CN1371421A - 坦科聚合酶2材料和方法 - Google Patents
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Abstract
本发明提供了被称为坦科聚合酶2的新型坦科聚合酶多肽,编码这种多肽的多核苷酸,含有该多核苷酸的表达结构,以及用该表达结构转化过的宿主细胞。还提供了生产坦科聚合酶2多肽的方法,能够与坦科聚合酶2多肽发生免疫反应的抗体。另外,提供了鉴定坦科聚合酶2的专一性结合配偶体的方法,尤其是鉴定能调节坦科聚合酶2生物学活性的结合配偶体的方法。还提供了体外和体内调节坦科聚合酶2生物学活性的方法。
Description
本申请要求申请日为1999年6月29日的美国临时申请流水号00/141582的优先权。
本申请总体上涉及具有聚ADP-核糖基化活性的新型坦科聚合酶2(tankyrase)多肽,涉及编码这种多肽的多核苷酸,并且涉及使用所述材料的方法。
发明背景
真核生物染色体的末端(端粒)的特征是简单重复的DNA序列。不同物种之间该重复序列的长度不同,但端粒在具有线性染色体的生物中的重要性是共同的。端粒能保护染色体的末端,并且显然起着预防染色体末端重组的作用,这种重组会导致染色体融合及不稳定。另外,有大量的证据表明,端粒重复的产物决定着细胞分裂的能力或者甚至决定着细胞存活的能力。
在缺乏再生端粒长度的有效机制的条件下,培养的原始人类成纤维细胞的端粒随每一次细胞分裂而逐渐变短[Harley等,自然345:458-60,1990]。在端粒缩短的某个关键时期,所述细胞不再能够分裂,并进入被称为细胞衰老的状态。因此,至少在人类原始成纤维细胞中,端粒长度起着监测细胞衰老和调控寿命的生物钟的作用。
端粒长度调节细胞衰老的发现促进了端粒调控对生物衰老也起着关键作用的假设。不能够复制端粒的小鼠在第3代之后表现出早衰的特征。这些特征包括早衰,细胞分裂能力下降,伤口愈合能力受到损害,以及癌症的发病率增加等。因此,端粒结构的调控对于与衰老相关的某些特征来说可能是重要的。因此,能调节端粒结构的调控作用的药物可以用来治疗与衰老相关的综合征或用于治疗由遗传因素决定的早衰综合征。
只是在最近从对重复端粒的结构做过一些报导。该结构被统称为端粒复合体,该复合体包括若干种蛋白,这些蛋白能复制端粒并保护端粒结构不进行DNA修复,否则的话,有可能将端粒作为受损伤的DNA进行处理,并影响末端接合或重组,因而破坏染色体的完整性。端粒酶是所述端粒复合体的复制成分,并且是一种DNA聚合酶,它的特征是以完整的RNA分子为模板添加重复的序列[有关综述可以参见Greider,生物化学年度综述65:337-65,1996]。有关端粒酶活性对连续细胞分裂是必要的这一发现表明,在某些场合下,不适当的端粒酶活性可能是细胞致癌转化的成因。端粒酶的强制表达其本身不会导致致癌转化,但超量表达端粒酶的细胞具有显著的无限复制的能力这一事实表明,端粒酶的不适当的表达可能是致癌转化的多步骤过程的一个步骤。另外,有很多研究已经证实肿瘤组织中的端粒酶活性比大多数正常组织中的高,这表明较高的端粒酶活性对于肿瘤生长来说可能是必要的[有关综述可以参见Bacchetti,Cancer Surv 28:197-216,1996;和Harley等,Cold Spring Harbor Symp Quant Biol 59:307-15,1994]。
另外,还证实了被称为TRF1和TRF2的两种端粒专一性DNA结合蛋白对于端粒的保持是重要的[Chong等,科学270:1663-7,1995;van Steensel等,细胞92:401-13,1998]。TRF1在端粒长度的调控方面起着关键作用,而TRF2似乎对于保护染色体末端起着重要作用。这两种分子都含有DNA结合结构域和二聚体化结构域,它们似乎都以同源二聚体形式起作用。TRF1与端粒重复的接合能抑制端粒酶的作用,因此在复制期间导致端粒缩短[van Steensel和de Lange,自然385:740-3,1997]。
业已鉴定了另一种分子坦科聚合酶,它能通过添加ADP-核糖聚合物修饰TRF1[Smith等,科学282:1484-7,1998]。坦科聚合酶在结构上和功能上与聚(ADP-核糖)聚合酶(PARP)分子相关,它能够通过添加ADP-核糖聚合酶修饰蛋白[有关综述可参见Alvarez-Zonzalez等,分子细胞生物化学138:33-7,1994]。与PARP的结构相关性存在于坦科聚合酶的推测的催化结构域上,它与RARP有很高的氨基酸序列相似性。另外,坦科聚合酶含有不育性α基序(SAM)和24个锚蛋白重复。这些结构通常与蛋白/蛋白相互作用相关,并且坦科聚合酶上锚蛋白重复的至少一部分已经被证实决定着与TRF1的相互作用。业已证实坦科聚合酶在体外能使TRF1聚ADP-核糖基化,这表明坦科聚合酶在体内的作用是使TRF1 ADP-核糖基化,导致TRF1与端粒重复分离,并因此使得端粒酶能复制端粒。因此,可以预期能抑制坦科聚合酶活性的药物能抑制端粒的复制,并因此导致分裂的细胞群体,例如,癌细胞或增殖的免疫系统细胞的最终衰老。
坦科聚合酶或与坦科聚合酶相关的基因产物可以是药物设计的有吸引力的目标。在本领域中需要鉴定具有相关功能和/或结构的其他分子。所述分子可以用做坦科聚合酶定向药物的专一性对照或者它本身就可以作为药物发现方案的合适目标。综述所述,很显然目前对细胞DNA修复机制、信号传导、以及细胞复制诱导,肿瘤形成机制,和癌症疾病的治疗的了解是缺乏的。因此,本领域存在着鉴定其他坦科聚合酶样分子的必要性。将所述分子用于测定为了调节坦科聚合酶功能而设计的治疗剂的选择性,并用做人类疾病的治疗介入的目标。对其他坦科聚合酶基因产物的坦科聚合酶抑制剂的鉴定可以获得坦科聚合酶选择性药物,这些药物对于特定指标、不希望的副作用的减弱、或治疗剂对特定组织的定向都是有利的。本发明的其他目的和优点对于本领域普通技术人员来说是显而易见的。
发明概述
业已发现,通过本发明可以实现上述目的和其他目的,一方面,本发明提供了纯化的和分离的坦科聚合酶2多肽,优选人坦科聚合酶2多肽。具体地讲,本发明提供了含有SEQ ID NO:133所示氨基酸序列的纯化和分离的坦科聚合酶2多肽(被命名为TANK2-LONG)或含有SEQ ID NO:135所示氨基酸序列的纯化和分离的坦科聚合酶2多肽(被命名为TANK2-SHORT)。本发明还提供了编码坦科聚合酶2多肽的多核苷酸。例如,所述多核苷酸可以包括SEQ ID NO:132或SEQ IDNO:134所示核苷酸序列的编码区。
本发明还提供了互补于TANK2-编码多核苷酸的多核苷酸,以及能在中等严格杂交条件下与所述坦科聚合酶2多核苷酸的编码或非编码链杂交的多核苷酸。在优选方案中,所述多核苷酸能在严格杂交条件下与SEQ ID NO:132或SEQ ID NO:134所示多核苷酸的互补体杂交,并且编码这样一种蛋白:(a)具有polyA(ADP)聚合酶活性,(b)与受损的DNA相互作用,或(c)结合于端粒重复接合因子和/或调节其活性。
所述多核苷酸可以是DNA分子或RNA分子。本发明的某些理想的多核苷酸,例如寡核苷酸探针还可以包括一个可检测的标记部分。
另一方面,本发明提供了一种含有坦科聚合酶2编码多核苷酸的表达结构,以及用该表达结构转化或转染过的宿主细胞。所述多核苷酸能够可操作地连接于异源启动子上。
在另一方面,本发明提供了一种在宿主细胞中生产坦科聚合酶2多肽的方法,该宿主细胞做过修饰,以便能表达坦科聚合酶2多肽,该方法包括以下步骤:
a)在适合表达坦科聚合酶2多肽的条件下生长宿主细胞;和
b)从所述宿主细胞或宿主细胞生长的培养基中分离坦科聚合酶2多肽。
在另一方面,本发明提供了能与坦科聚合酶2多肽发生免疫反应的抗体。例如,所述抗体可选自下列一组:单克隆抗体、多克隆抗体、单链抗体(scFv抗体)、嵌合抗体、双功能/双专一性抗体、人源化抗体、人抗体、CDR-移植抗体、Fab片段、Fab’片段、F(ab’)2片段、和Fv片段。还提供了能产生所述抗体的细胞系。还提供了能与坦科聚合酶2专一性抗体发生免疫反应的抗独特型抗体。
另一方面,本发明提供了一种鉴定坦科聚合酶2多肽的结合配偶体的方法,包括:
a)在容许坦科聚合酶2多肽和测试化合物结合的条件下,让坦科聚合酶2多肽与一种测试化合物接触;
b)检测测试化合物和坦科聚合酶2多肽的结合;和
c)鉴定作为坦科聚合酶2多肽结合配偶体的测试化合物。
例如,所述方法可用于鉴定能选择性地或专一性调节,即抑制或增强坦科聚合酶2多肽的生物学活性的结合配偶体。
另一方面,还提供了一种鉴定坦科聚合酶2多核苷酸的结合配偶体的方法,包括:
a)在容许坦科聚合酶2多核苷酸和测试化合物结合的条件下,让坦科聚合酶2多肽与一种测试化合物接触;
b)检测测试化合物和坦科聚合酶2多核苷酸的结合;和
c)鉴定作为坦科聚合酶2多核苷酸结合配偶体的测试化合物。
可将所述方法用于鉴定能选择性地或专一性的调节,即抑制或增强坦科聚合酶2多肽表达的结合配偶体。
本发明还提供了一种治疗具有由聚(ADP-核糖)聚合酶活性介导的医学症状的人或动物对象的方法,包括以能有效抑制受治对象的坦科聚合酶2的量给该对象服用坦科聚合酶2抑制化合物。另一方面,本发明提供了一种治疗具有由聚(ADP-核糖)聚合酶活性介导的医学症状的人或动物对象的方法,包括以能有效抑制受治对象的poly(ADP-核糖酶)聚合酶活性的量给该对象服用一种能抑制坦科聚合酶2表达或活性的化合物。所述方法特别适用于治疗与肿瘤组织相关的医学症状。例如,所述方法可用于治疗癌症,如癌肿、肉肿、白血病和淋巴瘤。更具体地讲,所述方法可用于治疗选自下列一组的癌症:ACTH生成肿瘤,急性淋巴细胞性白血病、急性非淋巴细胞性白血病、肾上腺皮质癌、膀胱癌、脑癌、乳腺癌、宫颈癌、慢性淋巴细胞性白血病、慢性髓细胞性白血病、结肠直肠癌、皮肤T细胞淋巴瘤,子宫内膜癌、食道癌、Ewing’s肉瘤、胆囊癌、毛细胞白血病、头颈癌、Hodgkin’s淋巴瘤、Kaposi’s肉瘤、肾癌、肝癌、肺癌、小细胞和非小细胞),恶性腹膜渗出、恶性胸膜渗出、黑素瘤、间皮瘤、多发性骨髓瘤、成神经细胞瘤、神经胶质瘤、非Hodgkin’s淋巴瘤、骨肉瘤、卵巢癌、卵巢(生殖细胞)癌、胰腺癌、阴茎癌、前列腺癌、成视网膜细胞瘤、皮肤癌、软组织肉瘤、鳞状细胞癌、胃癌、睾丸癌、甲状腺癌、滋养层肿瘤、子宫癌、阴道癌、外阴癌和Wilm’s肿瘤。
通过本文所提供的详细说明和实施例可以理解本发明的上述及其他特征和优点。提供这些详细说明和实施例是为了增强对本发明的理解,而不是要限定本发明范围。
优选实施方案详述
本发明总体上涉及以前没有鉴定过的编码被命名为坦科聚合酶2的新型人蛋白的核酸(以下还称之为TANK2)。正如在本文所披露的,坦科聚合酶2与已知的坦科聚合酶蛋白和其他具有聚(ADP-核糖)聚合酶活性的蛋白不同。本发明就是基于编码坦科聚合酶2蛋白的新型基因,及其核酸序列、寡核苷酸、片段、和反义分子的发现。
由本发明所提供的核苷酸序列信息使得用本领域众所周知的并且经常使用的技术大规模表达其所编码的TANK2多肽变成可能。本发明还可以通过已知技术鉴定并分离编码相关TANK2多肽的多核苷酸,这些技术包括Southern(DNA)和/或Northern(mRNA)杂交,以及诸如聚合酶链式反应(PCR)、连接酶链式反应(LCR)等的扩增技术。有关多核苷酸的例子包括人和非人tank2基因组序列,包括等位变体,以及编码与TANK2同源的多肽和具有TANK2的一种或多种生物学、免疫学和/或物理学特性的在结构上相关的多肽的多核苷酸。
本发明包括天然存在的和非天然存在的坦科聚合酶2多核苷酸及其多肽产物。天然存在的坦科聚合酶2产物包括人体内所存在的特殊的多核苷酸和多肽坦科聚合酶2。不过,本发明包括通过序列同源性分析所确定的其他人坦科聚合酶2多核苷酸和多肽。本发明还包括在其他物种的细胞中表达的人tank2多肽和tank2多核苷酸的相应的同系物,优选哺乳动物同系物,更优选灵长类动物同系物。在每一种坦科聚合酶2内部,本发明还提供了剪接变体,该变体是由相同的基因组DNA编码的,但是来自不同的mRNA转录物。非天然存在的坦科聚合酶2产物包括天然存在的坦科聚合酶2产物的变体,如多核苷酸和多肽类似物(即其中的一个或多个核苷酸或氨基酸是添加的、取代的或缺失的)。非天然存在的TANK2多肽产物还包括经过共价修饰的TANK2产物,例如,水溶性聚合物修饰和糖基化变体等。
坦科聚合酶2多肽和编码这种多肽的核酸构成了用于精确检测和/或量化TANK2表达和与TANK2活性过高或不足相关的医学症状的诊断方法。另外,本文所披露的核苷酸序列可用于检测发生在编码TANK2的基因上的异常,如突变和缺失。例如,本文所披露的核苷酸序列可用于鉴定并分离编码tank2的基因组序列。可以用基因组tank2核酸序列的内含子和外显子的各个部分设计PCR引物,用于检测该基因组序列上的异常。
本发明还提供了利用TANK2和遗传工程产生的能表达重组TANK2的宿主细胞评估并筛选所述酶的聚(ADP-核糖)聚合酶活性的调节剂的方法。所述筛选方法可用于鉴定TANK2活性的别构兴奋剂、拮抗剂和抑制剂以及鉴定该活性的指导(例如,竞争性抑制剂)。TANK2蛋白拮抗剂和抑制剂,如抗TANK2抗体和tank2反义分子构成了用于治疗和缓解与过高的聚(ADP-核糖)聚合酶活性相关的症状的药用组合物的基础。TANK2的兴奋剂构成了治疗和缓解与不足的聚(ADP-核糖)聚合酶活性相关的症状的基础。
坦科聚合酶2多核苷酸
本发明特别提供了编码人TANK2多肽的新型纯化的和分离的多核苷酸。本发明的多核苷酸包括DNA序列和RNA转录物,包括有义链和互补的反义链,及其剪切变体。本发明的DNA序列包括,但不限于cDNA和基因组序列。在本文中,小写的“tank2”表示坦科聚合酶2核酸序列,而大写的“TANK2”表示坦科聚合酶2氨基酸序列。
本文所使用的“核酸”表示寡核苷酸或多核苷酸序列,及其片段或部分,并且还指来源于基因组的或合成的DNA或RNA,它可以是双链的或单链的,可以是有义链或反义链。本发明的代表性的双链多核苷酸可以具有第一股链(即编码链),它具有编码TANK2多肽的序列,同时具有第二股链(即互补链或非编码链),它具有可以按照Watson-Crick的DNA碱基配对原则推测出来的序列。双链或“双链体”结构可以是DNA:DNA,DNA:RNA或RNA:RNA核酸,优选的双链多核苷酸是含有SEQ ID NO:132或SEQ ID NO:134所示核苷酸序列的编码区的cDNA。本发明的代表性单链多核苷酸是编码TANK2多肽的mRNA。另一种代表性的单链多核苷酸是寡核苷酸探针或引物,它能与选自SEQ ID NO:132和SEQ ID NO:134所示序列的多核苷酸的编码链或非编码链杂交。还涉及其他核酸结构,例如三链体。
本发明的基因组DNA包括TANK2多肽的蛋白编码区,并且包括本发明优选多核苷酸的等位变体,如单链多核苷酸多态性。本发明的基因组DNA与编码除了TANK2以外的多肽的基因组DNA的不同之处在于,它包括存在于本发明的tank2 cDNA上的TANK2-编码区。基因组DNA可以转录成RNA,所得到的RNA转录物可以进行一次或多次剪接过程,其中,该转录物的一个或多个内含子(即非编码区)被除去或“剪接掉”。可以通过其他机制剪接并因此除去不同的非编码RNA序列的、但仍然编码TANK2多肽的RNA转录物在本领域中被称为“剪接变体”,本发明包括这种剪接变体。因此,本发明所包括剪接变体是由相同的DNA序列编码的,但能产生不同的氨基酸序列。所述剪接变体可以包括读框发生移动的区域,其中,所述RNA序列的下游部分以不同的方式翻译,使所得到的多肽具有不同的氨基酸序列。在本领域中等位变体是野生型(优势)及其序列的修饰形式。所述修饰是在染色体分离期间所发生的重组或者接触会导致遗传突变的条件的结果。与野生型基因一样,等位变体是天然存在的序列,与非天然存在的变体不同,非天然存在的变体是通过体外操作产生的。
本发明还包括通过编码TANK2的RNA多核苷酸的逆转录,然后在合成第二股互补链以便形式双链DNA而获得的cDNA。例如,本发明提供了编码具有选自SEQ ID NO:133或SEQ ID NO:135所示序列的氨基酸序列的多肽的DNA序列。在一种优选实施方案中,本发明提供了含有选自SEQ ID NO:132和SEQ ID NO:134所示序列的核苷酸序列的编码区的多核苷酸。
正如所指出的,本发明的高度优选的核酸序列如SEQ ID NO:132或SEQ ID NO:134所示。不过,由于在其信息编码特性方面的遗传密码的丰余或“简并性”,不同的核苷酸序列可能编码相同的多肽序列。因此,本发明包括编码本发明的TANK2多肽及其功能等同物的其他(简并性)核苷酸序列。例如,本发明包括含有基本上与SEQ ID NO:132或SEQ ID NO:134所示核苷酸序列的TANK2-编码区同源的核苷酸序列的多核苷酸。更具体地讲,本发明包括这样的多核苷酸,其相应的核苷酸序列与SEQ ID NO:132或SEQ ID NO:134所示核苷酸序列的相同性至少为90%,优选至少95%,更优选至少98%,更优选至少99%。
本发明的变体多核苷酸还包括SEQ ID NO:132和SEQ ID NO:134所示tank2核苷酸序列的片段及其同系物。编码TANK2多肽的完整长度多核苷酸的披露使得本领域普通技术人员能够方便地获得完整长度多核苷酸的每一种可能的片段。本发明的多核苷酸片段优选包括TANK2编码核苷酸序列所特有的序列,因此,在高度严格或中等严格条件下只能够(即专一性地)与含有所述特有序列的编码TANK2的核苷酸或其片段杂交。本发明基因组序列的多核苷酸片段不仅包括所述编码区所特有的序列,而且包括源于内含子、调控区和/或其他非翻译序列的完整长度序列的片段。本发明多核苷酸序列的特有序列可以通过与已知的多核苷酸进行序列比较识别,并且可以通过使用本领域常用的计算机软件鉴定,例如,在公开的序列数据库中可以获得的排比程序。
本发明还提供了在编码TANK2多肽家族的一种或多种多核苷酸之间保守的多核苷酸片段。所述片段包括作TANK2多肽家族特征的序列,该序列被称为“标记”序列。所述保守的标记序列可以方便地理解TANK2家族的多核苷酸编码成员的以下简单的序列比较。可以能对其进行检测的方式标记本发明的多核苷酸片段,包括放射性和非放射性标记。
杂交可以被定义为包括通过互补单链核酸分子的序列专一性结合形成部分或完整双链核酸分子的过程。因此,本发明还包括能与编码TANK2蛋白的多核苷酸的编码或非编码链杂交的核酸的用途。优选的杂交核酸能够与SEQ ID NO:132或SEQ ID NO:134所示核苷酸序列的编码或非编码链杂交。本发明还包括由于遗传密码的丰余性而能够与TANK2编码多核苷酸杂交的核酸,即根据不同的密码子利用编码相同氨基酸序列的多核苷酸。
例如,杂交类型包括核酸杂交或扩增探针(寡核苷酸),它能检测编码TANK2的核苷酸序列(例如,基因组序列)或密切相关的分子,如等位基因。所述探针的专一性,即它是原因高度保守的、保守的、或是非保守的区域或结构域,以及杂交或扩增条件的严格性(高、中等或低)将决定该探针是仅能够识别天然存在的Tank2,或是相关的序列。用于检测相关序列的探针选自tank2家族成员的保守的或高度保守的区域,所述探针可用于简并性探针库中。为了检测特有的核苷酸序列,或者是否需要最大的专一性,从tank2多核苷酸的非保守核苷酸区或特有区选择寡核苷酸探针。在本文中,术语“非保守核苷酸区”是指本文所披露的tank2所特有的,但在相关的tank2家族成员中不存在的核苷酸区。
杂交的专一性通常是根据进行杂交时的条件的严格程度决定的。杂交条件的严格程度可以被称为核酸结合复合体的解链温度(Tm)[例如,参见Berzer和Kimmel,分子克隆技术指南,酶学方法,152卷,学术出版社,San Diego,CA,1987]。“最高严格性”通常是指大约Tm-5℃(低于探针Tm5℃);“高严格性”为低于Tm5-10℃;“中等严格性”低于Tm大约10-20℃;“低严格性”低于Tm大约20-25℃。
另外,杂交的严格性可以是指在该方法中所使用的物理化学条件。为了说明,代表性的中等严格杂交条件为:杂交是在65℃下,在3倍柠檬酸钠盐溶液(SSC),0.1%sarkosyl,和20mM磷酸钠,pH6.8中进行;而洗涤是在65℃下在2倍SSC,0.1%SDS中进行。代表性的高严格杂交条件为:杂交是在42℃下在50%甲酰胺,5倍SSC中进行一夜,而洗涤是在50℃下在0.5倍SSC和0.1%SDS中进行。本领域可以理解的是,通过改变披露于Ausubel等所披露的温度和缓冲液或盐浓度能够达到同等严格性的条件(当代分子生物学方法,John Wiley&Sons,1994,6.0.3-6.4.10页)。杂交条件的改进可以凭经验确定,或者根据寡核苷酸探针的长度和探针中G/C碱基对的百分比精确计算。杂交条件可以按照Sambrook等所披露的方法计算(分子克隆,实验室手册,冷泉港实验室出版社,冷泉港,纽约,1989,9.47-9.51页)。
技术人员可以理解的是,在较高条件下的杂交能够鉴定与靶序列具有较高程度同源性或序列相同性的序列。相反,在较低严格条件下的杂交,能够鉴定与目标序列具有较低的,但仍然是显著程度的同源性或序列相同性的序列。因此,本发明的范围还包括能中等至最高严格条件下与SEQ ID NO:132或SEQ ID NO:134的核苷酸序列杂交的核酸。所述杂交序列优选能在高严格条件下与SEQ ID NO:132或SEQ ID NO:134所示多核苷酸的编码或非编码链杂交。
本发明的多核苷酸包括能与编码TANK2氨基酸序列的核酸的编码链或非编码链杂交的寡核苷酸(即核酸寡聚体,长度通常为大约10-60个核苷酸)。具体地讲,本发明包括能与SEQ ID NO:132或SEQ ID NO:134所示多核苷酸的编码或非编码链杂交的寡核苷酸。寡核苷酸的长度并不重要只要它能够与目标核酸分子杂交就行。不过,核酸分子越长就越难于制备,并且需要较长的杂交时间。因此,寡核苷酸不要超过必须的长度。因此,寡核苷酸应当含有至少10个核苷酸,优选至少15个核苷酸,更优选至少20个核苷酸。通常,寡核苷酸不能包含超过60个核苷酸,优选不超过30个核苷酸,更优选不超过25个核苷酸。所述寡核苷酸在本发明中可以用做DNA合成的引物(例如,用做PCR的引物;“扩增引物”),用做检测样品中目标DNA存在的探针(例如,Northern或Southern印迹和原位杂交),用做治疗剂(例如,反义治疗),或用于其他目的。寡核苷酸可以是单链或双链的,双链形式的一端或两端是平的或阶梯形的。
所述寡核苷酸可以通过已知方法由天然来源获得或产生。另外,所述寡核苷酸可以按照本领域公知的方法合成产生。例如,所述方法包括合适序列的克隆和限制或通过任何合适方法直接化学合成。用于制备寡核苷酸的各种化学方法为本领域所公知,包括磷酸三酯方法、磷酸二酯方法;二乙基亚磷酰胺方法;固体支持方法,和H-磷酸方法[有关综述可以参见Caruthers,科学230:281-5,1985;Caruthers等,酶学方法211:3-20,1992]。通常,寡核苷酸的制备是通过在聚合物支持物上通过自动亚磷酰胺合成完成的。还可以用上述方法生产包括100或更多个核苷酸的核酸分子。
本发明的tank2多核苷酸包括变体,它是编码hAPRP2或其功能性等同物的多核苷酸,并且可以包括核苷酸残基的缺失、插入或取代。在本文中,“缺失”是核苷酸或氨基酸序列所发生的一种变化,其中,分别缺少一个或多个核苷酸或氨基酸残基。在本文中,“插入”或“添加”是核苷酸或氨基酸序列所发生的一种变化,它会分别导致一个或多个核苷酸或氨基酸残基的增加。在本文中,“取代”是核苷酸或氨基酸序列所发生的一种变化,其中,有一个或多个核苷酸或氨基酸分别被不同的核苷酸或氨基酸所取代。
在本发明的范围内还包括多核苷酸变体,它是tank2天然存在形式的等位基因或替代形式,等位基因是由于天然发生的突变,即在基因组核苷酸序列上发生的缺失、插入或取代所产生的,它会或者不会改变所表达的多肽的结构或功能。上述每一种类型的突变变化可以单独发生,或者与其他变化组合发生,在特定等位序列上发生一次或多次。可能出现单一核苷酸多态性(SNPs),其中,有单一的碱基突变确定一种改变了的多肽,而这种改变可能与明显的表型差异相关。当然,SNPs可以是沉默的,因为它可能不会改变所编码的多肽,或者对表型没有影响的它所编码的任何改变。
本发明还包括存在于诸如其他哺乳动物的其他动物物种中的人坦科聚合酶2 DNA的天然同系物。例如,哺乳动物同系物包括存在于小鼠、大鼠和豚鼠体内的同系物,更优选存在于其他灵长类动物体内的同系物。一般,所述物种的同系物在核苷酸水平上,在蛋白编码区内具有显著的同源性。因此,本发明包括与编码人TANK2多肽的多核苷酸的编码区,例如,由SEQ ID NO:132或SEQ ID NO:134所示多核苷酸具有至少75%,至少80%,至少85%,至少90%,至少95%,至少98%,或至少99%的核苷酸相同性的多核苷酸。相对本发明多核苷酸的百分序列“同源性”可以被定义为候选序列上与TANK2-编码序列上的核苷酸相同的核苷酸碱基的百分比,所述百分比是在排比序列并在必要时引入缺口之后得出的,以便获得最大的百分序列相同性。已经有计算机软件(从商业渠道和公开的结构域来源获得)用于以自动化形式计算百分相同性(例如,FASTA)。
本发明包括作过工程改造以便选择性地改变TANK2基因产物的克隆、加工和/或表达的多核苷酸。可以用本领域众所周知的技术引入突变,例如,通过定向诱变插入新的限制位点,改变糖基化形式或改变某些表达系统所固有的密码子使用偏倚性,而同时保持对所表达多肽产物的氨基酸序列的控制。例如,可以选择特定原核或真核宿主细胞优选的密码子(密码子优化),以便提高TANK2表达速度或产生具有所需特征,较长的半衰期的重组RNA转录物。可以用本领域所公知的化学方法完全或部分合成tank2多核苷酸。在本文中,“化学合成”在本领域中被理解为与酶促方法相对的用于生产多核苷酸的纯粹的化学方法。因此,“完全”化学合成的DNA序列是完全通过化学方法产生的;“部分” 化学合成的DNA包括这样的DNA,其中,所得到的DNA只有一部分是通过化学方法产生的。
可以对DNA分子进行修饰,以便提高细胞内稳定性和半衰期。可能的修饰包括,但不限于,在该分子的5’和/或3’末端添加旁侧序列或者在该分子的主链内使用硫代磷酸酯或2’O-甲基连链而不是磷酸二酯键。
本发明还提供了TANK2肽核酸(PNA)分子。所述TANK2 PNAS是具有中性“肽样”主链的信息分子,它所具有的核碱基使得该分子能以高于相应寡核苷酸的亲和力和专一性与互补的TANK2编码DNA或RNA杂交(PerSeptive Biosystems)。
多肽表达系统
对TANK2编码DNA序列的了解,使得技术人员能够对细胞进行修饰,以便能够进行或加强TANK2的表达。因此,提供了宿主细胞,包括原核或真核细胞,这些细胞是通过导入本发明的多核苷酸稳定地或瞬时地修饰过的,以便能够表达所编码的TANK2多肽。还提供了整合了TANK2编码序列的自主复制的重组表达结构,如质粒和病毒DNA载体。
还提供了含有可操作地连接在内源或外源表达控制DNA序列和转录终止子上的TANK2编码多核苷酸的表达结构。表达控制DNA序列包括启动子、增强子和操纵子,并且通常是根据要使用该表达结构的表达系统进行选择的。优选的启动子和增强子序列通常是针对提高基因表达能力进行选择的,而操纵子序列通常是针对调控基因表达的能力进行选择的。本发明的优选结构还包括在宿主细胞中复制所必需的序列。优选用表达结构生产所编码的TANK2多肽,但也可用于扩增该结构本身。
本发明的多核苷酸可以作为环状质粒的一部分或作为含有分离的蛋白编码区的线性DNA或病毒载体形式导入宿主细胞。将DNA导入宿主细胞的方法包括转化、转染、电穿孔、细胞核注射、或与载体融合,如脂质体、微团、血影细胞和原生质体。例如,本发明的表达系统包括细菌、酵母、真菌、植物、昆虫、无脊椎动物、两栖动物和哺乳动物细胞系统。例如,某些合适的原核宿主细胞包括大肠杆菌菌株SG-536、HB101、W3110、X1776、X2282、DHI、和MRC1,假单胞菌,芽孢杆菌,如枯草芽孢杆菌,和链霉菌。合适的真核宿主细胞包括酵母,如酿酒酵母、粟酒酵母、巴斯德毕赤酵母和其他真菌,昆虫细胞,如sf9或sf21细胞(草地滩夜蛾),动物细胞,如中国仓鼠卵巢(CHO)细胞,人细胞,如JY,293和NIH3T3,以及植物细胞,如拟南芥细胞。为了生产mRNA探针,可以将tank2核苷酸序列或其任何部分克隆到一种载体上。所述载体为本领域所公知,并且可以通过商业渠道获得,可以通过添加标记过的核苷酸和合适的RNA聚合酶,如T7、T3或SP6将其用于体外合成RNA探针。
本领域技术人员可以按照公知的标准筛选宿主细胞的类型\表达TANK2产物的形式、和生长条件等。哺乳动物宿主细胞的使用预计能提供所述翻译后修饰(例如,糖基化、截短、脂化和磷酸化),这些修饰可能是使本发明的重组表达产物具有最佳生物学活性所必需的,包括糖基化和非糖基化形式的TANK2多肽。根据所使用的序列和/或载体,由重组细胞所产生的蛋白能够被分泌或者包含在细胞内。正如本领域技术人员所理解的,含有tank2的表达载体可以被设计成具有信号序列,该序列能指导TANK2通过特定的原核细胞或真核细胞膜进行分泌。
表达结构可以包括有利于,并且优选能促进在宿主细胞中进行同源重组的序列。这一目的可以通过用一种异源启动子的全部或部分取代天然存在的tank2启动子的全部或部分而实现,以便该细胞以较高水平表达TANK2。应当插入异源启动子,以便它可操纵地连接于TANK2编码序列上。例如,参见PCT国际公开号WO94/12650、WO92/20808和WO91/09955。
本发明的宿主细胞可用于大规模生产TANK2多肽产物的方法中。例如,本发明的宿主细胞是可用于开发能与TANK2多肽发生免疫反应的抗体的有价值的免疫原的来源。作为另一个例子,重组TANK2可以在宿主细胞产生并从宿主细胞中分离,以便用于体外结合测定,如药物筛选测定。在所述方法中,在合适的培养基中生长宿主细胞,并且从所述细胞中或所述细胞所生长的培养基中分离所需要的多肽产物。
多肽产物可以用本领域已知的纯化方法分离,如常规的层析方法,包括免疫亲和层析、受体亲和层析、疏水相互作用层析、凝集素亲和层析、大小排阻过滤、阳离子或阴离子交换层析、高效液相层析(HPLC)、和反相HPLC等。
纯化的另一种方法包括以融合蛋白形式表达并纯化所需要的蛋白,其中,TANK2多肽连接于异源氨基酸序列上。合适的异源序列可以包括特殊标志、标记或能够被特殊结合配偶体或试剂识别的螯合部分。例如,为了筛选TANK2活性的调节物的肽文库,可以表达与特定异源蛋白融合的TANK2蛋白,所述异源蛋白是经过选择的,以便利用探针抗体进行专一性识别。还可通过工程方法使融合蛋白在TANK2序列和异源蛋白序列之间包含一个裂解位点(例如,因子XA或肠激酶敏感序列),使得TANK2蛋白能够与异源蛋白分离,并随后进行纯化。所述融合成分的裂解可以产生所需形式的蛋白,它具有由于裂解过程所产生的额外氨基酸残基。
代表性的异源肽结构域包括金属螯合肽,如组氨酸-色氨酸组件,通过它可以在固定化的金属上进行纯化[Porath,蛋白表达纯化3,263-281,1992],和蛋白A结构域,通过它可以在固定化的免疫球蛋白上进行纯化。另一种可以使用的系统是二价阳离子结合结构域,以及对它专一的用于肽延伸/免疫亲和纯化系统中的抗体,该系统披露于US4703004;4782137;4851431和5011912中。该系统由Immunex公司(Seattle WA)以FLAG系统为商标出售。另一种合适的异源融合配偶体是谷胱甘肽S-转移酶(GST),它能够利用固定化的谷胱甘肽进行亲和纯化。其他有用的融合配偶体包括免疫球蛋白及其片段,例如,Fc片段。
表达重组TANK2的宿主细胞的鉴定对于鉴定合适表达系统来说可能是关键的。因此本发明的表达结构还可以包括编码一种或多种选择标记的序列,通过该标记可以鉴定具有处在工作状态下的该结构的宿主细胞。除了插入异源启动子DNA之外,还可将可扩增的标记DNA(例如,ada、dhefr、和多功能cad基因,它们编码氨甲酰基磷酸合成酶、天冬氨酸转氨甲酰基酶,和二氢乳清酸酶)还希望将内含子DNA连同异源启动子DNA一起插入。如果与TANK2编码序列连接的话,通过标准筛选方法扩增标记DNA,会导致该细胞中TANK2编码序列的同时扩增。检测标记基因针对诱导或选择表达,通常也能表示TANK2的表达。另外,如果将tank2多核苷酸序列插入标记基因序列内部,可以通过标记基因功能的缺乏鉴定含有tank2的重组细胞。
含有TANK2编码序列并且表达TANK2的宿主细胞还可以通过多种本领域技术人员所公知的其他方法鉴定。这些方法包括,但不限于DNA-DNA或DNA-RNA杂交和蛋白生物测定或免疫测定技术,其中包括用于检测和/或定量核酸或蛋白的基于膜、基于溶液、或基于芯片的技术。
可以通过DNA-DNA或DNA-RNA杂交或用tank2多核苷酸的片段,例如,用SEQ ID NO:132或SEQ ID NO:134所示序列的片段做探针进行扩增检测tank2多核苷酸序列的存在。基于核酸扩增的测定包括利用以tank2序列为基础的寡核苷酸检测含有tank2 DNA或RNA的转化体。可以用各种方法制备用于检测tank2多核苷酸序列的标记过的杂交或PCR探针,包括寡聚标记,缺口翻译,末端标记或用标记过的核苷酸进行PCR扩增。在本发明的一种实施方案中,可以利用TANK2或其变体和/或能表达TANK2或其变体的宿主细胞系筛选能作为TANK2的生物学或免疫学活性调节物的抗体、肽、或其他分子,如有机或无机分子。例如,可将能够中和TANK2的聚合酶或DNA结合活性的抗TANK2抗体用于抑制TANK2介导的细胞死亡。另外,通过组合化学方法筛选的能重组表达TANK2或其变体的肽文库或有机文库或能表达TANK2或其变体的细胞系可用于鉴定通过调节TANK2的生物学或免疫学活性而起作用的治疗分子。合成化合物,天然产物,和其他具有潜在的生物学活性材料的来源可以用本领域技术人员所公知的多种常规方法筛选。例如,编码TANK2的DNA结合结构域的核苷酸序列可以在宿主细胞中表达,可将该细胞用于筛选TANK2活性的别构调节物,该调节物是兴奋剂或拮抗剂。另外,编码TANK2的保守的催化结构域的核苷酸序列可以在宿主细胞中表达,用于筛选VDP-核糖多聚化抑制剂。
TANK2多肽
本发明还提供了纯化的和分离的哺乳动物TANK2多肽。典型的TANK2多肽具有SEQ ID NO:133或SEQ ID NO:135所示氨基酸序列。本发明的TANK2多肽可以从天然细胞来源中分离或者化学合成,但优选通过使用本发明宿主细胞的重组方法产生。本发明的TANK2产物可以是完整长度的多肽或多肽产物的变体,如片段、截短物、缺失变体,及其保留了TANK2特有的生物学活性的其他变体。在本文中,“生物学活性”是指一种TANK2多肽具有天然存在的TANK2蛋白的结构、调控或生物化学功能。具体地讲,本发明的TANK2具有结合DNA并且在细胞中根据DNA损伤聚合ADP核糖亚基的能力。
本发明的蛋白和片段可以通过本领域已知方法制备,所述方法包括直接从细胞中分离蛋白,分离或合成编码所述蛋白的DNA并用该DNA生产重组蛋白,以及用单个氨基酸化学合成所述蛋白。
所述TANK2多肽可以用标准方法从诸如溶解的细胞级份的生物学样品中分离。某些合适的方法包括沉淀和液相层析方法,如离子交换、疏水性相互作用和凝胶过滤[例如,参见Deutscher著,酶学方法(蛋白质化学指南,第VII节)182:309,1990和Scopes,蛋白纯化,Springer-Verlag,纽约,1987]。另外,通过本领域已知方法获得纯化材料:在制备SDS-PAGE凝胶上分离蛋白,切除感兴趣的带,并从聚丙烯酰胺基质中电洗脱所述蛋白。通过已知方法从蛋白中除去洗涤剂SDS,如通过用合适的柱透析,如Pierce化学公司的Extracti-Gel柱(Rockford,IL)。
本发明的TANK2多肽还可以通过本领域已知方法完全或部分化学合成[例如,参见Stuart和Young,固相肽合成,第2版,Pierce化学公司,1984]。例如,可以通过固相技术合成肽,从树脂上裂解下来,并通过制备HPLC纯化[例如,参见Roberge等,科学269:202-4,1995]。例如,自动化合成可以用ABI431A肽合成仪(Perkin Elmer,Norwalk,CT)按照生产商提供的说明进行。合成肽的组成可以通过氨基酸分析或测序证实(例如,Edman降解方法)。
重组TANK2蛋白可以在用含有tank2核苷酸序列的表达载体转化过并且在细胞培养基中生长的宿主细胞中生产的并且从该细胞中分离。在本文中,原核或真核宿主细胞是用本发明的TANK2编码多核苷酸稳定地或瞬时地转染(真核细胞)或转化(原核细胞)过的,使其能够直接表达TANK2多肽。在上述方法中,让宿主细胞在合适的培养基中生长,并且从所述细胞或者从所述细胞所生长的培养基中分离所需要的多肽产物。例如,所述多肽的分离可以通过免疫亲和纯化实现。优选利用转化过的宿主细胞大规模生产TANK2多肽。
本发明包括含有大体上与本文所披露的TANK2多肽的序列同源的氨基酸序列的多肽。例如,本发明包括其相应的氨基酸序列与SEQ IDNO:133或SEQ ID NO:135所示多肽序列具有至少90%、优选至少95%、更优选至少98%、更优选至少99%的相同性的多肽。
与本发明优选多肽相关的百分比序列“相同性”被定义为候选序列中与参考TANK2序列中的氨基酸残基相同的氨基酸残基的百分比,上述百分比是在排比并且必要时引入缺口之后得出的,以便获得最大的百分比序列相同性。并且不将任何保守性取代作为序列相同性的因素。
与本发明优选多肽相关的百分比序列“同源性”被定义为候选序列中与参考TANK2序列中的氨基酸残基相同的氨基酸残基的百分比,上述百分比是在排比并且必要时引入缺口之后得出的,以便获得最大的百分比序列相同性。并且也将所有保守性取代作为序列相同性的因素。
