Leu Lys His Leu Ala Ser Tyr Ser Gly Ser Cys Leu545550555gcc tag cag gga egg gta gtg tgg cag gag gta tag agt cta gag aag 1776Ala Gln Gly Arg Val Val Trp Gln Glu Val Ser Leu Glu Lys560565570ate ccc tta tgt ctg agg ate ctc tcc caa ate cac ctc ctg agg tgt 1824lie Pro Leu Cys Leu Arg lie Leu Ser Gln lie His Leu Leu Arg Cys575580585tcc cag cac tgg act ctg cag age tgg get tcc ttt ccc ctc cct ttc 1872Ser Gln His Trp Thr Leu Gln Ser Trp Ala Ser Phe Pro Leu Pro Phe590595600tcc ata tgc tgc aag att tag tta gta caa ate ccc age age tgc age 1920Ser lie Cys Cys Lys lie Leu Val Gln lie Pro Ser Ser Cys Ser[4701]605610615[4702]ctg agg ctc act ttc agg tgc age tgg age aac tgc ggt cca tgg get 1968Leu Arg Leu Thr Phe Arg Cys Ser Trp Ser Asn Cys Gly Pro Trp Ala620625630ttc tga ate gtg aag cca ate ttc agg ccc tea ttg cta egg ggg gcg 2016Phe lie Val Lys Pro lie Phe Arg Pro Ser Leu Leu Arg Gly Ala635640645acg tgg atg ctg ctg tgg aga age tga gac agt cgt agg age ctt att 2064Thr Trp Met Leu Leu Trp Arg Ser Asp Ser Arg Arg Ser Leu lie650655660cat tea aac cat acg ttt tcc tct gtg cct ttt tcc cat ate cta gtt 2112His Ser Asn His Thr Phe Ser Ser Val Pro Phe Ser His lie Leu Val665670675ccc tag ctc tcc cat ttt tga ata cag ctg cat tat 3-3-3- CCS aat tta 2160Pro Leu Ser His Phe lie Gln Leu His Tyr Lys Pro Asn Leu680685690cta tga agt cct ttg ctg tgg agg caatgt tgt tcc aga gtc aac gag 2208Leu Ser Pro Leu Leu Trp Arg Gln Cys Cys Ser Arg Val Asn Glu695700705gaa gac taa tgg cca aaa cat agt gga ggt get gtg tgt gag tea acc 2256Glu Asp Trp Pro Lys His Ser Gly Gly Ala Val Cys Glu Ser Thr710715720act tgt acc act ata cca ctg ggg ggc ccc agt cta age tct get tat 2304Thr Cys Thr Thr lie Pro Leu Gly Gly Pro Ser Leu Ser Ser Ala Tyr725730735gcc tat ctt gag atg caa tta cac ccaatt tcc aat gtg aa aa 2347Ala Tyr Leu Glu Met Gln Leu His Pro lie Ser Asn Val740745750SEQUENCE LISTING21<110>北京思安创新生物技术有限公司段，陵浔<120>20种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗<130>NP_001455<160>1<170>PatentIn version 3. 5<210>1<211>735<212>PRT<213>Homo sapiens[4740]<220><221>SITE<222>(313). . (314) [4743]<220><221>SITE<222>(317). . (317)<220><221>SITE<222>(323). . (323)<400>1Met Trp Arg Val Leu Phe Leu Leu Ser Gly Leu Gly Gly Leu Arg Met151015Asp Ser Asn Phe Asp Ser Leu Pro Val Gln lie Thr Val Pro Glu Lys202530lie Arg Ser lie lie Lys Glu Gly lie Glu Ser Gln Ala Ser Tyr Lys354045lie Val lie Glu Gly Lys Pro Tyr Thr Val Asn Leu Met Gln Lys Asn505560Phe Leu Pro His Asn Phe Arg Val Tyr Ser Tyr Ser Gly Thr Gly lie65707580Met Lys Pro Leu Asp Gln Asp Phe Gln Asn Phe Cys His Tyr Gln Gly859095Tyr lie Glu Gly Tyr Pro Lys Ser Val Val Met Val Ser Thr Cys Thr100105110Gly Leu Arg Gly Val Leu Gln Phe Glu Asn Val Ser Tyr Gly lie Glu115120125Pro Leu Glu Ser Ser Val Gly Phe Glu His Val lie Tyr Gln Val Lys130135140His Lys Lys Ala Asp Val Ser Leu Tyr Asn Glu Lys Asp lie Glu Ser145150155160Arg Asp Leu Ser Phe Lys Leu Gln Ser Val Glu Pro Gln Gln Asp Phe165170175Ala Lys Tyr lie Glu Met His Val lie Val Glu Lys Gln Leu Tyr Asn180185190His Met Gly Ser Asp Thr Thr Val Val Ala Gln Lys Val Phe Gln Leu195200205lie Gly Leu Thr Asn Ala lie Phe Val Ser Phe Asn lie Thr lie lie210215220Leu Ser Ser Leu Glu Leu Trp lie Asp Glu Asn Lys lie Ala Thr Thr[4779]225230235240Gly Glu Ala Asn Glu Leu Leu His Thr Phe Leu Arg Trp Lys Thr Ser245250255Tyr Leu Val Leu Arg Pro His Asp Val Ala Phe Leu Leu Val Tyr Arg260265270Glu Lys Ser Asn Tyr Val Gly Ala Thr Phe Gln Gly Lys Met Cys Asp275280285 [4786]Ala Asn Tyr Ala Gly Gly Val Val Leu His Pro Arg Thr lie Ser Leu290295300Glu Ser Leu Ala Val lie Leu Ala Gln Leu Leu Ser Leu Ser Met Gly305310315320lie Thr Tyr Asp Asp lie Asn Lys Cys Gln Cys Ser Gly Ala Val Cys325330335lie Met Asn Pro Glu Ala lie His Phe Ser Gly Val Lys lie Phe Ser340345350Asn Cys Ser Phe Glu Asp Phe Ala His Phe lie Ser Lys Gln Lys Ser355360365Gln Cys Leu His Asn Gln Pro Arg Leu Asp Pro Phe Phe Lys Gln Gln370375380Ala Val Cys Gly Asn Ala Lys Leu Glu Ala Gly Glu Glu Cys Asp Cys385390395400Gly Thr Glu Gln Asp Cys Ala Leu lie Gly Glu Thr Cys Cys Asp lie405410415Ala Thr Cys Arg Phe Lys Ala Gly Ser Asn Cys Ala Glu Gly Pro Cys420425430Cys Glu Asn Cys Leu Phe Met Ser Lys Glu Arg Met Cys Arg Pro Ser435440445Phe Glu Glu Cys Asp Leu Pro Glu Tyr Cys Asn Gly Ser Ser Ala Ser450455460Cys Pro Glu Asn His Tyr Val Gln Thr Gly His Pro Cys Gly Leu Asn465470475480Gln Trp lie Cys lie Asp Gly Val Cys Met Ser Gly Asp Lys Gln Cys485490495Thr Asp Thr Phe Gly Lys Glu Val Glu Phe Gly Pro Ser Glu Cys Tyr500505510Ser His Leu Asn Ser Lys Thr Asp Val Ser Gly Asn Cys Gly lie Ser515520525Asp Ser Gly Tyr Thr Gln Cys Glu Ala Asp Asn Leu Gln Cys Gly Lys530535540[4818]Leu lie Cys Lys Tyr Val Gly Lys Phe Leu Leu Gln lie Pro Arg Ala545550555560Thr lie lie Tyr Ala Asn lie Ser Gly His Leu Cys lie Ala Val Glu [4821]565570575Phe Ala Ser Asp His Ala Asp Ser Gln Lys Met Trp lie Lys Asp Gly580585590Thr Ser Cys Gly Ser Asn Lys Val Cys Arg Asn Gln Arg Cys Val Ser595600605Ser Ser Tyr Leu Gly Tyr Asp Cys Thr Thr Asp Lys Cys Asn Asp Arg610615620Gly Val Cys Asn Asn Lys Lys His Cys His Cys Ser Ala Ser Tyr Leu625630635640Pro Pro Asp Cys Ser Val Gln Ser Asp Leu Trp Pro Gly Gly Ser lie645650655Asp Ser Gly Asn Phe Pro Pro Val Ala lie Pro Ala Arg Leu Pro Glu660665670Arg Arg Tyr lie Glu Asn lie Tyr His Ser Lys Pro Met Arg Trp Pro675680685Phe Phe Leu Phe lie Pro Phe Phe lie lie Phe Cys Val Leu lie Ala690695700lie Met Val Lys Val Asn Phe Gln Arg Lys Lys Trp Arg Thr Glu Asp705710715720Tyr Ser Ser Asp Glu Gln Pro Glu Ser Glu Ser Glu Pro Lys Gly725730735SEQUENCE LISTING22<110>北京思安创新生物技术有限公司段，陵浔<120>20种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗<130>NP_076957<160>1<170>PatentIn version 3. 5<210>1<211>300<212>PRT<213>Homo sapiens<220><221>D0MAIN<222>(1). . (300)<400>1[4857]Met Cys Ala Arg Gly Gln Val Gly Arg Gly Thr Gln Leu Arg Thr Gly151015Arg Pro Cys Ser Gln Val Pro Gly Ser Arg Trp Arg Pro Glu Arg Leu202530Leu Arg Arg Gln Arg Ala Gly Gly Arg Pro Ser Arg Pro His Pro Ala354045Arg Ala Arg Pro Gly Leu Ser Leu Pro Ala Thr Leu Leu Gly Ser Arg505560 [4865]Ala Ala Ala Ala Val Pro Leu Pro Leu Pro Pro Ala Leu Ala Pro Gly65707580Asp Pro Ala Met Pro Val Arg Thr Glu Cys Pro Pro Pro Ala Gly Ala859095Ser Ala Ala Ser Ala Ala Ser Leu lie Pro Pro Pro Pro lie Asn Thr100105110Gln Gln Pro Gly Val Ala Thr Ser Leu Leu Tyr Ser Gly Ser Lys Phe115120125Arg Gly His Gln Lys Ser Lys Gly Asn Ser Tyr Asp Val Glu Val Val130135140Leu Gln His Val Asp Thr Gly Asn Ser Tyr Leu Cys Gly Tyr Leu Lys145150155160lie Lys Gly Leu Thr Glu Glu Tyr Pro Thr Leu Thr Thr Phe Phe Glu165170175Gly Glu lie lie Ser Lys Lys His Pro Phe Leu Thr Arg Lys Trp Asp180185190Ala Asp Glu Asp Val Asp Arg Lys His Trp Gly Lys Phe Leu Ala Phe195200205Tyr Gln Tyr Ala Lys Ser Phe Asn Ser Asp Asp Phe Asp Tyr Glu Glu210215220Leu Lys Asn Gly Asp Tyr Val Phe Met Arg Trp Lys Glu Gln Phe Leu225230235240Val Pro Asp His Thr lie Lys Asp lie Ser Gly Ala Ser Phe Ala Gly245250255Phe Tyr Tyr lie Cys Phe Gln Lys Ser Ala Ala Ser lie Glu Gly Tyr260265270Tyr Tyr His Arg Ser Ser Glu Trp Tyr Gln Ser Leu Asn Leu Thr His275280285Val Pro Glu His Ser Ala Pro lie Tyr Glu Phe Arg290295300SEQUENCE LIST ING23[4896]<110>北京思安创新生物技术有限公司段，陵浔<120>20种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗<130>NP_659473<160>1<170>PatentIn version 3. 5<210>1<211>955<212>PRT<213>Homo sapiens<220><221>D0MAIN<222>(1). . (955)<400>1Met Glu Val Gly Ser Glu Glu Glu Lys Trp Glu Lys Leu Asp Ala Glu151015Phe Asp His Phe Val Val Asp Met Lys Pro Phe Val Leu Lys Leu Pro202530His Arg Thr Glu Arg Gln Arg Cys Ala Leu Trp lie Arg Lys Leu Cys354045Glu Pro Ser Gly Thr Gly Ala Gly lie Met Gly Arg Lys Asn Arg Asn505560Leu Tyr Ala Lys Leu Leu Leu His Met Leu Lys Arg Gly Ala Leu Glu65707580Gly Pro Phe Thr His Arg Pro Glu Pro Gly Thr Leu Lys lie Leu Pro859095Ser Tyr Met Ser lie Tyr Phe Asp Glu Pro Asn Pro Ala Arg Ala Lys100105110Gly Ser Ser Pro Glu Gly Leu Pro Ala Trp Val Leu Gly Glu Leu Glu115120125Thr Ser Glu His Lys Leu Asn Glu Ser Trp Lys Leu Ser Ser Gly Glu130135140Asp Asn Thr Leu Val Gln Ser Pro Thr Asp Val Tyr Ser Arg Glu Gln145150155160Tyr Thr Gly Lys Leu Arg Val Arg Ser His Ser Leu Ser Pro Thr His165170175Arg Glu Asp Gly Gln Asn lie Thr Pro Lys lie Cys Glu Val Tyr Ser180185190[4934]Lys Lys Ser Pro Val Ser Leu Asp Asp Ser Asp lie Glu Ala Arg Leu[4935]195200205[4936]Asn Ser Trp Asn Leu Gly lie Glu Asn Pro Arg Tyr Leu Arg Gln Lys210215220Pro lie Pro Val Ser Leu Met Thr Pro Lys Phe Ser Leu Arg Lys Ser225230235240Ser Ser Phe His Asp Asp His Phe Leu Ser Arg lie Arg Glu Lys Glu245250255Leu Asp Met Lys Thr Lys Met Met Glu Ala Lys Phe His Glu Glu Lys260265270Leu Lys Leu Gln Gln Lys His Asp Ala Asp Val Gln Lys lie Leu Glu275280285Arg Lys Asn Asn Glu lie Glu Glu Leu Lys Thr Leu Tyr Arg Ser Lys290295300Gln His Glu Thr Glu Glu Thr lie Arg Lys Leu Glu Lys Lys Val Gln305310315320Thr Leu lie Arg Asp Cys Gln Val lie Arg Glu Thr Lys Glu Asp Gln325330335lie Ala Glu Leu Lys Lys lie Cys Glu Gln Ser Thr Glu Ser Leu Asn340345350Asn Asp Trp Glu Lys Lys Leu His Asn Ala Val Ala Glu Met Glu Gln355360365Glu Lys Phe Asp Leu Gln Lys Gln His Thr Glu Asn lie Gln Glu Leu370375380Leu Glu Asp Thr Asn Val Arg Leu Asn Lys Met Glu Ser Glu Tyr Met385390395400Ala Gln Thr Gln Ser Thr Asn His Met lie Lys Glu Leu Glu Ala Arg405410415Val Gln Gln Leu Thr Gly Glu Ala Glu Asn Ser Asn Leu Gln Arg Gln420425430Lys Leu lie Gln Glu Lys Ala Glu Leu Glu Arg Cys Tyr Gln lie Thr435440445Cys Ser Glu Leu Gln Glu Val Lys Ala Arg Arg Asn Thr Leu His Lys450455460Glu Lys Asp His Leu Val Asn Asp Tyr Glu Gln Asn Met Lys Leu Leu465470475480Gln Thr Lys Tyr Asp Ala Asp lie Asn Leu Leu Lys Gln Glu His Ala485490495Leu Ser Ala Ser Lys Ala Ser Ser Met lie Glu Glu Leu Glu Gln Asn500505510[4974]Val Cys Gln Leu Lys Gln Gln Leu Gln Glu Ser Glu Leu Gln Arg Lys515520525Gln Gln Leu Arg Asp Gln Glu Asn Lys Phe Gln Met Glu Lys Ser His530535540Leu Lys His lie Tyr Glu Lys Lys Ala His Asp Leu Gln Ser Glu Leu545550555560 [4980]Asp Lys Gly Lys Glu Asp Thr Gln Lys Lys lie His Lys Phe Glu Glu565570575Ala Leu Lys Glu Lys Glu Glu Gln Leu Thr Arg Val Thr Glu Val Gln580585590Arg Leu Gln Ala Gln Gln Ala Asp Ala Ala Leu Glu Glu Phe Lys Arg595600605Gln Val Glu Leu Asn Ser Glu Lys Val Tyr Ala Glu Met Lys Glu Gln610615620Met Glu Lys Val Glu Ala Asp Leu Thr Arg Ser Lys Ser Leu Arg Glu625630635640Lys Gln Ser Lys Glu Phe Leu Trp Gln Leu Glu Asp lie Arg Gln Arg645650655Tyr Glu Gln Gln lie Val Glu Leu Lys Leu Glu His Glu Gln Glu Lys660665670Thr His Leu Leu Gln Gln His Asn Ala Glu Lys Asp Ser Leu Val Arg675680685Asp His Glu Arg Glu lie Glu Asn Leu Glu Lys Gln Leu Arg Ala Ala690695700Asn Met Glu His Glu Asn Gln lie Gln Glu Phe Lys Lys Arg Asp Ala705710715720Gln Val lie Ala Asp Met Glu Ala Gln Val His Lys Leu Arg Glu Glu725730735Leu lie Asn Val Asn Ser Gln Arg Lys Gln Gln Leu Val Glu Leu Gly740745750Leu Leu Arg Glu Glu Glu Lys Gln Arg Ala Thr Arg Glu His Glu lie755760765Val Val Asn Lys Leu Lys Ala Glu Ser Glu Lys Met Lys lie Glu Leu770775780Lys Lys Thr His Ala Ala Glu Thr Glu Met Thr Leu Glu Lys Ala Asn785790795800Ser Lys Leu Lys Gln lie Glu Lys Glu Tyr Thr Gln Lys Leu Ala Lys805810815Ser Ser Gln lie lie Ala Glu Leu Gln Thr Thr lie Ser Ser Leu Lys[5013]820825830Glu Glu Asn Ser Gln Gln Gln Leu Ala Ala Glu Arg Arg Leu Gln Asp835840845 Val Arg Gln Lys Phe Glu Asp Glu Lys Lys Gln Leu lie Arg Asp Asn850855860Asp Gln Ala lie Lys Val Leu Gln Asp Glu Leu Glu Asn Arg Ser Asn865870875880Gln Val Arg Cys Ala Glu Lys Lys Leu Gln His Lys Glu Leu Glu Ser885890895Gln Glu Gln lie Thr Tyr lie Arg Gln Glu Tyr Glu Thr Lys Leu Lys900905910Gly Leu Met Pro Ala Ser Leu Arg Gln Glu Leu Glu Asp Thr lie Ser915920925Ser Leu Lys Ser Gln Val Asn Phe Leu Gln Lys Arg Ala Ser lie Leu930935940Gln Glu Glu Leu Thr Thr Tyr Gln Gly Arg Arg945950955SEQUENCE LISTING24<110>北京思安创新生物技术有限公司段，陵浔<120>20种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗<130>NP_001295<160>1<170>PatentIn version 3. 5<210>1<211>1380<212>PRT<213>Homo sapiens<220><221>SITE<222>(172). . (172)<220><221>SITE<222>(811). . (811)<220><221>SITE<222>(955). . (955)<400>1Met Ala Ser Gly Arg Asp Glu Arg Pro Pro Trp Arg Leu Gly Arg Leu[5052]151015Leu Leu Leu Met Cys Leu Leu Leu Leu Gly Ser Ser Ala Arg Ala Ala202530 [5055]His lie Lys Lys Ala Glu Ala Thr Thr Thr Thr Thr Ser Ala Gly Ala354045Glu Ala Ala Glu Gly Gln Phe Asp Arg Tyr Tyr His Glu Glu Glu Leu505560Glu Ser Ala Leu Arg Glu Ala Ala Ala Ala Gly Leu Pro Gly Leu Ala65707580Arg Leu Phe Ser lie Gly Arg Ser Val Glu Gly Arg Pro Leu Trp Val859095Leu Arg Leu Thr Ala Gly Leu Gly Ser Leu lie Pro Glu Gly Asp Ala100105110Gly Pro Asp Ala Ala Gly Pro Asp Ala Ala Gly Pro Leu Leu Pro Gly115120125Arg Pro Gln Val Lys Leu Val Gly Asn Met His Gly Asp Glu Thr Val130135140Ser Arg Gln Val Leu lie Tyr Leu Ala Arg Glu Leu Ala Ala Gly Tyr145150155160Arg Arg Gly Asp Pro Arg Leu Val Arg Leu Leu Asn Thr Thr Asp Val165170175Tyr Leu Leu Pro Ser Leu Asn Pro Asp Gly Phe Glu Arg Ala Arg Glu180185190Gly Asp Cys Gly Phe Gly Asp Gly Gly Pro Ser Gly Ala Ser Gly Arg195200205Asp Asn Ser Arg Gly Arg Asp Leu Asn Arg Ser Phe Pro Asp Gln Phe210215220Ser Thr Gly Glu Pro Pro Ala Leu Asp Glu Val Pro Glu Val Arg Ala225230235240Leu lie Glu Trp lie Arg Arg Asn Lys Phe Val Leu Ser Gly Asn Leu245250255His Gly Gly Ser Val Val Ala Ser Tyr Pro Phe Asp Asp Ser Pro Glu260265270His Lys Ala Thr Gly lie Tyr Ser Lys Thr Ser Asp Asp Glu Val Phe275280285Lys Tyr Leu Ala Lys Ala Tyr Ala Ser Asn His Pro lie Met Lys Thr290295300Gly Glu Pro His Cys Pro Gly Asp Glu Asp Glu Thr Phe Lys Asp Gly305310315320[5091]lie Thr Asn Gly Ala His Trp Tyr Asp Val Glu Gly Gly Met Gln Asp325330335Tyr Asn Tyr Val Trp Ala Asn Cys Phe Glu lie Thr Leu Glu Leu Ser [5094]340345350Cys Cys Lys Tyr Pro Pro Ala Ser Gln Leu Arg Gln Glu Trp Glu Asn355360365Asn Arg Glu Ser Leu lie Thr Leu lie Glu Lys Val His lie Gly Val370375380Lys Gly Phe Val Lys Asp Ser lie Thr Gly Ser Gly Leu Glu Asn Ala385390395400Thr lie Ser Val Ala Gly lie Asn His Asn lie Thr Thr Gly Arg Phe405410415Gly Asp Phe Tyr Arg Leu Leu Val Pro Gly Thr Tyr Asn Leu Thr Val420425430Val Leu Thr Gly Tyr Met Pro Leu Thr Val Thr Asn Val Val Val Lys435440445Glu Gly Pro Ala Thr Glu Val Asp Phe Ser Leu Arg Pro Thr Val Thr450455460Ser Val lie Pro Asp Thr Thr Glu Ala Val Ser Thr Ala Ser Thr Val465470475480Ala lie Pro Asn lie Leu Ser Gly Thr Ser Ser Ser Tyr Gln Pro lie485490495Gln Pro Lys Asp Phe His His His His Phe Pro Asp Met Glu lie Phe500505510Leu Arg Arg Phe Ala Asn Glu Tyr Pro Asn lie Thr Arg Leu Tyr Ser515520525Leu Gly Lys Ser Val Glu Ser Arg Glu Leu Tyr Val Met Glu lie Ser530535540Asp Asn Pro Gly Val His Glu Pro Gly Glu Pro Glu Phe Lys Tyr lie545550555560Gly Asn Met His Gly Asn Glu Val Val Gly Arg Glu Leu Leu Leu Asn565570575Le lie Glu Tyr Leu Cys Lys Asn Phe Gly Thr Asp Pro Glu Val Thr580585590Asp Leu Val His Asn Thr Arg lie His Leu Met Pro Ser Met Asn Pro595600605Asp Gly Tyr Glu Lys Ser Gln Glu Gly Asp Ser lie Ser Val lie Gly610615620Arg Asn Asn Ser Asn Asn Phe Asp Leu Asn Arg Asn Phe Pro Asp Gln[5130]625630635640[5131]Phe Val Gln lie Thr Asp Pro Thr Gln Pro Glu Thr lie Ala Val Met645650655Ser Trp Met Lys Ser Tyr Pro Phe Val Leu Ser Ala Asn Leu His Gly660665670Gly Ser Leu Val Val Asn Tyr Pro Phe Asp Asp Asp Glu Gln Gly Leu675680685Ala Thr Tyr Ser Lys Ser Pro Asp Asp Ala Val Phe Gln Gln lie