还可以根据FASTA检索确定两种氨基酸序列是否基本上同源[Perearson等,美国科学院院报85:2444-8,1988],另外,百分比同源性是作为与比较的序列中的相同氨基酸排比的两个序列中较小一个中的氨基酸残基中的百分比计算的,此时,可以引入长度为100个氨基酸的4个缺口,以便使排比最大化[参见Dayhoff,蛋白质序列和结构图,5卷,国家生物化学研究基金,华盛顿特区,1972,124页]。如果多核苷酸编码两种能够相互杂交的多肽的话,可将这两种多肽视为与本发明的TANK2多肽同源。如果杂交是在较大严格性的杂交条件下进行,则表明具有较高的同源程度。杂交条件的控制和杂交条件与同源性程度之间的关系为本领域技术人员所理解[例如,参见Sambrook等,同上]。因此,同源性多肽可以是由这样一种多核苷酸所编码的多肽,该多核苷酸在具有特定严格程度的杂交条件下能够与编码本发明多肽的多核苷酸杂交。
可能希望所述结构上同源的多肽同样具有功能上的同源性,以便所述同源性多肽与本发明的多肽具有基本上相同的功能。例如,如果结构上同源的多肽具有相似的配体结合或相似的免疫活性等的话,则认为在功能上是同源的。
不过,众所周知,两种多肽或两种多核苷酸在结构上可能被认为是基本上同源的,但在功能上明显不同。例如,等位基因中的单一核苷酸多态性(SNPs)可能表达为在功能上具有显著差异的多肽。伴随着一种或多种可检测的参数,如抗体或配体结合亲和力或酶促底物专一性等。其他结构差异,如取代、缺失、剪接变体等有可能影响其他结构上相同或同源的多肽的功能。
本发明的TANK2多肽包括SEQ ID NO:133或SEQ ID NO:135所示TANK2多肽的功能性衍生物。所述功能性衍生物包括基本上类似于TANK2蛋白的结构特征或生物学活性的结构特征或生物学活性的多肽产物。因此,功能性衍生物包括亲本TANK2蛋白的变体、片段和化学衍生物。
在本文中,“变体”是指在结构上和功能上基本上类似完整TANK2分子或其片段的分子。如果两种分子具有基本上相似的结构或者两种分子具有类似的生物学活性的话则将一种分子说成是“基本上类似”于另一种分子。因此,如果两种分子具有相似的活性,即可将其视为变体,正如本文所使用的术语,即使一个分子具有另一种分子所没有的结构,或者其氨基酸残基的序列不同。
本发明所提供的变体多肽是在TANK2多肽的氨基酸序列含有一个或多个改变的变体。所述基于序列的改变包括所述TANK2序列上的缺失、取代或插入,及其组合。
TANK2多肽的缺失变体是去掉了该序列上的至少一个氨基酸残基的多肽。缺失可以在该蛋白的一端或两端进行,或者去掉TANK2氨基酸序列内部的一个或多个残基。例如,缺失变体包括本发明TANK2多肽所有不完整的片段。在本文中,“片段”是指TANK2多肽的任何多肽亚类。
需要具有TANK2所特有的生物学活性并且是可溶的(即不是膜结合的)的TANK2片段。可溶性片段优选是通过缺失亲本分子的所有跨膜区或者通过缺失亲水性氨基酸残基或者用疏水性氨基酸取代亲水性氨基酸残基而制备的。所述残基的鉴定在本领域是众所周知的。
提供了取代变体,包括TANK2多肽的至少一个氨基酸被另一种残基所取代的多肽。可以产生任何取代,优选保守性取代。可以根据已经明确的规范和其他氨基酸的物理化学参数(例如,大小、形状、极性、电荷、亲结合能力、溶解度、化学活性、疏水性、亲水性或该残基的两亲性特征)及其对二级和三级蛋白结构的影响进行直接氨基酸取代。取代变体可以包括含有一个或多个保守氨基酸取代的多肽,即用另一种具有所需相似物理化学特征的氨基酸取代一种氨基酸。为了说明,规范氨基酸可以按照以下类型分组:脂族侧链 甘氨酸,丙氨酸,缬氨酸,亮氨酸,异亮氨酸芳族侧链 苯丙氨酸,酪氨酸,色氨酸脂族羟基侧链 丝氨酸,苏氨酸碱性侧链 赖氨酸,精氨酸,组氨酸酸性侧链 天冬氨酸,谷氨酸酰胺侧链 天冬酰胺,谷氨酰胺含硫的侧链 半胱氨酸,甲硫氨酸仲氨基团 脯氨酸
当需要可操作地确定TANK2分子的特征时,优选按照下面的表1进行取代。
表1
原始残基 代表性保守取代
丙氨酸 甘氨酸;丝氨酸
精氨酸 赖氨酸
天冬酰胺 谷氨酰胺;组氨酸
天冬氨酸 谷氨酸
半胱氨酸 丝氨酸
谷氨酰胺 天冬酰胺
谷氨酸 天冬氨酸
甘氨酸 丙氨酸;脯氨酸
组氨酸 天冬酰胺;谷氨酰胺
异亮氨酸 亮氨酸;缬氨酸
亮氨酸 异亮氨酸;缬氨酸
赖氨酸 精氨酸;谷氨酰胺;谷氨酸
甲硫氨酸 亮氨酸;酪氨酸;异亮氨酸
苯丙氨酸 甲硫氨酸;亮氨酸;酪氨酸
丝氨酸 苏氨酸
苏氨酸 丝氨酸
色氨酸 酪氨酸
酪氨酸 色氨酸;苯丙氨酸
缬氨酸 异亮氨酸;亮氨酸
通过筛选比表1所示更具有挑战性的取代实现功能或免疫学相同性的显著变化,即选择对保持以下特征具有更大不同影响的残基:(a)多肽主链在取代部位的结构,例如,折叠或螺旋构象;(b)该分子在靶位点的电荷或疏水性或(c)侧链的体积。一般,所述取代在以下方面更具挑战性:(a)用其他氨基酸取代甘氨酸和/或脯氨酸或将其缺失或插入;(b)用亲水性残基取代疏水性残基;(c)用半胱氨酸残基取代任何其他残基(或被其他残基所取代);(d)用具有带正电侧链的残基取代具有负电荷的残基(或者被后者所取代);或(e)用具有大的侧链的残基取代不具有所述侧链的残基(或者被后者所取代)。能影响TANK2溶解度的氨基酸取代是最优选的。这一目的最优选是通过用亲水性取代疏水性氨基酸而实现。
不过,取代变体可以包括用非规范或非天然存在的氨基酸残基取代主要序列上的氨基酸残基。取代变体包括这样的多肽,其中,氨基酸取代是通过修饰编码TANK2多肽的多核苷酸而引入的。
提供了插入变体,其中,在TANK2氨基酸序列上添加了至少一个氨基酸残基。插入可位于所述多肽的任一端或两端,或者可以位于TANK2氨基酸序列内部。插入变体还包括融合蛋白,其中,TANK2多肽的氨基或羧基末端与另一种多肽融合。所述融合蛋白的例子包括免疫原性多肽,具有长的环化半衰期的蛋白(例如,免疫球蛋白恒定区),标记蛋白(例如,绿色荧光蛋白)和有利于所需TANK2多肽纯化的蛋白或多肽(例如,FLAG标记或聚组氨酸序列)。末端插入的另一个例子是宿主细胞的异源或同源信号序列与该分子的N-末端融合,以利于该衍生物从重组宿主中分离。内部序列插入(即插入TANK2分子序列内部)通常为大约1-10个残基,更优选1-5个。
本发明的多肽变体还可以包括成熟的TANK2产物,即去掉了前导序列或信号序列的TANK2产物,以及具有附加氨基末端残基的产物。涉及在-1位置上具有附加甲硫氨酸残基的TANK2产物(Met-1-TANK2),还涉及在-2和-1位置上分别有附加甲硫氨酸和赖氨酸残基的TANK2产物(Met-2-Lys-1-TANK2)。其他这样的变体特别适用于在细菌宿主细胞中生产重组蛋白。
本发明还包括利用特殊表达系统所得到的具有附加氨基酸残基的TANK2变体。例如,使用商业化的载体,该载体能表达所需要的多肽谷桄甘肽-S-转移酶(GST)融合产物,在从所需多肽上裂解掉GST成分之后得到了在-1位置上具有附加甘氨酸残基的所需多肽(Lys-1-TANK2)。还涉及在其他载体系统中表达的变体。
本发明还提供了作为SEQ ID NO:133或SEQ ID NO:135所示TANK2多肽的化学衍生物的TANK2多肽产物。在本文中,术语“化学衍生物”是指含有正常情况下不是天然存在的分子的一部分的附加化学部分的分子。所述部分可以使衍生的分子具有所需特征,如较高的溶解度、吸收性、生物半衰期等。所述部分还可以降低衍生分子的毒性,或消除或减弱衍生分子的任何不希望的副作用。因此,TANK2多肽的化学衍生物包括具有除了氨基酸残基的插入、缺失或取代之外的(或作为其补充)的修饰的多肽。所述修饰的性质优选是共价的,并且包括,例如,与聚合物、脂类、非天然存在的氨基酸和其他有机和无机部分的化学结合。本发明的衍生物可以制备成增加TANK2多肽的循环半衰期,或者设计成改善该多肽对所需细胞、组织或器官的定向能力。
例如,对多肽进行修饰以便包括一个或多个水溶性聚合物结合体,如聚乙二醇、聚氧乙烯二醇或聚丙二醇的方法在本领域中是公知的。特别优选的是用聚乙二醇(PEG)亚基共价修饰过的TANK2产物。水溶性聚合物可以连接在特定位点,例如,连接在TANK2产物的氨基末端,或者随机连接在该多肽的一个或多个侧链上。其他衍生物包括固定在固体支持物,针状微颗粒或层析树脂上的TANK2,以及被修饰成包括一种或多种可检测标记、标志、和螯合剂等的TANK2。
利用双功能试剂衍生可以将TANK2交联到水不溶性基质上。另外,可将活性水不溶性基质,如溴化氢活化的碳水化合物和活性底物用于蛋白固定[例如,参见US3969287;3691016;4195128;4247642;4229537;和4330440]。
预计,TANK2变体的表达可应用于研究野生型TANK2多肽的生物学活性特征。可将TANK2变体设计成保留了TANK2所特有的所有生物学或免疫学特征,或者专门使其丧失TANK2的一种或多种特定生物学或免疫学特性。例如,可以设计缺失了与一种特定特征相关的结构域的片段和截短物,或者将取代和缺失设计成能使与一个特定结构域相关的特征失活。所述变体(显性负突变体)的强制表达(超量表达)可用于通过观察与该突变相关的表现型来研究该蛋白在体内的功能。
通过体外合成可以方便地制备具有大约100个残基的TANK2的功能性衍生物。如果需要,可以用本领域已知的方法修饰所述片段,这一目的是通过让能够与特定侧链或末端残基起反应的有机衍生试剂与纯化或粗制蛋白的目标氨基酸残基反应而实现的。所得到的共价衍生物可用于鉴定对生物学活性重要的残基。
还可以通过诱变编码TANK2的DNA制备具有改变了的氨基酸序列的TANK2的功能性衍生物。可将氨基酸缺失、插入和取代的任意组合用于制备最终的结构。只要最终结构具有所需活性就行。很显然,所需诱变可以在编码功能性衍生物的DNA内进行,但一定不能将该序列排出读框,并且优选不产生会形成二级mRNA结构的互补区[参见EP专利公开号75444]。
尽管在氨基酸序列上引入变异的位点是预定的,但诱变本身不一定是预定的。例如,为了优化特定位点上突变的性能,可以在目标密码子或目标区域实施随机诱变,如接头分区诱变,以便产生大量衍生物,这些衍生物随后可以表达并且筛选所需活性的最佳组合。另外,可以采用定向诱变或其他已知技术在已知序列的DNA的预定位点上产生突变。
定向诱变技术在本领域是众所周知的[例如,参见Sambrook等,同上,McPherson著,定向诱变:实践方法,IRL出版社,牛津,1991]。通过利用编码所需突变的DNA序列的特殊寡核苷酸序列进行定向诱变,可以产生TANK2功能性衍生物。定向诱变方法和材料可以从商业渠道获得,例如,由Stratagene(La Jolla,CA)出售的QuikChangeTM试剂盒。可以利用该方法选择性地产生精确的氨基酸缺失、插入或取代。氨基酸序列缺失通常为大约1-30个残基,更优选大约1-10个残基,并且通常是连续的。最优选的缺失能产生催化性片段或DNA结合片段。
为了提高TANK2的亲和力而设计的突变可以通过引入存在于其他聚(ADP-核糖)聚合酶蛋白上的同源位点上的氨基酸残基而完成。类似的,可以制备缺乏功能结构域,例如,催化结构域残基的突变型TANK2分子,以便形成显性负蛋白。
要事先预测特定修饰,例如,取代、缺失、插入等对TANK2的生物学活性所产生的确切影响是困难的。不过,本领域技术人员可以理解的是,通过常规筛选测定可以评估上述影响。例如,衍生物通常是通过编码天然TANK2分子的DNA的接头分区定向诱变产生的。然后在重组宿主中表达该衍生物,并选择性地从细胞培养物中纯化,例如,通过免疫亲和层析纯化。然后用合适的筛选测定筛选细胞裂解物或纯化衍生物活性的所需特征。例如,功能性衍生物的免疫特征的改变,如对特定抗体亲和力的改变是通过竞争型免疫测定测定的。可以通过合适的测定方法测定表达产物的其他参数的改变。
抗体
本发明提供了能专一性结合TANK2多肽的抗体。在本文中,“抗体”被广义定义为一种能与TANK2多肽所特有的表位(抗原性决定子)发生特殊的免疫反应的蛋白。在本文中,如果一种抗体能够通过该抗体的一个或多个可变区或一个或多个互补性决定区(CDRs)专一性地识别并结合抗原所特有的表位的话,就认为该抗体与诸如多肽的这种抗原“免疫反应”。
如果两种多肽各自包括形成相同特殊的表位的共同结构特征的话,能与一种特定多肽发生免疫反应的抗体可能与另一种多肽有交叉反应性。对于相关的多肽来说,交叉反应性可能存在于相关多肽之间的诸如序列相同性、同源性或相似性的共同结构特征相关。因此,多肽家族通常可以通过交叉反应抗体鉴定,即能够与拥有共同表位的多肽家族的某些或所有成员发生免疫反应的抗体。因此,本发明包括能与TANK2多肽家族的特定成员发生免疫反应的抗体,例如,所述多肽是含有SEQID NO:133或SEQ ID NO:135所示氨基酸序列的多肽。本发明包括能与TANK2多肽家族的某些或所有成员发生免疫反应的抗体。
确定一种抗体的结合专一性的筛选测定方法是在本领域是众所周知的并且被常规使用[例如,参见Harlow著,抗体:实验室手册,第6章,冷泉实验室,冷泉港,NY,1988]。一种抗体结合特定多肽抗原的免疫反应专一性有别于与其他蛋白的相互作用,例如,金色链球菌蛋白A或ELISA技术中的其他抗体,这种相互作用是通过所述抗体的除了可变区之外的部分介导的,特别是该抗体的恒定区。
例如,抗体包括,单克隆抗体、多克隆抗体、单链抗体(scFv抗体)、嵌合抗体、多功能/多专一性(例如,双功能或双专一性抗体、人源化抗体、人抗体和CDR移植抗体(包括含有能与本发明多肽发生专一性免疫反应的CDR序列的部分)。本发明的的抗体还包括抗体片段,只要它具有所需的生物学活性就行。“抗体片段”包括完整长度抗体的一部分,通常是它的抗原结合区或可变区。抗体片段的例子包括Fab、Fab’、F(ab’)2,和Fv片段;diabodies;线性抗体;单链抗体分子;和由抗体片段形成多专一性抗体。
本发明的抗体可以用本领域公知的方法生产。例如,多克隆抗体是从用所述蛋白或功能性类似物按照本领域已知方法免疫过的哺乳动物体内分离的。简单地讲,多克隆抗体可以通过将免疫原性TANK2多肽(免疫原)注射到宿主哺乳动物(例如,兔、小鼠、大鼠或山羊)体内生产的。可以使用佐剂以便增强免疫反应。从所述宿主哺乳动物体内提取血清,并筛选,以便获得对TANK2多肽有专一性(已知免疫反应)的多克隆抗体。优选单克隆抗体(本文又称之为mAbs)。在本文中,“单克隆抗体”是指从一群基本上同源的抗体中获得的抗体,即构成该群体的单个抗体是相同的,所不同的是在该群体中可能存在少量天然出现的突变。单克隆抗体对单一的抗原位点是高度专一的(单一专一性)。另外,与常规(多克隆抗体制剂)不同,多克隆抗体通常包括针对不同决定子(表位)的不同抗体,而每一种单克隆抗体是针对抗原上的单一决定子的。
修饰“单克隆”表示从基本上同源的抗体群体中获得的抗体特征,并且不被视为通过任何特殊方法生产这种抗体。单克隆抗体可以用任何能够得到能产生同源抗体的连续细胞系的任何合适技术制备。所述方法包括免疫学方法[Kohler和Milstein,自然256:495-7,1975;Campbell,单克隆抗体技术,啮齿类和人类杂交瘤生产和鉴定,Burdon等著,生物化学和分子生物学实验室技术,13卷,Elsevier科学出版社,Amsterdam,1985]或任何类似方法。单克隆抗体还可以从噬菌体抗体文库中分离[Clackson等,自然352:624-8,1991;Marks等,分子生物学杂志222:81-97,1991]。
为了说明,通过注射免疫原性TANK2多肽对宿主哺乳动物进行免疫,然后强化免疫,以便生产单克隆抗体。在最终的强化免疫之后数天,从免疫的哺乳动物体内采集脾脏。让来自脾脏的细胞悬浮液与肿瘤细胞系融合,以便形成永生化杂交细胞系或“杂交瘤”。通过限制稀释可以分离单克隆,然后测试它所产生的抗体专一性。然后生长所筛选的细胞,例如,通过腹水方法生长,以便提供所需同源抗体的连续资源。
可以用遗传学技术对抗体进行功能改造,以便产生含有来自两种或两种以上物种的蛋白成分的嵌合抗体。为了在人类受治对象的体内使用,可以将抗体“人源化”,即将其修饰成含有源于一种生物,例如,啮齿类的抗原结合区,大量的抗体被源于人类免疫球蛋白的序列所取代。在一种方法中,将诸如小鼠或兔的非人CDR插入另一种生物,例如人的框架序列内,或者插入共有框架序列内。然后可以将其他改变引入抗体框架,以便调节工程改造过的抗体的亲和性或免疫原性。用于诱导工程抗体在各种类型细胞,如哺乳动物和微生物细胞中表达的方法也是已知的。本领域公开了多种用于制备工程抗体的技术[例如,Owens和Young,免疫方法杂志168:149-65,1994]。
抗体还包括重组多克隆或单克隆Fab片段[例如,Huse等科学246:1275-81,1989]。另外,所披露的用于生产单链抗体的技术[例如,US4946778]可用于生产TANK2专一性单链抗体(例如,单链Fv片段;简写为scFv)。可以使用制备抗体的快速、大规模重组方法,如噬菌体展示或核糖体展示方法,随后选择性地进行亲和成熟[例如,参见Ouwehand等,Vox Sang74(Suppl2):223-32,1998;Rader等,美国科学院院报95:8910-5,1998;Dall’Acqua等,当代结构生物学观点8:443-50,1998]。在用于人类治疗时特别优选完整的人类抗体,但要生产这种抗体是困难的。例如,当抗原是人的半抗原时,人类通常不能对这种抗原产生任何免疫反应。业已开发了制备完整人抗体的方法,并且为本领域所公知。因此,完整人抗体可以用免疫原性TANK2多肽对一种动物(例如,小鼠)进行免疫来生产,所述动物做过转基因修饰,以便能表达免疫球蛋白基因的人的所有成分的至少主要部分[例如,参见Bruggemann等,当代免疫学17:391-7,1996]。
正如本文所指出的,本发明的宿主细胞是用于开发能与TANK2发生专一性免疫反应的抗体的有价值的来源。为了可以用做制备多克隆或单克隆抗体的免疫原,TANK2肽片段必须含有足于形成一个免疫原性表位的氨基酸残基。如果该片段太短,以至于它本身没有免疫原性,可以结合一个载体分子。例如,合适的载体分子包括匙孔槭*血蓝蛋白(KLH)和牛血清白蛋白(BSA)。结合可以通过本领域已知方法进行。所述方法之一是让该片段半胱氨酸残基与载体分子上的半胱氨酸结合。
本发明的抗体可用于治疗方法(通过调节TANK2的活性)、诊断方法(通过检测样品中的TANK2)、以及用于纯化TANK2。所述抗体特别适用于检测和/或定量细胞、组织、器官、及其裂解物和提取物、以及诸如血清、血浆、脑脊液、尿液、唾液、腹水、胸积水或支气管肺泡灌洗液中TANK2表达。还涉及含有本发明抗体的用于本文所述任何目的的试剂盒。一般,本发明的试剂盒还包括一种对照抗原,所述抗体能与它发生免疫反应,并且还可以包括其他试剂、容器和包装插入件。
另外,本发明包括中和抗体,即能明显抑制或损害本发明蛋白或功能性类似物的生物学活性的抗体。具体地讲,中和抗体能抑制或损害TANK2的聚(ADP-核糖)聚合酶活性。将中和抗体用于治疗和诊断用途是特别理想的。
功能性等同物还包括具有与完整抗体相同的结合特征或者与完整抗体相当的结合特征的抗体片段。所述片段可以包括一个或多个Fab片段或F(ab)2片段。所述抗体片段优选含有完整抗体的所有6个互补决定区(CDRs),不过,含有少于所有决定区,如三个、四个或五个CDRs的片段可能也具有功能。可以用本领域公开的方法制备片段[例如,Lamoyi等,免疫学方法杂志56:235-43,1983;Parham,免疫学杂志131:2895-902,1983]。
另外,可以用分离的或重组的TANK2产物、TANK2变体或表达所述产物的细胞开发特殊结合蛋白。结合蛋白可用于纯化TANK2产物,并用于通过已知免疫学方法检测或定量体液和组织样品中的TANK2产物。还证实了结合蛋白可用于调节(即封闭、抑制或刺激TANK2多肽的生物学活性,特别是信号传导相关的活性。因此,提供了能抑制TANK2多肽活性的中和抗体。还涉及对抗TANK2抗体专一的抗独特型抗体。
可检测的多核苷酸和多肽探针
本发明还提供了一种检测样品中TANK2编码多核苷酸或TANK2多肽存在的方法。该方法包括使用能识别样品中特定目标存在的标记探针。所述探针可以是能识别TANK2多肽的抗体,或者是能识别编码TANK2多肽的多核苷酸的寡核苷酸。
本发明的探针可以用本领域已知方法进行可检测的标记。一般,可以通过在探针上连接可检测标记(报导基因)部分对该探针进行修饰,或者以在它上面整合有可检测标记部分的形式生产可检测探针。所述可检测的标记部分可以是任何可检测的部分,很多可检测的部分为本领域所公知,包括放射性原子、电子密集原子、酶、色原和有色化合物,荧光原和荧光化合物,以及特殊结合对的成员等。
现有技术中业已披露了对寡核苷酸探针进行标记的方法[例如,参见Leary等,美国科学院院报80:4045-49;1983;Renz和Kurz,核酸研究12:3435-44,1984;Richardson和Gumport,核酸研究11:6167-84,1983;Smith等,核酸研究13:2399-412,1985;Meinkoth和Wahl,分析生物化学138:267-84,1984;和US4711955;4687732;5241060;5244787;5328824;5580990和5714327]。
还披露用于标记抗体的方法[例如,参见Hunter等,自然144:495-6,1962;David等,生物化学13:1014-21,1974;和US3940475和3645090]。
所述标记部分可以是放射性的。有用的放射性标记的某些例子包括32P、125I、131I和3H。业已披露了放射性标记的使用[例如,UK专利文件2034323和US4358535和4302204]。
非放射性标记的某些例子包括酶、色原、可以通过电子显微镜检测的原子和分子,以及可以通过其磁性检测的金属离子。
某些有用的酶促标记包括能导致底物发生可检测的变化的酶。例如,某些有用的酶(及其底物)包括辣根过氧化物酶(pyrogallol和o-苯乙二胺)、β-半乳糖苷酶(荧光素β-D-半乳糖吡喃糖苷)和碱性磷酸酶(5-溴-4-氯-3-吲哚-磷酸-氮兰四唑)。本领域业已披露了酶促标记的使用[例如,参见UK专利文件2019404,欧洲专利文件EP63879和Rotman,美国科学院院报47:1981-91,1961]。
有用的报导部分报导,例如,荧光分子、磷光分子、化学发光分子和生物发光分子,以及染料。例如,可用于本发明的某些特殊有色或荧光化合物包括荧光素、香豆素、罗丹明、Tesas红、藻红蛋白、7-羟基香豆素、和鲁米洛等。还可将色原或荧光原,即能够被修饰(例如,氧化)成有色或有荧光或者改变其颜色或发色光谱的分子结合到探针中,以便在特殊条件下起着报导部分的作用。
标记部分可以通过本领域众所周知的方法结合到探针上。探针部分可以通过探针上的功能团直接连接。所述探针含有这样的功能团或者使其含有这样的功能团。例如,合适的功能团的某些例子包括氨基、羧基、硫氢基、马来酰亚胺、异氰酸、异硫氰酸。
另外,可将诸如酶和色原的标记部分通过偶合剂连接到抗体或核苷酸上,如二醛、碳二亚胺、和二马来酰亚胺等。
可以通过连接在探针上的配体将标记部分连接在该探针上,所述配体是通过上述方法和连接标记部分的配体的受体连接在探针上的。任何已知的配体-受体结合对组合都适用。例如,某些合适的配体-受体对包括生物素-亲和素或-链霉素,和抗体-抗原。优选生物素-链霉亲和素组合。
使用tamkyrase2多核苷酸和多肽的方法
证实了由于披露本发明的DNA和氨基酸序列的信息所产生的科学价值。作为一系列的例子,对编码tank2的cDNA序列的了解可以通过使用Southern杂交或聚合酶链式反应鉴定编码TANK2的基因组DNA序列和TANK2表达控制调控序列。预计,在中等至高严格条件下用本发明的DNA序列进行的DNA-DNA杂交方法还可以分离编码TANK2的等位变体的DNA。类似的,还可以通过Southern和/或PCR分子鉴定编码与TANK2同源的蛋白的非人基因。作为一种替代方案,互补性研究可用于鉴定其他人类TANK2产物以及非人蛋白,编码所述蛋白的DNA,拥有TANK2的一种或多种生物学特征。本发明的寡核苷酸还可用于杂交测定方法,用来鉴定细胞表达TANK2的能力。本发明的多核苷酸还可用做诊断方法的基础,用于确定tank2基因做上的遗传学改变,这种改变是疾病状态的基础。例如,TANK2-LONG和TANK2-SHORT的表达或活性的差别可能与特殊的疾病状态相关,使得这两种形式的TANK2的一种或两种适合作为诊断标记或作为本文所披露的治疗目标。因此,选择性的试剂,例如,能够与一种形式tank2选择性杂交的寡核苷酸或能够与一种形式的TANK2选择性免疫反应的抗体是特别有用的。
正如本文所披露的,本发明的寡核苷酸可用于为了各种目的而扩增DNA的方法中。本发明方法的“扩增”是指通过指数扩增模板核酸序列检测特定核酸序列的微量水平的任何分子生物学技术。具体地讲,合适的扩增技术包括诸如PCR、连接酶链式反应(LCR)及其变体的技术。已知PCR是高度灵敏的技术,并且具有广泛的用途[例如,参见Innis等,PCR方法:方法和应用指南,学术出版社公司,San Diego,1990;Dieffenbach和Dveksler,PCR引物:实验室手册,冷泉港实验室出版社,Plainview NY,1995;和US4683195;4800195;4965188]。已知最近开发的LCR技术是高度专一的,并且能够检测点突变[例如,参见Landegren等,科学241:1077-80,1988和Barany等,PCR方法和扩增1:5-16,1991]。LCR试剂盒可以从Stratagene获得。在某些场合下,需要将PCR和LCR技术结合在一起,以便提高检测的准确性。在本发明中可以采用其他扩增技术。
寡核苷酸扩增引物通常是以匹配的单链寡核苷酸对形式提供;其中的一条链具有正义方向(5’→3’)而另一个为反义方向(3’→5’)。所述特殊的引物对可以在优化条件下使用,用于鉴定特定的基因或条件。另外,可以在严格性较低的条件下使用相同的引物对、寡聚体的嵌套组、甚至是寡聚体的简并库,用于检测和/或定量密切相关的DNA或RNA序列。
所述寡核苷酸可用于本领域已知的各种方法中,用于延长特定的核苷酸序列。所述方法可以使用已知的序列测定未知的相邻序列,从而能够检测并测定诸如启动子和调控因子的上游序列。
例如,限制位点聚合酶链式反应是利用通用引物获得靠近已知基因座的未知序列的直接方法[例如,参见Gobinda等,PCR方法应用2:318-22,1993]。在该方法中,首先在存在对连接序列专一的引物和对已知片段专一的引物的条件下扩增基因组DNA。然后用相同的接头引物和另一种作为第一种引物的内部引物的特殊引物对扩增的序列进行第二轮PCR。用合适的RNA聚合酶对每一轮PCR的产物进行转录,并用逆转录酶测序。
可以利用基于已知片段的差异引物通过反PCR扩增或延伸序列[Triglia等,核酸研究16:8186,1988]。可以用Oligo4.0(国家生物科学公司,Plymouth,MN)和另一种合适的程序将引物设计成程度为22-30个核苷酸,GC含量为50%或更高,并且在大约68-72℃的温度下与目标序列退火。该方法使用若干种限制酶产生一种基因的已知区域的合适片段。然后通过分子间连接将该片段环化,并用做PCR模板。
捕获PCR是一种用于PCR扩增人和酵母人工染色体(YAC)DNA上的靠近已知序列的DNA片段的方法[Lagerstrom等,PCR方法应用1:111-9,1991]。捕获PCR还需要多种限制酶消化和连接,在PCR之前将工程产生的双链序列插入该DNA分子的未知部分。步行PCR是一种定向基因步行的方法,它能够获得未知序列[Parker等,核酸研究19:3055-60,1991]。PromoterFinderTM试剂盒(Clontech,Palo Alto,CA)使用PCR、嵌套引物、和特殊文库在基因组DNA内“步行”。该方法没有必要筛选文库,并且可用于发现内含子/外显子接头。
所述方法可用于研究基因组文库,以便延伸5’序列并获得内源tank2基因组序列,包括诸如启动子、内含子、操纵子、增强子和抑制子的因子。用于筛选完整长度cDNA的优选文库是已经做过大小筛选的包括较大cDNA的文库。另外,优选随机启动的文库,因为它含有更多的包括基因的5’和上游区的序列。
所述寡核苷酸探针还可用于对内源基因组序列进行作图。可以用已知技术将所述序列作图于特定染色体上或作图于该染色体的特定部分上。其中包括与染色体展开样品的原位杂交[Venna等,人类染色体:基础技术手册,Pergamon出版社,NY,1988],流式分选染色体制剂,或人工染色体结构,如YACs细菌人工染色体(BACs),细菌P1结构或单一染色体cDNA文库。
染色体制剂杂交和使用已确定的染色体标记进行物理作图的技术,如连锁分析在扩展遗传学图谱方面是无价的。在本领域可以找到遗传学图谱的例子[例如,Hodgkin等,科学270:410-4,1995和Murray等,科学265:2049-54,1994]。通常,确定一个基因在另一种哺乳动物的染色体上的位置可以发现相关的标记,即使特定人类染色体的数量或臂是未知的。通过物理作图可以确定所述序列赋予染色体的特殊结构特征。这样可以为研究人员利用定位克隆或其他基因发现技术寻找致病基因提供有价值的信息。一旦通过遗传学连锁将一种疾病或综合征大致定位于一个特定的基因组区间上,作图于该部位的任何序列都可能相关或者对调控基因作进一步的研究。例如,参见Gatti等,自然336:577-80,1988。本发明的多核苷酸可用于在正常的、载体或受感染的个体之间检测由于转位、颠换等所导致的染色体定位的差别。还可以开发其他类型的遗传学图谱,例如,基于序列标记位点(STS)的基因组的物理图谱[例如,参见Hudson等,科学270:1945-54,1995]。
本发明所提供的DNA序列信息还可用于通过同源重组或“剔除”方法[Capecchi,科学244:1288-92,1989]开发不能表达有功能的TANK2或表达TANK2的变体的动物。所述动物可以用做研究TANK2及其调节物的体内活性的模型。
如本文所述,本发明提供了能识别并且与编码TANK2的多核苷酸杂交的反义核酸序列。通过设计针对tank2基因的调控区,如启动子、增强子和内含子的反义序列可以实现对基因表达的修饰。优选源于转录起始位点的寡核苷酸,例如位于前导序列的-10和+10之间。还可以设计反义RNA和DNA分子,通过抑制转录物与核糖体结合抑制mRNA的翻译。普通技术人员可以理解的是,本发明的反义分子包括能专一性地识别并与tank2 DNA杂交的分子(通过对tank2 DNA和编码其他已知分子的DNA进行序列比较测定)。本发明的反义分子还包括能识别并与编码TANK2蛋白家族的其他成员的DNA杂交的分子。还可以通过序列比较鉴定能够与编码TANK2蛋白家族的其他成员的多种DNA杂交的反义多核苷酸,以便确定TANK2蛋白家族的特有或标记序列。因此,所述反义分子与目标tank2序列的相同性优选为至少95%,更优选至少98%,更优选至少99%。
反义多核苷酸与通过表达tank2 mRNA的细胞调控TANK2的表达有特别密切的关系。反义多核苷酸(优选10-20bp寡核苷酸)能够专一性地结合tank表达控制序列或将tank2RNA导入细胞,例如,通过病毒载体或诸如脂质体的胶体分散系统导入。所述反义寡核苷酸在细胞中结合tank2靶核苷酸序列,并抑制靶序列的转录或翻译。硫代磷酸酯和甲基磷酸酯反义寡核苷酸特别适用于本发明的治疗用途。还可以通过位于其5’末端的聚-L-赖氨酸、运铁蛋白聚赖氨酸或胆甾醇部分对反义寡核苷酸作进一步的修饰[有关反义技术的最新综述可以参见Delihas等,自然生物技术15:751-3,1997]。
本发明还包括通过核糖酶技术调控TANK2表达的方法[有关综述可以参见Gibson和Shillitoe,分子生物技术7:125-37,1997]。可以用核糖酶技术以序列专一性方式抑制tank2mRNA的翻译。这一目的是通过(i)互补RNA与目标mRNA的杂交和(ii)通过互补RNA所固有的内切核酸酶活性裂解杂交的mRNA而实现的。核糖酶可以通过经验方法确定,如通过核糖核酸酶保护测定用互补寡核苷酸确定,但更优选的是根据对能接触核糖酶裂解位点的靶分子的扫描专门设计[Bramlage等,生物技术趋势16:434-8,1998]。可以用本领域所公知并且采用的外源或内源输送技术将核糖酶输送到靶细胞。外源技术可以包括使用定向脂质体或直接局部注射。内源方法包括使用病毒载体和非病毒质粒。
当核糖酶被设计成互补于编码TANK2的多核苷酸的特定区域时,它可以专一性调节TANK2的表达。因此,“专一性地调节”是指本发明的核糖酶只能识别编码TANK2的多核苷酸。类似地,可将核糖酶设计成能调节TANK2蛋白家族的所有或部分成员的表达。将这种类型的核糖酶设计成能识别在编码TANK2家族成员的多核苷酸的全部或某些之间保守的核苷酸序列。
本发明还包括通过使用寡核苷酸定向三螺旋形成调节tank2转录的方法(又被称为Hogeboom碱基配对方法[有关综述可以参见Lavrovsky等,生物化学分子医学62:11-22,1997]。三螺旋的形成是通过使用能够与大沟中的双链DNA杂交序列专一性寡核苷酸实现的,所述大沟是按照Watson-Crick模型定义的。这种三螺旋杂交破坏了原始双螺旋充分展开以便结合聚合酶、转录因子或调控因子的能力。用于杂交的优选靶细胞包括启动子和增强子序列,以便可以进行TANK2表达的转录调控。还可以将能够形成三螺旋的寡核苷酸连接在DNA损伤试剂上,然后将其用于对靶DNA序列进行位点专一性共价修饰[参见Lavrovsky等,同上]。
本发明的反义RNA和DNA分子以及核糖酶可以用本领域所公知的用于合成RNA分子的任何方法制备。其中包括用于化学合成寡核苷酸的技术,如固相亚磷酰胺化学合成方法。另外,RNA分子还可以通过编码反义RNA分子的DNA序列的体外或体内转录制备。所述DNA序列可以插入具有诸如T7或SP6的合适的RNA聚合酶启动子的多种载体上。另外,可将能组成性地或诱导型地合成反义RNA的反义cDNA结构导入细胞系、细胞或组织。
发生在一个基因上的会导致基因产物的正常功能丧失的突变可能是TANK2相关疾病状态的原因。本发明包括恢复TANK2活性的基因治疗,即治疗以缺少或缺乏与TANK2酶相关的聚(ADP-核糖)聚合酶活性的疾病状态。将有功能的tank2基因输送到合适的细胞中是利用载体,特别是病毒载体(例如,腺病毒、腺相关病毒或逆转录病毒)通过自体内、原位、或体内方式进行的,或者通过利用物理DNA转移方法(例如,脂质体或化学处理)自体内进行的[例如,参见Anderson,自然392(6679Suppl):25-30,1998]。另外,对于其他疾病状态来说,抑制TANK2表达或抑制其活性可用于治疗这种疾病。反义治疗或基因治疗可用于对TANK2的表达进行负调控。
由本发明所提供的DNA和氨基酸信息还可用于TANK2蛋白结构和功能小系统分析。TANK2的DNA和氨基酸序列信息还可用于鉴定能与TANK2多肽相互作用的分子。通过将一种推测的TANK2调节物一起培养,并测定推测的调节物对TANK2活性的影响,可以鉴定能够调节(即增强、减弱或抑制)TANK2活性的试剂。通过比较一种化合物对TANK2的活性以及它对其他蛋白的活性,可以评估这种化合物调节TANK2多肽活性的选择性。
通过采用本发明的TANK2多肽或tank2多核苷酸可以对多种方法进行改进,包括基于细胞的方法,如用于检测结合配偶体的双杂交和三杂交筛选,以及用于检测能分解结合配偶体结合的分解杂合体筛选。其他方法包括体外方法,如将TANK2多肽、tank2多核苷酸或其结合配偶体固定的测定方法,以及溶液测定方法,这些方法都属于本发明范围。上述方法可以用一种通用方法加以说明,该方法包括以下步骤:让TANK2多肽与一种推测的结合配偶体化合物接触,检测或测定TANK2多肽与该化合物的结合,以及选择性地分离和/或鉴定结合配偶体化合物。
基于细胞的测定方法包括筛选基因组DNA或cDNA文库,以便鉴定TANK2多肽的结合配偶体的方法。代表性的方法包括双杂交或三杂交筛选[Fields和Song,自然340:245-6,1989;Fields,方法:酶学方法手册5:116-24,1993],所述方法可用于鉴定编码结合配偶体的DNA。披露了双杂交测定方法的改进和改良[Colas和Brent,生物技术趋势16:355-63,1998]。还可以采用三杂交筛选[Fuller等,生物技术25:85-8,90-2,1998]。
本发明的基于细胞的方法还可用于鉴定由TANK2生物学活性介导的生物学途径的成分,在一方面,该方法是在含有可溶形式的TANK2多肽以及可溶形式的它的结合配偶体的宿主细胞中进行的,其中,通过测定宿主细胞中与一种报导基因产物的表达改变相关的取决于结合的表型变化对结合的减弱或增强进行定量。
另外,用于鉴定TANK2多肽的抑制剂与已知结合配偶体的相互作用的基于细胞的测定方法,可以以诸如分解杂合体测定[PCT专利申请WO98/13502]及其改进[PCT专利公开WO95/20652]的方法为基础。
体外方法可以包括以下步骤:(a)让固定化的TANK2多肽与一种候选结合配偶体化合物接触,和(b)检测该候选化合物与TANK2多肽的结合。在另一种实施方案中,候选结合配偶体化合物是固定化的,而检测TANK2多肽的结合。固定化可以用本领域已知的任何方法完成,包括与支持物、小球、或层析树脂的结合,以及高亲和力相互作用,如抗体结合,或者采用亲和素:生物素类型的系统。例如,配体结合的检测可以这样进行:(i)在没有固定化的配体上采用一种可检测的(例如,放射性或荧光)标记,(ii)使用对非固定化配体具有免疫专一性到抗体,(iii)在非固定化的配体上使用一种标记,该标记能促进固定化配体与之结合的荧光支持物激发,以及本领域所常用的其他技术。