Ala690695700Leu Ser Tyr Ser Lys Glu Asn Ser Gln Met Phe Gln Gly Arg Pro Cys705710715720Lys Asn Met Tyr Pro Asn Glu Tyr Phe Pro His Gly lie Thr Asn Gly725730735Ala Ser Trp Tyr Asn Val Pro Gly Gly Met Gln Asp Trp Asn Tyr Leu740745750Gln Thr Asn Cys Phe Glu Val Thr lie Glu Leu Gly Cys Val Lys Tyr755760765Pro Leu Glu Lys Glu Leu Pro Asn Phe Trp Glu Gln Asn Arg Arg Ser770775780Leu lie Gln Phe Met Lys Gln Val His Gln Gly Val Arg Gly Phe Val785790795800Leu Asp Ala Thr Asp Gly Arg Gly lie Leu Asn Ala Thr lie Ser Val805810815Ala Glu lie Asn His Pro Val Thr Thr Tyr Lys Thr Gly Asp Tyr Trp820825830Arg Leu Leu Val Pro Gly Thr Tyr LysIle Thr Ala Ser Ala Arg Gly835840845Tyr Asn Pro Val Thr Lys Asn Val Thr Val Lys Ser Glu Gly Ala lie850855860Gln Val Asn Phe Thr Leu Val Arg Ser Ser Thr Asp Ser Asn Asn Glu865870875880Ser Lys Lys Gly Lys Gly Ala Ser Ser Ser Thr Asn Asp Ala Ser Asp885890895Pro Thr Thr Lys Glu Phe Glu Thr Leu lie Lys Asp Leu Ser Ala Glu900905910Asn Gly Leu Glu Ser Leu Met Leu Arg Ser Ser Ser Asn Leu Ala Leu915920925Ala Leu Tyr Arg Tyr His Ser Tyr Lys Asp Leu Ser Glu Phe Leu Arg930935940[5169]Gly Leu Val Met Asn Tyr Pro His lie Thr Asn Leu Thr Asn Leu Gly945950955960Gln Ser Thr Glu Tyr Arg His lie Trp Ser Leu Glu lie Ser Asn Lys [5172]965970975Pro Asn Val Ser Glu Pro Glu Glu Pro Lys lie Arg Phe Val Ala Gly980985990lie His Gly Asn Ala Pro Val Gly Thr Glu Leu Leu Leu Ala Leu Ala99510001005Glu Phe Leu Cys Leu AsnTyr Lys Lys Asn Pro Ala Val Thr Gln101010151020Leu Val Asp Arg Thr Arglie Val lie Val Pro Ser Leu Asn Pro102510301035Asp Gly Arg Glu Arg AlaGln Glu Lys Asp Cys Thr Ser Lys lie104010451050Gly Gln Thr Asn Ala ArgGly Lys Asp Leu Asp Thr Asp Phe Thr105510601065Asn Asn Ala Ser Gln ProGlu Thr Lys Ala lie lie Glu Asn Leu107010751080lie Gln Lys Gln Asp PheSer Leu Ser Val Ala Leu Asp Gly Gly108510901095Ser Met Leu Val Thr TyrPro Tyr Asp Lys Pro Val Gln Thr Val110011051110Glu Asn Lys Glu Thr LeuLys His Leu Ala Ser Leu Tyr Ala Asn111511201125Asn His Pro Ser Met His Met Gly Gln Pro Ser Cys Pro Asn Lys113011351140Ser Asp Glu Asn lie ProGly Gly Val Met Arg Gly Ala Glu Trp114511501155His Ser His Leu Gly SerMet Lys Asp Tyr Ser Val Thr Tyr Gly116011651170His Cys Pro Glu lie ThrVal Tyr Thr Ser Cys Cys Tyr Phe Pro117511801185Ser Ala Ala Arg Leu ProSer Leu Trp Ala Asp Asn Lys Arg Ser119011951200Leu Leu Ser Met Leu ValGlu Val His Lys Gly Val His Gly Phe120512101215Val Lys Asp Lys Thr GlyLys Pro lie Ser Lys Ala Val lie Val122012251230Leu Asn Glu Gly lie LysVal Gln Thr Lys Glu Gly Gly Tyr Phe[5208]123512401245His Val Leu Leu Ala Pro Gly Val His Asn lie lie Ala lie Ala125012551260 Asp Gly Tyr Gln Gln Gln His Ser Gln Val Phe Val His His Asp126512701275Ala Ala Ser Ser Val Val lie Val Phe Asp Thr Asp Asn Arg lie128012851290Phe Gly Leu Pro Arg Glu Leu Val Val Thr Val Ser Gly Ala Thr129513001305Met Ser Ala Leu lie Leu Thr Ala Cys lie lie Trp Cys lie Cys131013151320Ser lie Lys Ser Asn Arg His Lys Asp Gly Phe His Arg Leu Arg132513301335Gln His His Asp Glu Tyr Glu Asp Glu lie Arg Met Met Ser Thr134013451350Gly Ser Lys Lys Ser Leu Leu Ser His Glu Phe Gln Asp Glu Thr135513601365Asp Thr Glu Glu Glu Thr Leu Tyr Ser Ser Lys His137013751380SEQUENCE LISTING25<110>北京思安创新生物技术有限公司段，陵浔<120>20种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗<130>NP_000746<160>1<170>PatentIn version 3. 5<210>1<211>626<212>PRT<213>Homo sapiens<220><221>D0MAIN<222>(1). . (626)<400>1Met Leu Ala Phe Ala Ala Arg Thr Val Val Lys Pro Leu Gly Phe Leu151015Lys Pro Phe Ser Leu Met Lys Ala Ser Ser Arg Phe Lys Ala His Gln202530Asp Ala Leu Pro Arg Leu Pro Val Pro Pro Leu Gln Gln Ser Leu Asp[5247]354045His Tyr Leu Lys Ala Leu Gln Pro lie Val Ser Glu Glu Glu Trp Ala505560His Thr Lys Gln Leu Val Asp Glu Phe Gln Ala Ser Gly Gly Val Gly65707580Glu Arg Leu Gln Lys Gly Leu Glu Arg Arg Ala Arg Lys Thr Glu Asn859095Trp Leu Ser Glu Trp Trp Leu Lys Thr Ala Tyr Leu Gln Tyr Arg Gln100105110Pro Val Val lie Tyr Ser Ser Pro Gly Val Met Leu Pro Lys Gln Asp115120125Phe Val Asp Leu Gln Gly Gln Leu Arg Phe Ala Ala Lys Leu lie Glu130135140Gly Val Leu Asp Phe Lys Val Met lie Asp Asn Glu Thr Leu Pro Val145150155160Glu Tyr Leu Gly Gly Lys Pro Leu Cys Met Asn Gln Tyr Tyr Gln lie165170175Leu Ser Ser Cys Arg Val Pro Gly Pro Lys Gln Asp Thr Val Ser Asn180185190Phe Ser Lys Thr Lys Lys Pro Pro Thr His lie Thr Val Val His Asn195200205Tyr Gln Phe Phe Glu Leu Asp Val Tyr His Ser Asp Gly Thr Pro Leu210215220Thr Ala Asp Gln lie Phe Val Gln Leu Glu Lys lie Trp Asn Ser Ser225230235240Leu Gln Thr Asn Lys Glu Pro Val Gly lie Leu Thr Ser Asn His Arg245250255Asn Ser Trp Ala Lys Ala Tyr Asn Thr Leu lie Lys Asp Lys Val Asn260265270Arg Asp Ser Val Arg Ser lie Gln Lys Ser lie Phe Thr Val Cys Leu275280285Asp Ala Thr Met Pro Arg Val Ser Glu Asp Val Tyr Arg Ser His Val290295300Ala Gly Gln Met Leu His Gly Gly Gly Ser Arg Leu Asn Ser Gly Asn305310315320Arg Trp Phe Asp Lys Thr Leu Gln Phe lie Val Ala Glu Asp Gly Ser325330335Cys Gly Leu Val Tyr Glu His Ala Ala Ala Glu Gly Pro Pro lie Val[5285]340345350[5286]Thr Leu Leu Asp Tyr Val lie Glu Tyr Thr Lys Lys Pro Glu Leu Val355360365Arg Ser Pro Met Val Pro Leu Pro Met Pro Lys Lys Leu Arg Phe Asn370375380lie Thr Pro Glu lie Lys Ser Asp lie Glu Lys Ala Lys Gln Asn Leu385390395400Ser lie Met lie Gln Asp Leu Asp lie Thr Val Met Val Phe His His 405410415Phe Gly Lys Asp Phe Pro Lys Ser Glu Lys Leu Ser Pro Asp Ala Phe420425430lie Gln Met Ala Leu Gln Leu Ala Tyr Tyr Arg lie Tyr Gly Gln Ala435440445Cys Ala Thr Tyr Glu Ser Ala Ser Leu Arg Met Phe His Leu Gly Arg450455460Thr Asp Thr lie Arg Ser Ala Ser Met Asp Ser Leu Thr Phe Val Lys465470475480Ala Met Asp Asp Ser Ser Val Thr Glu His Gln Lys Val Glu Leu Leu485490495Arg Lys Ala Val Gln Ala His Arg Gly Tyr Thr Asp Arg Ala lie Arg500505510Gly Glu Ala Phe Asp Arg His Leu Leu Gly Leu Lys Leu Gln Ala lie515520525Glu Asp Leu Val Ser Met Pro Asp lie Phe Met Asp Thr Ser Tyr Ala530535540lie Ala Met His Phe His Leu Ser Thr Ser Gln Val Pro Ala Lys Thr545550555560Asp Cys Val Met Phe Phe Gly Pro Val Val Pro Asp Gly Tyr Gly Val565570575Cys Tyr Asn Pro Met Glu Ala His lie Asn Phe Ser Leu Ser Ala Tyr580585590Asn Ser Cys Ala Glu Thr Asn Ala Ala Arg Leu Ala His Tyr Leu Glu595600605Lys Ala Leu Leu Asp Met Arg Ala Leu Leu Gln Ser His Pro Arg Ala610615620Lys Leu625SEQUENCE LISTING26<110>北京思安创新生物技术有限公司段，陵浔[5325]<120>20种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗<130>NP_001887<160>1<170>PatentIn version 3. 5<210>1<211>350<212>PRT<213>Homo sapiens [5333]<220><221>SITE<222> (46) · · (47)<220><221>SITE<222> (49) · · (49)<220><221>SITE<222>(52). . (52)<220><221>SITE<222> (54) · · (54)<220><221>SITE<222>(67). . (67)<220><221>SITE<222>(114). . (114)<220><221>SITE<222>(119). . (119)<220><221>SITE<222>(121). . (121)<220><221>SITE<222>(155). . (164)<220><221>SITE<222>(159). . (159)<220>[5364]<221>SITE<222>(162). . (162)<220><221>SITE <222>(164). . (164)<220><221>SITE<222>(175). . (177)<220><221>SITE<222>(176). . (185)<220><221>SITE<222>(189). . (202)<220><221>SITE<222>(191). . (192)<220><221>SITE<222>(194). . (194)<220><221>SITE<222>(196). . (197)<400>1Met Pro Gly Pro Ala Ala Gly Ser Arg Ala Arg Val Tyr Ala Glu Val151015Asn Ser Leu Arg Ser Arg Glu Tyr Trp Asp Tyr Glu Ala His Val Pro202530Ser Trp Gly Asn Gln Asp Asp Tyr Gln Leu Val Arg Lys Leu Gly Arg354045Gly Lys Tyr Ser Glu Val Phe Glu Ala lie Asn lie Thr Asn Asn Glu505560Arg Val Val Val Lys lie Leu Lys Pro Val Lys Lys Lys Lys lie Lys65707580Arg Glu Val Lys lie Leu Glu Asn Leu Arg Gly Gly Thr Asn lie lie859095Lys Leu lie Asp Thr Val Lys Asp Pro Val Ser Lys Thr Pro Ala Leu100105110Val Phe Glu Tyr lie Asn Asn Thr Asp Phe Lys Gln Leu