在溶液测定中,本发明的方法包括以下步骤:(a)让TANK2多肽与一种或多种候选配偶体化合物接触,和(b)鉴定与TANK2多肽结合的化合物。结合TANK2的化合物的鉴定可以通过以下方式完成:分离TANK2:结合配偶体复合体,以及从分离结合配偶体化合物中分离TANK2多肽。本发明还包括一个鉴定结合配偶体化合物的物理、生物学、或生物化学特征的步骤。在一种方法中,TANK2:结合配偶体复合体的分离是用第二种结合配偶体化合物(例如,抗体或其他蛋白)完成的,这种化合物能够与复合体中的任一种主要配体相互作用。
例如,选择性地调节物包括能选择性地或专一性地结合TANK2多肽或TANK2编码多核苷酸的抗体和其他蛋白或肽,能选择性地或专一性地结合TANK2多肽或TANK2编码多核苷酸的寡核苷酸,以及能够选择性地或专一性地与TANK2多肽或与TANK2编码多核苷酸起反应的其他非肽化合物(例如,分离的或合成的有机分子)。调节物还包括上述化合物,但它能与TANK2多肽的专一性结合配偶体相互作用。本发明还涉及TANK2的突变形式,如能影响野生型TANK2的生物学活性或细胞定位的突变形式。例如,本发明所优选的用于开发选择性调节物的目标包括:
(1)TANK2多肽的接触其他蛋白和/或TANK2定位于细胞内,例如,定位于端粒的细胞质或跨膜区;
(2)TANK2多肽的能结合专一性结合配偶体的胞外区;
(3)TANK2多肽的能结合底物,即ADP-核糖的区域;
(4)TANK2多肽的别构调控位点;
(5)TANK2多肽的能介导多聚化的区域;
(6)TANK2或其他蛋白(TRF1或TRF2)的起着受体ADP核糖化作用的区域。
其他的选择性调节物包括能识别,特别是调控TANK2编码核苷酸序列的化合物。TANK2活性的选择性和专一性调节具有治疗用途,可用于治疗多种疾病和生理学症状,其中,与这些症状TANK2活性的异常相关。TANK2编码多核苷酸序列可用于诊断由于TANK2表达或活性所导致的或者与之相关的疾病。例如,编码TANK2多肽(例如,TANK2-LONG或TANK2-SHORT)的多核苷酸序列可用于生物学样品的杂交或PCR测定,例如,来自活组织或尸体组织的样品或提取物或液体或组织,以便检测TANK2表达的异常性。所述定向或定量方法可以包括Southern或Northern分析,斑点印迹或其他基于膜的技术;PCR技术;量杆、针或芯片技术;以及ELISA或其他多样品方案技术。这种类型的技术为本领域所公知,并且业已以商业化诊断试剂盒形式采用。
可以对所述测定方法加以改进,以便评估特殊治疗方案的效力,并可用于动物研究,临床实验,或用于监测个体患者的治疗。为了提供疾病诊断的基础,必须建立TANK2表达的正常或标准曲线。这一目的是通过在适合杂交或扩增的条件下将从正常对象体内采集的生物学样品与tank2多核苷酸混合而实现的。通过将所获得的正常对象的值与在相同实验中所做的阳性对照的稀释系列进行比较可以对标准杂交进行定量,在所述对照实验中,使用已知量的纯化tank2多核苷酸。可将来自正常样品的标准值与获自有可能患有TANK2表达相关的失调或疾病的对象的样品所获得的值进行比较。标准值和对象值之间的差异可以确立疾病状态的存在。如果确定了疾病的存在,就服用一种现有的治疗剂,并制备治疗曲线或值,该测定可以按照一定的规则重复进行,以便评估所述值是朝向正常或标准形式发展或回到正常或标准形式。连续的治疗曲线可用于表现在为期数天或数月的时间内治疗的效力。
抗TANK2抗体可用于诊断以TANK2多肽的异常表达为特征或与之相关的症状、失调或疾病。TANK2多肽的诊断测定包括采用标记过的抗体检测诸如体液、细胞、组织、切片或所述材料的提取物的生物学样品中TANK2多肽的方法。可以使用修饰过的或未修饰过的本发明的多肽和抗体。优选通过将所述多肽或抗体与本文所披露的可检测的标记部分共价或非共价连接对其进行标记。
本发明可以采用的用于检测生物学样品中TANK2多肽存在的基于抗体的方法包括差异检测TANK2-LONG与TANK2-SHORT的测定方法。以前已经披露了用于检测蛋白与抗体存在的测定方法,并且遵循已知方案,如酶联免疫吸附测定(ELISA)、放射性免疫测定(RIA)和荧光活性的细胞分选(FACS)和流式细胞测定,Western印迹、和三明知测定等。通常,上述方法的基础是:将一种抗体与被怀疑含有TANK2蛋白的样品一起培养,并检测该抗体和蛋白之间的复合体的存在。在所述培养步骤之前、期间或之后对抗体进行标记。抗体的具体浓度、培养温度和时间、以及其他诸如此类的测定条件可以根据包括样品中抗原的浓度、样品的性质等在内的各种因素而变化。本领域技术人员可以通过采用常规实验来确定每一种测定的工作和最佳测定条件[例如,参见Hampton等,血清学方法:实验室手册,APS出版社,Sd PaulMN,1990]。为了提供定量样品中TANK2蛋白或诊断疾病的基础,必须建立TANK2多肽表达的正常或标准值。这一目的是通过取自正常样品或取自正常对象(动物或人)的体液或细胞提取物与TANK2多肽的抗体混合而实现的。标准复合体的形成量可以通过将其与阳性对照的稀释系列比较而进行定量,其中,将已知量的抗体与已知浓度的纯化TANK2多肽混合。然后,可将从正常样品获得的标准值与从测试样品,例如,有可能患有与TANK2表达相关的失调或疾病的样品所获得的值进行比较。标准值和测定值之间的差异可以确定疾病状态的存在。
鉴定TANK2活性调节物的方法
TANK2蛋白及其具有生物学活性的片段可用于在多种药物筛选技术的任一种中筛选推测的调节化合物。本文所使用的术语“调节物”是指起着TANK2活性的兴奋剂或拮抗剂作用的化合物。本发明的调节物包括活性的别构调节物以及活性的抑制剂。TANK2的“兴奋剂”是能增强或提高TANK2执行其任何生物学功能的能力。所述兴奋剂的一个例子是能提高TANK2结合受损的DNA或聚合ADP-核糖的能力的试剂。TANK2的“拮抗剂”是能减弱或破坏TANK2执行其任何生物学功能的能力的化合物。所述拮抗剂的一个例子是抗TANK2抗体。因此,本发明提供了用于筛选对TANK2多肽有专一性结合亲和力的多种实验化合物的方法,包括提供多种实验化合物;在合适的调节让TANK2多肽与所述多种实验化合物混合足于使其结合的时间;并检测TANK2多肽与所述多种实验化合物的每一种的结合,从而确定能专一性地结合TANK2多肽的实验化合物。
本发明还提供了鉴定TANK2多肽的生物学活性调节物的方法,包括以下步骤:a)让所述化合物与TANK2多肽结合,b)在适合于其生物学活性的条件下将步骤a)的混合物与底物一起培养,c)测定所述生物学活性的量,和d)将步骤c)的生物学活性量与用不使用该化合物培养的TANK2多肽获得的生物学活性的量进行比较,从而确定该化合物是促进或是抑制所述生物学活性。在该方法的一种实施方案中,所述TANK2多肽是从TANK2的非裂解区分离的,并且提供了鉴定TANK2的别构调节物的方法。在另一种实施方案中,TANK2多肽是从TANK2的裂解区分离的,并且提供了一种鉴定所述生物学活性的抑制剂的方法。可以使用TANK2-LONG和TANK2-SHORT多肽或其特定片段。
因此,用于上述方法的多肽可以在溶液是游离的,固定在固体支持物上,展示在细胞表面上,或者位于细胞内。可以测定活性的调节或TANK2多肽与测试试剂之间的测试复合体的形成。TANK2多肽可以按照本领域所公知的和采用的方法用于生物化学或基于细胞的高处理量筛选(HTS)测定方法,包括用于研究受体-配体相互作用的黑素细胞测定系统,基于酵母的测定系统和哺乳动物细胞表达系统[有关综述可以参见Jayawickreme和Kost,当代生物技术观点8:629-34,1997]。还包括自动化和微型化HTS测定[例如,Houston和Banks,当代生物技术观点8:734-40,1997]。
将所述HTS测定用于筛选化合物文库,以便确定具有所需特征的特定化合物。任何化合物文库都可以使用,包括化学文库、天然产物文库、含有随机的或设计的寡肽、寡核苷酸、或其他有机化合物的组合文库。
化学文库可以含有已知化合物,已知化合物的专有结构类似物,或通过天然产物筛选鉴定的化合物。
天然产物文库从天然来源,通常为微生物、动物、植物、海洋生物中分离的材料的总称。天然产物是通过以下方法从其来源中分离的:对微生物进行发酵,然后从发酵液中分离和提取或者直接从微生物或组织(植物或动物)中提取。天然产物文库包括聚酮化合物、非核糖肽及其变体(包括非天然存在的变体)[有关综述可以参见Cane等,科学282:63-8,1998]。组成文库是由大量的相关化合物组成的混合物,如肽、核苷酸或其他有机化合物。所述化合物可以通过传统自动化合成方法、BCR、克隆或专用合成方法比较简单地设计和制备。特别感兴趣的是肽和寡核苷酸的组合文库。
其他感兴趣的文库包括肽、蛋白、肽模拟物、多重合成综合,重组、和多肽文库[有关组合化学以及由此产生的文库的综述可以参见Myers,生物技术当代观点8:701-7,1997]。
但鉴定化合物具有作为TANK2功能调节物的活性,就可以采取优化方案,以便改善该活性的效力和/或选择性。这种结构-活性关系(SAR)的分析包括对化合物结构及其与生物化学或生物学活性的相关性的一系列重复的选择性修饰。可以将相关化合物家族设计成均具有所需活性,使该家族的某些成员可潜在的定性为治疗候选物。
本发明还包括竞争性药物筛选测定的使用,其中,能够专一性结合TANK2多肽的中和抗体与一种测试化合物竞争结合TANK2多肽。以这种方式可将所述抗体用于检测任何化合物的存在,例如,拥有与TANK2多肽相同的一个或多个抗原性决定子的另一种肽。
TANK2编码多核苷酸和TANK2多肽的治疗用途
本发明提供了一种用于在人体上或其他动物上治疗性地或预防性地抑制TANK2表达或活性的方法。该方法包括以能有效抑制TANK2表达或活性的量服用TANK2拮抗剂。因此,本发明提供了一种用于治疗由于细胞损伤所导致的组织损伤或由于坏死或程序死亡导致的死亡的方法,包括给所述动物服用能抑制TANK2活性的有效量的化合物。该方法可用于治疗这样的动物,该动物是或者可能患有其症状或病理学是由TANK2表达或活性介导的任何疾病。对TANK2-LONG或TANK2-SHORT具有专一性的拮抗剂特别适用于其病理或症状是由特殊形式的TANK2介导的疾病。
所述方法还包括服用另一种聚(ADP-核糖)聚合酶活性,如酶PARP、坦科聚合酶1等相关的活性的拮抗剂。例如,适用于本实施方案的代表性的PARP拮抗剂包括Banasik等所披露的化合物[生物化学杂志267:1569-75,1992]。其他代表性化合物包括披露于PCT专利公开号WO99/11623和WO99/11649中的化合物。另外,所述TANK2抑制方法可能需要使用能同时拮抗TANK2和另一种具有聚(ADP-核糖)聚合酶活性的酶的化合物。
本文所使用的“治疗”是指预防一种动物发病,这种动物可能有发生这种疾病的倾向,但尚未诊断出具有这种病;抑制所述疾病,即抑制其发展;解除该疾病,即导致其消退,或缓解该疾病,即减轻与该疾病相关的症状的严重程度。“疾病”包括医学失调、疾病、症状、和综合征等没有限制。
本发明的方法包括治疗表达TANK2的动物的各种模式,其中,可以治疗TANK2介导的失调。可以用本发明治疗的动物包括哺乳动物(包括人)和非哺乳动物,例如,禽类、鱼、爬行类和两栖动物。可以治疗的非人哺乳动物包括伴侣动物(宠物),包括狗、猫;农场动物,包括牛、马、绵羊、猪和山羊;实验室动物,包括大鼠、小鼠、兔、豚鼠和灵长类动物。该方法最优选用于治疗人类的由TANK2介导的疾病。
具体地讲,本发明的方法可用于治疗性地或预防性地治疗患有或有可能患有与过量的或不希望的端粒酶活性相关的疾病的动物。本发明的一个方面源于TANK2及其功能性衍生物与受损的DNA相互作用并调节端粒重复结合因子(例如,TRF1和TRF2)的活性的能力。
业已证实,细胞中过高的端粒酶活性与明显的细胞无限繁殖的能力的诱导相关。另外,有证据表明肿瘤组织中端粒酶的活性比大多数正常组织中的活性高,这表明较高的端粒酶活性可能是肿瘤生长所必需的。因此,本发明提供了一种抑制致癌转化或抑制肿瘤组织生长,例如,动物的癌症的方法,包括给所述动物服用有效量的能抑制TANK2活性的化合物。在该实施方案中,所述方法还包括辅助服用化疗或抗癌药物和/或放射治疗。
肿瘤或赘生物包括组织的新的生长,其中,细胞的增殖不受控制并且是侵入性的。这种生长的某些是良性的,但另一些被称为“恶性的”,会导致生物死亡。恶性肿瘤或“癌”与良性生长的区别在于,除了表现出侵入性细胞繁殖之外,癌还会侵入周围组织并转移。另外,恶性肿瘤的特征是其表现出分化的较大的丧失(较大的“脱分化”),以及它们彼此之间和相对周围组织的组构。这种特征被称为“退行发育”。
可以通过本发明治疗的肿瘤包括实体瘤,即癌和肉瘤。癌包括由上皮细胞产生的恶性肿瘤,这种肿瘤偏向于侵入周围的组织,并发生转移。腺癌是由腺组织产生的癌或者其中的肿瘤细胞形成可以识别的腺结构。另一种较大类型的癌症包括肉瘤,它是这样的肿瘤,其细胞被包埋在纤维状或同源物质中,如胚性结缔组织。本发明还能够治疗骨髓或淋巴系统的癌症,包括白血病、淋巴瘤和其他癌症,所述其他癌症通常不是以肿块存在,而是分布于血管或淋巴网状细胞系统中。
例如,可以用本发明治疗的癌症或肿瘤包括:ACTH生成肿瘤,急性淋巴细胞性白血病、急性非淋巴细胞性白血病、肾上腺皮质癌、膀胱癌、脑癌、乳腺癌、宫颈癌、慢性淋巴细胞性白血病、慢性髓细胞性白血病、结肠直肠癌、皮肤T细胞淋巴瘤,子宫内膜癌、食道癌、Ewing’s肉瘤、胆囊癌、毛细胞白血病、头颈癌、Hodgkin’s淋巴瘤、Kaposi’s肉瘤、肾癌、肝癌、肺癌、小细胞和非小细胞),恶性腹膜渗出、恶性胸膜渗出、黑素瘤、间皮瘤、多发性骨髓瘤、成神经细胞瘤、神经胶质瘤、非Hodgkin’s淋巴瘤、骨肉瘤、卵巢癌、卵巢(生殖细胞)癌、胰腺癌、阴茎癌、前列腺癌、成视网膜细胞瘤、皮肤癌、软组织肉瘤、鳞状细胞癌、胃癌、睾丸癌、甲状腺癌、滋养层肿瘤、子宫癌、阴道癌、外阴癌和Wilm’s肿瘤。
如上文所述,端粒结构的调控似乎与老化有关。预计,能调节端粒结构的调控的药物可用于治疗与衰老相关的综合征或用于治疗遗传决定的早衰和早老综合征,例如,早老(Hutchinson-Gilford早老综合征),Werner’s综合征、和其他诸如此类的疾病。因此,本发明提供了提高患有上述综合征的动物体内TANK2活性的方法。预计该方法能减弱TRF与端粒的结合,从而促进提高端粒酶活性。
当端粒缩短到超过临界长度时会导致很多类型细胞的衰老的诱导。因此端粒酶的活性通常是维持端粒长度所必需的,并且,由于TANK2抑制可能减弱端粒酶功能,本发明提供了对非肿瘤增殖性疾病的治疗,其中,可将TANK2拮抗剂用于诱导缩短的端粒和细胞衰老。增殖性疾病包括,但不限于andrestenosis、糖尿病性视网膜病、肾小球增殖病、增殖性肾小球性肾炎、红细胞增多症、骨髓纤维化、移植后淋巴增殖病、子宫内膜异位、颅缝早闭、免疫增殖性小肠病、胸腺淋巴增殖病、脊髓发育不良病、骨髓及外骨髓增殖病、von Willebrand’s病、和增殖性肾炎。
另外,TANK2抑制剂可用于所有炎性疾病,包括自身免疫病,其中,淋巴细胞的增殖起作用。本文所说的“炎性疾病”可以指任何疾病、失调或综合征,其中,过度的或失调的炎性反应会导致过度的炎性症状,宿主组织损伤或组织功能的丧失。
“炎性疾病”还可以指由输入白细胞和/或嗜中性白细胞趋化性所介导的病理学状态。
本文所使用的“发炎”是指由于组织的损伤或破坏所引起的局部的保护性反应,它起着杀伤、稀释或屏蔽(隔离)有害试剂和受损伤的组织的作用。发炎明显与白细胞和/或嗜中性白细胞趋化性的输入相关。发炎可能是由于感染致病性生物和病理所导致的,以及由于由非感染方式所导致的,如创伤或在心肌梗塞或中风之后的再灌注、对外源抗原的免疫反应、和自身免疫反应。本发明所适用的炎性疾病包括与专一防御系统的反应相关的疾病以及与非专一防御系统的反应相关的疾病。
因此,本发明提供了治疗诸如关节病的炎性疾病的方法,如类风湿关节炎,骨关节炎,痛风关节炎,脊椎炎;Behcet病;脓毒症,脓度性休克,内毒性休克,革兰氏阴性脓度症,革兰氏阳性脓度症,以及中毒性休克综合征;继之以白血症的多器官损伤综合征,创伤,或出血;眼病,如过敏性结膜炎,春季结膜炎,葡萄膜炎,和与甲状腺相关的眼病;嗜曙红肉芽肿;肺或呼吸道疾病,如哮喘,慢性支气管炎,过敏性鼻炎,ARDS,慢性肺炎病(例如,慢性障碍性肺病),硅肺病,肺肉样瘤病,胸膜炎,肺泡炎,脉管炎,肺炎,支气管扩张,和肺氧气中毒;心肌、大脑、或肢端的再灌注损伤;纤维化,如囊性纤维化;蟹状肿瘤形成或瘢痕组织形成;动脉硬化;自身免疫病,如系统性红斑狼疮(FLE),自身免疫甲状腺炎,多发性硬化,某些形式的糖尿病,和Reynaud’s综合征;以及移植排斥病,如GVHD和同种异体移植排斥;慢性肾小球肾炎;炎性胃病,如Crohn’s病,溃疡性大肠炎和坏死性小肠结肠炎;炎性皮肤病,如接触性皮炎,过敏性皮炎,牛皮癣,或风疹;由于感染引起的发热和肌痛;中枢或外周神经系统炎症,如脑膜炎,脑炎,和由于轻微创伤导致的大脑或脊髓损伤;Sjogren’s综合征;与白细胞白血球渗出相关的疾病;酒精性肝炎;细菌性肺炎;抗原-抗体复合体介导的疾病;血容量减少性休克;I型糖尿病;急性和延迟性过敏;由于白细胞dyscrasia和转移所导致的疾病状态;热损伤;与粒细胞输入相关的综合征;和细胞因子诱导的毒性。
本发明所提供的tank2多核苷酸还具有治疗用途,将所述多核苷酸用于治疗本文所述的疾病和失调,这些疾病和失调的病因与TANK2或活性相关。例如,tank2反义分子可以提供用于治疗与过高的或不希望的聚(ADP-核糖)聚合酶活性水平相关的各种异常症状。另外,编码TANK2的多核苷酸序列能提供用于治疗与聚(ADP-核糖)聚合酶活性的缺乏相关的各种异常症状。对tank2-LONG和tank2-SHORT中的一种或两种有专一性的多核苷酸特别适用于某些疾病。
源于逆转录病毒、腺病毒、疱疹或痘苗病毒,或源于各种细菌质粒的表达载体可用于将重组的tank2有义或反义分子输送到靶细胞群体中。可以用本领域技术人员所公知的方法构建含有tank2的重组载体,例如,参见披露于以下文献中的技术:Sambrook等,同上,和Ausubel等,同上。另外,可以通过脂质体将重组tank2输入靶细胞。
cDNA序列和/或其调控因子使得研究人员能够用tank2多核苷酸作为工具用于基因功能的有义研究[Youssoufian和Lodish,分子细胞生物学13:98-104,1993]或反义研究[Eguchi等,生物化学年度综述60:631-52,1991]。根据由基因组DNA所获得的cDNA或控制序列设计的寡核苷酸可用于在体外或体内抑制表达。所述技术为本领域所公知,并且可以根据各种位点和编码或控制区设计有义或反义寡核苷酸或较大的片段。同样,tank2-LONG和tank2-SHORT的专一性序列根据感兴趣的是哪一种形式的tank2而具有不同的用途。
另外,可以通过用表达高水平tank2多核苷酸片段的表达载体转染细胞或组织调节TANK2表达,优选在能抑制TANK2生物学活性的条件下表达。可将所述结构输入具有不可翻译的有义或反义序列的细胞。即使在不能整合到DNA中的情况下,所述载体也可以继续转录RNA分子,直到该载体的所有拷贝被内源核酸酶所破坏。所述瞬时表达可以通过非复制载体或整合了合适的复制因子的载体完成。
用于将载体导入细胞或组织的方法包括本文所披露的方法。另外,若干上述转化或转染方法同样适用于自体内治疗。另外,本文所披露的tank2多核苷酸序列可用于尚未开发的分子生物学技术中,其前提是,所述新技术依赖于核苷酸序列的目前已知的特征,包括但不限于诸如三联体遗传密码和特殊碱基对相互作用的特征。
药用组合物
本发明还涉及含有作为TANK2表达或活性的调节物的化学或生物学化合物(试剂),以及生物兼容的药用载体、佐剂或酶介物。药用组合物中的活性剂选自所有或部分下列化合物:tank2多核苷酸序列,tank2反义分子,TANK2多肽,蛋白,肽,或TANK2生物活性的有机调节物,如抑制剂,拮抗剂(包括抗体)或兴奋剂。所述试剂在用于治疗由TANK2表达或活性介导的或以此为特征的医学症状时是有活性的。该组合物可以包括所述试剂作为唯一的活性成分,或者与其他核苷酸序列、多肽、药物或激素组合,与赋形剂或其他药用载体混合。
制备和服用药用组合物的技术可以参见Remington’s药理科学,第18版,Mack出版公司,Easton,PA,1990。本发明的药用组合物可以用任何常规方法生产,例如,混合、溶解、粒化、制备糖衣丸、研磨、乳化、胶囊化、entrapping,熔纺、喷雾干燥、或冷冻干燥方法。不过,最佳的药用制剂可以由本领域技术人员根据服用途径和所需的剂量确定。所述制剂可能影响所服用的制剂的物理学状态、稳定性、体内释放速度、以及体内清除速度。根据要治疗的症状,可将所述药用组合物制备并且系统性地或局部服用。
所述药用组合物可以通过任何常规方法给受治对象服用,包括肠胃外和肠内技术。肠胃外服用方法包括通过除了胃肠道以下的途径服用该组合物的方法,例如,静脉内、动脉内、腹膜内、髓内、肌内、关节内、鞘内和心室内注射。肠内服用方法包括,例如,口服(包括口腔和舌下)和直肠内使用。例如,经皮服用方法包括经黏膜服用和经表皮服用。例如,经黏膜服用包括肠内服用以及鼻腔、吸入、和深入肺部的服用;阴道使用;和直肠使用。经皮服用包括被动和主动经皮和经肌肉方法,例如,包括膏贴和离子电渗装置,以及表皮涂敷糊剂、软膏或油膏。手术方法包括置入储存组合物、和渗透泵等。用于治疗炎症的优选服用途径是局部或表皮输送,用于诸如关节炎的局部炎症,以及用于再灌注损伤或用于诸如白血症的系统性症状的静脉内输送。
将所述药用组合物制成含有合适的可以药用的载体,并可以选择性地含有赋形剂和有利于将活性化合物加工成能够药用的制剂的佐剂。载体的性质通常由服用方法决定。例如,用于肠胃外服用的制剂可以含有水可溶形式的所述活性化合物的水溶液。适用于肠胃外服用的载体可选自盐水、缓冲过的盐水、葡萄糖、水、和其他生理上兼容的溶液。肠胃外服用的优选的常用载体是生理上兼容的缓冲液,如Hank’s溶液、Ringer’s溶液。或生理缓冲盐溶液。对于组织或细胞服用来说,将适合穿透特定屏障的渗透剂用于该制剂中。所述渗透剂一般为本领域所公知。对于含有蛋白的制剂来说,该制剂可以含有稳定材料,如多元醇(例如,蔗糖)和/或表面活性剂(例如,非离子表面活性剂等)。
用于肠胃外使用的制剂可以含有包括以合适的油性注射悬浮液形式制备的活性化合物的悬浮液。合适的亲脂性溶剂或媒介物包括脂肪油,例如,芝麻油以及合成的脂肪酸酯,如油酸乙酯或甘油三酯或脂质体。含水的注射悬浮液可以含有能提高该悬浮液稠度的物质,如羧甲基纤维素钠、山梨醇或葡聚糖。所述悬浮液还可选择性地含有合适的稳定剂或能提高该化合物溶解度的制剂,以便能制备高浓度的溶液。还可以使用乳液,例如,水包油和油包水分散剂,并选择性地用乳化剂或分散剂(表面活性材料;表面活性剂)稳定。还可将含有活性剂的脂质体用于肠胃外服用。能够提供pH敏感型溶解和/或活性剂的缓释含水聚合物也可用作包衣或基质结构,例如,异丁烯酸聚合物,例如由RohmVmerica公司(Piscataway,NJ)出售的Eudragit系列。
另外,可以用本领域众所周知的可以药用的载体制备适合于口服的剂型的含有所述活性剂的药用组合物。被制备用于口服的制剂可以是片剂、丸剂、胶囊、扁形胶囊、糖衣丸、酊剂、液体、凝胶、糖浆、糊剂、悬浮液或粉剂形式。为了说明,用于口服的药用制剂可以通过将活性化合物与一种固体赋形剂混合而获得,并选择性地研磨所得到的混合物,并在添加合适的佐剂之后(如果必要的话)对颗粒的混合物进行加工以便获得片剂或糖衣丸。注意,口服制剂可以采用类似于上述肠胃外使用的类型的液体载体,例如,缓冲过的水溶液和悬浮液等。
优选的口服制剂包括片剂、糖衣丸和凝胶胶囊。这些制剂可以含有一种或几种赋形剂,其中,但不限于:
a)诸如糖的稀释剂,包括乳糖,葡萄糖,蔗糖,甘露糖醇或山梨醇;
b)粘结剂,如硅铝酸镁,源于玉米、小麦、水稻、马铃薯等的淀粉;
c)纤维素材料,如甲基纤维素,羟丙基甲基纤维素,和羧甲基纤维素钠,聚依稀吡咯烷酮,胶,如阿拉伯树胶,如黄蓍胶,以及蛋白,如凝胶和胶原;
d)分散剂或增溶剂,如交联的聚乙烯吡咯烷酮,淀粉,琼脂,藻酸或其盐,如藻酸钠,或疱腾组合物;
e)润滑剂,如硅石、滑石、硬脂酸或其镁盐或钙盐,和聚乙二醇;
f)芳香剂和甜味剂;
g)颜料或色素,例如,用于鉴定产物或鉴定活性化合物的量(剂量);和
h)其他成分,如防腐剂,稳定剂,膨胀剂,乳化剂,溶液促进剂,用于调节渗透压的盐和缓冲剂。
明胶胶囊包括由凝胶制成的插入结合式胶囊,以及由明胶制成的软的密封胶囊,以及诸如甘油或山梨醇的包衣。插入结合式胶囊可以含有与填充剂、粘结剂、润滑剂和/或稳定剂等混合的活性成分。在软胶囊中,活性化合物可以溶解或悬浮在合适的液体中,如脂肪油、液态石蜡、或液态聚乙二醇,有或没有稳定剂。
糖衣丸可以具有合适的包衣,如浓缩的糖溶液,它也可以含有阿拉伯树胶、滑石、聚乙烯吡咯烷酮、carbopol凝胶、聚乙二醇和/或二氧化钛、漆用溶液,和合适的溶剂或溶剂混合物。
药用组合物可以活性剂的盐的形式提供,它可以是用多种酸制成的盐,包括,但不限于盐酸,硫酸,乙酸,乳酸,酒石酸,苹果酸,琥珀酸等。盐倾向于更容易溶解在水或其他质子溶剂中,这种溶剂是相应的无碱形式。
为了有效治疗对中枢神经系统目标的调节,用于本发明方法中的试剂在外周使用时应当便于穿透血脑屏障。不过,不能够穿透血脑屏障的化合物也可以通过静脉内途径有效使用。
如上文所述,所述制剂本身的特征以及该试剂的制备可能影响所服用的试剂的物理状态、稳定性、体内释放速度、和体内清除速度。所述药物动力学和药物动态信息可以通过预先的临床体外和体内研究收集,然后在临床试验期间在人体上加以证实。因此,对用于本发明的任何化合物来说,治疗有效的剂量最初可以根据基于生物化学和/或细胞的测定估计。然后制备用于动物模型的剂型,以便达到能调节TANK2表达或活性的所需循环浓度范围。在进行人体研究时,可以得到有关各种疾病和症状的合适的剂量水平和治疗持续时间的其他信息。
所述化合物的毒性和治疗效力可以通过标准药理学方法用细胞培养物或实验动物确定,例如,测定LD50(50%群体的致死剂量)和ED50(50%群体的治疗有效剂量)。毒性和治疗效果之间的剂量比例是“治疗指数”,它通常被表示为LD50/ED50。优选具有大的治疗指数的化合物。从所述细胞培养测定和其他动物研究中所获得的数据可用于制备供人使用的多种剂量。所述化合物的剂量优选落入包括LD50在内的,少有或没有毒性的循环浓度范围。
对于本发明的方法来说,调节剂量的时间和顺序的任何有效的服用方法都可以采用。所述制剂的剂量优选包括含有有效量的该制剂的药用剂量单位。在本文中,“有效量”是指通过服用一个或多个药用剂量单位足于调节TANK2表达或活性和/或使受治对象的生理学参数发生可检测的变化的量。
用于人体对象的代表性剂量水平在大约0.001-100毫克活性剂每千克体重(毫克/千克)水平上。通常,活性剂的剂量单位包括大约0.01-大约10000毫克,优选大约0.1-大约1000毫克,取决于症状、服用的途径等。根据服用途径,可以按照体重、身体表面积或器官大小计算出合适的剂量。最终的剂量方案由主治医生根据良好的医学经验确定,要坚固到会影响药物作用的各种因素,例如,制剂的比活性,疾病症状的严重程度,患者的反应,年龄,状态,体重,性别,以及患者的饮食,所有感染的严重程度等。需要考虑的其他因素包括服用的时间和次数,药物组合,反应敏感性,和对治疗的耐受力/反应。对剂量作进一步的微调以便适合治疗包括本文所提到的制剂是由有经验的医生根据常规方法完成的,不需要作过多的实验,特别是考虑到所纰漏的剂量信息和测定,以及在人体临床实验上所观察到药物动力学数据。可以通过使用确立的测定方法确定合适的剂量,以便测定制剂在体液或其他样品中的浓度,同时确定剂量反应数据。
用药次数取决于该制剂的药物动力学参数和服用途径。对剂量和服用方法进行调整,以便提供足够水平的活性成分或保持所需的效果。因此,所述药用组合物可以单一剂量服用,多个独立的剂量服用,连续输液,缓式储存,或其组合,以便满足保持所需的最低药剂水平的要求。短效药用组合物(即短半衰期)可以每日服用一次或每日服用一次以上(例如,每日两次、三次或四次)长效药用组合物可以每3-4天服用一次,每周一次,或每2周一次。优选将诸如皮下、腹膜或硬膜下的泵用于连续输液。可以制备在可以药用的载体中配置的含有本发明化合物的组合物,将其放入合适的容器中,并标明用于治疗特定症状。在标签所标明的症状可以包括治疗炎性疾病、癌症、神经组织损伤等。还涉及试剂盒,其中,该试剂盒含有药用组合物的剂型,和含有指导该组合物用于治疗医学症状时的使用方法的包装说明。
提供以下实施例是为了更好地理解本发明。所采用的特定材料和条件是用于说明本发明的特定方面的,不应当被理解为对其合理范围的限定。
这些实施例的先决条件是了解实施例所属领域的普通技术人员众所周知的常规方法,例如,载体和质粒的构建,将编码多肽的基因插入所述载体和质粒,或将载体和质粒导入宿主细胞。所述方法详细披露于多种出版物中。例如,包括Sambrook等,分子克隆,实验室手册,冷泉港实验室出版社,1989,Ausubel等著,当代分子生物学方法,JonnWiley&Sons公司,1994;和Ausubel等著,分子生物学简明方法,第4版,Jonn Wiley&Sons公司,1999。
实施例1
与人坦科聚合酶1相关的EST的鉴定和坦科聚合酶2多核苷酸的分离
用人坦科聚合酶1(SEQ ID NO:3)的核苷酸序列[Smith等,1998,同上]对国家生物技术信息中心(NCBI)表达序列标记(EST)数据库进行检索,以便鉴定与坦科聚合酶1同源的基因。EST数据库提供了源于多种组织来源的cDNA克隆的5’和/或3’核苷酸序列。使用NCBIBLASTn程序[Altschul等,核酸研究25:3389-402,1997]将人坦科聚合酶1的核苷酸查询序列与核苷酸序列数据库进行比较,以便鉴定EST序列数据库中与人坦科聚合酶1有明显同源性的DNA序列。上述BLASTn检索鉴定了两个感兴趣的EST序列:从被称为HCC的人结肠癌细胞系中克隆的AA307492(SEQ ID NO:5),和由人大脑克隆的人17748(SEQ ID NO:7)。
比较AA307492和坦科聚合酶1多核苷酸发现了AA307492(SEQID NO:5)的从307到432(nt307-432)的一个片段与坦科聚合酶1(SEQ ID NO:3)的nt3313-3438的一段序列具有明显的同源性;在126个核苷酸中有105个相同;相同性为83%。翻译AA307492的核苷酸307-432,并将推测的蛋白(SEQ ID NO:6)与坦科聚合酶1蛋白(SEQID NO:4的1105-1146号氨基酸)进行比较,发现上述两种蛋白在42个氨基酸位置上有36个相同(相同性86%)。比较H17748和坦科聚合酶1多核苷酸发现H17748的nt3-356(SEQ ID NO:7)与坦科聚合酶1的nt3544-3897具有明显的同源性(SEQ ID NO:3;354个核苷酸中有280个相同;相同性为79%)。翻译H17748的nt3-356,并将推测的蛋白(SEQ ID NO:8)与坦科聚合酶1的相应部分进行比较(SEQID NO:4的1182-1199号氨基酸),发现这两种蛋白在118个氨基酸位置上有111个相同(相同性为94%)。AA307492和H17748的推测的氨基酸序列与坦科聚合酶1蛋白同源,但又不同于该蛋白,这表明它们是从新的坦科聚合酶基因翻译得到的蛋白产物。
将AA307492和H17748用于通过NCBI UniGene程序检索GenBank数据库,以便鉴定源于相同基因的其他EST序列。所述UniGene程序将GenBank序列组合成基因定向类型的非丰余类型,每一类含有源于相同基因一组序列。用AA307492对人GenBank数据库进行UniGene检索,在与AA307492相同的基因片段上没有发现任何其他人EST序列丛。相反,用AA307492对人GenBank数据库进行UniGene检索,在与H17748对人GenBank数据库进行UniGene检索,鉴定了16个与H17748属于相同的基因丛的人EST序列,这些序列如下:AA305587(SEQ ID NO:9),AA371079(SEQ ID NO:10),AA970617(SEQ ID NO:11),AI247608(SEQ ID NO:12),H11505(SEQ ID NO:13),H11865(SEQ ID NO:14),H17635(SEQ ID NO:15),N29528(SEQ ID NO:16),N57467(SEQ ID NO:17),R06902(SEQ ID NO:18),R06946(SEQ ID NO:19),R14158(SEQ ID NO:20),R33944(SEQ ID NO:21),R63031(SEQ ID NO:22),R63337(SEQ ID NO:23)和T17118(SEQ ID NO:24)。EST H17748和ESTH17635含有来自被称为50806的同一个克隆的相反两端的序列。ESTH11505和EST H11865含有来自被称为47912的同一个克隆的相反两端的序列。EST R06902和EST R06946含有来自被称为126654的同一个克隆的相反两端的序列。获得了cDNA克隆50806、47912、和126654的大肠杆菌菌株是从美国典型培养物保藏中心(ATCC Rockville,MD)购买的,该保藏中心保藏了由I.M.A.G.E.鉴定并测序的ESTs的保藏物并可以向公众提供(Lawrence Livermore国家实验室,Livermore,CA)。按以下方法对三个克隆进行测序:
使用能与载体DNA杂交的引物(SEQ ID NO:25-26)和设计用于与人cDNA杂交的引物(SEQ ID NO:27-34)对克隆50806的两股链进行全面测序。
M13正向 TGTAAAACGACGGCCAGT(SEQ ID NO:25)
M13反向 GGAAACAGCTATGACCATG(SEQ ID NO:26)
NT-7 TTTGCCGGGTAACCTTGG(SEQ ID NO:27)
NT-8 CCAAGGTTACCCGGCAAA(SEQ ID NO:28)
NT-9 GTAGGCCCAGTGTAAATG(SEQ ID NO:29)
NT-10 CATTTACACTGGGCCTAC(SEQ ID NO:30)
NT-11 GAGTAAGTTGCAGGGCATGT(SEQ ID NO:31)
NT-12 ACATGCCCTGCAACTTACTC(SEQ ID NO:32)
NT-13 GAATCACCGCAGTTACTAAA(SEQ ID NO:33)
NT-14 TTTAGTAACTGCGGTGATTC(SEQ ID NO:34)
使用能与载体DNA杂交的引物(SEQ ID NO:25-26,同上)和设计用于与人cDNA杂交的引物(SEQ ID NO:27-34,同上,和SEQ IDNO:35-37)对克隆47912的两股链进行全面测序
NT-15 GGCCTGAAGGTATGGTCOAT(SEQ ID NO:35)
NT-16 ATCGACCATACCTTCAGGCC(SEQ ID NO:36)
NT-18 TGAGGGCATTACAGTTTGTT(SEQ ID NO:37)
使用能与载体DNA杂交的引物:M13正向(SEQ ID NO:25,同上)和T7启动子(SEQ ID NO:38),和设计用于与人cDNA杂交的引物(SEQ ID NO:27-30,同上,和SEQ ID NO:39-40)对克隆126654的两股链进行全面测序。
T7启动子TAATACGAACTCACTATAGGG(SEQ ID NO:38)
NT-5 ATACACTCACCGGAGAAA(SEQ ID NO:39)
NT-6 TTTCTCCGGTGAGTGTAT(SEQ ID NO:40)
通过测序发现50806、47912和126654与EST数据库中报导的序列一致。50806、47912和126654的多核苷酸序列分别示于SEQ ID NO:41、43和45中。50806、47912和126654的推测的氨基酸序列分别示于SEQ ID NO:42、44和46中。50806和47912的序列表明这两种克隆是相同的,并且仅对50806作进一步的考虑。50806和1266544含有重叠的核苷酸序列,但126654在5’末端多出63个碱基对,而50806在3’末端多出大约400个碱基对。
50806被确定具有一个开放读框(ORF),该读框始于1号核苷酸位置,在nt358-1138处有一个潜在的内含子序列,有一个从nt1999开始的终止密码子和一个距离终止密码子的3’末端474个碱基对的潜在的聚腺苷酸化尾巴。当比较50806的nt1-357和坦科聚合酶1的nt3528-3897时,在翻译50806的1-357号核苷酸并将所得到的蛋白与坦科聚合酶1(1181-1299号氨基酸)进行比较时,发现两种蛋白在120个氨基酸位置上有116个相同(相同性为97%)。在50806中鉴定了一个推测的内含子,包括nt358-1138,它可能是cDNA克隆的伪迹。位于推测的内含子(AG)前面并位于推测的内含子(CAG)3’末端的DNA序列与外显子/内含子/外显子接合的共有序列具有很高的相似性[Lewin,GENES IV,牛津大学出版社:纽约,1997,88页]。在外显子的3’末端最常见的序列是AG,而在内含子的3’末端是CAG。为了确定在50806序列上是否包含内含子,通过对基因组DNA的PCR分析证实该推测。
比较50806和坦科聚合酶1发现了50806的由nt1139-1198组成的一个小的片段与坦科聚合酶1的nt3896-3956具有显著的同源性(在60个核苷酸中有40个相同,相同性为67%)。当翻译50806的nt1139-1198并将所得到的蛋白与坦科聚合酶1(1300-1319号氨基酸)进行比较时,这两种蛋白在20个氨基酸位置上有14个相同(相同性为70%)。
确定126654有一个从1号核苷酸位置开始的ORF,一个从481号位置开始的终止密码子,和一个距离终止密码子3’末端81个碱基对的潜在的聚腺苷酸化尾巴。比较126654和坦科聚合酶1发现了一个由126654的nt1-480组成的片段与坦科聚合酶1的nt3478-3957具有明显的同源性(在481个核苷酸中有367个相同,相同性为76%),当翻译126654的该片段并将所得到的蛋白与坦科聚合酶1蛋白的相应片段(即1160-1319号氨基酸)进行比较时,这两种蛋白在160个氨基酸位置上有149个相同(相同性为97%)。有可能50806和126654的推测的聚腺苷酸化尾巴是cDNA克隆的伪迹,或者50806和126654代表一个使用不同的聚腺苷酸化位点的mRNA群体。50806具有一个距离终止密码子的3’末端81个碱基对的8A残基的片段,表明126654的推测的聚腺苷酸化尾巴最有可能是克隆伪迹。与SequencherTM程序(基因编码公司,Ann Arbor,MI)对AA307492和126654和人坦科聚合酶1进行排比,发现AA307492是126654的上游,并有11个核苷酸分割AA307492和126654。为了证实AA307492和126654是来自相同基因的多核苷酸序列,合成了相应于AA307492的有义链的引物(SEQ ID NO:47)和相应于126654的反义引物(SEQ ID NO:48),用于以人Marathon-Ready脾和睾丸cDNA(Clontech)为模板的PCR反应中。
AA307492有义CTCCGGACAACAAGGTCTTAACC(SEQ IDNO:47)
126654反义CCACCTATGTACGCATGCC(SEQ ID NO:48)
PCR反应含有2.5微升人脾Marathon-ReadycDNA,2.5微升睾丸Marathon-ReadycDNA,每一种引物250nM,0.25mM dNTPs,1×PCR缓冲液,1.8mM氯化镁,和5单位Taq聚合酶(Perkin Elmer)。该反应是在GeneAmpPCR系统9700仪中进行(以下称之为GeneAmpPCR系统9700;PE应用生物系统,Norwalk CT),应首先在94℃下加热2分钟,然后进行35轮94℃30秒,55℃30秒,和72℃30秒,最后72℃7分钟结束。按照生产商的说明通过凝胶电泳和QIAquick凝胶提取试剂盒(以下称之为QIAquick试剂盒;Qiagen,Valencia,CA)分离PCR片段。按照生产商的说明将PCR片段直接克隆到pCR2.1-TOPO载体上(Invitrogen,Carlsbad,CA)。用能与载体DNA杂交的引物(SEQID NO:25和26,同上)对PCR片段进行测序,并将AA307492/126654PCR片段的序列示于SEQ ID NO:49中。该序列证实AA307492是126654的上游,并且这两个EST由11个核苷酸隔开,并且AA307492和126654是来自被称为坦科聚合酶2的新型基因序列。
为了鉴定完整长度的坦科聚合酶2基因,用126654制备探针,并用于筛选cDNA文库,使用本领域常用的方法。用XhoI和BglII消化126654,并通过凝胶电泳和QIAquick试剂盒分离被称为NT-5’的大约有260个核苷酸的片段。通过随机启动的DNA标记试剂盒(BoehringerMannheim/Roche分子生物化学,Indivnapolis IN)按照生产商的说明用32P标记NT-5’,并用于筛选来自人胎儿大脑文库的106cDNA(Stratagene)。与标记探针的杂交在65℃下在含有以下成分的缓冲液中进行一夜:3×SSC,0.1%sarkosyl,20mM乙酸钠,pH6.8,10×Denhardt’s溶液,和50微克/毫升鲑鱼精DNA。在放射性自显影之前在65℃下在含有2×SSC和0.1%SDS的缓冲液中洗涤滤膜。用NT-5’探针获得了46个阳性,其中有15个是使用相应于NT-5’的杂交强度首次鉴定的。限制性消化作图和部分测序导致了两个克隆被挑选出来,它们是FB2B.1和FB2D.1,对其作进一步鉴定。
用包括T7启动子(SEQ ID NO:38,同上)和T3启动子(SEQ IDNO:50)在内的能与载体DNA杂交的启动子和涉及用于与cDNA序列退火的引物(SEQ ID NO:51-69)对FB2B.1的两条链进行全面测序。
T3启动子ATTTAACCCTCACTAAAGGG(SEQ ID NO:50)
2B.1 F1 AAAGGCTCCCATCGGCAAAT(SEQ ID NO:51)
2B.1 F2 GTTGAGGGCATTACAGTTTG(SEQ ID NO:52)
2B.1 F3 AAAACGTAGAGGCCACTGCT(SEQ ID NO:53)
2B.1 P4 TGGTGTAGACTGACGCCCTT(SEQ ID NO:54)
2B.1 P5 TCCGGTGAGTGTATCTTTCC(SEQ ID NO:55)
2B.1 P6 CTCCTTTGTCTTGGGCATTC(SEQ ID NO:56)
2B.1 P9 ATCTGCTCTGCCCTCTTGTT(SEQ ID NO:57)
2B.1 F10 GGGTATCGCGGCAATTTACA(SEQ ID NO:58)
2B.1 F11 AACAAGAGGGCAGAGCAGAT(SEQ ID NO:59)
2B.1 F12 TGCCCCATCTCAACTAATAC(SEQ ID NO:60)
2B.1 R2 GTAATGCCCTCAACAGAACT(SEQ ID NO:61)
2B.1 R3 GGCGTCAGTCTACACCACTT(SEQ ID NO:62)
2B.1 R4 TAAATTGCCCGCGATACCCA(SEQ ID NO:63)
2B.1 R5 CACTCAGTCACTGGTAGGCC(SEQ ID NO:64)
2B.1 R6 ATCTGCTCTGCCCTCTTGTT(SEQ ID NO:65)
2B.1 R7 TAGTTGAGATGGGGCACAAG(SEQ ID NO:66)
2B.1 R8 AAACGTAGAGGCCACTGCTG(SEQ ID NO:67)
2B.1 R9 CGGGTAACCTTGGGAAAGTC(SEQ ID NO:68)
2B.1&2D.1 GGGCTTTACTGCTTTACAGA(SEQ ID NO:69)
用能与载体DNA杂交的引物(SEQ ID NO:38和50,同上)和涉及用于与cDNA序列杂交的引物,包括2B.1和2D.1(SEQ ID NO:69)和SEQ ID NO:70-87对FB2D.1的两股链进行全面测序。
2D.1 F1 GTAAGGGCTGCTGACAGTGA(SEQ ID NO:70)
2D.1 F2 TTACTCCA(3CAGAGGGCACT(SEQ ID NO:71)
2D.1 F3 CTGACGCCCTTCAATGTCTC(SEQ ID NO:72)
2D.1 F4 GGTACTAAGGCCACAATTCA(SEQ ID NO:73)
2D.1 F5 GGGTATCGCGGCAATTTACA(SEQ ID NO:74)
2D.1 F6 GTTGAGGGCATTACAGTTTG(SEQ ID NO:75)
2D.1 F7 TAACAAGAGGGCAGAGCAGA(SEQ ID NO:76)
2D.1 F8 AGTTCTGTTGAGGGCATTAC(SEQ ID NO:77)
2D.1 F9 GGCCTACCAGTGACTGAGTG(SEQ ID NO:78)
2D.1 F10 GGGCTAGAGGACCTGAAGAG(SEQ ID NO:79)
2D.1 R2 AGTGCCCTCTGCTGGAGTAA(SEQ ID NO:80)
2D.1 R3 GGCGTCAGTCTACACCACTT(SEQ ID NO:81)
2D.1 R4 TGAATTGTGOCCTTAGTACC(SEQ ID NO:82)
2D.1 R5 ATGCCCAAGACAAAGGAGGA(SEQ ID NO:83)
2D.1 R6 GTAATGCCCTCAACAGAACT(SEQ ID NO:84)
2D.1 R7 ATCTGCTCTGCCCTCTTGTT(SEQ ID NO:85)
2D.1 R8 CGGGTAACCTTGGGAAAGTC(SEQ ID NO:86)
2D.1 R9 CCGGACAACAAGGTCTTAAC(SEQ ID NO:87)
FB2B.1和FB2D.1的多核苷酸序列分别如SEQ ID NO:88和90所示,而FB2B.1和FB2D.1的推测的氨基酸序列分别如SEQ ID NO:89和91所示。
比较FB2B.1和坦科聚合酶1核苷酸和氨基酸序列,以便确定所述序列之间的相关性程度。发现FB2B.1(SEQ ID NO:88)的由nt4-279所组合的一个片段与坦科聚合酶1(SEQ ID NO:3)的nt1624-1899具有明显的相同性,其中,276个核苷酸中有203个相同(相同性为73%)。FB2B.1的402-1254号核苷酸与坦科聚合酶1的2022-2874号核苷酸有显著的相同性,853个核苷酸中有630个相同(相同性为73%)。另外,FB2B.1的1507-2338号核苷酸与坦科聚合酶1的3112-3943号核苷酸有同源性,832个核苷酸中有634个相同(相同性为76%)。确定FB2B.1具有一个始于1号核苷酸位置的ORF,一个始于2353号位置的终止密码子,大约1kb的3’非翻译序列,但没有明显的聚腺苷酸化尾巴。FB2B.1的1-2352号核苷酸的翻译表明由推测的氨基酸序列组成的一个片段(SEQ ID NO:89)与坦科聚合酶1的相应部分同源(SEQ ID NO:4的540-1327号氨基酸),在该片段上,这两种蛋白的777个氨基酸位置上有623个相同(相同性为80%)。
将FB2B.1与坦科聚合酶1作类似的比较,在这种情况下,FB2B.1(SEQ ID NO:90)的由6-197号核苷酸组成的一个片段与坦科聚合酶1的1708-1899号核苷酸明显相关,在192个核苷酸中有137个相同(相同性为71%)。发现FB2B.1的320-1172号核苷酸与坦科聚合酶1的相应的2022-2874号核苷酸有显著的同源性,在853个核苷酸中有630个相同(相同性为73%)。FB2B.1的1425-2256号核苷酸与坦科聚合酶1的3112-3943号核苷酸有显著的同源性,在832个核苷酸中有634个相同(相同性为76%)。确定FB2B.1具有一个始于3号核苷酸位置的ORF,一个始于2271号核苷酸位置的终止密码子,大约1.5kb的3’非翻译序列,但没有明显的聚腺苷酸化尾巴。在翻译FB2B.1时(SEQ IDNO:91),由3-2270号核苷酸预测的一个区域与坦科聚合酶1(SEQID NO:4)的569-1327号氨基酸具有同源性。在这里,这两种蛋白在749个氨基酸位置上有602个相同(相同性为80%)。
用SequencherTM对FB2B.1和FB2D.1进行排比。FB2B.1和FB2D.1含有重叠的多核苷酸序列,但FB2B.1在5’末端长出82个碱基对,而FB2D.1在3’末端长出大约0.5kb。FB2B.1和FB2D.1的核苷酸序列在FB2B.1的83-2971号核苷酸部分和FB2D.1的1-2889号核苷酸部分相同。不过,FB2B.1的剩余的382个核苷酸和FB2D.1剩余的910个核苷酸不同。有可能FB2B.1和FB2D.1是从3’非翻译区的不同位点随机启动的,和/或这种错排是在一个或两个克隆中出现克隆伪迹的结果。由于FB2B.1和FB2D.1没有表现出具有聚腺苷酸化尾巴,ESTs50806和126654的聚腺苷酸化尾巴很可能是克隆伪迹,而坦科聚合酶2的聚腺苷酸化尾巴极有可能距离终止密码子超过0.5kb。通过排比FB2B.1和FB2D.1产生了一个被称为2B.1/2D.1的共有多核苷酸序列,并且在SEQID NO:92中示出,2B.1/2D.1含有FB2B.1的1-2971号核苷酸和FB2D.1的1-2889号核苷酸。
用SequencherTM将FB2B.1和FB2D.1与坦科聚合酶2进行排比,发现FB2B.1和FB2D.1都不是完整长度的基因,并且缺少坦科聚合酶2的5’末端的核苷酸序列。因此,用EcoRI和SphI消化FB2B.1,并通过凝胶电泳和QIAquick试剂盒分离仅靠着FB2B.1的5’末端的大约466bp的核苷酸片段(SEQ ID NO:88的49-515号核苷酸)。用随机启动的DNA标记试剂盒用32P标记该片段,并用作探针(被命名为NT-37/38)采用在第一次筛选中所使用的条件和方法筛选胎儿脑文库(Stratagene)的106cDNA文库。用NT-37/38探针获得了16个阳性克隆,其中的一个(被命名为30B.2A)还能与NT-5’探针杂交,但此时不选择它作进一步的鉴定。限制作图和部分测序导致了选择30B.2A作进一步的鉴定。
用能与载体DNA杂交的引物(SEQ ID NO:38和50,同上)和被设计用于与cDNA序列退火的引物,包括2B.1F4(SEQ ID NO:54,同上和SEQ ID NO:93-97)对克隆FB2B.1的30B.2A上游部分进行测序。
30B.2A#1 GGGCGGAAAGACGTAGTTGA(SEQ ID NO:93)
30B.2A#2 GCGGCTGTTCACCTTCTCAG(SEQ ID NO:94)
30B.2A#5 ACGCAAGTGATGGCAGAAAG(SEQ ID NO:95)
30B.2A#6 TCACTTGCGTGGCAGTTGAC(SEQ ID NO:96)
30B.2A#7 GCGGCAGGTTTGTAGATGAC(SEQ ID NO:97)
30B.2A的部分多核苷酸序列如SEQ ID NO:98所示,而部分推测的氨基酸序列如SEQ ID NO:99所示。将30B.2A和坦科聚合酶1的核苷酸序列进行比较发现显著的同源性出现在30B.2A的由167-1435号核苷酸组成的片段上,它相当于坦科聚合酶1的631-1899号核苷酸。在该片段上,1269个核苷酸中有953个相同(相同性为75%),确定30B.2A的具有一个始于2号核苷酸位置的ORF。在推测的30B.2A的推测的385个氨基酸序列(基于2-1156号核苷酸)和坦科聚合酶1的相应部分(160-539号氨基酸)之间发现了显著的氨基酸序列相同性。在该部分,所述蛋白序列的385个氨基酸位置上有319个相同(相同性为83%)。
用SequencherTM对2B.1/2D.1和30B.2A进行排比。30B.2A含有1.157kb的新的序列位于开始2B.1/2D.1的5’末端重叠部分的前面,并从1158号位置开始与2B.1/2D.1重叠。根据2B.1/2D.1和30B.2A的排比形成了一个被命名为2B.1/2D.1/30B.2A的共有多核苷酸序列,并且如SEQ ID NO:100所示。2B.1/2D.1/30B.2A含有30B.2的1-1157号核苷酸和2B.1/2D.1的1-2971号核苷酸。由SEQ ID NO:100的2-3508号核苷酸所编码的推测的氨基酸序列如SEQ ID NO:101所示。TANK2编码区的核苷酸序列如SEQ ID NO:1所示,而相应的TANK2多肽序列如SEQ ID NO:2所示。
实施例2
克隆坦科聚合酶2的5’末端
用SequencherTM程序排比30B.2A和坦科聚合酶1发现,仍然缺乏坦科聚合酶2基因的5’序列。为了克隆人坦科聚合酶2的5’末端,用Marathon-Ready人脾cDNA(Clontech)作模板进行5’RACE分析。合成相当于2B.1/2D.1/30B.2A多核苷酸序列(SEQ ID NO:100的337-367号核苷酸)反义链的引物(NT-Marathon;SEQ ID NO:102),用于聚合酶链式反应中,将为了与MarathoncDNA接头退火而设计的AP1引物(Clontech;SEQ ID NO:103)连接到该文库的cDNA的末端。
NT-Marathon GAGCATTGGGGTCTGCACCATGTCGCAAAAGG(SEQ ID NO:102)
AP1 CCATCCTAATACGACTCACTATAGGGC(SEQ ID NO:103)
PCR反应含有5微升人脾Marathon-ReadycDNA,每一种引物0.20微摩尔,0.20mMdNTPs,1×ClontechGC2PCR缓冲液,ClontechGC-解链缓冲液(0,0.5,1.0或1.5摩尔),和1微升Clontech Advantage-GC2聚合酶缓冲物,该反应在GeneAmpPCR系统9700中进行,采用以下四个步骤:1)1轮94℃1分钟;2)5轮94℃30秒和12℃30秒;3)5轮94℃30秒和70℃30秒;和4)25轮94℃30秒和60℃30秒。然后在以下条件下在GeneAmpPCR系统9700中继续上述反应:1)1轮94℃1分钟;2)5轮94℃30秒和72℃3分钟;3)5轮94℃30秒和70℃3分钟;和4)25轮94℃30秒和60℃3分钟。按照说明通过凝胶电泳和QIAquick试剂盒分离PCR片段。按照说明将该PCR片段直接克隆到pCR2.1-TOPO载体上。因为Taq聚合酶具有8.0×106突变/碱基对的错误率(Cline等,核酸研究24:3546-51),对从4个独立的PCR反应中分离的4个克隆进行测序,并比较,以便消除Taq聚合酶在5’RACE序列中引起的误差的可能性。使用能与载体DNA杂交的M13正向和M13反向引物(SEQ ID NO:25和26)对这4个5’RACE克隆进行测序。将4个独立的核苷酸序列编辑成共有核苷酸序列,命名5’-RACEtank2,如SEQ ID NO:104所示,而推测的氨基酸序列如SEQ IDNO:105所示。在5’-RACEtank2的共有核苷酸序列上,每一个碱基对出现在用于编辑该共有序列的4个独立克隆的至少3个的相应位置上。用SequencherTM程序对5’-RACEtank2和坦科聚合酶进行比较。5’-RACEtank2(SEQ ID NO:104)的1-279号核苷酸与坦科聚合酶相比没有明显的相似性。确定5’-RACEtank2具有一个始于2号核苷酸位置的ORF。当翻译5’-RACEtank2的2-277号核苷酸并将所得到的蛋白与坦科聚合酶进行比较时,没有发现明显的相似性。
用SequencherTM程序对5’-RACEtank2和2B.1/2D.1/30B.2A进行排比。5’-RACEtank2在开始与FB2B.1/2D.1/30B.2A的5’末端重叠之前有279bp的新的序列,并在280号位置上开始与2B.1/2D.1/30B.2A重叠。通过比较5’-RACEtank2和2B.1/2D.1/30B.2A产生了被称为和2B.1/2D.1/30B.2A/5’RACE的共有多核苷酸序列,并且如SEQ ID NO:106所示。和2B.1/2D.1/30B.2A/5’RACE含有5’RACEtank2的1-279号核苷酸和2B.1/2D.1/30B.2A的1-4140号核苷酸。2B.1/2D.1/30B.2A/5’RACE的推测的氨基酸序列如SEQ ID NO:107所示。
连续的ORF在5’RACEtank序列中的存在表明,仍然缺乏坦科聚合酶2基因的5’序列。用5’RACE分析获得坦科聚合酶2的其他5’序列的进一步的尝试没有成功。使用NCBIBLASTn程序比较FB2B.1/2D.1/30B.2A的核苷酸查询序列和核苷酸序列标记数据库(GenBank+EMBL+DDBJSTS Divisions)。该BLASTn检索鉴定了一个被称为stWI-16054的STS标记序列(GenBank保藏号24639;SEQID NO:108)。当比较2B.1/2D.1/30B.2A的3608-3985号核苷酸和stWI-16054的反义互补核苷酸8-397时,在378个核苷酸有361个相同(相同性为96%)。使用Sanger中心(剑桥,英国)人类基因组克隆检索程序(http://www.sanger.ac.uk/cgi-bin/humace/searcher.cgi)鉴定含有stWI-16054的BAC克隆。BAC克隆bA329B8被鉴定为含有STS标记stWI-16054。BAC克隆bA329B8源于基因组RPCI-11.2雄性白血细胞库(Pieter DeJong,Roswell Park癌症研究所,Buffalo,NJ)购自Research Genetics,Inc.(Huntsville,AL)。用大型结构试剂盒(Qiagen)分离bA329B8DNA,将其用做模板用于反PCR扩增反应[Ochman等,通过反PCR扩增旁侧序列,参见PCR方法:方法和应用指南(Innis等著),219-27页,学术出版社,San Diego,CA,1990]。所述反PCR技术可以扩增已知序列两侧的未知DNA序列。简单地讲,用限制酶消化模板DNA(优选使用能够识别个或5个碱基对共有位点的限制酶),然后将限制片段环化。将环化片段用做模板用于PCR反应,该反应使用被设计用于与已知旁侧序列退火的两个引物,但所述旁侧序列的朝向相反。在20微升反应物中消化1微克Ba329B8,该反应物含有1倍合适的反应缓冲液和10单位的下列限制酶之一:RsaI(Promega,Madison,WI),BfaI(新英格兰实验室,Beverly,MA),或Tru9I(Promega)。该限制酶消化在37℃(RsaI和BfaI)或65℃(Tru9I)下进行1小时。将RsaI和BfaI消化物在68℃下加热20分钟,以便使限制酶失活。用QIAquick试剂盒使Tru9I消化物中的限制酶失活。连接反应物含有以下成分:20微升Tru9I,RsaI或BfaI反应物,448微升蒸馏水,50微升10倍反应缓冲液,和2微升T4DNA连接酶(5单位/微升;Boehringer Mannheim,Indianapolis,IN)。连接物在15℃下培养一夜。然后通过添加129.26微升7摩尔乙酸铵和2.3mM95%乙醇使连接反应物中的DNA沉淀。使DNA沉淀,用75%乙醇洗涤,重新悬浮在15微升蒸馏水中,并用做PCR扩增反应的模板。合成相应于5’-RACEtank2的正义链(SEQ ID NO:104的423-443号核苷酸)的引物(5-Inv-1;SEQ ID NO:109)和相应于5’RACEtank2的反义链(SEQ IDNO:104的364-383号核苷酸)的引物(3-Inv-1;SEQ ID NO:110),用于PCR扩增反应。5-Inv-1 CGCCTGAGAAGGTGAACAGCC(SEQID NO:109)
3-Inv-1 ACGCTCGAACAGCTCTCGG(SEQ ID NO:110)
PCR反应物(最终反应体积为20微升)含有5微升Tru9I,RsaI或BfaI DNA模板,0.20微摩尔每一种引物,0.20mMdNTPs,1×clonetech,GC2PCR缓冲液,1.0mMclontechGC-解链缓冲液,和0.4微升clontch advantage-R-GC2聚合酶。该反应是在GeneAmpPCR系统9700中进行,采用以下四个步骤:1)1轮94℃1分钟;2)5轮94℃30秒和65℃3分30秒;3)5轮94℃30秒和65℃3分30秒;和4)25轮94℃30秒和58℃3分30秒。按照说明通过凝胶电泳和QIAquick试剂盒分离PCR片段,按照说明将该PCR片段直接克隆到pCR2.1-TOPO载体上。用能与载体DNA杂交的M13引物(SEQ ID NO:25和26)和被设计用于与cDNA序列退火的引物(SEQ ID NO:109-112)对Tru9I,RsaI和BfaI克隆进行测序。
5-Inv-2 GCGTGGGCGCGGCCATGGGACTG(SEQ ID NO:111)
3-Inv-2 CAGCGCGAATCCGCCGTCCG(SEQ ID NO:112)
Tru9I,RsaI和BfaI多核苷酸序列分别如SEQ ID NO:113、115和117所示。Tru9I,RsaI和BfaI推测的氨基酸序列分别如SEQ ID NO:114、166和118所示。
用SequencherTM程序对克隆Tru9I和5’-RACEtank2进行排比。克隆Tru9I(SEQ ID NO:113)在开始与5’-RACEtank2(SEQ ID NO:104)的5’末端重叠之前有235bp的新的序列,并从236号位置开始与5’-RACEtank2重叠。在比较克隆Tru9I的1-235号核苷酸和坦科聚合酶时没有发现明显的相似性。确定克隆Tru9I具有一个始于3号核苷酸位置的ORF。当翻译克隆Tru9I的3-236号核苷酸并将所得到的蛋白与坦科聚合酶比较时没有发现明显的相似性。
用SequencherTM程序对克隆RsaI和5’-RACEtank2进行排比。克隆RsaI(SEQ ID NO:115)在开始与5’-RACEtank2(SEQ ID NO:104)的5’末端重叠之前有654bp的新的序列,并从655号位置开始与5’-RACEtank2重叠。在比较克隆RsaI的1-654号核苷酸和坦科聚合酶时没有发现明显的相似性。确定克隆RsaI具有一个始于106号核苷酸位置的ORF,和一个始于287号核苷酸的ATG起始密码子。当翻译克隆RsaI的287-655号核苷酸并将所得到的蛋白与坦科聚合酶比较时没有发现明显的相似性。
用SequencherTM程序对克隆BfaI(SEQ ID NO:117)和5’-RACEtank2进行排比。克隆BfaI在开始与5’-RACEtank2(SEQ ID NO:104)的5’末端重叠之前含有88bp的新的序列,并从89号位置开始与5’-RACEtank2重叠。在比较克隆BfaI的1-88号核苷酸和坦科聚合酶时没有发现明显的相似性。确定克隆BfaI具有一个始于3号核苷酸位置的ORF。当翻译克隆BfaI的3-89号核苷酸并将所得到的蛋白与坦科聚合酶比较时没有发现明显的相似性。
为了证实从Tru9I,RsaI和BfaI克隆所获得的新的核苷酸序列并确定在所述新序列中是否存在内含子,对cDNA进行PCR扩增。合成相应于克隆RsaI的有义链(SEQ ID NO:115的59-84号核苷酸)的引物(5-RSA-1;SEQ ID NO:119)和相应于克隆RsaI的反义链(SEQ IDNO:115的708-727号核苷酸)的引物(3-Inv-1;SEQ ID NO:110),并用于PCR扩增反应中。
5-RSA-1 GTTCCTCTAATCAATCCTGAGC(SEQ ID NO:109)
进行6个独立的PCR反应(被命名为18、19、20、24、25和26),以便鉴定上述Taq聚合酶诱导的误差。每20微升反应物含有5微升人脾、胎盘、或睾丸Clontech Marathon-ReadycDNA DNA模板,0.20微摩尔每一种引物,0.20mMdNTPs,1×ClontechGC2PCR缓冲液,1.0摩尔ClontechGC-解链缓冲液,和0.4微升Clontech Advantage-GC2聚合酶。该反应在GeneAmpPCR系统9700中进行,采用以下四个步骤:1)1轮94℃1分钟;2)5轮94℃30秒和65℃2.5分钟;3)5轮94℃30秒和60℃2.5分钟;和4)25轮94℃30秒和58℃2.5分钟。按照说明通过凝胶电泳和QIAquick试剂盒分离PCR片段。按照说明将该PCR片段直接克隆到pCR2.1-TOPO载体上。用能与载体DNA杂交的M13引物(SEQ ID NO:25和26)和涉及用于与cDNA序列退火的引物(SEQID NO:112、120、121和122)对克隆18、19、20、24、25和26进行测序。
5-RSA-2 GGAAAGAGTAATTGATCAGAGCCATC
(SEQ ID NO:120)
5-RSA4 CGCCGAAGCCTCTCGCCTCACATTTCC
(SEQ ID NO:121)
3-RSAA GGAAATGTGAGGCGAGAGGCTTCGGCG
(SEQ ID NO:122)
克隆18、19、20、24、25和26的多核苷酸序列分别示于SEQ ID NO:123-128中。
用SequencherTM程序将克隆18、19、20、24、25和26与克隆RsaI进行排比。CDNA克隆的多核苷酸序列证实在RsaI克隆序列上不存在内含子。将克隆18、19、20、24、25和26的碱基对1-596编辑成共有核苷酸序列,该序列具有克隆RsaI的59-596bp,它被命名为5’-RSA/cDNA,并且如SEQ ID NO:129所示。5’-RSA/cDNA的多核苷酸序列不包括克隆18、19、20、24、25和26的碱基对597的3’序列,下面将加以讨论。5’-RSA/cDNA的多核苷酸序列还不包括克隆RsaI的bp1-58,没有证实该核苷酸序列存在于所述cDNA克隆序列上。在5’-RSA/cDNA的共有核苷酸序列,每个碱基对存在于这7个克隆中6个的相应的位置上。只有47号位置上的核苷酸例外,其中,该共有碱基对存在于7个克隆中4个的相应位置上。
对克隆18、19、20、24、25和26进行的排比确定了碱基对597 3’的核苷酸序列上的差别(参见SEQ ID NQ:123-128上的位置)。所有排比的克隆含有一个拷贝的位于588-597号核苷酸的(SEQ ID NO:123-128)的10个碱基对的序列(GAGCTGGCAG;SEQ ID NO:130)。克隆19和26具有第二个拷贝的序列GAGCTGGCAG重复,它直接与所述第一个拷贝相邻(589-607号核苷酸)(SEQ ID NO:124和128)。克隆RsaI、Tru9I和BfaI也具有两个拷贝的序列GAGCTGGCAG,这两个拷贝彼此直接相邻(在克隆RsaI上为646-665号核苷酸(SEQ IDNO:115);在克隆Tru9I上为227-246号核苷酸(SEQ ID NO:113);在克隆BfaI上为80-99号核苷酸(SEQ ID NO:117))。克隆18、20、24和25没有第二个拷贝的序列GAGCTGGCAG。第二个拷贝的序列GAGCTGGCAG的存在与否可能是由于Taq聚合酶引起的PCR扩增的误差所导致的。通过基因组DNA的直接测序可以证实这一推测。第二个序列GAGCTGGCAG的存在与否还可能是由于存在于细菌中的用于繁殖克隆DNA的复制和/或修复蛋白所导致的。可以通过对PCR产物直接测序证实这一推测。第二个序列GAGCTGGCAG的存在与否还可能是由于其他3’剪接受体的使用导致的。这种可能性似乎不大可能,因为在GAGCTGGCAG周围的序列与外显子/内含子/外显子边界的共有序列没有很高的相似性[Lewin,同上]。另外,业已分离了由基因组DNA的PCR扩增所产生的克隆,它仅含有一个拷贝的GAGCTGGCAG序列(基因组1X;SEQ ID NO:131),以及含有两个拷贝GAGCTGGCAG序列的克隆(克隆RsaI(SEQ ID NO:115),Tru9I(SEQ ID NO:113)和(SEQ ID NO:117))。第二个拷贝的序列GAGCTGGCAG的存在与否还可能是存在于人群中的多态性。在这种情况下,长的和短的形式的TANK2蛋白的表达是可能的,如下文所述。
第二个拷贝的序列GAGCTGGCAG的存在产生了长的形式的TANK2蛋白。用SequencherTM程序将克隆19、26、RsaI、Tru9I和BfaI与5’-RSA/cDNA和2B.1/2D.1/30B.2A/5’-RACE进行排比。通过该排比得到了一个被称为坦科聚合酶2-LONG的共有多核苷酸序列,并且如SEQ ID NO:132所示。确定坦科聚合酶2-LONG的序列具有一个从103-4386号核苷酸的ORF,第一个甲硫氨酸始于229号核苷酸。在推测的起始甲硫氨酸上游存在一个框内终止密码子(始于100号核苷酸)。假设该残基是起始甲硫氨酸,坦科聚合酶2-LONG的ORF编码一种具有1385个氨基酸的蛋白(被命名为TANK2-LONG;SEQ ID NO:133),其推测的分子量为149892Da。
一个拷贝的序列GAGCTGGCAG的存在能产生短的形式的TANK2蛋白。用SequencherTM程序将克隆10、20、24和25与5’-RSA/cDNA和2B.1/2D.1/30B.2A/5’-RACE进行排比。通过该排比得到了一个被称为坦科聚合酶2-SHORT的共有多核苷酸序列,并且如SEQ ID NO:134所示。确定坦科聚合酶2-SHORT的序列具有一个从513-4376号核苷酸的ORF,第一个甲硫氨酸始于876号核苷酸。在推测的起始甲硫氨酸上游存在一个框内终止密码子(始于510号核苷酸)。假设该残基是起始甲硫氨酸,坦科聚合酶2-SHORT的ORF编码一种具有1166个氨基酸的蛋白(被命名为TANK2-SHORT;SEQ ID NO:135),其推测的分子量为126908Da。TANK2-SHORT在其氨基末端比TANK2-LONG短219个氨基酸。TANK2-SHORT的推测的起始甲硫氨酸相当于TANK2-LONG的120号位置的甲硫氨酸。除了TANK2-LONG的头219个氨基酸之外,TANK2-LONG和TANK2-SHORT是相同的。
坦科聚合酶1基因(SEQ ID NO:3)编码一种含有羧基末端催化结构域,该结构域与人PARPI的催化结构域同源。parp1的多核苷酸序列如SEQ ID NO:136所示,PARP1的氨基酸序列如SEQ ID NO:137所示。TANK1的催化结构域(SEQ ID NO:4的1176-1314号氨基酸)与PARP1的催化结构域(SEQ ID NO:137的854-1014号氨基酸)同源,并且具有PARP催化活性(Smith等,同上)。类似的,TANK2-LONG的推测的催化结构域(SEQ ID NO:133的1242-1382号氨基酸)和TANK2-SHORT的推测的催化结构域(SEQ ID NO:135的1023-1161号氨基酸)与TANK1的催化结构域高度同源(在139个氨基酸中有130个相同;相同性为94%)。
TANK1的中央结构域含有24个锚蛋白重复,表明TANK1可能属于锚蛋白家族,它能将整合膜蛋白连接到细胞骨骼上[Bennett,生物化学杂志267:8703-6,1992]。TANK1的锚蛋白重复结构域(SEQ ID NO:4的181-1110号氨基酸)与TANK2-LONG的中央结构域(SEQ ID NO:133的242-1078号氨基酸)和TANK2-SHORT的中央结构域(SEQ IDNO:135的23-859号氨基酸)明显同源(在837个氨基酸中有692个相同;相同性为83%)。
在TANK1的锚蛋白重复结构域内有一个用于结合端粒重复结合因子(TRF1)的结合位点(Smith等,同上),它起着调节端粒长度的作用[van Steensel和de-Lange,自然385:740-3,1997]。TANK1的TRF1结合域(SEQ ID NO:4的435-797号氨基酸)与TANK2-LONG的一部分(SEQ ID NO:133的497-858号氨基酸)和TANK2-SHORT的一部分(SEQ ID NO:135的278-639号氨基酸)明显同源(在364个氨基酸中有297个相同;相同性为82%)。TANK1还含有一个不育性α组件(SAM)结构域[Smith等,同上],它被认为与蛋白-蛋白相互作用有关[Ponting,蛋白质科学4:1928-30,1995;Schultz等,蛋白质科学6:249-53,1997]。TANK2-LONG的一部分(SEQ ID NO:133的1089-1154号氨基酸)和TANK2-SHORT的一部分(SEQ ID NO:135的870-935号氨基酸)与TANK1的SAM结构域(SEQ ID NO:4的1023-1088号氨基酸)同源(在66个氨基酸中有50个相同;相同性为76%)。
TANK2与TANK1的若干推测的功能域的(催化结构域、锚蛋白重复、TRF1-结合域、和SAM结构域)的比较如上文所述。TANK2-LONG所包含其他氨基序列(锚蛋白重复的氨基末端的所有残基,即SEQ IDNO:133的1-241号氨基酸)可以与TANK1的氨基末端进行比较。TANK1的氨基末端含有组氨酸、脯氨酸和丝氨酸的同源聚合排列(HPS结构域,即SEQ ID NO:4的1-180号氨基酸)[Smith等,同上]。TANK2-LONG的氨基末端既不含有HPS结构域又不与TANK1的氨基末端明显同源。TANK2-LONG的氨基末端也比TANK1长61个氨基酸残基,并且包括48.6%的非极性残基,32.4%的极性残基,和19.5%的带电荷的残基。
TANK2-SHORT比TANK2-LONG短219个氨基酸残基,并且仅含有锚蛋白重复的氨基末端的22个氨基酸残基。有趣的是,果蝇坦科聚合酶(GenBank保藏号AF132196;SEQ ID NO:138)编码一种被称为dTANK的推测的蛋白(SEQ ID NO:139),它仅含有其锚蛋白重复的氨基末端的21个氨基酸残基。TANK2-SHORT和dTANK的氨基末端不是明显同源的,不过这两种蛋白在上文所讨论的其他推测的功能域上具有同源性。TANK2-SHORT的催化结构域(SEQ ID NO:135的1023-1161号氨基酸)与dTANK的一部分(SEQ ID NO:139的1033-1171号氨基酸)同源(139个氨基酸中有113个相同;相同性为81%)。TANK2-SHORT的推测的锚蛋白重复结构域(SEQ ID NO:135的23-859号氨基酸)与dTANK的中央结构域(SEQ ID NO:139的22-895号氨基酸)明显同源。TANK2-SHORT的推测的TRF1结合域与dTANK的一部分(SEQ ID NO:139的277-633号氨基酸)明显同源(在324个氨基酸中有241个相同;相同性为66%)。TANK2-SHORT的推测的SAM结合域(SEQ ID NO:135的870-935号氨基酸)与dTANK的一部分(SEQ ID NO:139的886-951号氨基酸)明显同源(在66个氨基酸中有31个相同;相同性为66%)。
实施例3
制备能与TANK2多肽发生免疫反应的抗体