Tyr Gln lie[5403]115120125[5404]Leu Thr Asp Phe Asp lie Arg Phe Tyr Met Tyr Glu Leu Leu Lys Ala130135140Leu Asp Tyr Cys His Ser Lys Gly lie Met His Arg Asp Val Lys Pro145150155160His Asn Val Met lie Asp His Gln Gln Lys Lys Leu Arg Leu lie Asp165170175Trp Gly Leu Ala Glu Phe Tyr His Pro Ala Gln Glu Tyr Asn Val Arg180185190Val Ala Ser Arg Tyr Phe Lys Gly Pro Glu Leu Leu Val Asp Tyr Gln195200205Met Tyr Asp Tyr Ser Leu Asp Met Trp Ser Leu Gly Cys Met Leu Ala210215220Ser Met lie Phe Arg Arg Glu Pro Phe Phe His Gly Gln Asp Asn Tyr225230235240Asp Gln Leu Val Arg lie Ala Lys Val Leu Gly Thr Glu Glu Leu Tyr245250255Gly Tyr Leu Lys Lys Tyr His lie Asp Leu Asp Pro His Phe Asn Asp260265270lie Leu Gly Gln His Ser Arg Lys Arg Trp Glu Asn Phe lie His Ser275280285Glu Asn Arg His Leu Val Ser Pro Glu Ala Leu Asp Leu Leu Asp Lys290295300Leu Leu Arg Tyr Asp His Gln Gln Arg Leu Thr Ala Lys Glu Ala Met305310315320Glu His Pro Tyr Phe Tyr Pro Val Val Lys Glu Gln Ser Gln Pro Cys325330335Ala Asp Asn Ala Val Leu Ser Ser Gly Leu Thr Ala Ala Arg340345350SEQUENCE LISTING27<110>北京思安创新生物技术有限公司段，陵浔<120>20种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗<130>NP_003453<160>1<170>PatentIn version 3. 5<210>1<211>280<212>PRT[5442]<213>Homo sapiens<220><221>D0MAIN<222>(1). . (280)<400>1Met Val Thr Ala Asn Lys Ala His Thr Gly Gln Gly Ser Cys Trp Val151015Ala Thr Leu Ala Ser Ala Met lie Pro Pro Ala Asp Ser Leu Leu Lys202530Tyr Asp Thr Pro Val Leu Val Ser Arg Asn Thr Glu Lys Arg Ser Pro354045Lys Ala Arg Leu Leu Lys Val Ser Pro Gln Gln Pro Gly Pro Ser Gly505560Ser Ala Pro Gln Pro Pro Lys Thr Lys Leu Pro Ser Thr Pro Cys Val65707580Pro Asp Pro Thr Lys Gln Ala Glu Glu lie Leu Asn Ala lie Leu Pro859095Pro Arg Glu Trp Val Glu Asp Thr Gln Leu Trp lie Gln Gln Val Ser100105110Ser Thr Pro Ser Thr Arg Met Asp Val Val His Leu Gln Glu Gln Leu115120125Asp Leu Lys Leu Gln Gln Arg Gln Ala Arg Glu Thr Gly lie Cys Pro130135140Val Arg Arg Glu Leu Tyr Ser Gln Cys Phe Asp Glu Leu lie Arg Glu145150155160Val Thr lie Asn Cys Ala Glu Arg Gly Leu Leu Leu Leu Arg Val Arg165170175Asp Glu lie Arg Met Thr lie Ala Ala Tyr Gln Thr Leu Tyr Glu Ser180185190Ser Val Ala Phe Gly Met Arg Lys Ala Leu Gln Ala Glu Gln Gly Lys195200205Ser Asp Met Glu Arg Lys lie Ala Glu Leu Glu Thr Glu Lys Arg Asp210215220Leu Glu Arg Gln Val Asn Glu Gln Lys Ala Lys Cys Glu Ala Thr Glu225230235240Lys Arg Glu Ser Glu Arg Arg Gln Val Glu Glu Lys Lys His Asn Glu245250255Glu lie Gln Phe Leu Lys Arg Thr Asn Gln Gln Leu Lys Ala Gln Leu[5480]260265270[5481]Glu Gly lie lie Ala Pro Lys Lys275280SEQUENCE LISTING28 <110>北京思安创新生物技术有限公司段，陵浔<120>20种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗<130>NP_149991<160>1<170>PatentIn version 3. 5<210>1<211>553<212>PRT<213>Homo sapiens<220><221>D0MAIN<222>(1). . (553)<400>1Met Ala Ala Pro Lys Thr Ala Ala Val Gly Pro Leu Val Gly Ala Val151015Val Gln Gly Thr Asn Ser Thr Arg Phe Leu Val Phe Asn Ser Lys Thr202530Ala Glu Leu Leu Ser His His Lys Val Glu Leu Thr Gln Glu Phe Pro354045Lys Glu Gly Trp Val Glu Gln Asp Pro Lys Glu lie Leu Gln Ser Val505560Tyr Glu Cys lie Ala Arg Thr Cys Glu Lys Leu Asp Glu Leu Asn lie65707580Asp lie Ser Asn lie Lys Ala Val Gly Val Ser Asn Gln Arg Glu Thr859095Thr Val lie Trp Asp Lys Leu Thr Gly Glu Pro Leu Tyr Asn Ala Val100105110Val Trp Leu Asp Leu Arg Thr Gln Thr Thr Val Glu Asp Leu Ser Lys115120125Lys lie Pro Gly Asn Ser Asn Phe Val Lys Ser Lys Thr Gly Leu Pro130135140Leu Ser Thr Tyr Phe Ser Ala Val Lys Leu Arg Trp Met Leu Asp Asn145150155160Val Arg Asn Val Gln Lys Ala Val Glu Glu Gly Arg Ala Leu Phe Gly165170175[5520]ThrlieAspSerTrpLeulieTrpSerLeuThrGlyGlyValAsnGly[5521]180185190[5522]GlyValHisCysThrAspValThrAsnAlaSerArgThrMetLeuPhe[5523]195200 205[5524]AsnlieHisSerLeuGluTrpAspLysGluLeuCysAspPhePheGlu[5525]210215220[5526]lieProMetAspLeuLeuProAsnValPheSerSerSerGlulieTyr[5527]225230235240[5528]GlyLeulieLysThrGlyAlaLeuGluGlyValProlieSerGlyCys[5529]245250255[5530]LeuGlyAspGinCysAlaAlaLeuValGlyGinMetCysPheGinGlu[5531]260265270[5532]GlyGinAlaLysAsnThrTyrGlyThrGlyCysPheLeuLeuCysAsn[5533]275280285[5534]ThrGlyArgLysCysValPheSerGluHisGlyLeuLeuThrThrVal[5535]290295300[5536]AlaTyrLysLeuGlyArgGluLysProAlaTyrTyrAlaLeuGluGly[5537]305310315320[5538]SerValAlalieAlaGlyAlaVallieArgTrpLeuArgAspAsnLeu[5539]325330335[5540]GlylielieGluThrSerGlyAsplieGluArgLeuAlaLysGluVal[5541]340345350[5542]GlyThrSerTyrGlyCysTyrPheValProAlaPheSerGlyLeuTyr[5543]355360365[5544]AlaProTyrTrpGluProSerAlaArgGlylieLeuCysGlyLeuThr[5545]370375380[5546]GinPheThrAsnLysCysHislieAlaPheAlaAlaLeuGluAlaVal[5547]385390395400[5548]CysPheGinThrArgGlulieLeuGluAlaMetAsnArgAspCysGly[5549]405410415[5550]lieProLeuArgHisLeuGinValAspGlyGlyMetThrAsnAsnLys[5551]420425430[5552]ValLeuMetGinLeuGinAlaAsplieLeuHislieProVallieLys[5553]435440445[5554]ProPheMetProGluThrThrAlaLeuGlyAlaAlaMetAlaAlaGly[5555]450455460[5556]AlaAlaGluGlyValSerValTrpSerLeuGluProGinAlaLeuSer[5557]465470475480[5558]ValLeuArgMetGluArgPheGluProGinlieGinAlaThrGluSer
165[5559]485490495[5560]Glu lie Arg TyrAlaThrTrpLysLysAlaValMetLysSerMet Gly[5561]500505510[5562]Trp Val Thr SerGinSerProGluGlyGlyAspProSerliePheSer[5563]515520525[5564]Ser Leu Pro LeuGlyPhePhelieValSerSerMetValMetLeulie[5565]530535540[5566]Gly Ala Arg TyrlieSerGlyValPro[5567]545550[5568]SEQUENCE LISTING29[5569]<110>北京思安创新生物技术有限公司[5570]段，陵浔[5571]<120>20种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗[5572]<130>NP_722545[5573]<160>1[5574]<170>PatentIn version 3.5[5575]<210>1[5576]<211>326[5577]<212>PRT[5578]<213>Homo sapiens[5579]<220>[5580]<221>D0MAIN[5581]<222>⑴(326)[5582]<400>1[5583]Met Ala Lys MetGluLeuSerLysAlaPheSerGlyGinArgThrLeu[5584]151015[5585]Leu Ser Ala lieLeuSerMetLeuSerLeuSerPheSerThrThrSer[5586]202530[5587]Leu Leu Ser AsnTyrTrpPheValGlyThrGinLysValProLysPro[5588]354045[5589]Leu Cys Glu LysGlyLeuAlaAlaLysCysPheAspMetProValSer[5590]505560[5591]Leu Asp Gly AspThrAsnThrSerThrGinGluValValGinTyrAsn[5592]65707580[5593]Trp Glu Thr GlyAspAspArgPheSerPheArgSerPheArgSerGly[5594]859095[5595]Met Trp Leu SerCysGluGluThrValGluGluProGlyGluArgCys[5596]100105110[5597]Arg Ser Phe lieGluLeuThrProProAlaLysArgGlulieLeuTrp[5598]115120125[5599]Leu Ser LeuGlyThrGinlieThrTyrlieGlyLeuGinPhelieSer[5600]130135140[5601]Phe Leu LeuLeuLeuThrAspLeuLeuLeuThrGlyAsnProAlaCys[5602]145150155160[5603]Gly Leu LysLeuSerAlaPheAlaAlaValSerSerValLeuSerGly[5604]165170175[5605]Leu Leu GlyMetValAlaHisMetMetTyrSerGinValPheGinAla[5606]180185190[5607]Thr Val AsnLeuGlyProGluAspTrpArgProHisValTrpAsnTyr[5608]195200205[5609]Gly Trp AlaPheTyrMetAlaTrpLeuSerPheThrCysCysMetAla[5610]210215220[5611]Ser Ala ValThrThrPheAsnThrTyrThrArgMetValLeuGluPhe[5612]225230235240[5613]Lys Cys LysHisSerLysSerPheLysGluAsnProAsnCysLeuPro[5614]245250255[5615]His His HisGinCysPheProArgArgLeuSerSerAlaAlaProThr[5616]260265270[5617]Val Gly ProLeuThrSerTyrHisGinTyrHisAsnGinProlieHis[5618]275280285[5619]Ser Val SerGluGlyValAspPheTyrSerGluLeuArgAsnLysGly[5620]290295300[5621]Phe Gin ArgGlyAlaSerGinGluLeuLysGluAlaValArgSerSer[5622]305310315320[5623]Val Glu GluGluGinCys[5624]325[5625]SEQUENCE LISTING30[5626]<110>北京思安创新生物技术有限公司[5627]段，陵浔[5628]<120>20 种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗[5629]<130>NP_000840[5630]<160>1[5631]<170>PatentIn version 3.5[5632]<210>1[5633]<211>225[5634]<212>PRT[5635]<213>Homo sapiens[5636]<220>
167[5637]<221>D0MAIN[5638]<222>(1) . (225)[5639]<400>1[5640]MetSerCysGluSerSerMetValLeuGlyTyrTrpAsplieArgGly[5641]151015[5642]LeuAlaHisAlalieArgLeuLeuLeuGluPheThrAspThrSerTyr[5643]202530[5644]GluGluLysArgTyrThrCysGlyGluAlaProAspTyrAspArgSer[5645]354045[5646]GinTrpLeuAspValLysPheLysLeuAspLeuAspPheProAsnLeu[5647]505560[5648]ProTyrLeuLeuAspGlyLysAsnLyslieThrGinSerAsnAlalie[5649]65707580[5650]LeuArgTyrlieAlaArgLysHisAsnMetCysGlyGluThrGluGlu[5651]859095[5652]GluLyslieArgValAsplielieGluAsnGinValMetAspPheArg[5653]100105110[5654]ThrGinLeulieArgLeuCysTyrSerSerAspHisGluLysLeuLys[5655]115120125[5656]ProGinTyrLeuGluGluLeuProGlyGinLeuLysGinPheSerMet[5657]130135140[5658]PheLeuGlyLysPheSerTrpPheAlaGlyGluLysLeuThrPheVal[5659]145150155160[5660]AspPheLeuThrTyrAsplieLeuAspGinAsnArgliePheAspPro[5661]165170175[5662]LysCysLeuAspGluPheProAsnLeuLysAlaPheMetCysArgPhe[5663]180185190[5664]GluAlaLeuGluLyslieAlaAlaTyrLeuGinSerAspGinPheCys[5665]195200205[5666]LysMetProlieAsnAsnLysMetAlaGinTrpGlyAsnLysProVal[5667]210215220[5668]Cys[5669]225[5670]SEQUENCE LISTING31[5671]<110>北京思安创新生物技术有限公司[5672]段，陵浔[5673]<120>20 种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗[5674]<130>NP_000631[5675]<160>15676]<170>PatentIn version 3.55677]<210>15678]<211>3805679]<212>PRT5680]<213>Homo sapiens5681]<220>5682]<221>D0MAIN5683]<222>(1) . (380)5684]<400>15685]Met AlaPheValCysLeuAlalieGlyCysLeuTyrThrPheLeulie5686]1510155687]Ser ThrThrPheGlyCysThrSerSerSerAspThrGlulieLysVal5688]2025305689]Asn ProProGinAspPheGlulieValAspProGlyTyrLeuGlyTyr5690]3540455691]Leu TyrLeuGinTrpGinProProLeuSerLeuAspHisPheLysGlu5692]5055605693]Cys ThrValGluTyrGluLeuLysTyrArgAsnlieGlySerGluThr5694]657075805695]Trp LysThrlielieThrLysAsnLeuHisTyrLysAspGlyPheAsp5696]8590955697]Leu AsnLysGlylieGluAlaLyslieHisThrLeuLeuProTrpGin5698]1001051105699]Cys ThrAsnGlySerGluValGinSerSerTrpAlaGluThrThrTyr5700]1151201255701]Trp lieSerProGinGlylieProGluThrLysValGinAspMetAsp5702]1301351405703]Cys ValTyrTyrAsnTrpGinTyrLeuLeuCysSerTrpLysProGly5704]1451501551605705]lie GlyValLeuLeuAspThrAsnTyrAsnLeuPheTyrTrpTyrGlu5706]1651701755707]Gly LeuAspHisAlaLeuGinCysValAspTyrlieLysAlaAspGly5708]1801851905709]Gin AsnlieGlyCysArgPheProTyrLeuGluAlaSerAspTyrLys5710]1952002055711]Asp PheTyrlieCysValAsnGlySerSerGluAsnLysProlieArg5712]2102152205713]Ser SerTyrPheThrPheGinLeuGinAsnlieValLysProLeuPro5714]225230235240[5715]Pro Val Tyr Leu Thr Phe Thr Arg Glu Ser Ser Cys Glu lie Lys Leu245250255 Lys Trp Ser lie Pro Leu Gly Pro lie Pro Ala Arg Cys Phe Asp Tyr260265270Glu lie Glu lie Arg Glu Asp Asp Thr Thr Leu Val Thr Ala Thr Val275280285Glu Asn Glu Thr Tyr Thr Leu Lys Thr Thr Asn Glu Thr Arg Gln Leu290295300Cys Phe Val Val Arg Ser Lys Val Asn lie Tyr Cys Ser Asp Asp Gly305310315320lie Trp Ser Glu Trp Ser Asp Lys Gln Cys Trp Glu Gly Glu Asp Leu325330335Ser Lys Lys Thr Leu Leu Arg Phe Trp Leu Pro Phe Gly Phe lie Leu340345350lie Leu Val lie Phe Val Thr Gly Leu Leu Leu Arg Lys Pro Asn Thr355360365Tyr Pro Lys Met lie Pro Glu Phe Phe Cys Asp Thr370375380SEQUENCE LISTING32<110>北京思安创新生物技术有限公司段，陵浔<120>20种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗<130>NP_057452<160>1<170>PatentIn version 3. 5<210>1<211>558<212>PRT<213>Homo sapiens<220><221>D0MAIN<222>(1). . (558)<400>1Met Glu Ala Ala Ala Gln Phe Phe Val Glu Ser Pro Asp Val Val Tyr151015Gly Pro Glu Ala lie Glu Ala Gln Tyr Glu Tyr Arg Thr Thr Arg Val202530Ser Arg Glu Gly Gly Val Leu Lys Val His Pro Thr Ser Thr Arg Phe354045[5754]Thr Phe Arg Thr Ala Arg Gln Val Pro Arg Leu Gly Val Met Leu Val505560Gly Trp Gly Gly Asn Asn Gly Ser Thr Leu Thr Ala Ala Val Leu Ala 65707580Asn Arg Leu Arg Leu Ser Trp Pro Thr Arg Ser Gly Arg Lys Glu Ala859095Asn Tyr Tyr Gly Ser Leu Thr Gln Ala Gly Thr Val Ser Leu Gly Leu100105110Asp Ala Glu Gly Gln Glu Val Phe Val Pro Phe Ser Ala Val Leu Pro115120125Met Val Ala Pro Asn Asp Leu Val Phe Asp Gly Trp Asp lie Ser Ser130135140Leu Asn Leu Ala Glu Ala Met Arg Arg Ala Lys Val Leu Asp Trp Gly145150155160Leu Gln Glu Gln Leu Trp Pro His Met Glu Ala Leu Arg Pro Arg Pro165170175Ser Val Tyr lie Pro Glu Phe lie Ala Ala Asn Gln Ser Ala Arg Ala180185190Asp Asn Leu lie Pro Gly Ser Arg Ala Gln Gln Leu Glu Gln lie Arg195200205Arg Asp lie Arg Asp Phe Arg Ser Ser Ala Gly Leu Asp Lys Val lie210215220Val Leu Trp Thr Ala Asn Thr Glu Arg Phe Cys Glu Val lie Pro Gly225230235240Leu Asn Asp Thr Ala Glu Asn Leu Leu Arg Thr lie Glu Leu Gly Leu245250255Glu Val Ser Pro Ser Thr Leu Phe Ala Val Ala Ser lie Leu Glu Gly260265270Cys Ala Phe Leu Asn Gly Ser Pro Gln Asn Thr Leu Val Pro Gly Ala275280285Leu Glu Leu Ala Trp Gln His Arg Val Phe Val Gly Gly Asp Asp Phe290295300Lys Ser Gly Gln Thr Lys Val Lys Ser Val Leu Val Asp Phe Leu lie305310315320Gly Ser Gly Leu Lys Thr Met Ser lie Val Ser Tyr Asn His Leu Gly325330335Asn Asn Asp Gly Glu Asn Leu Ser Ala Pro Leu Gln Phe Arg Ser Lys340345350Glu Val Ser Lys Ser Asn Val Val Asp Asp Met Val Gln Ser Asn Pro[5793]355360365Val Leu Tyr Thr Pro Gly Glu Glu Pro Asp His Cys Val Val lie Lys [5795]370375380Tyr Val Pro Tyr Val Gly Asp Ser Lys Arg Ala Leu Asp Glu Tyr Thr385390395400Ser Glu Leu Met Leu Gly Gly Thr Asn Thr Leu Val Leu His Asn Thr405410415Cys Glu Asp Ser Leu Leu Ala Ala Pro lie Met Leu Asp Leu Ala Leu420425430Leu Thr Glu Leu Cys Gln Arg Val Ser Phe Cys Thr Asp Met Asp Pro435440445Glu Pro Gln Thr Phe His Pro Val Leu Ser Leu Leu Ser Phe Leu Phe450455460Lys Ala Pro Leu Val Pro Pro Gly Ser Pro Val Val Asn Ala Leu Phe465470475480Arg Gln Arg Ser Cys lie Glu Asn lie Leu Arg Ala Cys Val Gly Leu485490495Pro Pro Gln Asn His Met Leu Leu Glu His Lys Met Glu Arg Pro Gly500505510Pro Ser Leu Lys Arg Val Gly Pro Val Ala Ala Thr Tyr Pro Met Leu515520525Asn Lys Lys Gly Pro Val Pro Ala Ala Thr Asn Gly Cys Thr Gly Asp530535540Ala Asn Gly His Leu Gln Glu Glu Pro Pro Met Pro Thr Thr545550555SEQUENCE LISTING33<110>北京思安创新生物技术有限公司段，陵浔<120>20种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗<130>NP_004914<160>1<170>PatentIn version 3. 5<210>1<211>508<212>PRT<213>Homo sapiens<220><221>D0MAIN<222>(1). . (508)[5832]<400>1Met Glu Glu Gly Pro Leu Pro Gly Gly Leu Pro Ser Pro Glu Asp Ala151015Met Val Thr Glu Leu Leu Ser Pro Glu Gly Pro Phe Ala Ser Glu Asn202530lie Gly Leu Lys Ala Pro Val Lys Tyr Glu Glu Asp Glu Phe His Val354045 [5839]Phe Lys Glu Ala Tyr Leu Gly Pro Ala Asp Pro Lys Glu Pro Val Leu505560His Ala Phe Asn Pro Ala Leu Gly Ala Asp Cys Lys Gly Gln Val Lys65707580Ala Lys Leu Ala Gly Gly Asp Ser Asp Gly Gly Glu Leu Leu Gly Glu859095Tyr Pro Gly lie Pro Glu Leu Ser Ala Leu Glu Asp Val Ala Leu Leu100105110Gln Ala Pro Gln Pro Pro Ala Cys Asn Val His Phe Leu Ser Ser Leu115120125Leu Pro Ala His Arg Ser Pro Ala Val Leu Pro Leu Gly Ala Trp Val130135140Leu Glu Gly Ala Ser His Pro Gly Val Arg Met lie Pro Val Glu lie145150155160Lys Glu Ala Gly Gly Thr Thr Thr Ser Asn Asn Pro Glu Glu Ala Thr165170175Leu Gln Asn Leu Leu Ala Gln Glu Ser Cys Cys Lys Phe Pro Ser Ser180185190Gln Glu Leu Glu Asp Ala Ser Cys Cys Ser Leu Lys Lys Asp Ser Asn195200205Pro Met Val lie Cys Gln Leu Lys Gly Gly Thr Gln Met Leu Cys lie210215220Asp Asn Ser Arg Thr Arg Glu Leu Lys Ala Leu His Leu Val Pro Gln225230235240Tyr Gln Asp Gln Asn Asn Tyr Leu Gln Ser Asp Val Pro Lys Pro Met245250255Thr Ala Leu Val Gly Arg Phe Leu Pro Ala Ser Thr Lys Leu Asn Leu260265270lie Thr Gln Gln Leu Glu Gly Ala Leu Pro Ser Val Val Asn Gly Ser275280285Ala Phe Pro Ser Gly Ser Thr Leu Pro Gly Pro Pro Lys lie Thr Leu290295300[5871]Ala Gly Tyr Cys Asp Cys Phe Ala Ser Gly Asp Phe Cys Asn Asn Cys305310315320Asn Cys Asn Asn Cys Cys Asn Asn Leu His His Asp lie Glu Arg Phe 325330335Lys Ala lie Lys Ala Cys Leu Gly Arg Asn Pro Glu Ala Phe Gln Pro340345350Lys lie Gly Lys Gly Gln Leu Gly Asn Val Lys Pro Gln His Asn Lys355360365Gly Cys Asn Cys Arg Arg Ser Gly Cys Leu Lys Asn Tyr Cys Glu Cys370375380Tyr Glu Ala Gln lie Met Cys Ser Ser lie Cys Lys Cys lie Gly Cys385390395400Lys Asn Tyr Glu Glu Ser Pro Glu Arg Lys Thr Leu Met Ser Met Pro405410415Asn Tyr Met Gln Thr Gly Gly Leu Glu Gly Ser His Tyr Leu Pro Pro420425430Thr Lys Phe Ser Gly Leu Pro Arg Phe Ser His Asp Arg Arg Pro Ser435440445Ser Cys lie Ser Trp Glu Val Val Glu Ala Thr Cys Ala Cys Leu Leu450455460Ala Gln Gly Glu Glu Ala Glu Lys Glu His Cys Ser Lys Cys Leu Ala465470475480Glu Gln Met lie Leu Glu Glu Phe Gly Arg Cys Leu Ser Gln lie Leu485490495His Thr Glu Phe Lys Ser Lys Gly Leu Lys Met Glu500505SEQUENCE LISTING34<110>北京思安创新生物技术有限公司段，陵浔<120>20种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗<130>NP_004289<160>1<170>PatentIn version 3. 