本发明提供了对TANK2多肽有专一性的抗体。TANK2的抗体可以用本领域所公知的任何方法生产,例如,这些方法通常包括用纯化的TANK2、纯化的重组TANK2、纯化的重组TANK2片段或源于TANK2预测氨基酸序列的合成肽的制剂对实验动物进行免疫。为了增加获得对TANK2有合适的专一性的抗体的可能性,可以根据TANK21和TANK2之间的差异选择所述多肽的片段用做免疫原。例如,通过排比TANK21和TANK2发现了TANK1的由969-974号氨基酸组成的一个片段(SEQID NO:4)基本上不同于TANK2-LONG的相应部分(1030-1042号氨基酸)(SEQ ID NO:133)。另外,TANK1的氨基末端结构域(SEQID NO:4的1-180号氨基酸)和TANK2-LONG的氨基末端结构域(SEQID NO:133的1-241号氨基酸)明显不同,如上文所述。所述部分可以在合适的表达系统中以截短的多肽形式表达,以便用作免疫原或用于检测多克隆或单克隆抗体制剂。类似的方法可用于TANK2多肽的其他部分。同样,可以制备相应于有差别的各部分的合成肽,并可将这种肽用于通过本领域已知的方法制备专一性多克隆或单克隆抗体。例如,参见HarloW(1988)所述,同上。
比较TANK1和TANK2发现,TANK2-LONG的由1030-1042号氨基酸组成的一个片段(SEQ ID NO:133)明显不同于TANK1的相应部分(SEQ ID NO:4的969-974号氨基酸)。AnaSpec公司(San Jose,CA)合成了一种被称为ICEC#2的具有该TANK2序列的肽,用作抗体开发的免疫原。按照生产商的方法用Imject马来酰亚胺活化的载体蛋白(Pierce,#77106)将肽ICEC#2与KLH连接。
在第0天对4只6-12周龄Balb-c小鼠中的每一只进行预先放血,并通过给每只小鼠皮下注射50微克溶解在弗氏完全佐剂中的KLH-ICEC-2肽进行免疫。随后在第21天和42天用弗氏不完全佐剂进行强化免疫。在第52天对小鼠进行检测放血,并使用标准方法在用KLH-ICEC-2肽包衣的平板上通过ELISA对血液进行筛选。用山羊抗小鼠IgG(fc)辣根过氧化物酶(HRP)接合物检测专一性抗体。在第118和119天用溶解在PBS中的50微克KLH-ICEC-2肽对#3616小鼠进行融合前强化免疫。在第112天取出脾脏并融合。
用聚乙二醇1500(Boehringer Mannheim/Roche分子生物化学)通过标准方法以5∶1的比例将脾细胞与NS-1细胞融合[HarloW等,1988,同上]。将融合的细胞重新悬浮在250毫升RPMI中,该溶液含有15%的FBS,100mM次黄嘌呤钠,0.4mM氨基蝶呤,16mM胸苷(HAT)(Gibco BRL,Rockville,MD),10单位/毫升IL-6(BoehringerMannheim/Roche分子生物化学)和1.5×106鼠胸腺细胞/毫升。将该悬浮液分配到12个半96孔组织培养平板(Corning,英国)上,用量为200微升/孔。在融合之后第4、5和6天对平板上的细胞进行饲养,从每一个孔中吸出大约100微升,并添加100微升/孔上述铺平板培养基,所不同的是培养基中缺少胸腺细胞。
在第7-12天筛选来自融合细胞的上清液,按上述方法首先通过ELISA在所述免疫原上进行。为了确保克隆能力,将从融合中挑选的阳性孔通过有限稀释亚克隆3次,使用缺少氨基蝶呤的培养基。完成了一种融合体345C的克隆,它仍然能够与免疫蛋白反应。用山羊抗小鼠IgG1,IgG2a,IgG2b和IgG3HRP结合物作检测抗体通过标准ELISA方法对抗体进行分型。克隆345C是IgG1。
还通过Western分析检测345C对TANK2的免疫反应性。通过离心将1×107非增殖人PBL细胞沉淀,并通过添加0.5毫升缓冲液D进行裂解[0.1%NP40,0.1TX-100,100mM氯化钾,20mMHEPES,pH7.9,0.2mMEDTA,0.2mMEGTA,1.0mM二硫苏糖醇(DTT),和蛋白酶抑制剂混合片剂,Boehringer Mannheim/Roche分子生物化学]。以20%的功率输出对裂解物进行超声处理30秒(Sonifier250,Branson超声波公司,Danbury,CT),并在4℃下在离心机中离心5分钟进行澄清,并弃掉沉淀。将小鼠IgG(2.5微克)或0.5毫升345CmAb培养上清液添加到该裂解液中,在4℃下培养90分钟。通过用30微升蛋白G-琼脂浆(Pierce)在4℃下轻揉摇动30分钟使免疫复合体沉淀收集复合体。用4倍缓冲液D洗涤沉淀,重新悬浮在25微升1×SDS样品缓冲液中[50mMTris-HCl,pH6.8,2%SDS,0.1%溴酚兰,10%甘油,和100mMDDT],并在100℃下加热5分钟。
在8%Tris-甘氨酸聚丙烯酸酰胺凝胶(Novex,San Diego,CA)上以60mA的电流将样品电泳30分钟,按照生产商的说明进行。将凝胶转移到Immobilon-P转移膜(Millipore,Bedford,MA)上,使用Bio-Rad(Hercules,CA)半干燥吸印装置,按照生产商的说明在250mA的电流下进行90分钟。然后在含有5.0%脱脂奶粉的TBST缓冲液(Tris缓冲的盐溶液,pH7.5和0.5%Tween)中在室温下将印迹封闭20-30分钟。然后以1∶2的稀释比例将最初的mAb345C培养上清液假如含有0.1%脱脂奶粉的TBST中,并在室温下培养印迹90分钟。然后用含有1.0%脱脂奶粉的TBST稀释液以1∶3000的稀释比例洗涤4次以后(山羊抗小鼠IgG HRP结合物,Bio-Rad),所述稀释液是含有1.0%脱脂奶粉的TBST,并在室温下培养印迹30分钟。再次用TBST洗涤印迹4次,然后按照生产商的说明在ECL检测试剂中培养(Amersham生命科学,Uppsala,瑞典),然后对X光胶片进行曝光,获得了大致相当于预期的TANK2-LONG和TANK2-SHORT的大小的阳性信号。重复以上整个过程,以便获得更强的免疫活性单克隆抗体。
实施例4
通过Northern印迹杂交分析TANK2表达
为了证实能表达坦科聚合酶2mRNA的细胞和组织类型,用化学制备的多组织Northern印迹(Clontech)进行Northern印迹分析。用相当于FB2B.1多核苷酸序列的有义链(SEQ ID NO:88的2330-2349号核苷酸)的引物(5-TANK2-15;SEQ ID NO:140)和相当于FB2B.1多核苷酸序列的反义链(SEQ ID NO:88的2656-2675号核苷酸)的引物(3-TANK2-18;SEQ ID NO:141)通过PCR扩增DNA探针模板。
5-TANK2-15 GGCCTGAAGGTATGGTCGAT(SEQ ID NO:140)
3-TANK2-18 TGAGGGCATTACAGTTTGTT(SEQ ID NO:141)
PCR反应物含有100纳克FB2B.1DNA,每一种引物0.25微摩尔,0.20mMdNTPs,1×PCR缓冲液,和1微升Clontech Advantage-GC2聚合酶缓冲物。该反应在GeneAmpPCR系统9700中进行,采用以下四个步骤:1)1轮94℃1分钟;2)30轮94℃30秒,60℃30秒,72℃30秒;和3)1轮72℃7分钟。按照说明通过凝胶电泳和QIAquick试剂盒分离PCR片段(被命名为TANK2-Nprobe;SEQ ID NO:142)。按照说明用随机启动的DNA标记试剂盒(Boehringer Mannheim/Roche分子生物化学)用32P标记(TANK2-Nprobe),并用于检测Clontech多组织Northern印迹。与Clontech’s ExpressHYBTMDNA杂交溶液预杂交是在68℃下进行30分钟。与标记探针的杂交是在68℃下在ExpressHYBTM进行1小时。在室温下用含有2×SSC和0.05%SDS的缓冲液洗涤印迹3次,然后在50℃下用含有0.1×SSC和0.1%SDS的缓冲液洗涤2次,然后进行放射性自显影。
组织Northern印迹含有一个大约6.3kb的带,它的信号在胎盘、PBL、卵巢和脾脏中最强,并且存在于胰腺、肾、骨骼肌、肝脏、肺、大脑、心脏、结肠、小肠、睾丸、前列腺和胸腺中。
实施例5
通过原位杂交分析TANK2表达
按照以下方法通过原位杂交检查坦科聚合酶2在组织切片中的表达。
探针制备
用本领域常用方法制备用于坦科聚合酶2原位杂交的探针。合成相当于FB2B.1多核苷酸序列的有义链(SEQ ID NO:88的2330-2349号核苷酸)的引物(5-TANK2-15p;SEQ ID NO:143)和相当于FB2B.1多核苷酸序列的反义链(SEQ ID NO:88的2656-2675号核苷酸的引物(3-TANK2-18p;SEQ ID NO:144),用于以FB2B.1作模板的PCR反应中。
5-Tank2-1 5p GCCGAATTCGGCCTGAAGGTATGGTCGAT
(SEQ ID NO:143)
3-Tank2-1 8p GCCGAATTCTAGATGAGGGCATTACAGTITGTT
(SEQ ID NO:144)
PCR反应物含有100纳克FB2B.1cDNA,每一种引物0.5微摩尔,0.25mMdNTPs,1×PCR缓冲液,和2.5单位PfuTurbo聚合酶混合物(Stratagene)。该反应在GeneAmpPCR系统9700中进行,采用以下四个步骤:1)1轮94℃1分钟;2)25轮94℃30秒,55℃1分钟,72℃1分钟;和3)1轮72℃7分钟。用EcoRI消化PCR片段,通过凝胶电泳和QIAquick试剂盒分离,并亚克隆到Bluescript载体(Stratagene)上。该克隆被命名为TANK2-Isprobe,被设计用于与载体退火的M13引物(SEQ ID NO:25和26)进行测序,其序列如SEQID NO:145所示。用XhoI消化TANK2-ISprobe,并用T3聚合酶转录(见下文),以便制备反义探针。通过用BamHI消化TANK2-Isprobe并用T7聚合酶制备有义探针。
为了比较TANK2和TANK1的组织表达,制备了TANK1探针。TANK1相当于TANK1基因的3’非翻译序列部分。TANK1的3’非翻译序列被命名3-Tank1UT,如SEQ ID NO:146所示。合成相当于3-Tank1UT多核苷酸序列的有义链(SEQ ID NO:146的407-426号核苷酸)的引物(5-Tank1-7p;SEQ ID NO:149)和相当于3-Tank1UT多核苷酸序列的反义链(SEQ ID NO:146的72-767号核苷酸)的引物(3-Tank1-13p;SEQ ID NO:148),并用于以3-Tank1UT作模板的PCR反应中。
5-Tank1-7p GCCGAATTCCTTGTTTTTGATTTGCCAGA
(SEQ ID NO:147)
3-Tank1-13p GCCGAATTCCGGCTTTGACTTCTCTGAATTTAGG
(SEQ ID NO:148)
PCR反应物含有100纳克3-Tank1UTcDNA,每一种引物0.5微摩尔,0.25mMdNTPs,1×PCR缓冲液,和2.5单位PfuTurbo聚合酶混合物(Stratagene)。该反应在GeneAmpPCR系统9700中进行,采用以下四个步骤:1)1轮94℃1分钟;2)30轮94℃30秒,55℃1分钟,72℃1分钟;和3)1轮72℃7分钟。用EcoRI消化PCR片段,通过凝胶电泳和QIAquick试剂盒分离,并亚克隆到Bluescript载体(Stratagene)上。该克隆被命名为Tank1-ISprobe,用M13引物(SEQID NO:25和26)测序,其序列如SEQ ID NO:149所示。用BamHI消化Tank1-ISprobe,并用T7聚合酶转录以便制备反义探针。通过用XhoI消化Tank1-ISprobe,并用T3聚合酶转录制备有义探针。
用RNA转录试剂盒(Stratagene)在一种反应物中转录Tank1-ISprobe和Tank2-Isprobe,该反应物含有5微升5倍转录缓冲液,30mMDTT,ATP、CTP、GTP各0.8mM,40单位RNase BlockII,12.5单位T3或T7聚合酶,300纳克线性化的质粒模板,和50μCi35S-UTP(大于1000Ci/mM,Amersham,Arlington Heights,IL)。在37℃下培养该混合物1小时,然后通过添加1微升无RNase的DNaseI(Stratagene)并在37℃下培养15分钟清除模板DNA。按照生产商推荐的方法制备Quick Spin G50RNA柱(5’→3’公司,Boulder,CO)。将25微升蒸馏水添加到探针中,并将其放入柱的中央,以1100rpm的速度在吊篮式离心机中离心该柱4分钟。将该柱的流通液与50微升蒸馏水,2微升10毫克/毫升tRNA溶液,10微升3M乙酸钠,和200微升100%乙醇(VWR,So.Plainfield,NJ)混合,并将所得到的混合物在-20℃下培养一夜。在4℃下将探针溶液离心15分钟,除去上清液,将沉淀重新悬浮在含有1微升0.1MDTT的40微升1×TBE[90mMTris硼酸和2mMEDTA,pH8.0]中。将探针在-70℃下保存,直到进行原位杂交。
制备组织样品和原位杂交
将组织(国家疾病研究交流中心,费城,PA和人类组织协作网,费城,PA)切成6微米的厚度,并放在SuperfrostPlus载玻片(VWR)上。在4℃下用4%的低聚甲醛(Sigma,St.Louis,MO)固定切片20分钟。用1×CMF-FBS漂洗载玻片3次,在连续3次洗涤之后用70%、95%和100%的乙醇脱水,并在室温下干燥30分钟。将载玻片放入2×SSC制成的70%甲酰胺(J.T.Baker,Phillpsburg,NJ)中在70℃下放置2分钟,在4℃下用2×SSC漂洗,通过用70%、95%和100%的乙醇洗涤脱水,并在室温下干燥30分钟。将载玻片放入装有一片滤纸的空气密封箱中,该滤纸用4×SSC制备的甲酰胺的box缓冲液饱和。上述探针是这样单独制备的,将4×105cpm/组织切片与5微升10毫克/毫升tRNA溶液/切片混合,并将该混合物在95℃下加热3分钟。将冰镇的tHB2缓冲液[10%硫酸葡聚糖酯(Sigma),50%甲酰胺,100mMDTT(Boehringer Mannheim/Roche分子生物化学),0.3M氯化钠(Sigma),20mMTris,pH7.5,5mMEDTA(Sigma),和1×denhardt’s溶液(Sigma)]添加到探针混合物中,将最终体积调整到60微升/切片。然后将该探针溶液添加到组织切片中。在50℃下培养载玻片12-16小时,在杂交之后,用含有10mMDTT4×SSC洗涤载玻片一次,在室温下进行1小时,用50%去离子甲酰胺、1×SSC、和1mMDTT洗涤1次,在60℃下进行40分钟,用2×SSC洗涤1次,在室温下进行30分钟。并用0.1×SSC洗涤1次,在室温下进行30分钟。通过用70%、95%和100%的乙醇洗涤将切片脱水,并风干30分钟。将载玻片浸入柯达(Rochester,NY)NTP2核溶液中、在45℃下保持3小时,在室温下黑暗中保存,并在4℃下在黑暗中在有干燥剂的条件下保存,直到显影时间。用蒸馏水漂洗载玻片,并用苏木金和洋红染色,是通过下列一系列步骤转移载玻片完成的:在甲醛/醇(100毫升甲醇,900毫升80%乙醇)中5分钟;用水漂洗3次,共2分钟;在0.75%Harris苏木金(Sigma)中5分钟;用水漂洗3次,共2分钟;在1%HCl/50%乙醇中浸泡1次,用水漂洗1次,用1%碳酸锂浸泡4次;用自来水浸泡10分钟;用0.5%洋红(Sigma)浸泡2分钟,用水漂洗3次,2分钟;用70%的乙醇浸泡2分钟,用50%的乙醇漂洗3次1分钟;用100%漂洗2次1分钟;用二甲苯漂洗2次2分钟。用cytoseal60(Stephens Scientific;Riverdale,NJ)安装载玻片。
将用反义坦科聚合酶1或反义坦科聚合酶2探针所获得的信号与用相应的有义探针所获得的对照信号进行比较,将对反义坦科聚合酶1或反义坦科聚合酶2探针专一的任何信号认为是分别代表坦科聚合酶1或坦科聚合酶2表达。在人睾丸的大部分,包括精原细胞和精母细胞中检测到了坦科聚合酶1和坦科聚合酶2信号。在人脾的红色浆体中检测到坦科聚合酶1的信号,而在人脾的白色浆体中检测到坦科聚合酶2的信号。用类似方法将坦科聚合酶1和坦科聚合酶2的探针用于检测在其他组织中的表达。在小鼠胚胎中统一检测到坦科聚合酶1信号,最强的信号出现在皮肤中。在小鼠胚胎中还统一检测到坦科聚合酶2信号,最强的信号出现在大脑的间充质部分。
实施例6
鉴定坦科聚合酶2结合配偶体
如上文所述,TANK1能与端粒专一性DNA结合蛋白TRF1相互作用[Smith等,1998,同上]。TRF1的多核苷酸序列如SEQ ID NO:150所示,TRF1的氨基酸序列如SEQ ID NO:151所示。用酵母双杂交系统[Hollenburg等,分子细胞生物学15:3813-22,1995]测定TANK2是否也与TRF1相互作用。在这种酵母双杂交系统中,通过工程方法使酵母菌株L40含有位于HIS3上游的多个LexA结合位点和β-半乳糖苷酶基因。与LexA融合的蛋白(在BTM116载体上产生)和与VP16活化域融合的第二种蛋白(在VP16载体上产生)的相互作用导致了HIS3的表达,使得酵母能在缺乏组氨酸的培养基中生长。这两种蛋白的相互作用还导致了β-半乳糖苷酶基因的表达,这种表达可以通过比色测定检测[Breeden和Nasmyth,Cold Spring Harbor Symp Quant Biol643-650,1985]。
TRF1的TANK1结合域在这里被命名为TRF1-TankBD,业已将其作图于TRF1的氨基末端部分。用相当于TRF1多核苷酸序列的有义链(SEQ ID NO:150的1-24号核苷酸)的引物(5-TRF1;SEQ ID NO:152)和相当于TRF1多核苷酸序列的反义链(SEQ ID NO:150的184-201号核苷酸)的引物(3-TRF1;SEQ ID NO:153)通过PCR扩增TRF1-TankBD。
5-TRF1 GCCCCGGGGATCCTCATGGCGGAGGATGTTTCCTCAGCG
(SEQ ID NO:152)
3-TRF1 TCCCGGGGATCCTCACACCAGGCCCGCGTCCTC
(SEQ ID NO:153)
PCR反应物含有5微升Clontech人睾丸Marathon-ReadycDNA,每一种引物0.20微摩尔,0.20mMdNTPs,1×PCR缓冲液,和1微升Clontech Advantage-GC2聚合酶混合物(Stratagene)。用被设计用于与载体退火的M13反向引物(SEQ ID NO:26)和被设计用于与cDNA序列退火的引物(SEQ ID NO:153)对TRF1-TankBD进行测序。TRF1-TankBD的多核苷酸序列如SEQ ID NO:154所示,而氨基酸序列如SEQ ID NO:155所示。
如上文所述,TANK1的TRF1结合域在TANK2的由SEQ ID NO:133的497-858号氨基酸所组成的一个片段高度同源,用相当于tank2多核苷酸序列的有义链(SEQ ID NO:132的1717-1742号核苷酸)的引物(5-T2/TRF1BD;SEQ ID NO:156)和相当于tank2多核苷酸序列的反义链(SEQ ID NO:132的2765-2805号核苷酸)的引物(3-T2/TRF1BD;SEQ ID NO:157)通过PCR扩增相当于被称为Tank2-TRF1BD的TANK2的该结构域的多核苷酸片段。
5-T2/RF1BD CGCAGGATCCCCTTCACTCCTCTTCATGAGGCAGCTTC
(SEQ ID NO:156)
3-T2/RF1BD GGATCCGCTAAATATCTGTATCTCCATCTTTAACAA
GATCCAAAGGAG (SEQ ID NO:157)
PCR反应物含有5微升Clontech人睾丸Marathon-ReadycDNA,每一种引物0.5微摩尔,0.25mMdNTPs,1×PCR缓冲液,和2.5单位PfuTurbo聚合酶混合物(Stratagene)。该反应在GeneAmpPCR系统9700中进行,采用以下四个步骤:1)1轮94℃1分钟;2)30轮94℃30秒,55℃2分钟,72℃2分钟;和3)1轮72℃7分钟。按照说明通过凝胶电泳和QIAquick试剂盒分离PCR片段,并亚克隆到Pcr-BluntIITM-载体(Invitrogen)上。用BamHI将Tank2-TRF1BD从Pcr-BluntIITM-上消化下来,并亚克隆到VP16载体上。用被设计用于与载体序列结合的引物:M13正向(SEQ ID NO:25)和009(SEQ IDNO:158)对Tank2-TRF1BD克隆进行测序。
009 GCCGACTTCGAGTTTGAGCAG(SEQ ID NO:158)
该核苷酸序列如SEQ ID NO:159所示,而氨基酸序列如SEQ IDNO:160所示。
用TRF1-TankBD和Tank2-TRF1 BD质粒对L40进行共转化发现与TANK1一样,TANK2能结合TRF1。
实施例7
测定TANK2生物学活性
构建表达质粒
坦科聚合酶2多肽的一级结构表明,TANK2与TANK1一样,具有聚(ADP-核糖)聚合酶活性。TANK2的PARP活性或其某些亚结构可以通过该成分将ADP-核糖单元从NAD整合到与蛋白底物结合的ADP-核糖的聚合物上的能力测定。例如,在分子测定中,TANK1将ADP-核糖聚合物添加到TRF1蛋白上[Smith等,同上]。预计TANK2也具有这种功能和/或对其他底物进行ATP-核糖化。本领域技术人员很容易实现在特定底物上验证上述活性[例如,参见Smith等,同上]。
TANK21和TANK2的结构差异表明了这样的可能性,即TANK2可能具有不同于TANK1的蛋白质底物专一性。正如有关TANK1能结合TRF-1并将TRF-1聚ADP-核糖化的结果所证实的那样,预计TANK2的蛋白底物可以通过其结合TANK2的能力加以鉴定。能结合TANK2的其他底物可以通过在本申请的其他部分所披露的多种方法鉴定。
将含有与TANK2的996-1385号氨基酸上游(SEQ ID NO:133)融合的PARP1的1-662号氨基酸(SEQ ID NO:137)的被命名为PARP1A/TANK2B的融合蛋白用于测定TANK2聚(ADP-核糖)聚合酶活性。PARP1A/TANK2含有PARP1的DNA结合域(SEQ ID NO:137的1-373号氨基酸)和自我修饰结构域(SEQ ID NO:137的373-525号氨基酸)以及TANK2的推测的催化结构域(SEQ ID NO:133的1242-1382号氨基酸)。用相当于parp1多核苷酸序列的有义链(SEQID NO:136的1-30号核苷酸)的引物(Sal-PARP1;SEQ ID NO:161)和相当于parp1多核苷酸序列的反义链(SEQ ID NO:136的1957-1985号核苷酸)的引物(revMlu-PARP1;SEQ ID NO:162)通过PCR扩增该融合蛋白的PARP1A部分。
Sal-PARP1 CGTCGACCCATGGCGGAGTCTTCGGATAAGCTCTATCGA
(SEQ ID NO:161)
revMlu-PARP1 GGAAACGCGTTTGGTGCCAGGATTTACTGTCAGCTTCTT
(SEQ ID NO:162)
PCR反应物含有0.5微升人胸腺和睾丸QUICK-LoneTMcDNA(Clontech),每一种引物0.25微摩尔,0.20mMdNTPs,1×PCR缓冲液,和1微升Clontech Advantage-GC2聚合酶混合物。该反应在GeneAmp中进行(PE应用生物系统),采用以下四个步骤:1)1轮94℃1分钟;2)30轮94℃30秒,55℃2分钟,72℃2分钟;和3)1轮72℃7分钟。按照说明通过凝胶电泳和QIAquick试剂盒分离PCR片段(被称为parp1A)。按照说明将Parp1A亚克隆到pTrcHis2TM-TOPO载体(Invitrogen)上。用SalI和MluI将Parp1A从pTrcHis2TM-TOPO上消化下来,通过凝胶电泳和QIAquick试剂盒分离该片段,并保存用于下文所述的进一步的亚克隆。
用相当于tank2多核苷酸序列的有义链(SEQ ID NO:132的3214-3240号核苷酸)的引物(forMlu-TANK2;SEQ ID NO:163)和相当于tank2多核苷酸序列的反义链(SEQ ID NO:132的4350-4383号核苷酸)的引物(TANK2Strep-Not;164)通过PCR扩增该融合蛋白的TANK2B部分。
ForMIu-TANK2 CTTAAACGCGTTGAAGGACAAACACCTTTAGATTTAGTT
(SEQ ID NO:163)
TANK2-Strep-Not GTCGAAAGCGGCCGCTTAGCCTCCGAACTGTGGATGCC
TCCACGCTCCATCGACCATACCTTCAGGCCTCATAATCTGG
(SEQ ID NO:164)
PCR反应物含有100纳克2B.1Cdna,每一种引物0.25微摩尔,0.20mMdNTPs,1×PCR缓冲液,和1微升Clontech Advantage-GC2聚合酶混合物。该反应在GeneAmpPCR系统9700中进行,采用以下四个步骤:1)1轮94℃1分钟;2)30轮94℃30秒,60℃2分钟,72℃2分钟;和3)1轮72℃7分钟。按照说明通过凝胶电泳和QIAquick试剂盒分离PCR片段(被称为tank2B)。按照说明将tank2B亚克隆到pCDNA3.1/NT-GFP-TOPO载体(Invitrogen)上。用MluI和NotI将tank2B从pCDNA3.1/NT-GFP-TOPO上消化下来,并与SalI/MluI消化过的parp1A(见上文)一起亚克隆到pFASTBAC载体(Gibco BRL)上,该载体预先用SalI和NotI消化过。所得到的质粒被命名为pFB-PARP1A/TANK2B。
用被设计用于与载体序列退火的引物(SEQ ID NO:165-166)和被设计用于与cDNA序列退火的引物(SEQ ID NO:55、60和66,同上,和SEQ ID NO:167和176)对pFB-PARP1A/TANK2B进行测序。
载体引物
FastBac for TTTGTTCGCCCAOACTC(SEQ ID NO:165)
FastBac rev TATGTTTCAGGTTCAGGGGGAG(SEQ ID NO:166)
cDNA引物
P1 GCGGAAGCTGGAGGAGTGAC(SEQ ID NO:167)
P2 GTCACTCCTCCAGCTTCCGC(SEQ ID NO:168)
P3 AAGCCCTGAAGAAGCAGCTC(SEQ ID NO:169)
P4 GAGCTGCTTCTTCAGGOCTT(SEQ ID NO:170)
P5 CAGACACCCAACCGGAAGGA(SEQ ID NO:171)
P6 TCCTTCCGGTTGGGTGTCTG(SEQ ID NO:172)
P7 TCCGCCTCCACCAAGAGCCT(SEQ ID NO:173)
P8 AGGCTCTFGGTGGAGGCGGA(SEQ ID NO:174)
P9 TGGCCTGGTGGACATCGTTA(SEQ ID NO:175)
P10 TAACGAT(3TCCACCAGGCCA(SEQ ID NO:176)
PARP1A/TANK2B的核苷酸序列如SEQ ID NO:177所示,而PARP1A/TANK2B的氨基酸序列如SEQ ID NO:178所示。包括以下部分:位于1-36号氨基酸的HIS标记前导部分;位于37-698号氨基酸的PARP1;位于699-700号氨基酸的间隔序列;位于701-1090号氨基酸的TANK2部分;和位于1091-1099号氨基酸的Strep-tag部分。
重组病毒原种的生产和蛋白质纯化
按照生产商推荐的方法用FastBac系统(Gibco BRL)生产PARP1A/TANK2B重组病毒原种。并按以下方法进行蛋白表达。在27℃下让Sf9细胞在含有50单位/毫升青霉素和50微克/毫升硫酸链霉素(Gibco BRL)的CCM3培养基(Hyclone,Logan,UT)中生长。以大约每个细胞0.5个病毒的感染复数感染指数生长的细胞,并培养48小时。通过以1000×g离心15分钟收集细胞,并将沉淀冷冻在-80℃下包村待用。
用于蛋白纯化的试剂是从Sigma公司获得,除非另有说明。通过超声处理在裂解缓冲液[25mMTris-HCl,pH9.0,50mM葡萄糖,10mMEDTA,1mM2-巯基乙醇,1mMPMSF,100μM抗蛋白酶和2微克/毫升抑蛋白酶肽]中裂解细胞。将Igepal CA-630(最终浓度为0.2%)、Tween-20(最终浓度为0.2%)、和氯化钠(最终浓度为0.5M)添加到裂解缓冲液中,并在4℃下搅拌样品30分钟。在4℃下以20000×g的速度离心,然后收集上清液,此时用1毫克/毫升鱼精蛋白硫酸酯处理上清液,并在4℃下搅拌1小时。通过在4℃下以4000×g的速度离心20分钟收集上清液,此时通过用70%的硫酸铵使蛋白沉淀。通过在4℃下以20000×g的速度离心15分钟收集蛋白沉淀,并重新悬浮在再悬浮缓冲液[100mMTris-HCl,pH7.4,0.5mMEDTA,10%甘油,1mMPMSF,和12mM 2-巯基乙醇中]。按照说明首先用TaloSuperflow金属亲和树脂(Clontech)通过HIS标记纯化蛋白,并用200mM咪唑(Clontech)洗脱。然后用3-氨基苯甲酰胺Affi-Gel基质(Bio-Rad实验室)纯化蛋白洗脱液,该基质是按照在其他文献中披露的方法制备的[D’Amours等,分析生物化学249:106-8,1997]。用溶解在洗脱缓冲液[50mM Tris-HCl,pH7.5,0.3M氯化钠,10mM 2-巯基乙醇,1mMPMSF,100μM抗蛋白酶和2微克/毫升抑蛋白酶肽]中的10mM 3-甲氧基苯甲酰胺洗脱蛋白。用1升透析缓冲液[50mM Tris-HCl,pH8.0,1mMDTT,4M氯化镁,10mM EDTA,1mM PMSF,2微克/毫升抑蛋白酶肽]透析蛋白4次。以10%的最终浓度添加甘油,并在-80℃下保存该蛋白。
聚(ADP-核糖)聚合酶活性
用于聚(ADP-核糖)聚合酶活性测定的试剂是从Sigma公司获得,除非另有说明。在室温下在测定缓冲液(总体积为20微升)[100mMTris-HCl,pH8.0,10M氯化镁,10%甘油,1.5DTT(BoehringerMannheim/Roche分子生物化学),2.5μM未标记过的NAD+,16.7微克/毫升大肠杆菌菌株B DNA,0.33μCiγ-[32P]-NAD+(NEN,Boston,MA)]将PARP1A/TANK2B(250纳克)蛋白培养10分钟。通过在SDS电泳缓冲液中煮沸终止反应,并通过SDS-PAGE电泳分离。通过放射性自显影观察标记过的蛋白。将聚(ADP-核糖)聚合物添加到蛋白底物中会导致该蛋白分子量的增加,并因此导致该蛋白在SDS-PAGE上走的更高。另外,添加到蛋白底物中的聚(ADP-核糖)聚合物的量可能随着每一种蛋白分子而不同,导致标记的蛋白具有不同的分子量,它以梯子或条带形式出现在放射性自显影胶片上[例如,参见Smith等,科学282:2484-7,1998]。PARP1A/TANK2B具有内在的聚(ADP-核糖)聚合酶活性,表现在具有产生聚(ADP-核糖)聚合物的能力。PARP1A/TANK2B聚(ADP-核糖)聚合酶反应能产生从大约136-250kDa的标记过的蛋白梯。
在本说明书中所引用的所有出版物和专利文献以其全文形式收作本文参考。
尽管为了清楚和便于理解起见,结合特定优选实施方案对本发明进行了说明,但本领域技术人员可以理解的是,在下面所提供的 书所限定的本发明的范围内可以对这些实施方案作进一步的改变和改进。因此,除了在权利要求书中所专门限定的范围之外,不应当对本发明进行其他限定。
序列表<110>E.克里斯坦森(Christenson,Erik)
A.J.德马吉奥(DeMaggio,Anthony J)
P.S.戈德曼(Goldman,Phyllis S)
D.L.策里戈特(McElligott,David L)<120>坦科聚合酶2材料和方法<130>36559<140><141><150>60/141,582<151>1999-06-29<160>178<170>PatentIn Ver.2.1<210>1<211>3507<212>DNA<213>人(Homo sapiens)<220><221>CDS<222>(2)..(3508)<400>1g gcc agg atc atg tcg ggt cgc cgc tgc gcc ggc ggg gga gcg gcc tgc 49Ala Arg Ile Met Ser Gly Arg Arg Cys Ala Gly Gly Gly Ala Ala Cys
1 5 10 15gcg agc gcc gcg gcc gag gcc gtg gag ccg gcc gcc cga gag ctg ttc 97Ala Ser Ala Ala Ala Glu Ala Val Glu Pro Ala Ala Arg Glu Leu Phe
20 25 30gag gcg tgc cgc aac ggg gac gtg gaa cga gtc aag agg ctg gtg acg 145Glu Ala Cys Arg Asn Gly Asp Val Glu Arg Val Lys Arg Leu Val Thr
35 40 45cct gag aag gtg aac agc cgc gac acg gcg ggc agg aaa tcc acc ccg 193Pro Glu Lys Val Asn Ser Arg Asp Thr Ala Gly Arg Lys Ser Thr Pro
50 55 60ctg cac ttc gcc gca ggt ttt ggg cgg aaa gac gta gtt gaa tat ttg 241Leu His Phe Ala Ala Gly Phe Gly Arg Lys Asp Val Val Glu Tyr Leu65 70 75 80ctt cag aat ggt gca aat gtc caa gca cgt gat gat ggg ggc ctt att 289Leu Gln Asn Gly Ala Asn Val Gln Ala Arg Asp Asp Gly Gly Leu Ile
85 90 95cct ctt cat aat gca tgc tct ttt ggt cat gct gaa gta gtc aat ctc 337Pro Leu His Asn Ala Cys Ser Phe Gly His Ala Glu Val Val Asn Leu
100 105 110ctt ttg cga cat ggt gca gac ccc aat gct cga gat aat tgg aat tat 385Leu Leu Arg His Gly Ala Asp Pro Asn Ala Arg Asp Asn Trp Asn Tyr
115 120 125act cct ctc cat gaa gct gca att aaa gga aag att gat gtt tgc att 433Thr Pro Leu His Glu Ala Ala Ile Lys Gly Lys Ile Asp Val Cys Ile
130 135 140gtg ctg tta cag cat gga gct gag cca acc atc cga aat aca gat gga 481Val Leu Leu Gln His Gly Ala Glu Pro Thr Ile Arg Asn Thr Asp Gly145 150 155 160agg aca gca ttg gat tta gca gat cca tct gcc aaa gca gtg ctt act 529Arg Thr Ala Leu Asp Leu Ala Asp Pro Ser Ala Lys Ala Val Leu Thr
165 170 175ggt gaa tat aag aaa gat gaa ctc tta gaa agt gcc agg agt ggc aat 577Gly Glu Tyr Lys Lys Asp Glu Leu Leu Glu Ser Ala Arg Ser Gly Asn
180 185 190gaa gaa aaa atg atg gct cta ctc aca cca tta aat gtc aac tgc cac 625Glu Glu Lys Met Met Ala Leu Leu Thr Pro Leu Asn Val Asn Cys His
195 200 205gca agt gat ggc aga aag tca act cca tta cat ttg gca gca gga tat 673Ala Ser Asp Gly Arg Lys Ser Thr Pro Leu His Leu Ala Ala Gly Tyr
210 215 220aac aga gta aag att gta cag ctg tta ctg caa cat gga gct gat gtc 721Asn Arg Val Lys Ile Val Gln Leu Leu Leu Gln His Gly Ala Asp Val225 230 235 240cat gct aaa gat aaa ggt gat ctg gta cca tta cac aat gcc tgt tct 769His Ala Lys Asp Lys Gly Asp Leu Val Pro Leu His Asn Ala Cys Ser