5<210>1<211>1332<212>PRT<213>Homo sapiens<220><221>D0MAIN[5910]<222>(1). . (1332)<400>1Met Ser Asp Met Asp Tyr Pro Leu Gln Gly Pro Gly Leu Leu Ser Val151015Pro Asn Leu Pro Glu lie Ser Ser lie Arg Arg Val Pro Leu Pro Pro202530 [5916]Glu Leu Val Glu Gln Phe Gly His Met Gln Cys Asn Cys Met Met Gly354045Val Phe Pro Pro lie Ser Arg Ala Trp Leu Thr lie Asp Ser Asp lie505560Phe Met Trp Asn Tyr Glu Asp Gly Gly Asp Leu Ala Tyr Phe Asp Gly65707580Leu Ser Glu Thr lie Leu Ala Val Gly Leu Val Lys Pro Lys Ala Gly859095lie Phe Gln Pro His Val Arg His Leu Leu Val Leu Ala Thr Pro Val100105110Asp lie Val lie Leu Gly Leu Ser Tyr Ala Asn Leu Gln Thr Gly Ser115120125Gly Val Leu Asn Asp Ser Leu Ser Gly Gly Met Gln Leu Leu Pro Asp130135140Pro Leu Tyr Ser Leu Pro Thr Asp Asn Thr Tyr Leu Leu Thr lie Thr145150155160Ser Thr Asp Asn Gly Arg lie Phe Leu Ala Gly Lys Asp Gly Cys Leu165170175Tyr Glu Val Ala Tyr Gln Ala Glu Ala Gly Trp Phe Ser Gln Arg Cys180185190Arg Lys lie Asn His Ser Lys Ser Ser Leu Ser Phe Leu Val Pro Ser195200205Leu Leu Gln Phe Thr Phe Ser Glu Asp Asp Pro lie Leu Gln lie Ala210215220lie Asp Asn Ser Arg Asn lie Leu Tyr Thr Arg Ser Glu Lys Gly Val225230235240lie Gln Val Tyr Asp Leu Gly Gln Asp Gly Gln Gly Met Ser Arg Val245250255Ala Ser Val Ser Gln Asn Ala lie Val Ser Ala Ala Gly Asn lie Ala260265270Arg Thr lie Asp Arg Ser Val Phe Lys Pro lie Val Gln lie Ala Val275280285lie Glu Asn Ser Glu Ser Leu Asp Cys Gln Leu Leu Ala Val Thr His[5949]290295300[5950]Ala Gly Val Arg Leu Tyr Phe Ser Thr Cys Pro Phe Arg Gln Pro Leu305310315320Ala Arg Pro Asn Thr Leu Thr Leu Val His Val Arg Leu Pro Pro Gly325330335Phe Ser Ala Ser Ser Thr Val Glu Lys Pro Ser Lys Val His Arg Ala340345350Leu Tyr Ser Lys Gly lie Leu Leu Met Ala Ala Ser Glu Asn Glu Asp355360365Asn Asp lie Leu Trp Cys Val Asn His Asp Thr Phe Pro Phe Gln Lys370375380Pro Met Met Glu Thr Gln Met Thr Ala Gly Val Asp Gly His Ser Trp385390395400Ala Leu Ser Ala lie Asp Glu Leu Lys Val Asp Lys lie lie Thr Pro405410415Leu Asn Lys Asp His lie Pro lie Thr Asp Ser Pro Val Val Val Gln420425430Gln His Met Leu Pro Pro Lys Lys Phe Val Leu Leu Ser Ala Gln Gly435440445Ser Leu Met Phe His Lys Leu Arg Pro Val Asp Gln Leu Arg His Leu450455460Leu Val Ser Asn Val Gly Gly Asp Gly Glu Glu lie Glu Arg Phe Phe465470475480Lys Leu His Gln Glu Asp Gln Ala Cys Ala Thr Cys Leu lie Leu Ala485490495Cys Ser Thr Ala Ala Cys Asp Arg Glu Val Ser Ala Trp Ala Thr Arg500505510Ala Phe Phe Arg Tyr Gly Gly Glu Ala Gln Met Arg Phe Pro Thr Thr515520525Leu Pro Pro Pro Ser Asn Val Gly Pro lie Leu Gly Ser Pro Val Tyr530535540Ser Ser Ser Pro Val Pro Ser Gly Ser Pro Tyr Pro Asn Pro Ser Phe545550555560Leu Gly Thr Pro Ser His Gly lie Gln Pro Pro Ala Met Ser Thr Pro565570575Val Cys Ala Leu Gly Asn Pro Ala Thr Gln Ala Thr Asn Met Ser Cys580585590Val Thr Gly Pro Glu lie Val Tyr Ser Gly Lys His Asn Gly lie Cys595600605[5988]lie Tyr Phe Ser Arg lie Met Gly Asn lie Trp Asp Ala Ser Leu Val610615620Val Glu Arg lie Phe Lys Ser Gly Asn Arg Glu lie Thr Ala lie Glu625630635640Ser Ser Val Pro Cys Gln Leu Leu Glu Ser Val Leu Gln Glu Leu Lys645650655Gly Leu Gln Glu Phe Leu Asp Arg Asn Ser Gln Phe Ala Gly Gly Pro660665670Leu Gly Asn Pro Asn Thr Thr Ala Lys Val Gln Gln Arg Leu lie Gly675680685Phe Met Arg Pro Glu Asn Gly Asn Pro Gln Gln Met Gln Gln Glu Leu690695700Gln Arg Lys Phe His Glu Ala Gln Leu Ser Glu Lys lie Ser Leu Gln705710715720Ala lie Gln Gln Leu Val Arg Lys Ser Tyr Gln Ala Leu Ala Leu Trp725730735Lys Leu Leu Cys Glu His Gln Phe Thr lie lie Val Ala Glu Leu Gln740745750Lys Glu Leu Gln Glu Gln Leu Lys lie Thr Thr Phe Lys Asp Leu Val755760765lie Arg Asp Lys Glu Leu Thr Gly Ala Leu lie Ala Ser Leu lie Asn770775780Cys Tyr lie Arg Asp Asn Ala Ala Val Asp Gly lie Ser Leu His Leu785790795800Gln Asp lie Cys Pro Leu Leu Tyr Ser Thr Asp Asp Ala lie Cys Ser805810815Lys Ala Asn Glu Leu Leu Gln Arg Ser Arg Gln Val Gln Asn Lys Thr820825830Glu Lys Glu Arg Met Leu Arg Glu Ser Leu Lys Glu Tyr Gln Lys lie835840845Ser Asn Gln Val Asp Leu Ser Asn Val Cys Ala Gln Tyr Arg Gln Val850855860Arg Phe Tyr Glu Gly Val Val Glu Leu Ser Leu Thr Ala Ala Glu Lys865870875880Lys Asp Pro Gln Gly Leu Gly Leu His Phe Tyr Lys His Gly Glu Pro885890895Glu Glu Asp lie Val Gly Leu Gln Ala Phe Gln Glu Arg Leu Asn Ser900905910Tyr Lys Cys lie Thr Asp Thr Leu Gln Glu Leu Val Asn Gln Ser Ly[6027]915920925Ala Ala Pro Gln Ser Pro Ser Val Pro Lys Lys Pro Gly Pro Pro Val930935940 Leu Ser Ser Asp Pro Asn Met Leu Ser Asn Glu Glu Ala Gly His His945950955960Phe Glu Gln Met Leu Lys Leu Ser Gln Arg Ser Lys Asp Glu Leu Phe965970975Ser lie Ala Leu Tyr Asn Trp Leu lie Gln Val Asp Leu Ala Asp Lys980985990Leu Leu Gln Val Ala Ser Pro Phe Leu Glu Pro His Leu Val Arg Met99510001005Ala Lys Val Asp Gln AsnArg Val Arg Tyr Met Asp Leu Leu Trp101010151020Arg Tyr Tyr Glu Lys AsnArg Ser PheSer Asn Ala Ala Arg Val102510301035Leu Ser Arg Leu Ala AspMet His SerThr Glu lie Ser Leu Gln104010451050Gln Arg Leu Glu Tyr lieAla Arg Ala lie Leu Ser Ala Lys Ser105510601065Ser Thr Ala lie Ser Serlie Ala Ala Asp Gly Glu Phe Leu His107010751080Glu Leu Glu Glu Lys MetGlu Val Ala Arg lie Gln Leu Gln lie108510901095Gln Glu Thr Leu Gln Arg Gln Tyr SerHis His Ser Ser Val Gln110011051110Asp Ala Val Ser Gln LeuAsp Ser GluLeu Met Asp lie Thr Lys111511201125Leu Tyr Gly Glu Phe Ala Asp Pro PheLys Leu Ala Glu Cys Lys113011351140Leu Ala lie lie His CysAla Gly TyrSer Asp Pro lie Leu Val114511501155Gln Thr Leu Trp Gln Asplie lie GluLys Glu Leu Ser Asp Ser116011651170Val Thr Leu Ser Ser SerAsp Arg MetHis Ala Leu Ser Leu Lys117511801185lie Val Leu Leu Gly Lyslie Tyr Ala Gly Thr Pro Arg Phe Phe119011951200Pro Leu Asp Phe lie ValGln Phe LeuGlu Gln Gln Val Cys Thr120512101215[6066]Leu Asn Trp Asp Val Gly Phe Val lie Gln Thr Met Asn Glu lie122012251230 [6068]Gly Val Pro Leu Pro Arg Leu Leu Glu Val Tyr Asp Gln Leu Phe123512401245Lys Ser Arg Asp Pro Phe Trp Asn Arg Met Lys Lys Pro Leu His125012551260Leu Leu Asp Cys lie His Val Leu Leu lie Arg Tyr Val Glu Asn126512701275Pro Ser Gln Val Leu Asn Cys Glu Arg Arg Arg Phe Thr Asn Leu128012851290Cys Leu Asp Ala Val Cys Gly Tyr Leu Val Glu Leu Gln Ser Met129513001305Ser Ser Ser Val Ala Val Gln Ala lie Thr Gly Asn Phe Lys Ser131013151320Leu Gln Ala Lys Leu Glu Arg Leu His13251330SEQUENCE LISTING35<110>北京思安创新生物技术有限公司段，陵浔<120>20种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗<130>NP_056404<160>1<170>PatentIn version 3. 5<210>1<211>717<212>PRT<213>Homo sapiens<220><221>D0MAIN<222>(1). . (717)<400>1Met Phe Arg Asp Gln Val Gly Val Leu Ala Gly Trp Phe Lys Gly Trp151015Ash Glu Cys Glu Gln Thr Val Ala Leu Leu Ser Leu Leu Lys Arg Val202530Ser Gln Thr Gln Ala Arg Phe Leu Gln Leu Cys Leu Glu His Ser Leu354045Ala Asp Cys Ala Glu Leu His Val Leu Glu Arg Glu Ala Asn Ser Pro505560[6105]Gly lie lie Asn Gln Trp Gln Gln Glu Ser Lys Asp Lys Val lie Ser65707580Leu Leu Leu Thr His Leu Pro Leu Leu Lys Pro Gly Asn Leu Asp Ala859095Lys Val Glu Tyr Met Lys Leu Leu Pro Lys lie Leu Ala His Ser lie100105110Glu His Asn Gln His lie Glu Glu Ser Arg Gln Leu Leu Ser Tyr Ala 115120125Leu lie His Pro Ala Thr Ser Leu Lys Asp Arg Ser Ala Leu Ala Met130135140Trp Leu Asn His Leu Glu Asp Arg Thr Ser Thr Ser Phe Gly Gly Gln145150155160Asn Arg Gly Arg Ser Asp Ser Val Asp Tyr Gly Gln Thr His Tyr Tyr165170175His Gln Arg Gln Asn Ser Asp Asp Lys Leu Asn Gly Trp Gln Asn Ser180185190Arg Asp Ser Gly lie Cys lie Asn Ala Ser Asn Trp Gln Asp Lys Ser195200205Met Gly Cys Glu Asn Gly His Val Pro Leu Tyr Ser Ser Ser Ser Val210215220Pro Thr Thr lie Asn Thr lie Gly Thr Ser Thr Ser Thr lie Leu Ser225230235240Gly Gln Ala His His Ser Pro Leu Lys Arg Ser Val Ser Leu Thr Pro245250255Pro Met Asn Val Pro Asn Gln Pro Leu Gly His Gly Trp Met Ser His260265270Glu Asp Leu Arg Ala Arg Gly Pro Gln Cys Leu Pro Ser Asp His Ala275280285Pro Leu Ser Pro Gln Ser Ser Val Ala Ser Ser Gly Ser Gly Gly Ser290295300Glu His Leu Glu Asp Gln Thr Thr Ala Arg Asn Thr Phe Gln Glu Glu305310315320Gly Ser Gly Met Lys Asp Val Pro Ala Trp Leu Lys Ser Leu Arg Leu325330335His Lys Tyr Ala Ala Leu Phe Ser Gln Met Thr Tyr Glu Glu Met Met340345350Ala Leu Thr Glu Cys Gln Leu Glu Ala Gln Asn Val Thr Lys Gly Ala355360365Arg His Lys lie Val lie Ser lie Gln Lys Leu Lys Glu Arg Gln Asn[6144]370375380[6145]Leu Leu Lys Ser Leu Glu Arg Asp lie lie Glu Gly Gly Ser Leu Arg385390395400lie Pro Leu Gln Glu Leu His Gln Met lie Leu Thr Pro lie Lys Ala405410415Tyr Ser Ser Pro Ser Thr Thr Pro Glu Ala Arg Arg Arg Glu Pro Gln420425430Ala Pro Arg Gln Pro Ser Leu Met Gly Pro Glu Ser Gln Ser Pro Asp435440445Cys Lys Asp Gly Ala Ala Ala Thr Gly Ala Thr Ala Thr Pro Ser Ala450455460Gly Ala Ser Gly Gly Leu Gln Pro His Gln Leu Ser Ser Cys Asp Gly465470475480Glu Leu Ala Val Ala Pro Leu Pro Glu Gly Asp Leu Pro Gly Gln Phe485490495Thr Arg Val Met Gly Lys Val Cys Thr Gln Leu Leu Val Ser Arg Pro500505510Asp Glu Glu Asn lie Ser Ser Tyr Leu Gln Leu lie Asp Lys Cys Leu515520525lie His Glu Ala Phe Thr Glu Thr Gln Lys Lys Arg Leu Leu Ser Trp530535540Lys Gln Gln Val Gln Lys Leu Phe Arg Ser Phe Pro Arg Lys Thr Leu545550555560Leu Asp lie Ser Gly Tyr Arg Gln Gln Arg Asn Arg Gly Phe Gly Gln565570575Ser Asn Ser Leu Pro Thr Ala Gly Ser Val Gly Gly Gly Met Gly Arg580585590Arg Asn Pro Arg Gln Tyr Gln lie Pro Ser Arg Asn Val Pro Ser Ala595600605Arg Leu Gly Leu Leu Gly Thr Ser Gly Phe Val Ser Ser Asn Gln Arg610615620Asn Thr Thr Ala Thr Pro Thr lie Met Lys Gln Gly Arg Gln Asn Leu625630635640Trp Phe Ala Asn Pro Gly Gly Ser Asn Ser Met Pro Ser Arg Thr His645650655Ser Ser Val Gln Arg Thr Arg Ser Leu Pro Val His Thr Ser Pro Gln660665670Asn Met Leu Met Phe Gln Gln Pro Glu Phe Gln Leu Pro Val Thr Glu675680685[6183]Pro Asp lie Asn Asn Arg Leu Glu Ser Leu Cys Leu Ser Met Thr Glu690695700His Ala Leu Gly Asp Gly Val Asp Arg Thr Ser Thr lie 705710715SEQUENCE LISTING36<110>北京思安创新生物技术有限公司段，陵浔<120>20种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗<130>NP_057086<160>1<170>PatentIn version 3.5<210>1<211>429<212>PRT<213>Homo sapiens<220><221>D0MAIN<222>(1). . (429)<400>1Met Ala Thr Glu Gln Arg Pro Phe His Leu Val Val Phe Gly Ala Ser151015Gly Phe Thr Gly Gln Phe Val Thr Glu Glu Val Ala Arg Glu Gln Val202530Asp Pro Glu Arg Ser Ser Arg Leu Pro Trp Ala Val Ala Gly Arg Ser354045Arg Glu Lys Leu Gln Arg Val Leu Glu Lys Ala Ala Leu Lys Leu Gly505560Arg Pro Thr Leu Ser Ser Glu Val Gly lie lie lie Cys Asp lie Ala65707580Asn Pro Ala Ser Leu Asp Glu Met Ala Lys Gln Ala Thr Val Val Leu859095Asn Cys Val Gly Pro Tyr Arg Phe Tyr Gly Glu Pro Val lie Lys Ala100105110Cys lie Glu Asn Gly Ala Ser Cys lie Asp lie Ser Gly Glu Pro Gln115120125Phe Leu Glu Leu Met Gln Leu Lys Tyr His Glu Lys Ala Ala Asp Lys130135140Gly Val Tyr lie lie Gly Ser Ser Gly Phe Asp Ser lie Pro Ala Asp145150155160[6222]Leu Gly Val lie Tyr Thr Arg Asn Lys Met Asn Gly Thr Leu Thr Ala165170175Val Glu Ser Phe Leu Thr lie His Ser Gly Pro Glu Gly Leu Ser lie [6225]180185190His Asp Gly Thr Trp Lys Ser Ala lie Tyr Gly Phe Gly Asp Gln Ser195200205Asn Leu Arg Lys Leu Arg Asn Val Ser Asn Leu Lys Pro Val Pro Leu210215220lie Gly Pro Lys Leu Lys Arg Arg Trp Pro lie Ser Tyr Cys Arg Glu225230235240Leu Lys Gly Tyr Ser lie Pro Phe Met Gly Ser Asp Val Ser Val Val245250255Arg Arg Thr Gln Arg Tyr Leu Tyr Glu Asn Leu Glu Glu Ser Pro Val260265270Gln Tyr Ala Ala Tyr Val Thr Val Gly Gly lie Thr Ser Val lie Lys275280285Leu Met Phe Ala Gly Leu Phe Phe Leu Phe Phe Val Arg Phe Gly lie290295300Gly Arg Gln Leu Leu lie Lys Phe Pro Trp Phe Phe Ser Phe Gly Tyr305310315320Phe Ser Lys Gln Gly Pro Thr Gln Lys Gln lie Asp Ala Ala Ser Phe325330335Thr Leu Thr Phe Phe Gly Gln Gly Tyr Ser Gln Gly Thr Gly Thr Asp340345350Lys Asn Lys Pro Asn lie Lys lie Cys Thr Gln Val Lys Gly Pro Glu355360365Ala Gly Tyr Val Ala Thr Pro lie Ala Met Val Gln Ala Ala Met Thr370375380Leu Leu Ser Asp Ala Ser His Leu Pro Lys Ala Gly Gly Val Phe Thr385390395400Pro Gly Ala Ala Phe Ser Lys Thr Lys Leu lie Asp Arg Leu Asn Lys405410415His Gly lie Glu Phe Ser Val lie Ser Ser Ser Glu Val420425SEQUENCE LISTING37<110>北京思安创新生物技术有限公司段，陵浔<120>20种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗<130>NP_061946[6261]<160>1<170>PatentIn version 3. 5<210>1<211>488<212>PRT<213>Homo sapiens<220> [6268]<221>D0MAIN<222>(1). . (488)<400>1Met Pro Lys Val Lys Ala Leu Gln Cys Ala Leu Ala Leu Glu lie Ser151015Ser Val Thr Cys Pro Gly Val Val Leu Lys Asp Lys Glu Asp lie Tyr202530Leu Ser lie Cys Val Phe Gly Gln Tyr Lys Lys Thr Gln Cys Val Pro354045Ala Thr Phe Pro Leu Val Phe Asn Ala Arg Met Val Phe Glu Lys Val505560Phe Pro Asp Ala Val Asp Pro Gly Asp Val Val Thr Gln Leu Glu Tyr65707580Asp Thr Ala Val Phe Glu Leu lie Gln Leu Val Pro Pro Val Gly Glu859095Thr Leu Ser Thr Tyr Asp Glu Asn Thr Arg Asp Phe Met Phe Pro Gly100105110Pro Asn Gln Met Ser Gly His His Asp Ser Asn Arg Gln Val Thr Met115120125Arg Arg lie Ser Gly Leu Arg Gly Asn Ala Pro Arg Leu Glu Phe Ser130135140Thr Thr Ser Val lie Thr Glu Cys Leu lie Ser Ser Arg Lys Cys His145150155160Thr Gln Asp Lys Phe lie Tyr His Leu Ala Pro Val Glu Lys Ser His165170175Gly Arg Leu Gln Asn Arg Thr Ser Arg Ser Gln Lys Lys Lys Ser Lys180185190Ser Pro Glu Arg Ser Lys Tyr Cys lie Asn Ala Lys Asn Tyr Glu Gln195200205Pro Thr lie Ser Ser Lys Ser His Ser Pro Ser Pro Tyr Thr Lys Arg210215220Arg Met Cys Glu Leu Ser Glu Asp Thr Arg Arg Arg Leu Ala His Leu[6300]225230235240Asn Leu Gly Pro Tyr Glu Phe Lys Lys Glu Thr Asp Lys Pro Pro Phe245250255 [6303]Val lie Arg His Val Asp Pro Pro Ser Pro Arg Ala Asp Thr Leu Leu260265270Gly Ser Ser Gly Arg Asp Cys Glu Arg Asp Gly Trp Ser Arg Val His275280285Asn Asp His Ser His Leu Gly Cys Cys Arg Pro Lys Asp Tyr Lys Val290295300lie Arg Thr Pro His Gly Arg Asp Phe Asp Asp Ser Leu Glu Lys Cys305310315320Glu Glu Tyr Leu Ser Pro Arg Ser Cys Ser Lys Pro Arg His Ser Ala325330335Arg Thr Leu Leu Val His Ser Ala Pro Ser Thr Met Pro Lys His Ser340345350Pro Ser Pro Val Leu Asn Arg Ala Ser Leu Arg Glu Arg Phe His Ser355360365Asp Trp Cys Ser Pro Ser Asn Cys Asp Glu lie His Asp Arg Val Lys370375380Asn Val Leu Lys Ser His Gln Ala His Gln Arg His Leu Tyr Asp Glu385390395400Arg Asp Leu Glu Lys Asp Asp Glu Leu Glu Leu Lys Arg Ser Leu Leu405410415Cys Arg Asp Ser Ala Tyr Asp Ser Asp Pro Glu Tyr Ser Ser Cys Gln420425430Gln Pro Arg Gly Thr Phe His Leu Asp Asp Gly Glu Tyr Trp Ser Asn435440445Arg Ala Ala Ser Tyr Lys Gly Lys Ser His Arg Pro lie Phe Glu Asn450455460Ser Met Asp Lys Met Tyr Arg Asn Leu Tyr Lys Lys Ala Cys Ser Ser465470475480Ala Ser His Thr Gln Glu Ser Phe485SEQUENCE LISTING38<110>北京思安创新生物技术有限公司段，陵浔<120>20种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗<130>NP_003275<160>1[6339]<170>PatentIn version 3. 