245 250 255tat ggt cat tat gaa gta act gaa ctt ttg gtc aag cat ggt gcc tgt 817Tyr Gly His Tyr Glu Val Thr Glu Leu Leu Val Lys His Gly Ala Cys
260 265 270gta aat gca atg gac ttg tgg caa ttc act cct ctt cat gag gca gct 865Val Asn Ala Met Asp Leu Trp Gln Phe Thr Pro Leu His Glu Ala Ala
275 280 285tct aag aac agg gtt gaa gta tgt tct ctt ctc tta agt tat ggt gca 913Ser Lys Asn Arg Val Glu Val Cys Ser Leu Leu Leu Ser Tyr Gly Ala
290 295 300gac cca aca ctg ctc aat tgt cac aat aaa agt gct ata gac ttg gct 961Asp Pro Thr Leu Leu Asn Cys His Asn Lys Ser Ala Ile Asp Leu Ala305 310 315 320ccc aca cca cag tta aaa gaa aga tta gca tat gaa ttt aaa ggc cac 1009Pro Thr Pro Gln Leu Lys Glu Arg Leu Ala Tyr Glu Phe Lys Gly His
325 330 335tcg ttg ctg caa gct gca cga gaa gct gat gtt act cga atc aaa aaa 1057Ser Leu Leu Gln Ala Ala Arg Glu Ala Asp Val Thr Arg Ile Lys Lys
340 345 350cat ctc tct ctg gaa atg gtg aat ttc aag cat cct caa aca cat gaa 1105His Leu Ser Leu Glu Met Val Asn Phe Lys His Pro Gln Thr His Glu
355 360 365aca gca ttg cat tgt gct gct gca tct cca tat ccc aaa aga aag caa 1153Thr Ala Leu His Cys Ala Ala Ala Ser Pro Tyr Pro Lys Arg Lys Gln
370 375 380ata tgt gaa ctg ttg cta aga aaa gga gca aac atc aat gaa aag act 1201Ile Cys Glu Leu Leu Leu Arg Lys Gly Ala Asn Ile Asn Glu Lys Thr385 390 395 400aaa gaa ttc ttg act cct ctg cac gtg gca tct gag aaa gct cat aat 1249Lys Glu Phe Leu Thr Pro Leu His Val Ala Ser Glu Lys Ala His Asn
405 410 415gat gtt gtt gaa gta gtg gtg aaa cat gaa gca aag gtt aat gct ctg 1297Asp Val Val Glu Val Val Val Lys His Glu Ala Lys Val Asn Ala Leu
420 425 430gat aat ctt ggt cag act tct cta cac aga gct gca tat tgt ggt cat 1345Asp Asn Leu Gly Gln Thr Ser Leu His Arg Ala Ala Tyr Cys Gly His
435 440 445cta caa acc tgc cgc cta ctc ctg agc tat ggg tgt gat cct aac att 1393Leu Gln Thr Cys Arg Leu Leu Leu Ser Tyr Gly Cys Asp Pro Asn Ile
450 455 460ata tcc ctt cag ggc ttt act gct tta cag atg gga aat gaa aat gta 1441Ile Ser Leu Gln Gly Phe Thr Ala Leu Gln Met Gly Asn Glu Asn Val465 470 475 480cag caa ctc ctc caa gag ggt atc tca tta ggt aat tca gag gca gac 1489Gln Gln Leu Leu Gln Glu Gly Ile Ser Leu Gly Asn Ser Glu Ala Asp
485 490 495aga caa ttg ctg gaa gct gca aag gct gga gat gtc gaa act gta aaa 1537Arg Gln Leu Leu Glu Ala Ala Lys Ala Gly Asp Val Glu Thr Val Lys
500 505 510aaa ctg tgt act gtt cag agt gtc aac tgc aga gac att gaa ggg cgt 1585Lys Leu Cys Thr Val Gln Ser Val Asn Cys Arg Asp Ile Glu Gly Arg
515 520 525cag tct aca cca ctt cat ttt gca gct ggg tat aac aga gtg tcc gtg 1633Gln Ser Thr Pro Leu His Phe Ala Ala Gly Tyr Asn Arg Val Ser Val
530 535 540gtg gaa tat ctg cta cag cat gga gct gat gtg cat gct aaa gat aaa 1681Val Glu Tyr Leu Leu Gln His Gly Ala Asp Val His Ala Lys Asp Lys545 550 555 560gga ggc ctt gta cct ttg cac aat gca tgt tct tat gga cat tat gaa 1729Gly Gly Leu Val Pro Leu His Asn Ala Cys Ser Tyr Gly His Tyr Glu
565 570 575gtt gca gaa ctt ctt gtt aaa cat gga gca gta gtt aat gta gct gat 1777Val Ala Glu Leu Leu Val Lys His Gly Ala Val Val Asn Val Ala Asp
580 585 590tta tgg aaa ttt aca cct tta cat gaa gca gca gca aaa gga aaa tat 1825Leu Trp Lys Phe Thr Pro Leu His Glu Ala Ala Ala Lys Gly Lys Tyr
595 600 605gaa att tgc aaa ctt ctg ctc cag cat ggt gca gac cct aca aaa aaa 1873Glu Ile Cys Lys Leu Leu Leu Gln His Gly Ala Asp Pro Thr Lys Lys
610 615 620aac agg gat gga aat act cct ttg gat ctt gtt aaa gat gga gat aca 1921Asn Arg Asp Gly Asn Thr Pro Leu Asp Leu Val Lys Asp Gly Asp Thr625 630 635 640gat att caa gat ctg ctt agg gga gat gca gct ttg cta gat gct gcc 1969Asp Ile Gln Asp Leu Leu Arg Gly Asp Ala Ala Leu Leu Asp Ala Ala
645 650 655aag aag ggt tgt tta gcc aga gtg aag aag ttg tct tct cct gat aat 2017Lys Lys Gly Cys Leu Ala Arg Val Lys Lys Leu Ser Ser Pro Asp Asn
660 665 670gta aat tgc cgc gat acc caa ggc aga cat tca aca cct tta cat tta 2065Val Asn Cys Arg Asp Thr Gln Gly Arg His Ser Thr Pro Leu His Leu
675 680 685gca gct ggt tat aat aat tta gaa gtt gca gag tat ttg tta caa cac 2113Ala Ala Gly Tyr Asn Asn Leu Glu Val Ala Glu Tyr Leu Leu Gln His
690 695 700gga gct gat gtg aat gcc caa gac aaa gga gga ctt att cct tta cat 2161Gly Ala Asp Val Asn Ala Gln Asp Lys Gly Gly Leu Ile Pro Leu His705 710 715 720aat gca gca tct tac ggg cat gta gat gta gca gct cta cta ata aag 2209Asn Ala Ala Ser Tyr Gly His Val Asp Val Ala Ala Leu Leu Ile Lys
725 730 735tat aat gca tgt gtc aat gcc acg gac aaa tgg gct ttc aca cct ttg 2257Tyr Asn Ala Cys Val Asn Ala Thr Asp Lys Trp Ala Phe Thr Pro Leu
740 745 750cac gaa gca gcc caa aag gga cga aca cag ctt tgt gct ttg ttg cta 2305His Glu Ala Ala Gln Lys Gly Arg Thr Gln Leu Cys Ala Leu Leu Leu
755 760 765gcc cat gga gct gac ccg act ctt aaa aat cag gaa gga caa aca cct 2353Ala His Gly Ala Asp Pro Thr Leu Lys Asn Gln Glu Gly Gln Thr Pro
770 775 780tta gat tta gtt tca gca gat gat gtc agc gct ctt ctg aca gca gcc 2401Leu Asp Leu Val Ser Ala Asp Asp Val Ser Ala Leu Leu Thr Ala Ala785 790 795 800atg ccc cca tct gct ctg ccc tct tgt tac aag cct caa gtg ctc aat 2449Met Pro Pro Ser Ala Leu Pro Ser Cys Tyr Lys Pro Gln Val Leu Asn
805 810 815ggt gtg aga agc cca gga gcc act gca gat gct ctc tct tca ggt cca 2497Gly Val Arg Ser Pro Gly Ala Thr Ala Asp Ala Leu Ser Ser Gly Pro
820 825 830tct agc cca tca agc ctt tct gca gcc agc agt ctt gac aac tta tct 2545Ser Ser Pro Ser Ser Leu Ser Ala Ala Ser Ser Leu Asp Asn Leu Ser
835 840 845ggg agt ttt tca gaa ctg tct tca gta gtt agt tca agt gga aca gag 2593Gly Ser Phe Ser Glu Leu Ser Ser Val Val Ser Ser Ser Gly Thr Glu
850 855 860ggt gct tcc agt ttg gag aaa aag gag gtt cca gga gta gat ttt agc 2641Gly Ala Ser Ser Leu Glu Lys Lys Glu Val Pro Gly Val Asp Phe Ser865 870 875 880ata act caa ttc gta agg aat ctt gga ctt gag cac cta atg gat ata 2689Ile Thr Gln Phe Val Arg Asn Leu Gly Leu Glu His Leu Met Asp Ile
885 890 895ttt gag aga gaa cag atc act ttg gat gta tta gtt gag atg ggg cac 2737Phe Glu Arg Glu Gln Ile Thr Leu Asp Val Leu Val Glu Met Gly His
900 905 910aag gag ctg aag gag att gga atc aat gct tat gga cat agg cac aaa 2785Lys Glu Leu Lys Glu Ile Gly Ile Asn Ala Tyr Gly His Arg His Lys
915 920 925cta att aaa gga gtc gag aga ctt atc tcc gga caa caa ggt ctt aac 2833Leu Ile Lys Gly Val Glu Arg Leu Ile Ser Gly Gln Gln Gly Leu Asn
930 935 940cca tat tta act ttg aac acc tct ggt agt gga aca att ctt ata gat 2881Pro Tyr Leu Thr Leu Asn Thr Ser Gly Ser Gly Thr Ile Leu Ile Asp945 950 955 960ctg tct cct gat gat aaa gag ttt cag tct gtg gag gaa gag atg caa 2929Leu Ser Pro Asp Asp Lys Glu Phe Gln Ser Val Glu Glu Glu Met Gln
965 970 975agt aca gtt cga gag cac aga gat gga ggt cat gca ggt gga atc ttc 2977Ser Thr Val Arg Glu His Arg Asp Gly Gly His Ala Gly Gly Ile Phe
980 985 990aac aga tac aat att ctc aag att cag aag gtt tgt aac aag aaa cta 3025Asn Arg Tyr Asn Ile Leu Lys Ile Gln Lys Val Cys Asn Lys Lys Leu
995 1000 1005tgg gaa aga tac act cac cgg aga aaa gaa gtt tct gaa gaa aac cac 3073Trp Glu Arg Tyr Thr His Arg Arg Lys Glu Val Ser Glu Glu Asn His1010 1015 1020aac cat gcc aat gaa cga atg cta ttt cat ggg tct cct ttt gtg aat 3121Asn His Ala Asn Glu Arg Met Leu Phe His Gly Ser Pro Phe Val Asn1025 1030 1035 1040gca att atc cac aaa ggc ttt gat gaa agg cat gcg tac ata ggt ggt 3169Ala Ile Ile His Lys Gly Phe Asp Glu Arg His Ala Tyr Ile Gly Gly
1045 1050 1055atg ttt gga gct ggc att tat ttt gct gaa aac tct tcc aaa agc aat 3217Met Phe Gly Ala Gly Ile Tyr Phe Ala Glu Asn Ser Ser Lys Ser Asn
1060 1065 1070caa tat gta tat gga att gga gga ggt act ggg tgt cea gtt cac aaa 3265Gln Tyr Val Tyr Gly Ile Gly Gly Gly Thr Gly Cys Pro Val His Lys
1075 1080 1085gac aga tct tgt tac att tgc cac agg cag ctg ctc ttt tgc cgg gta 3313Asp Arg Ser Cys Tyr Ile Cys His Arg Gln Leu Leu Phe Cys Arg Val1090 1095 1100acc ttg gga aag tct ttc ctg cag ttc agt gca atg aaa atg gca cat 3361Thr Leu Gly Lys Ser Phe Leu Gln Phe Ser Ala Met Lys Met Ala His1105 1110 1115 1120tct cct cca ggt cat cac tca gtc act ggt agg ccc agt gta aat ggc 3409Ser Pro Pro Gly His His Ser Val Thr Gly Arg Pro Ser Val Asn Gly
1125 1130 1135cta gca tta gct gaa tat gtt att tac aga gga gaa cag gct tat cct 3457Leu Ala Leu Ala Glu Tyr Val Ile Tyr Arg Gly Glu Gln Ala Tyr Pro
1140 1145 1150gag tat tta att act tac cag att atg agg cct gaa ggt atg gtc gat 3505Glu Tyr Leu Ile Thr Tyr Gln Ile Met Arg Pro Glu Gly Met Val Asp
1155 1160 1165gga 3508Gly<210>2<211>1169<212>PRT<213>人(Homo sapiens)<400>2Ala Arg Ile Met Ser Gly Arg Arg Cys Ala Gly Gly Gly Ala Ala Cys1 5 10 15Ala Ser Ala Ala Ala Glu Ala Val Glu Pro Ala Ala Arg Glu Leu Phe
20 25 30Glu Ala Cys Arg Asn Gly Asp Val Glu Arg Val Lys Arg Leu Val Thr
35 40 45Pro Glu Lys Val Asn Ser Arg Asp Thr Ala Gly Arg Lys Ser Thr Pro
50 55 60Leu His Phe Ala Ala Gly Phe Gly Arg Lys Asp Val Val Glu Tyr Leu65 70 75 80Leu Gln Asn Gly Ala Asn Val Gln Ala Arg Asp Asp Gly Gly Leu Ile
85 90 95Pro Leu His Asn Ala Cys Ser Phe Gly His Ala Glu Val Val Asn Leu
100 105 110Leu Leu Arg His Gly Ala Asp Pro Asn Ala Arg Asp Asn Trp Asn Tyr
115 120 125Thr Pro Leu His Glu Ala Ala Ile Lys Gly Lys Ile Asp Val Cys Ile
130 135 140Val Leu Leu Gln His Gly Ala Glu Pro Thr Ile Arg Asn Thr Asp Gly145 150 155 160Arg Thr Ala Leu Asp Leu Ala Asp Pro Ser Ala Lys Ala Val Leu Thr
165 170 175Gly Glu Tyr Lys Lys Asp Glu Leu Leu Glu Ser Ala Arg Ser Gly Asn
180 185 190Glu Glu Lys Met Met Ala Leu Leu Thr Pro Leu Asn Val Asn Cys His
195 200 205Ala Ser Asp Gly Arg Lys Ser Thr Pro Leu His Leu Ala Ala Gly Tyr
210 215 220Asn Arg Val Lys Ile Val Gln Leu Leu Leu Gln His Gly Ala Asp Val225 230 235 240His Ala Lys Asp Lys Gly Asp Leu Val Pro Leu His Asn Ala Cys Ser
245 250 255Tyr Gly His Tyr Glu Val Thr Glu Leu Leu Val Lys His Gly Ala Cys
260 265 270Val Asn Ala Met Asp Leu Trp Gln Phe Thr Pro Leu His Glu Ala Ala
275 280 285Ser Lys Asn Arg Val Glu Val Cys Ser Leu Leu Leu Ser Tyr Gly Ala
290 295 300Asp Pro Thr Leu Leu Asn Cys His Asn Lys Ser Ala Ile Asp Leu Ala305 310 315 320Pro Thr Pro Gln Leu Lys Glu Arg Leu Ala Tyr Glu Phe Lys Gly His
325 330 335Ser Leu Leu Gln Ala Ala Arg Glu Ala Asp Val Thr Arg Ile Lys Lys
340 345 350His Leu Ser Leu Glu Met Val Asn Phe Lys His Pro Gln Thr His Glu
355 360 365Thr Ala Leu His Cys Ala Ala Ala Ser Pro Tyr Pro Lys Arg Lys Gln
370 375 380Ile Cys Glu Leu Leu Leu Arg Lys Gly Ala Asn Ile Asn Glu Lys Thr385 390 395 400Lys Glu Phe Leu Thr Pro Leu His Val Ala Ser Glu Lys Ala His Asn
405 410 415Asp Val Val Glu Val Val Val Lys His Glu Ala Lys Val Asn Ala Leu
420 425 430Asp Asn Leu Gly Gln Thr Ser Leu His Arg Ala Ala Tyr Cys Gly His
435 440 445Leu Gln Thr Cys Arg Leu Leu Leu Ser Tyr Gly Cys Asp Pro Asn Ile
450 455 460Ile Ser Leu Gln Gly Phe Thr Ala Leu Gln Met Gly Asn Glu Asn Val465 470 475 480Gln Gln Leu Leu Gln Glu Gly Ile Ser Leu Gly Asn Ser Glu Ala Asp
485 490 495Arg Gln Leu Leu Glu Ala Ala Lys Ala Gly Asp Val Glu Thr Val Lys
500 505 510Lys Leu Cys Thr Val Gln Ser Val Asn Cys Arg Asp Ile Glu Gly Arg
515 520 525Gln Ser Thr Pro Leu His Phe Ala Ala Gly Tyr Asn Arg Val Ser Val
530 535 540Val Glu Tyr Leu Leu Gln His Gly Ala Asp Val His Ala Lys Asp Lys545 550 555 560Gly Gly Leu Val Pro Leu His Asn Ala Cys Ser Tyr Gly His Tyr Glu
565 570 575Val Ala Glu Leu Leu Val Lys His Gly Ala Val Val Asn Val Ala Asp
580 585 590Leu Trp Lys Phe Thr Pro Leu His Glu Ala Ala Ala Lys Gly Lys Tyr
595 600 605Glu Ile Cys Lys Leu Leu Leu Gln His Gly Ala Asp Pro Thr Lys Lys
610 615 620Asn Arg Asp Gly Asn Thr Pro Leu Asp Leu Val Lys Asp Gly Asp Thr625 630 635 640Asp Ile Gln Asp Leu Leu Arg Gly Asp Ala Ala Leu Leu Asp Ala Ala
645 650 655Lys Lys Gly Cys Leu Ala Arg Val Lys Lys Leu Ser Ser Pro Asp Asn
660 665 670Val Asn Cys Arg Asp Thr Gln Gly Arg His Ser Thr Pro Leu His Leu
675 680 685Ala Ala Gly Tyr Asn Asn Leu Glu Val Ala Glu Tyr Leu Leu Gln His
690 695 700Gly Ala Asp Val Asn Ala Gln Asp Lys Gly Gly Leu Ile Pro Leu His705 710 715 720Asn Ala Ala Ser Tyr Gly His Val Asp Val Ala Ala Leu Leu Ile Lys
725 730 735Tyr Asn Ala Cys Val Asn Ala Thr Asp Lys Trp Ala Phe Thr Pro Leu
740 745 750His Glu Ala Ala Gln Lys Gly Arg Thr Gln Leu Cys Ala Leu Leu Leu
755 760 765Ala His Gly Ala Asp Pro Thr Leu Lys Asn Gln Glu Gly Gln Thr Pro
770 775 780Leu Asp Leu Val Ser Ala Asp Asp Val Ser Ala Leu Leu Thr Ala Ala785 790 795 800Met Pro Pro Ser Ala Leu Pro Ser Cys Tyr Lys Pro Gln Val Leu Asn
805 810 815Gly Val Arg Ser Pro Gly Ala Thr Ala Asp Ala Leu Ser Ser Gly Pro
820 825 830Ser Ser Pro Ser Ser Leu Ser Ala Ala Ser Ser Leu Asp Asn Leu Ser
835 840 845Gly Ser Phe Ser Glu Leu Ser Ser Val Val Ser Ser Ser Gly Thr Glu
850 855 860Gly Ala Ser Ser Leu Glu Lys Lys Glu Val Pro Gly Val Asp Phe Ser865 870 875 880Ile Thr Gln Phe Val Arg Asn Leu Gly Leu Glu His Leu Met Asp Ile
885 890 895Phe Glu Arg Glu Gln Ile Thr Leu Asp Val Leu Val Glu Met Gly His
900 905 910Lys Glu Leu Lys Glu Ile Gly Ile Asn Ala Tyr Gly His Arg His Lys
915 920 925Leu Ile Lys Gly Val Glu Arg Leu Ile Ser Gly Gln Gln Gly Leu Asn
930 935 940Pro Tyr Leu Thr Leu Asn Thr Ser Gly Ser Gly Thr Ile Leu Ile Asp945 950 955 960Leu Ser Pro Asp Asp Lys Glu Phe Gln Ser Val Glu Glu Glu Met Gln
965 970 975Ser Thr Val Arg Glu His Arg Asp Gly Gly His Ala Gly Gly Ile Phe
980 985 990Asn Arg Tyr Asn Ile Leu Lys Ile Gln Lys Val Cys Asn Lys Lys Leu
995 1000 1005Trp Glu Arg Tyr Thr His Arg Arg Lys Glu Val Ser Glu Glu Asn His1010 1015 1020Asn His Ala Asn Glu Arg Met Leu Phe His Gly Ser Pro Phe Val Asn1025 1030 1035 1040Ala Ile Ile His Lys Gly Phe Asp Glu Arg His Ala Tyr Ile Gly Gly
1045 1050 1055Met Phe Gly Ala Gly Ile Tyr Phe Ala Glu Asn Ser Ser Lys Ser Asn
1060 1065 1070Gln Tyr Val Tyr Gly Ile Gly Gly Gly Thr Gly Cys Pro Val His Lys
1075 1080 1085Asp Arg Ser Cys Tyr Ile Cys His Arg Gln Leu Leu Phe Cys Arg Val1090 1095 1100Thr Leu Gly Lys Ser Phe Leu Gln Phe Ser Ala Met Lys Met Ala His1105 1110 1115 1120Ser Pro Pro Gly His His Ser Val Thr Gly Arg Pro Ser Val Asn Gly
1125 1130 1135Leu Ala Leu Ala Glu Tyr Val Ile Tyr Arg Gly Glu Gln Ala Tyr Pro
1140 1145 1150Glu Tyr Leu Ile Thr Tyr Gln Ile Met Arg Pro Glu Gly Met Val Asp
1155 1160 1165Gly<210>3<211>3984<212>DNA<213>人(Homo sapiens)<220><221>CDS<222>(1)..(3981)<400>3atg gcg gcg tcg cgt cgc tct cag cat cat cac cac cat cat caa caa 48Met Ala Ala Ser Arg Arg Ser Gln His His His His His His Gln Gln1 5 10 15cag ctc cag ccc gcc cca ggg gct tca gcg ccg ccg ccg cca cct cct 96Gln Leu Gln Pro Ala Pro Gly Ala Ser Ala Pro Pro Pro Pro Pro Pro
20 25 30ccc cca ctc agc cct ggc ctg gcc ccg ggg acc acc cca gcc tct ccc 144Pro Pro Leu Ser Pro Gly Leu Ala Pro Gly Thr Thr Pro Ala Ser Pro
35 40 45acg gcc agc ggc ctg gcc ccc ttc gcc tcc ccg cgg cac ggc cta gcg 192Thr Ala Ser Gly Leu Ala Pro Phe Ala Ser Pro Arg His Gly Leu Ala
50 55 60ctg ccg gag ggg gat ggc agt cgg gat ccg ccc gac agg ccc cga tcc 240Leu Pro Glu Gly Asp Gly Ser Arg Asp Pro Pro Asp Arg Pro Arg Ser65 70 75 80ccg gac ccg gtt gac ggt acc agc tgt tgc agt acc acc agc aca atc 288Pro Asp Pro Val Asp Gly Thr Ser Cys Cys Ser Thr Thr Ser Thr Ile
85 90 95tgt acc gtc gcc gcc gct ccc gtg gtc cca gcg gtt tct act tca tct 336Cys Thr Val Ala Ala Ala Pro Val Val Pro Ala Val Ser Thr Ser Ser
100 105 110gcc gct ggg gtc gct ccc aac cca gcc ggc agt ggc agt aac aat tca 384Ala Ala Gly Val Ala Pro Asn Pro Ala Gly Ser Gly Ser Asn Asn Ser
115 120 125ccg tcg tcc tct tct tcc ccg act tct tcc tca tct tcc tct cca tcc 432Pro Ser Ser Ser Ser Ser Pro Thr Ser Ser Ser Ser Ser Ser Pro Ser
130 135 140tcc cct gga tcg agc ttg gcg gag agc ccc gag gcg gcc gga gtt agc 480Ser Pro Gly Ser Ser Leu Ala Glu Ser Pro Glu Ala Ala Gly Val Ser145 150 155 160agc aca gca cca ctg ggg cct ggg gca gca gga cct ggg aca ggg gtc 528Ser Thr Ala Pro Leu Gly Pro Gly Ala Ala Gly Pro Gly Thr Gly Val
165 170 175cca gca gtg agc ggg gcc cta cgg gaa ctg ctg gag gcc tgt cgc aat 576Pro Ala Val Ser Gly Ala Leu Arg Glu Leu Leu Glu Ala Cys Arg Asn
180 185 190ggg gac gtg tcc cgg gta aag agg ctg gtg gac gcg gca aac gta aat 624Gly Asp Val Ser Arg Val Lys Arg Leu Val Asp Ala Ala Asn Val Asn
195 200 205gca aag gac atg gcc ggc cgg aag tct tct ccc ctg cac ttc gct gca 672Ala Lys Asp Met Ala Gly Arg Lys Ser Ser Pro Leu His Phe Ala Ala
210 215 220ggt ttt gga agg aag gat gtt gta gaa cac tta cta cag atg ggt gct 720Gly Phe Gly Arg Lys Asp Val Val Glu His Leu Leu Gln Met Gly Ala225 230 235 240aat gtc cac gct cgt gat gat gga ggt ctc atc ccg ctt cat aat gcc 768Asn Val His Ala Arg Asp Asp Gly Gly Leu Ile Pro Leu His Asn Ala
245 250 255tgt tct ttt ggc cat gct gag gtt gtg agt ctg tta ttg tgc caa gga 816Cys Ser Phe Gly His Ala Glu Val Val Ser Leu Leu Leu Cys Gln Gly
260 265 270gct gat cca aat gcc agg gat aac tgg aac tat aca cct ctg cat gaa 864Ala Asp Pro Asn Ala Arg Asp Asn Trp Asn Tyr Thr Pro Leu His Glu
275 280 285gct gct att aaa ggg aag atc gat gtg tgc att gtg ctg ctg cag cac 912Ala Ala Ile Lys Gly Lys Ile Asp Val Cys Ile Val Leu Leu Gln His
290 295 300gga gct gac cca aac att cgg aac act gat ggg aaa tca gcc ctg gac 960Gly Ala Asp Pro Asn Ile Arg Asn Thr Asp Gly Lys Ser Ala Leu Asp305 310 315 320ctg gca gat cct tca gca aaa gct gtc ctt aca ggt gaa tac aag aaa 1008Leu Ala Asp Pro Ser Ala Lys Ala Val Leu Thr Gly Glu Tyr Lys Lys
325 330 335gac gaa ctc cta gaa gct gct agg agt ggt aat gaa gaa aaa cta atg 1056Asp Glu Leu Leu Glu Ala Ala Arg Ser Gly Asn Glu Glu Lys Leu Met
340 345 350gct tta ctg act cct cta aat gtg aat tgc cat gca agt gat ggg cga 1104Ala Leu Leu Thr Pro Leu Asn Val Asn Cys His Ala Ser Asp Gly Arg
355 360 365aag tcg act cct tta cat cta gca gcg ggc tac aac aga gtt cga ata 1152Lys Ser Thr Pro Leu His Leu Ala Ala Gly Tyr Asn Arg Val Arg Ile
370 375 380gtt cag ctt ctt ctt cag cat ggt gct gat gtt cat gca aaa gac aaa 1200Val Gln Leu Leu Leu Gln His Gly Ala Asp Val His Ala Lys Asp Lys385 390 395 400ggt gga ctt gtg cct ctt cat aat gca tgt tca tat gga cat tat gaa 1248Gly Gly Leu Val Pro Leu His Asn Ala Cys Ser Tyr Gly His Tyr Glu
405 410 415gtc aca gaa ctg cta cta aag cat gga gct tgt gtt aat gcc atg gat 1296Val Thr Glu Leu Leu Leu Lys His Gly Ala Cys Val Asn Ala Met Asp
420 425 430ctc tgg cag ttt act cca ctg cac gag gct gct tcc aag aac cgt gta 1344Leu Trp Gln Phe Thr Pro Leu His Glu Ala Ala Ser Lys Asn Arg Val
435 440 445gaa gtc tgc tct ttg tta ctt agc cat ggc gct gat cct acg tta gtc 1392Glu Val Cys Ser Leu Leu Leu Ser His Gly Ala Asp Pro Thr Leu Val
450 455 460aac tgc cat ggc aaa agt gct gtg gat atg gct cca act ccg gag ctt 1440Asn Cys His Gly Lys Ser Ala Val Asp Met Ala Pro Thr Pro Glu Leu465 470 475 480agg gag aga ttg act tat gaa ttt aaa ggt cat tct tta cta caa gca 1488Arg Glu Arg Leu Thr Tyr Glu Phe Lys Gly His Ser Leu Leu Gln Ala
485 490 495gcc aga gaa gca gac tta gct aaa gtt aaa aaa aca ctc gct ctg gaa 1536Ala Arg Glu Ala Asp Leu Ala Lys Val Lys Lys Thr Leu Ala Leu Glu
500 505 510atc att aat ttc aaa caa ccg cag tct cat gaa aca gca ctg cac tgt 1584Ile Ile Asn Phe Lys Gln Pro Gln Ser His Glu Thr Ala Leu His Cys
515 520 525gct gtg gcc tct ctg cat ccc aaa cgt aaa caa gtg aca gaa ttg tta 1632Ala Val Ala Ser Leu His Pro Lys Arg Lys Gln Val Thr Glu Leu Leu
530 535 540ctt aga aaa gga gca aat gtt aat gaa aaa aat aaa gat ttc atg act 1680Leu Arg Lys Gly Ala Asn Val Asn Glu Lys Asn Lys Asp Phe Met Thr545 550 555 560ccc ctg cat gtt gca gcc gaa aga gcc cat aat gat gtc atg gaa gtt 1728Pro Leu His Val Ala Ala Glu Arg Ala His Asn Asp Val Met Glu Val
565 570 575ctg cat aag cat ggc gcc aag atg aat gca ctg gac acc ctt ggt cag 1776Leu His Lys His Gly Ala Lys Met Asn Ala Leu Asp Thr Leu Gly Gln
580 585 590act gct ttg cat aga gcc gcc cta gca ggc cac ctg cag acc tgc cgc 1824Thr Ala Leu His Arg Ala Ala Leu Ala Gly His Leu Gln Thr Cys Arg
595 600 605ctc ctg ctg agt tac ggc tct gac ccc tcc atc atc tcc tta caa ggc 1872Leu Leu Leu Ser Tyr Gly Ser Asp Pro Ser Ile Ile Ser Leu Gln Gly
610 615 620ttc aca gca gca cag atg ggc aat gaa gca gtg cag cag att ctg agt 1920Phe Thr Ala Ala Gln Met Gly Asn Glu Ala Val Gln Gln Ile Leu Ser625 630 635 640gag agt aca cct ata cgt act tct gat gtt gat tat cga ctc tta gag 1968Glu Ser Thr Pro Ile Arg Thr Ser Asp Val Asp Tyr Arg Leu Leu Glu
645 650 655gca tct aaa gct gga gac ttg gaa act gtg aag caa ctt tgc agc tct 2016Ala Ser Lys Ala Gly Asp Leu Glu Thr Val Lys Gln Leu Cys Ser Ser
660 665 670caa aat gtg aat tgt aga gac tta gag ggc cgg cat tcc acg ccc tta 2064Gln Asn Val Asn Cys Arg Asp Leu Glu Gly Arg His Ser Thr Pro Leu
675 680 685cac ttc gca gca ggc tac aac cgc gtg tct gtt gta gag tac ctg cta 2112His Phe Ala Ala Gly Tyr Asn Arg Val Ser Val Val Glu Tyr Leu Leu
690 695 700cac cac ggt gcc gat gtc cat gcc aaa gac aag ggt ggc ttg gtg ccc 2160His His Gly Ala Asp Val His Ala Lys Asp Lys Gly Gly Leu Val Pro705 710 715 720ctt cat aat gcc tgt tca tat gga cac tat gag gtg gct gag ctt tta 2208Leu His Asn Ala Cys Ser Tyr Gly His Tyr Glu Val Ala Glu Leu Leu
725 730 735gta agg cat ggg gct tct gtc aat gtg gcg gac tta tgg aaa ttt acc 2256Val Arg His Gly Ala Ser Val Asn Val Ala Asp Leu Trp Lys Phe Thr
740 745 750cct ctc cat gaa gca gca gct aaa gga aag tat gaa atc tgc aag ctc 2304Pro Leu His Glu Ala Ala Ala Lys Gly Lys Tyr Glu Ile Cys Lys Leu
755 760 765ctt tta aaa cat gga gca gat cca act aaa aag aac aga gat gga aat 2352Leu Leu Lys His Gly Ala Asp Pro Thr Lys Lys Asn Arg Asp Gly Asn
770 775 780aca cct ttg gat ttg gta aag gaa gga gac aca gat att cag gac tta 2400Thr Pro Leu Asp Leu Val Lys Glu Gly Asp Thr Asp Ile Gln Asp Leu785 790 795 800ctg aaa ggg gat gct gct ttg ttg gat gct gcc aag aag ggc tgc ctg 2448Leu Lys Gly Asp Ala Ala Leu Leu Asp Ala Ala Lys Lys Gly Cys Leu
805 810 815gca aga gtg cag aag ctc tgt acc cca gag aat atc aac tgc aga gac 2496Ala Arg Val Gln Lys Leu Cys Thr Pro Glu Asn Ile Asn Cys Arg Asp
820 825 830acc cag ggc aga aat tca acc cct ctg cac ctg gca gca ggc tat aat 2544Thr Gln Gly Arg Asn Ser Thr Pro Leu His Leu Ala Ala Gly Tyr Asn
835 840 845aac ctg gaa gta gct gaa tat ctt cta gag cat gga gct gat gtt aat 2592Asn Leu Glu Val Ala Glu Tyr Leu Leu Glu His Gly Ala Asp Val Asn
850 855 860gcc cag gac aag ggt ggt tta att cct ctt cat aat gcg gca tct tat 2640Ala Gln Asp Lys Gly Gly Leu Ile Pro Leu His Asn Ala Ala Ser Tyr865 870 875 880ggg cat gtt gac ata gcg gct tta ttg ata aaa tac aac acg tgt gta 2688Gly His Val Asp Ile Ala Ala Leu Leu Ile Lys Tyr Asn Thr Cys Val
885 890 895aat gca aca gat aag tgg gcg ttt act ccc ctc cat gaa gca gcc cag 2736Asn Ala Thr Asp Lys Trp Ala Phe Thr Pro Leu His Glu Ala Ala Gln
900 905 910aaa gga agg acg cag ctg tgc gcc ctc ctc cta gcg cat ggt gca gac 2784Lys Gly Arg Thr Gln Leu Cys Ala Leu Leu Leu Ala His Gly Ala Asp
915 920 925ccc acc atg aag aac cag gaa ggc cag acg cct ctg gat ctg gca aca 2832Pro Thr Met Lys Asn Gln Glu Gly Gln Thr Pro Leu Asp Leu Ala Thr
930 935 940gct gac gat atc aga gct ttg ctg ata gat gcc atg ccc cca gag gcc 2880Ala Asp Asp Ile Arg Ala Leu Leu Ile Asp Ala Met Pro Pro Glu Ala945 950 955 960tta cct acc tgt ttt aaa cct cag gct act gta gtg agt gcc tct ctg 2928Leu Pro Thr Cys Phe Lys Pro Gln Ala Thr Val Val Ser Ala Ser Leu
965 970 975atc tca cca gca tcc acc ccc tcc tgc ctc tcg gct gcc agc agc ata 2976Ile Ser Pro Ala Ser Thr Pro Ser Cys Leu Ser Ala Ala Ser Ser Ile
980 985 990gac aac ctc act ggc cct tta gca gag ttg gcc gta gga gga gcc tcc 3024Asp Asn Leu Thr Gly Pro Leu Ala Glu Leu Ala Val Gly Gly Ala Ser
995 1000 1005aat gca ggg gat ggc gcc gcg gga aca gaa agg aag gaa gga gaa gtt 3072Asn Ala Gly Asp Gly Ala Ala Gly Thr Glu Arg Lys Glu Gly Glu Val1010 1015 1020gct ggt ctt gac atg aat atc agc caa ttt cta aaa agc ctt ggc ctt 3120Ala Gly Leu Asp Met Asn Ile Ser Gln Phe Leu Lys Ser Leu Gly Leu1025 1030 1035 1040gaa cac ctt cgg gat atc ttt gaa aca gaa cag att aca cta gat gtg 3168Glu His Leu Arg Asp Ile Phe Glu Thr Glu Gln Ile Thr Leu Asp Val
1045 1050 1055ttg gct gat atg ggt cat gaa gag ttg aaa gaa ata ggc atc aat gca 3216Leu Ala Asp Met Gly His Glu Glu Leu Lys Glu Ile Gly Ile Asn Ala
1060 1065 1070tat ggg cac cgc cac aaa tta atc aaa gga gta gaa aga ctc tta ggt 3264Tyr Gly His Arg His Lys Leu Ile Lys Gly Val Glu Arg Leu Leu Gly
1075 1080 1085gga caa caa ggc acc aat cct tat ttg act ttt cac tgt gtt aat cag 3312Gly Gln Gln Gly Thr Asn Pro Tyr Leu Thr Phe His Cys Val Asn Gln1090 1095 1100gga acg att ttg ctg gat ctt gct cca gaa gat aaa gaa tat cag tca 3360Gly Thr Ile Leu Leu Asp Leu Ala Pro Glu Asp Lys Glu Tyr Gln Ser1105 1110 1115 1120gtg gaa gaa gag atg caa agt act att cga gaa cac aga gat ggt ggt 3408Val Glu Glu Glu Met Gln Ser Thr Ile Arg Glu His Arg Asp Gly Gly
1125 1130 1135aat gct ggc ggc atc ttc aac aga tac aat gtc att cga att caa aaa 3456Asn Ala Gly Gly Ile Phe Asn Arg Tyr Asn Val Ile Arg Ile Gln Lys
1140 1145 1150gtt gtc aac aag aag ttg agg gag cgg ttc tgc cac cga cag aag gaa 3504Val Val Asn Lys Lys Leu Arg Glu Arg Phe Cys His Arg Gln Lys Glu
1155 1160 1165gtg tct gag gag aat cac aac cat cac aat gag cgc atg ttg ttt cat 3552Val Ser Glu Glu Asn His Asn His His Asn Glu Arg Met Leu Phe His1170 1175 1180ggt tct cct ttc att aat gcc att att cat aaa ggg ttt gat gag cga 3600Gly Ser Pro Phe Ile Asn Ala Ile Ile His Lys Gly Phe Asp Glu Arg1185 1190 1195 1200cat gca tac ata gga gga atg ttt ggg gcc ggg att tat ttt gct gaa 3648His Ala Tyr Ile Gly Gly Met Phe Gly Ala Gly Ile Tyr Phe Ala Glu
1205 1210 1215aac tcc tca aaa agc aac caa tat gtt tat gga att gga gga gga aca 3696Asn Ser Ser Lys Ser Asn Gln Tyr Val Tyr Gly Ile Gly Gly Gly Thr
1220 1225 1230ggc tgc cct aca cac aag gac agg tca tgc tat ata tgt cac aga caa 3744Gly Cys Pro Thr His Lys Asp Arg Ser Cys Tyr Ile Cys His Arg Gln
1235 1240 1245atg ctc ttc tgt aga gtg acc ctt ggg aaa tcc ttt ctg cag ttt agc 3792Met Leu Phe Cys Arg Val Thr Leu Gly Lys Ser Phe Leu Gln Phe Ser1250 1255 1260acc atg aaa atg gcc cac gcg cct cca ggg cac cac tca gtc att ggt 3840Thr Met Lys Met Ala His Ala Pro Pro Gly His His Ser Val Ile Gly1265 1270 1275 1280aga ccg agc gtc aat ggg ctg gca tat gct gaa tat gtc atc tac aga 3888Arg Pro Ser Val Asn Gly Leu Ala Tyr Ala Glu Tyr Val Ile Tyr Arg
1285 1290 1295gga gaa cag gca tac cca gag tat ctt atc act tac cag atc atg aag 3936Gly Glu Gln Ala Tyr Pro Glu Tyr Leu Ile Thr Tyr Gln Ile Met Lys
1300 1305 1310cca gaa gcc cct tcc cag acc gca aca gcc gca gag cag aag acc tag 3984Pro Glu Ala Pro Ser Gln Thr Ala Thr Ala Ala Glu Gln Lys Thr
1315 1320 1325<210>4<211>1327<212>PRT<213>人(Homo sapiens)<400>4Met Ala Ala Ser Arg Arg Ser Gln His His His His His His Gln Gln1 5 10 15Gln Leu Gln Pro Ala Pro Gly Ala Ser Ala Pro Pro Pro Pro Pro Pro
20 25 30Pro Pro Leu Ser Pro Gly Leu Ala Pro Gly Thr Thr Pro Ala Ser Pro
35 40 45Thr Ala Ser Gly Leu Ala Pro Phe Ala Ser Pro Arg His Gly Leu Ala
50 55 60Leu Pro Glu Gly Asp Gly Ser Arg Asp Pro Pro Asp Arg Pro Arg Ser65 70 75 80Pro Asp Pro Val Asp Gly Thr Ser Cys Cys Ser Thr Thr Ser Thr Ile
85 90 95Cys Thr Val Ala Ala Ala Pro Val Val Pro Ala Val Ser Thr Ser Ser
100 105 110Ala Ala Gly Val Ala Pro Asn Pro Ala Gly Ser Gly Ser Asn Asn Ser
115 120 125Pro Ser Ser Ser Ser Ser Pro Thr Ser Ser Ser Ser Ser Ser Pro Ser
130 135 140Ser Pro Gly Ser Ser Leu Ala Glu Ser Pro Glu Ala Ala Gly Val Ser145 150 155 160Ser Thr Ala Pro Leu Gly Pro Gly Ala Ala Gly Pro Gly Thr Gly Val
165 170 175Pro Ala Val Ser Gly Ala Leu Arg Glu Leu Leu Glu Ala Cys Arg Asn
180 185 190Gly Asp Val Ser Arg Val Lys Arg Leu Val Asp Ala Ala Asn Val Asn
195 200 205Ala Lys Asp Met Ala Gly Arg Lys Ser Ser Pro Leu His Phe Ala Ala
210 215 220Gly Phe Gly Arg Lys Asp Val Val Glu His Leu Leu Gln Met Gly Ala225 230 235 240Asn Val His Ala Arg Asp Asp Gly Gly Leu Ile Pro Leu His Asn Ala
245 250 255Cys Ser Phe Gly His Ala Glu Val Val Ser Leu Leu Leu Cys Gln Gly
260 265 270Ala Asp Pro Asn Ala Arg Asp Asn Trp Asn Tyr Thr Pro Leu His Glu
275 280 285Ala Ala Ile Lys Gly Lys Ile Asp Val Cys Ile Val Leu Leu Gln His
290 295 300Gly Ala Asp Pro Asn Ile Arg Asn Thr Asp Gly Lys Ser Ala Leu Asp305 310 315 320Leu Ala Asp Pro Ser Ala Lys Ala Val Leu Thr Gly Glu Tyr Lys Lys
325 330 335Asp Glu Leu Leu Glu Ala Ala Arg Ser Gly Asn Glu Glu Lys Leu Met
340 345 350Ala Leu Leu Thr Pro Leu Asn Val Asn Cys His Ala Ser Asp Gly Arg
355 360 365Lys Ser Thr Pro Leu His Leu Ala Ala Gly Tyr Asn Arg Val Arg Ile
370 375 380Val Gln Leu Leu Leu Gln His Gly Ala Asp Val His Ala Lys Asp Lys385 390 395 400Gly Gly Leu Val Pro Leu His Asn Ala Cys Ser Tyr Gly His Tyr Glu
405 410 415Val Thr Glu Leu Leu Leu Lys His Gly Ala Cys Val Asn Ala Met Asp
420 425 430Leu Trp Gln Phe Thr Pro Leu His Glu Ala Ala Ser Lys Asn Arg Val
435 440 445Glu Val Cys Ser Leu Leu Leu Ser His Gly Ala Asp Pro Thr Leu Val
450 455 460Asn Cys His Gly Lys Ser Ala Val Asp Met Ala Pro Thr Pro Glu Leu465 470 475 480Arg Glu Arg Leu Thr Tyr Glu Phe Lys Gly His Ser Leu Leu Gln Ala
485 490 495Ala Arg Glu Ala Asp Leu Ala Lys Val Lys Lys Thr Leu Ala Leu Glu
500 505 510Ile Ile Asn Phe Lys Gln Pro Gln Ser His Glu Thr Ala Leu His Cys
515 520 525Ala Val Ala Ser Leu His Pro Lys Arg Lys Gln Val Thr Glu Leu Leu
530 535 540Leu Arg Lys Gly Ala Asn Val Asn Glu Lys Asn Lys Asp Phe Met Thr545 550 555 560Pro Leu His Val Ala Ala Glu Arg Ala His Asn Asp Val Met Glu Val
565 570 575Leu His Lys His Gly Ala Lys Met Asn Ala Leu Asp Thr Leu Gly Gln
580 585 590Thr Ala Leu His Arg Ala Ala Leu Ala Gly His Leu Gln Thr Cys Arg
595 600 605Leu Leu Leu Ser Tyr Gly Ser Asp Pro Ser Ile Ile Ser Leu Gln Gly
610 615 620Phe Thr Ala Ala Gln Met Gly Asn Glu Ala Val Gln Gln Ile Leu Ser625 630 635 640Glu Ser Thr Pro Ile Arg Thr Ser Asp Val Asp Tyr Arg Leu Leu Glu
645 650 655Ala Ser Lys Ala Gly Asp Leu Glu Thr Val Lys Gln Leu Cys Ser Ser
660 665 670Gln Asn Val Asn Cys Arg Asp Leu Glu Gly Arg His Ser Thr Pro Leu
675 680 685His Phe Ala Ala Gly Tyr Asn Arg Val Ser Val Val Glu Tyr Leu Leu
690 695 700His His Gly Ala Asp Val His Ala Lys Asp Lys Gly Gly Leu Val Pro705 710 715 720Leu His Asn Ala Cys Ser Tyr Gly His Tyr Glu Val Ala Glu Leu Leu
725 730 735Val Arg His Gly Ala Ser Val Asn Val Ala Asp Leu Trp Lys Phe Thr
740 745 750Pro Leu His Glu Ala Ala Ala Lys Gly Lys Tyr Glu Ile Cys Lys Leu
755 760 765Leu Leu Lys His Gly Ala Asp Pro Thr Lys Lys Asn Arg Asp Gly Asn
770 775 780Thr Pro Leu Asp Leu Val Lys Glu Gly Asp Thr Asp Ile Gln Asp Leu785 790 795 800Leu Lys Gly Asp Ala Ala Leu Leu Asp Ala Ala Lys Lys Gly Cys Leu
805 810 815Ala Arg Val Gln Lys Leu Cys Thr Pro Glu Asn Ile Asn Cys Arg Asp
820 825 830Thr Gln Gly Arg Asn Ser Thr Pro Leu His Leu Ala Ala Gly Tyr Asn
835 840 845Asn Leu Glu Val Ala Glu Tyr Leu Leu Glu His Gly Ala Asp Val Asn
850 855 860Ala Gln Asp Lys Gly Gly Leu Ile Pro Leu His Asn Ala Ala Ser Tyr865 870 875 880Gly His Val Asp Ile Ala Ala Leu Leu Ile Lys Tyr Asn Thr Cys Val
885 890 895Asn Ala Thr Asp Lys Trp Ala Phe Thr Pro Leu His Glu Ala Ala Gln
900 905 910Lys Gly Arg Thr Gln Leu Cys Ala Leu Leu Leu Ala His Gly Ala Asp
915 920 925Pro Thr Met Lys Asn Gln Glu Gly Gln Thr Pro Leu Asp Leu Ala Thr
930 935 940Ala Asp Asp Ile Arg Ala Leu Leu Ile Asp Ala Met Pro Pro Glu Ala945 950 955 960Leu Pro Thr Cys Phe Lys Pro Gln Ala Thr Val Val Ser Ala Ser Leu
965 970 975Ile Ser Pro Ala Ser Thr Pro Ser Cys Leu Ser Ala Ala Ser Ser Ile
980 985 990Asp Asn Leu Thr Gly Pro Leu Ala Glu Leu Ala Val Gly Gly Ala Ser
995 1000 1005Asn Ala Gly Asp Gly Ala Ala Gly Thr Glu Arg Lys Glu Gly Glu Val1010 1015 1020Ala Gly Leu Asp Met Asn Ile Ser Gln Phe Leu Lys Ser Leu Gly Leu1025 1030 1035 1040Glu His Leu Arg Asp Ile Phe Glu Thr Glu Gln Ile Thr Leu Asp Val
1045 1050 1055Leu Ala Asp Met Gly His Glu Glu Leu Lys Glu Ile Gly Ile Asn Ala
1060 1065 1070Tyr Gly His Arg His Lys Leu Ile Lys Gly Val Glu Arg Leu Leu Gly
1075 1080 1085Gly Gln Gln Gly Thr Asn Pro Tyr Leu Thr Phe His Cys Val Asn Gln1090 1095 1100Gly Thr Ile Leu Leu Asp Leu Ala Pro Glu Asp Lys Glu Tyr Gln Ser1105 1110 1115 1120Val Glu Glu Glu Met Gln Ser Thr Ile Arg Glu His Arg Asp Gly Gly
1125 1130 1135Asn Ala Gly Gly Ile Phe Asn Arg Tyr Asn Val Ile Arg Ile Gln Lys
1140 1145 1150Val Val Asn Lys Lys Leu Arg Glu Arg Phe Cys His Arg Gln Lys Glu
1155 1160 1165Val Ser Glu Glu Asn His Asn His His Asn Glu Arg Met Leu Phe His1170 1175 1180Gly Ser Pro Phe Ile Asn Ala Ile Ile His Lys Gly Phe Asp Glu Arg1185 1190 1195 1200His Ala Tyr Ile Gly Gly Met Phe Gly Ala Gly Ile Tyr Phe Ala Glu
1205 1210 1215Asn Ser Ser Lys Ser Asn Gln Tyr Val Tyr Gly Ile Gly Gly Gly Thr
1220 1225 1230Gly Cys Pro Thr His Lys Asp Arg Ser Cys Tyr Ile Cys His Arg Gln
1235 1240 1245Met Leu Phe Cys Arg Val Thr Leu Gly Lys Ser Phe Leu Gln Phe Ser1250 1255 1260Thr Met Lys Met Ala His Ala Pro Pro Gly His His Ser Val Ile Gly1265 1270 1275 1280Arg Pro Ser Val Asn Gly Leu Ala Tyr Ala Glu Tyr Val Ile Tyr Arg
1285 1290 1295Gly Glu Gln Ala Tyr Pro Glu Tyr Leu Ile Thr Tyr Gln Ile Met Lys
1300 1305 1310Pro Glu Ala Pro Ser Gln Thr Ala Thr Ala Ala Glu Gln Lys Thr