5<210>1<211>55<212>PRT<213>Homo sapiens<220><221>D0MAIN<222>(1). · (55)<400>1Met Ser Thr Ser Arg Lys Leu Lys Ser His Gly Met Arg Arg Ser Lys151015Ser Arg Ser Pro His Lys Gly Val Lys Arg Gly Gly Ser Lys Arg Lys202530Tyr Arg Lys Gly Asn Leu Lys Ser Arg Lys Arg Gly Asp Asp Ala Asn354045Arg Asn Tyr Arg Ser His Leu5055SEQUENCE LISTING39<110>北京思安创新生物技术有限公司段，陵浔<120>20种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗<130>NP_068379<160>1<170>PatentIn version 3. 5<210>1<211>592<212>PRT<213>Homo sapiens<220><221>D0MAIN<222>(1). . (592)<400>1Met Ala Ser Val Val Val Lys Thr lie Trp Gln Ser Lys Glu lie His151015Glu Ala Gly Asp Thr Pro Thr Gly Val Glu Ser Cys Ser Gln Leu Val202530Pro Glu Ala Pro Arg Arg Val Thr Ser Arg Ala Lys Gly lie Pro Lys[6376]354045Lys Lys Lys Ala Val Ser Phe His Gly Val Glu Pro Gln Met Ser His[6378]505560Gln Pro Met His Trp Cys Leu Asn Leu Lys Arg Ser Ser Ala Cys Thr65707580Asn Val Ser Leu Leu Asn Leu Ala Ala Met Glu Pro Thr Asp Ser Thr859095Gly Thr Asp Ser Thr Val Glu Asp Leu Ser Gly Gln Leu Thr Leu Ala [6384]100105110Gly Pro Pro Ala Ser Pro Thr Leu Pro Trp Asp Pro Asp Asp Ala Asp115120125lie Thr Glu lie Leu Ser Gly Val Asn Ser Gly Leu Val Arg Ala Lys130135140Asp Ser lie Thr Ser Leu Lys Glu Lys Thr Asn Arg Val Asn Gln His145150155160Val Gln Ser Leu Gln Ser Glu Cys Ser Val Leu Ser Glu Asn Leu Glu165170175Arg Arg Arg Gln Glu Ala Glu Glu Leu Glu Gly Tyr Cys lie Gln Leu180185190Lys Glu Asn Cys Trp Lys Val Thr Arg Ser Val Glu Asp Ala Glu lie195200205Lys Thr Asn Val Leu Lys Gln Asn Ser Ala Leu Leu Glu Glu Lys Leu210215220Arg Tyr Leu Gln Gln Gln Leu Gln Asp Glu Thr Pro Arg Arg Gln Glu225230235240Ala Glu Leu Gln Glu Pro Glu Glu Lys Gln Glu Pro Glu Glu Lys Gln245250255Glu Pro Glu Glu Lys Gln Lys Pro Glu Ala Gly Leu Ser Trp Asn Ser260265270Leu Gly Pro Ala Ala Thr Ser Gln Gly Cys Pro Gly Pro Pro Gly Ser275280285Pro Asp Lys Pro Ser Arg Pro His Gly Leu Val Pro Ala Gly Trp Gly290295300Met Gly Pro Arg Ala Gly Glu Gly Pro Tyr Val Ser Glu Gln Glu Leu305310315320Gln Lys Leu Phe Thr Gly lie Glu Glu Leu Arg Arg Glu Val Ser Ser325330335Leu Thr Ala Arg Trp His Gln Glu Glu Gly Ala Val Gln Glu Ala Leu340345350Arg Leu Leu Gly Gly Leu Gly Gly Arg Val Asp Gly Phe Leu Gly Gln355360365[6417]Trp Glu Arg Ala Gln Arg Glu Gln Ala Gln Thr Ala Arg Asp Leu Gln370375380Glu Leu Arg Gly Arg Ala Asp Glu Leu Cys Thr Met Val Glu Arg Ser385390395400 Ala Val Ser Val Ala Ser Leu Arg Ser Glu Leu Glu Gly Leu Gly Pro405410415Leu Lys Pro lie Leu Glu Glu Phe Gly Arg Gln Phe Gln Asn Ser Arg420425430Arg Gly Pro Asp Leu Ser Met Asn Leu Asp Arg Ser His Gln Gly Asn435440445Cys Ala Arg Cys Ala Ser Gln Gly Ser Gln Leu Ser Thr Glu Ser Leu450455460Gln Gln Leu Leu Asp Arg Ala Leu Thr Ser Leu Val Asp Glu Val Lys465470475480Gln Arg Gly Leu Thr Pro Ala Cys Pro Ser Cys Gln Arg Leu His Lys485490495Lys lie Leu Glu Leu Glu Arg Gln Ala Leu Ala Lys His Val Arg Ala500505510Glu Ala Leu Ser Ser Thr Leu Arg Leu Ala Gln Asp Glu Ala Leu Arg515520525Ala Lys Asn Leu Leu Leu Thr Asp Lys Met Lys Pro Glu Glu Lys Met530535540Ala Thr Leu Asp His Leu His Leu Lys Met Cys Ser Leu His Asp His545550555560Leu Ser Asn Leu Pro Leu Glu Gly Ser Thr Gly Thr Met Gly Gly Gly565570575Ser Ser Ala Gly Thr Pro Pro Lys Gln Gly Gly Ser Ala Pro Glu Gln580585590SEQUENCE LISTING40<110>北京思安创新生物技术有限公司段，陵浔<120>20种肿瘤特异相关的基因与产物的癌症的灵敏诊断与相关治疗<130>NP_059509<160>1<170>PatentIn version 3. 5<210>1<211>655<212>PRT<213>Homo sapiens[6456]<220><221>D0MAIN<222>(1). . (655) [6459]<400>1Met Ala Lys Gly Gly Glu Ala Leu Pro Gln Gly Ser Pro Ala Pro Val151015Gln Asp Pro His Leu lie Lys Val Thr Val Lys Thr Pro Lys Asp Lys202530Glu Asp Phe Ser Val Thr Asp Thr Cys Thr lie Gln Gln Leu Lys Glu354045Glu lie Ser Gln Arg Phe Lys Ala His Pro Asp Gln Leu Val Leu lie505560Phe Ala Gly Lys lie Leu Lys Asp Pro Asp Ser Leu Ala Gln Cys Gly65707580Val Arg Asp Gly Leu Thr Val His Leu Val lie Lys Arg Gln His Arg859095Ala Met Gly Asn Glu Cys Pro Ala Ala Ser Val Pro Thr Gln Gly Pro100105110Ser Pro Gly Ser Leu Pro Gln Pro Ser Ser lie Tyr Pro Ala Asp Gly115120125Pro Pro Ala Phe Ser Leu Gly Leu Leu Thr Gly Leu Ser Arg Leu Gly130135140Leu Ala Tyr Arg Gly Phe Pro Asp Gln Pro Ser Ser Leu Met Arg Gln145150155160His Val Ser Val Pro Glu Phe Val Thr Gln Leu lie Asp Asp Pro Phe165170175lie Pro Gly Leu Leu Ser Asn Thr Gly Leu Val Arg Gln Leu Val Leu180185190Asp Asn Pro His Met Gln Gln Leu lie Gln His Asn Pro Glu lie Gly195200205His lie Leu Asn Asn Pro Glu lie Met Arg Gln Thr Leu Glu Phe Leu210215220Arg Asn Pro Ala Met Met Gln Glu Met lie Arg Ser Gln Asp Arg Val225230235240Leu Ser Asn Leu Glu Ser lie Pro Gly Gly Tyr Asn Val Leu Cys Thr245250255Met Tyr Thr Asp lie Met Asp Pro Met Leu Asn Ala Val Gln Glu Gln260265270Phe Gly Gly Asn Pro Phe Ala Thr Ala Thr Thr Asp Asn Ala Thr Thr[6495]275280285Thr Thr Ser Gln Pro Ser Arg Met Glu Asn Cys Asp Pro Leu Pro Asn290295300 Pro Trp Thr Ser Thr His Gly Gly Ser Gly Ser Arg Gln Gly Arg Gln305310315320Asp Gly Asp Gln Asp Ala Pro Asp lie Arg Asn Arg Phe Pro Asn Phe325330335Leu Gly lie lie Arg Leu Tyr Asp Tyr Leu Gln Gln Leu His Glu Asn340345350Pro Gln Ser Leu Gly Thr Tyr Leu Gln Gly Thr Ala Ser Ala Leu Ser355360365Gln Ser Gln Glu Pro Pro Pro Ser Val Asn Arg Val Pro Pro Ser Ser370375380Pro Ser Ser Gln Glu Pro Gly Ser Gly Gln Pro Leu Pro Glu Glu Ser385390395400Val Ala lie Lys Gly Arg Ser Ser Cys Pro Ala Phe Leu Arg Tyr Pro405410415Thr Glu Asn Ser Thr Gly Gln Gly Gly Asp Gln Asp Gly Ala Gly Lys420425430Ser Ser Thr Gly His Ser Thr Asn Leu Pro Asp Leu Val Ser Gly Leu435440445Gly Asp Ser Ala Asn Arg Val Pro Phe Ala Pro Leu Ser Phe Ser Pro450455460Thr Ala Ala lie Pro Gly lie Pro Glu Pro Pro Trp Leu Pro Ser Pro465470475480Ala Tyr Pro Arg Ser Leu Arg Pro Asp Gly Met Asn Pro Ala Pro Gln485490495Leu Gln Asp Glu lie Gln Pro Gln Leu Pro Leu Leu Met His Leu Gln500505510Ala Ala Met Ala Asn Pro Arg Ala Leu Gln Ala Leu Arg Gln lie Glu515520525Gln Gly Leu Gln Val Leu Ala Thr Glu Ala Pro Arg Leu Leu Leu Trp530535540Phe Met Pro Cys Leu Ala Gly Thr Gly Ser Val Ala Gly Gly lie Glu545550555560Ser Arg Glu Asp Pro Leu Met Ser Glu Asp Pro Leu Pro Asn Pro Pro565570575Pro Glu Val Phe Pro Ala Leu Asp Ser Ala Glu Leu Gly Phe Leu Ser580585590[6534]Pro Pro Phe Leu His Met Leu Gln Asp Leu Val Ser Thr Asn Pro Gln595600605Gln Leu Gln Pro Glu Ala His Phe Gln Val Gln Leu Glu Gln Leu Arg610615620Ser Met Gly Phe Leu Asn Arg Glu Ala Asn Leu Gln Ala Leu lie Ala625630635640 [6540]Thr Gly Gly Asp Val Asp Ala Ala Val Glu Lys Leu Arg Gln Ser64565065权利要求
分离的多核苷酸，其选自(1)序列为SEQ ID NO1-20之一的多核苷酸；(2)序列为SEQID NO1-20之一的多核苷酸的互补序列；(3)序列为SEQ ID NO1-20之一的多核苷酸的反义序列。
2.权利要求1所述的多核苷酸编码的多肽。
3.权利要求2所述的多肽，其氨基酸序列为SEQID N0:21-40之一。
4.表达载体，其携带有权利要求1所述的分离的多核苷酸。
5.宿主细胞，其转化或转染了权利要求1所述的分离的多核苷酸或者权利要求4所述 的表达载体。
6.特异性结合权利要求2或3所述多肽的抗体或其片段。
7.权利要求1所述的分离的多核苷酸在制备制剂中的用途，该制剂用于刺激或扩增能 特异性识别肿瘤蛋白的T细胞。
8.权利要求2或3所述的多肽在制备制剂中的用途，该制剂用于刺激或扩增能特异性 识别肿瘤蛋白的T细胞。
9.用于刺激或扩增能特异性识别肿瘤蛋白的T细胞的制剂，其含有权利要求1所述的 分离的多核苷酸。
10.用于刺激或扩增能特异性识别肿瘤蛋白的T细胞的制剂，其含有权利要求2或3所 述的多肽。
11.由权利要求9或10所述的制剂制备的T细胞群。
12.权利要求1所述的分离的多核苷酸在制备预防和/或治疗肿瘤和/或癌症的疫苗 中的用途。
13.权利要求2或3所述的多肽在制备预防和/或治疗肿瘤和/或癌症的疫苗中的用途。
14.权利要求2或3所述的多肽在制备预防和/或治疗肿瘤和/或癌症的疫苗中的用途。
15.权利要求11所述的T细胞群在制备预防和/或治疗肿瘤和/或癌症的疫苗中的用途。
16.预防和/或治疗肿瘤和/或癌症的疫苗，其含有权利要求1所述的分离的多核苷酸 和药学上可接受的佐剂。
17.预防和/或治疗肿瘤和/或癌症的疫苗，其含有权利要求2或3所述的多肽和药学 上可接受的佐剂。
18.预防和/或治疗肿瘤和/或癌症的疫苗，其含有权利要求11所述的T细胞群和药 学上可接受的佐剂。
19.权利要求1所述的分离的多核苷酸在制备诊断肿瘤和/或癌症的试剂盒中的用途。
20.权利要求2或3所述的多肽在制备诊断肿瘤和/或癌症的试剂盒中的用途。
21.权利要求2或3所述的多肽在制备诊断肿瘤和/或癌症的试剂盒中的用途。
22.权利要求11所述的T细胞群在制备诊断肿瘤和/或癌症的试剂盒中的用途。
23.诊断肿瘤和/或癌症的试剂盒，其含有权利要求1所述的分离的多核苷酸。
24.诊断肿瘤和/或癌症的试剂盒，其含有权利要求2或3所述的多肽。
25.诊断肿瘤和/或癌症的试剂盒，其含有权利要求11所述的T细胞群。
全文摘要
本发明涉及通过定量检测肿瘤样品中20种肿瘤特异相关的基因及其产物如mRNA或蛋白质来诊断癌症的方法，以及它们在制备诊断试剂盒和治疗肿瘤的疫苗中的应用。本发明中20种肿瘤特异相关的基因及其产物的鉴定方法和试剂盒可有效地应用于包括结肠癌、乳腺癌、胃癌、肺癌和肝癌在内的癌症的灵敏诊断与相关治疗。
文档编号C07K16/18GK101845438SQ20091011965
公开日2010年9月29日 申请日期2009年3月25日 优先权日2009年3月25日
发明者段陵浔 申请人:北京锐健特生物技术有限公司