1315 1320 1325<210>5<21l>460<212>DNA<213>人(Homo sapiens)<400>5gaactgtctt cagtagttag ttcaagtgga acagagggtg cttccagttt ggagaaaaag 60gaggttccag gagtagattt tagcataact caattcgtaa ggaatcttgg acttgagcac 120ctaatggata tatttnagag agaacagatc actttggatg tattagttga gatggggcac 180aaggagctga aggagattgg aatcaatgct tatggacata ggcacaaact aattaaagga 240gtcgagagac ttatctccgg acaacaaggt cttaacccat atttaacttt gaacacctct 300ggtagtggaa caattcttat agatctgtct cctgatgata aagagtttca gtctgtggag 360gaagagatgc aaagtacagt tcgagagcac agagatggag gtcatgcagg tggaatcttc 420aacagataca atattctcaa gattcagaag gtttgtaaca 460<210>6<211>42<212>PRT<213>人(Homo sapiens)<400>6Gly Thr Ile Leu Ile Asp Leu Ser Pro Asp Asp Lys Glu Phe Gln Ser1 5 10 15Val Glu Glu Glu Met Gln Ser Thr Val Arg Glu His Arg Asp Gly Gly
20 25 30His Ala Gly Gly Ile Phe Asn Arg Tyr Asn
35 40<210>7<211>564<212>DNA<213>人(Homo sapiens)<400>7tgctatttca tgggtctcct tttgtgaatg caattatcca caaaggcttt gatgaaaggc 60atgcgtacat aggtggtatg tttggagctg gcatttattt tgctgaaaac tcttccaaaa 120gcaatcaata tgtatatgga attggaggag gtactgggtg tccagttcac aaagacagat 180cttgttacat ttgccacagg agnctgctct tttgccgggt aaccttggga aagtctttcc 240tgcagttcag tgcaatgaaa atggcacatt ctcctccagg tcatcactca gtcactggta 300ggcccagtgt aaatggccta gcattagctg aatatgttat ttacagagga gaacaggtaa 360tgtagtttta tttgttcatc ttcaaaantg ctaggggagg catactttaa ctttttatta 420atctcttgaa ttgacaagac ntttgcctta acgggntttt ttaaaatttt atttgggggt 480attttcagtt tgggaagtta caaatagtaa agagattttc ttattaccct tacccggntt 540ccnaatgtta tattttgttc cctt 564<210>8<211>118<212>PRT<213>人(Homo sapiens)<400>8Leu Phe His Gly Ser Pro Phe Val Asn Ala Ile Ile His Lys Gly Phe1 5 10 15Asp Glu Arg His Ala Tyr Ile Gly Gly Met Phe Gly Ala Gly Ile Tyr
20 25 30Phe Ala Glu Asn Ser Ser Lys Ser Asn Gln Tyr Val Tyr Gly Ile Gly
35 40 45Gly Gly Thr Gly Cys Pro Val His Lys Asp Arg Ser Cys Tyr Ile Cys
50 55 60His Arg Xaa Leu Leu Phe Cys Arg Val Thr Leu Gly Lys Ser Phe Leu65 70 75 80Gln Phe Ser Ala Met Lys Met Ala His Ser Pro Pro Gly His His Ser
85 90 95Val Thr Gly Arg Pro Ser Val Asn Gly Leu Ala Leu Ala Glu Tyr Val
100 105 110Ile Tyr Arg Gly Glu Gln
115<210>9<211>397<212>DNA<213>人(Homo sapiens)<400>9aacagagtta acttgaacct tttatatgtt atgcattgat tctaacaaac tgtaatgccc 60tcaacagaac taattttact aatacaatac tgtgttcttt aaaacacagc atttacactg 120aatacaattt catttgtaaa actgtaaata agagcttttg tactagccca gtatttattt 180acattgcttt gtaatataaa tctgttttag aactgcagcg gtttacaaaa ttttttcata 240tgtattgttc atctatactt catcttacat cgtcatgatt gagtgatctt tacatttgat 300tccagaggct atgttcagtt gttagttggg gaaagattga gttatcagat ttaatttgcc 360gatgggagcc tttatctgtc ataggaaatc tttctca 397<210>10<211>343<212>DNA<213>人(Homo sapiens)<400>10cttatcctga gtatttaatt acttaccaga ttatgaggcc tgaaggtatg gtcgatggat 60aaatagttat tttaagaaac taattccact gaacctaaaa tcatcaaagc agcagtggcc 120tctacgtttt actcctttgc tgaaaaaaaa tcatcttgcc cacaggcctg tggcaaaagg 180ataaaaatgt gaacgaagtt ttaacattct gacttgataa agctttaata atgtacagtg 240ttttctaaat atttnctgtt ttttcagcac tttaacagat gccattccag ggtaaactgg 300ggttgtctgt actaaattat aaacagggtt aactggaccc ttt 343<210>11<211>334<212>DNA<213>人(Homo sapiens)<400>11gcagttctaa aacagattta tattacaaag caatgtaaat aaatactggg ctagtacaaa 60agctcttatt tacagtttta caaatgaaat tgtattcagt gtaaatgctg tgttttaaag 120aacacagtat tgtattagta aaattagttc tgttgagggc attacagttt gttagaatca 180atgcataaca tataaaaggt tcaagttaac tctgtttata atttagtaca gacaacccag 240tttaacctgg aatggcatct gttaaagtgc tgaaaaaaca ggaaatattt agaaaacact 300gtacattatt aaagctttat caagtcagaa tgtt 334<210>12<211>353<212>DNA<213>人(Homo sapiens)<400>12cagcaaagga gtaaaacgta gaggccactg ctgctttgat gattttaggt tcagtggaat 60tagtttctta aaataactat ttatccatcg accatacctt caggcctcat aatctggtaa 120gtaattaaat actcaggata agcctgttct cctctgtaaa taacatattc agctaatgct 180aggccattta cactgggcct accagtgact gagtgatgac ctggaggaga atgtgccatt 240ttcattgcac tgaactgcag gaaagacttt cccaaggtta cccggcaaaa gagcagctgc 300ctgtggcaaa tgaacaagat ctgtctttgt gaactggaca cccagtacct tct 353<210>13<211>436<212>DNA<213>人(Homo sapiens)<400>13ttttttttgc agttctaaaa cagatttata ttacaaagca atgtaaataa atactgggct 60agtacaaaag ctcttattta cagttttaca aatgaaattg tattcagtgt aaatgctgtg 120ttttaaagaa cacagtattg tattagtaaa attagttctg ttgagggcat tacagtttgt 180tagaatcaat gcataacata taaaaggttc aagttaactc tgtttataat ttagtacaga 240caacccagtt taacctggga tgggcatctg ttaaagtgct ggaaaaaaca gggaaatatt 300taggaaaaca ctggtacatt atttaaaggc tttntccaag gtcaggantg tttaaacttc 360gtttcacatt tttatccntt tggccacggc ctgtggggcn aggatggatt ttttttccgg 420ccaagggtgt taaacg 436<210>14<211>392<212>DNA<213>人(Homo sapiens)<400>14tgctatttca tgggtctcct tttgtgaatg caattatcca caaaggcttt gatgaaaggc 60atgcgtacat aggtggtatg tttggagctg gcatttattt tgctgaaaac tcttccaaaa 120gcaatcaata tgtatatgga attggaggag gtactgggtg tccagttcac aaagacagat 180cttgttacat ttgccacagg cagctgctct tttgccgggt aaccttggga aagtctttcc 240tgcagttcag tgcaatgaaa atggcacatt ctcctccagg tcatcactca gtcactggta 300ggcccagtgt aaatggccta gcattagctg naatatgtta tttacagagg agaacaggta 360atgtagtttt aattttgttt catcttccaa aa 392<210>15<211>317<212>DNA<213>人(Homo sapiens)<400>15ttttttttgc agttctaaaa cagatttata ttacaaagca atgtaaataa atactgggct 60agtacaaaag ctcttattta cagttttaca aatgaaattg tattcagtgt aaatgctgtn 120ttttaaagaa cacagtattg tattagtaaa attagttctg ttgagggcat tacagtttgt 180taggaatcaa tgcataacat ataaaaggtt caagttaact ctgtttataa tttaggtaca 240gacaacccag tttaaccggg gaatgggcat ctgttaaagt gctgaaaaaa cnggganata 300tttaggaaaa cnctgta 317<210>16<211>485<212>DNA<213>人(Homo sapiens)<400>16tgcagttcta aaacagattt atattacaaa gcaatgtaaa taaatactgg gctagtacaa 60aagctcttat ttacagtttt acaaatgaaa ttgtattcag tgtaaatgct gtgttttaaa 120gaacacagta ttgtattagt aaaattagtt ctgttgaggg cattacagtt tgttagaatc 180aatgcataac atataaaagg ttcaagttaa ctctgtttat aatttagtac agacaaccca 240gtttaacctg gaatggcatc tgttaaagtg ctgaaaaaac aggaaatatt tacgaaaaca 300ctgtacatta ttaaagcttt atcaagtcag aatgttaaac ttcgttcaca tttttatcct 360tttgccacag gcctgtgggg caagatgatt ttttttcagc aaaggagtaa aacgtagagg 420gccactggct gctttgatga ttttagggtt cagtgggaat tagtttccta aaataacnat 480ttatc 485<210>17<211>291<212>DNA<213>人(Homo sapiens)<400>17ttncctgcag ttcagtgcaa tgaanatggc acattctcct ccaggtcatc actcagtcac 60tggtaggccc agtgtaaatg gcctagcatt agctgaatat gttatttaca gaggagaaca 120ggcttatcct gagtatttaa ttacttacca gattatgagg cctgaaggta tggtcgatgg 180ataaatagtt attttaagaa actaattcca ctgaacctaa aatcatcaaa gcagcagtgg 240cctctacgtt ttactccttt gctgaaaaaa aatcatcttg cccacaggcc t 291<210>18<211>371<212>DNA<213>人(Homo sapiens)<400>18cgtagaggcc actgctgctt tgatganttt tanggttcan gtggaattng tttcttaaaa 60taactattta tccatcgacc ataccttcag gcctcataat ctggtaagta attaaatact 120caggataagc ctgttctcct ctgtaaataa catattcagc taatgctagg ccatttacac 180tgggcctacc agtgactgaa gtgatgcctg gggggagaat gtgccatttt cattgcactg 240aactgcaggn aagactttcc caagggttac ccgggcaaaa gagcagctgc ctgtgggnaa 300tgttacaagg tcttgtcttt tgtngacctn gggcaccccg taccctcctc caattccata 360tacatatttg a 371<210>19<211>341<212>DNA<213>人(Homo sapiens)<400>19gaaagataca ctcaccggag aaaagaagtt tctgaagaaa accacaacca tgccaatgaa 60cgaatgctat ttcatgggtc tccttttgtg aatgcaatta tccacaaagg ctttgatgaa 120aggcatgcgt acataggtgg tatgtttgga gctggcattt attttgctgg aaaactcttc 180caaaaggcaa tcaatatgta tatgggaatt gggagggagg gtactggggt gtccagtttc 240acaaaggaca gatcttgttt acatttggcc acaggcaggc tggctctttt tgcccgggtn 300accttggggg aagtcttttc ctggcagttt cagttgccat g 341<210>20<211>385<212>DNA<213>人(Homo sapiens)<400>20tactaaatta taaacagagt taacttgaac cttttatatg ttatgcattg attctaacaa 60actgtaatgc cctcaacaga actaatttta ctaatacaat aangtgttct ttaaaacaca 120gcatttacac tgaatacaat ttcatttgta aaactgtaaa taagagcttt tgtactagcc 180cagtatttat ttacattgct ttgtaatata aatctgtttt aggaactgca ggcggtttac 240aaaatttttt catatgtatt gttcatttat acttcatctt acatcgtcat ggattgaggt 300gatctttaca tttggattcc ngggggctat ggttcaggtt gttaggttgg gggaaagggt 360tggggtttat ccgggnttta ntttg 385<210>21<211>335<212>DNA<213>人(Homo sapiens)<400>21gaaggtatgg tcgatggata aatagttatt ttaagaaact aattccactg aacctaaaat 60catcaaagca gcagtggcct ctacgtttta ctcctttgct gaaaaaaaat catcttgccc 120acaggcctgt ggcaaaagga taaaaatgtg aacgaagttt aacattctga cttgataaag 180ctttaataat gtacagtgtt ttctaaatat ttcctgtttt ttcagcactt taacagatgc 240cattccgggt taaactgggg ttgtctgtac taaattatta aacagngtta acttggaacc 300nttttatatg ttatggcctt ggttcttaac caana 335<210>22<211>388<212>DNA<213>人(Homo sapiens)<400>22gttttactcc tttgctgaaa aaaaatcatc ttgcccacag gcctgtggaa naaggataaa 60aatgtgaacg aagtttaaca ttctgacttg ataaagcttt aataatgtac agtgttttct 120aaatatttcc tgttttttca gcactttaac agatgccatt ccaggttaaa ctgggttgtc 180tgtactaaat tataaacaga gttaacttga accttttata tgttatgcat tgattctaac 240aaactgtaat gccctcaaca gaactaattt tactaataca atactgtgtt ctttaaaaca 300caggcattta cactggaata caatttcatt tgttaaaact ggtaantagg agcttttgta 360ctagcccagt atttatttac atgctttg 388<210>23<211>401<212>DNA<213>人(Homo sapiens)<400>23gttttactcc tttgctgaaa aaaaatcatc ttgcccacag gcctgtggaa naaggataaa 60aatgtgaacg aagttaacat tctgacttga taaagcttta ataatgtaca gtgttttcta 120aatatttcct gttttttcag cactttaaca gatgccattc caggttaaac tgggttgtct 180gtactaaatt ataaacagag ttaacttgaa ccttttatat gttatgcatt gattctaaca 240aactgtaatg ccctcaacag aactantttt acttaataca atactgtgtt ctttnaaaac 300acaggcattt acactggaat acaattttca ttttgttaaa actggttaaa ttaaggnggc 360tttttgtact nggccccgtn ttttatttta cattgctttg g 401<210>24<211>354<212>DNA<213>人(Homo sapiens)<400>24taattttact aatacaatac tgtgttcttt aaaacacagc atttacactg aatacaattt 60catttgtaaa actgtaaata agagcttttg tactagccca gtatttattt acattgcttt 120gtaatataaa tctgttttag aactgcagcg gtttacaaaa ttttttcata tgtattgttc 180atctatactt catcttacat cgtcatgatt gagtgatctt tacatttgat tccagaggct 240atgttcagtt gttagttggg aaagattgag ttatcagatt taatttgccg atgggagcct 300ttatctgtca ttagaaatct ttctnattta agaacttatg aatatgctga agat 354<210>25<211>18<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>25tgtaaaacga cggccagt 18<210>26<211>19<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>26ggaaacagct atgaccatg 19<210>27<211>18<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>27tttgccgggt aaccttgg 18<210>28<211>18<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>28ccaaggttac ccggcaaa 18<210>29<211>18<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>29gtaggcccag tgtaaatg 18<210>30<211>18<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>30catttacact gggcctac 18<210>31<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>31gagtaagttg cagggcatgt 20<210>32<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>32acatgccctg caacttactc 20<210>33<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>33gaatcaccgc agttactaaa 20<210>34<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>34tttagtaact gcggtgattc 20<210>35<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>35ggcctgaagg tatggtcgat 20<210>36<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>36atcgaccata ccttcaggcc 20<210>37<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>37tgagggcatt acagtttgtt 20<210>38<211>21<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>38taatacgaac tcactatagg g 21<210>39<211>18<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>39atacactcac cggagaaa 18<210>40<211>18<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>40tttctccggt gagtgtat 18<210>41<211>1691<212>DNA<213>人工序列<220><221>CDS<222>(1)..(357)<220><223>人工序列说明:非蛋白编码区序列是载体序列<400>41atg cta ttt cat ggg tct cct ttt gtg aat gca att atc cac aaa ggc 48Met Leu Phe His Gly Ser Pro Phe Val Asn Ala Ile Ile His Lys Gly1 5 10 15ttt gat gaa agg cat gcg tac ata ggt ggt atg ttt gga gct ggc att 96Phe Asp Glu Arg His Ala Tyr Ile Gly Gly Met Phe Gly Ala Gly Ile
20 25 30tat ttt gct gaa aac tct tcc aaa agc aat caa tat gta tat gga att 144Tyr Phe Ala Glu Asn Ser Ser Lys Ser Asn Gln Tyr Val Tyr Gly Ile
35 40 45gga gga ggt act ggg tgt cca gtt cac aaa gac aga tct tgt tac att 192Gly Gly Gly Thr Gly Cys Pro Val His Lys Asp Arg Ser Cys Tyr Ile
50 55 60tgc cac agg cag ctg ctc ttt tgc cgg gta acc ttg gga aag tct ttc 240Cys His Arg Gln Leu Leu Phe Cys Arg Val Thr Leu Gly Lys Ser Phe65 70 75 80ctg cag ttc agt gca atg aaa atg gca cat tct cct cca ggt cat cac 288Leu Gln Phe Ser Ala Met Lys Met Ala His Ser Pro Pro Gly His His
85 90 95tca gtc act ggt agg ccc agt gta aat ggc cta gca tta gct gaa tat 336Ser Val Thr Gly Arg Pro Ser Val Asn Gly Leu Ala Leu Ala Glu Tyr
100 105 110gtt att tac aga gga gaa cag gtaatgtagt tttatttgtt catcttcaaa 387Val Ile Tyr Arg Gly Glu Gln
115aatgctaggg aggcatactt taacttttta ttaatctctt gaattgacaa gacatattgc 447cttaactgga ttttttaaaa attttatttg gagataattt cagatttgga aagttacaaa 507aatagtaaag agaattttct tataaccttt acctagattt cctaaatgtt aatattttgt 567tctctttttt actcttacca ttctctcctt ctttccttgt gtgtgtacct atttttttgt 627gaactgtttg agagtaagtt gcagggcatg tccctttacc attaactatt tcaattgtaa 687atttcctaaa aacaagaaga ttttattcaa atttcgccag tcgttccgga tttttcttag 747ctcttataaa taattgaaat cttgtattta acagcctgtc catagcaaag aagtatataa 807ctgtgttttg ctctcagtga gagccaaaag tagttctaga gcagtgttgt gaactgggag 867taggtatcgg aatcaccgca gttactaaaa tcagacatga ttttagtctt atctgatact 927tatgaactta gtattcatct tagacttgct gattgaaaat ctgaagaact gtactcaggg 987taaagatgtt ttgagaaaat gtccctagat gattctgatc tacaacagta atttagaacc 1047tcctccctaa gattaggaat acttccggaa agtctgttta tctttcaaga aaatttttgt 1107accattattt gaatttatct ttctcttcca ggcttatcct gagtatttaa ttacttacca 1167gattatgagg cctgaaggta tggtcgatgg ataaatagtt attttaagaa actaattcca 1227ctgaacctaa aatcatcaaa gcagcagtgg cctctacgtt ttactccttt gctgaaaaaa 1287aatcatcttg cccacaggcc tgtggcaaaa ggataaaaat gtgaacgaag tttaacattc 1347tgacttgata aagctttaat aatgtacagt gttttctaaa tatttcctgt tttttcagca 1407ctttaacaga tgccattcca ggttaaactg ggttgtctgt actaaattat aaacagagtt 1467aacttgaacc ttttatatgt tatgcattga ttctaacaaa ctgtaatgcc ctcaacagaa 1527ctaattttag taatacaata ctgtgttctt taaaacacag catttacact gaatacaatt 1587tcatttgtaa aactgtaaat aagagctttt gtactagccc agtatttatt tacattgctt 1647tgtaatataa tcctgtttta gaagtgcaaa aaaaaaaaaa aaaa 1691<210>42<211>119<212>PRT<213>人工序列<223>人工序列说明:非蛋白编码区序列是载体序列<400>42Met Leu Phe His Gly Ser Pro Phe Val Asn Ala Ile Ile His Lys Gly1 5 10 15Phe Asp Glu Arg His Ala Tyr Ile Gly Gly Met Phe Gly Ala Gly Ile
20 25 30Tyr Phe Ala Glu Asn Ser Ser Lys Ser Asn Gln Tyr Val Tyr Gly Ile
35 40 45Gly Gly Gly Thr Gly Cys Pro Val His Lys Asp Arg Ser Cys Tyr Ile
50 55 60Cys His Arg Gln Leu Leu Phe Cys Arg Val Thr Leu Gly Lys Ser Phe65 70 75 80Leu Gln Phe Ser Ala Met Lys Met Ala His Ser Pro Pro Gly His His
85 90 95Ser Val Thr Gly Arg Pro Ser Val Asn Gly Leu Ala Leu Ala Glu Tyr
100 105 110Val Ile Tyr Arg Gly Glu Gln
115<210>43<211>1692<212>DNA<213>人工序列<220><221>CDS<222>(1)..(357)<220><223>人工序列说明:非蛋白编码区序列是载体序列<400>43atg cta ttt cat ggg tct cct ttt gtg aat gca att atc cac aaa ggc 48Met Leu Phe His Gly Ser Pro Phe Val Asn Ala Ile Ile His Lys Gly1 5 10 15ttt gat gaa agg cat gcg tac ata ggt ggt atg ttt gga gct ggc att 96Phe Asp Glu Arg His Ala Tyr Ile Gly Gly Met Phe Gly Ala Gly Ile
20 25 30tat ttt gct gaa aac tct tcc aaa agc aat caa tat gta tat gga att 144Tyr Phe Ala Glu Asn Ser Ser Lys Ser Asn Gln Tyr Val Tyr Gly Ile
35 40 45gga gga ggt act ggg tgt cca gtt cac aaa gac aga tct tgt tac att 192Gly Gly Gly Thr Gly Cys Pro Val His Lys Asp Arg Ser Cys Tyr Ile
50 55 60tgc cac agg cag ctg ctc ttt tgc cgg gta acc ttg gga aag tct ttc 240Cys His Arg Gln Leu Leu Phe Cys Arg Val Thr Leu Gly Lys Ser Phe65 70 75 80ctg cag ttc agt gca atg aaa atg gca cat tct cct cca ggt cat cac 288Leu Gln Phe Ser Ala Met Lys Met Ala His Ser Pro Pro Gly His His
85 90 95tca gtc act ggt agg ccc agt gta aat ggc cta gca tta gct gaa tat 336Ser Val Thr Gly Arg Pro Ser Val Asn Gly Leu Ala Leu Ala Glu Tyr
100 105 110gtt att tac aga gga gaa cag gtaatgtagt tttatttgtt catcttcaaa 387Val Ile Tyr Arg Gly Glu Gln
115aatgctaggg aggcatactt taacttttta ttaatctctt gaattgacaa gacatattgc 447cttaactgga ttttttaaaa attttatttg gagataattt cagatttgga aagttacaaa 507aatagtaaag agaattttct tataaccttt acctagattt cctaaatgtt aatattttgt 567tctctttttt actcttacca ttctctcctt ctttccttgt gtgtgtacct atttttttgt 627gaactgtttg agagtaagtt gcagggcatg tccctttacc attaactatt tcaattgtaa 687atttcctaaa aacaagaaga ttttattcaa atttcgccag tcgttccgga tttttcttag 747ctcttataaa taattgaaat cttgtattta acagcctgtc catagcaaag aagtatataa 807ctgtgttttg ctctcagtga gagccaaaag tagttctaga gcagtgttgt gaactgggag 867taggtatcgg aatcaccgca gttactaaaa tcagacatga ttttagtctt atctgatact 927tatgaactta gtattcatct tagacttgct gattgaaaat ctgaagaact gtactcaggg 987taaagatgtt ttgagaaaat gtccctagat gattctgatc tacaacagta atttagaacc 1047tcctccctaa gattaggaat acttccggaa agtctgttta tctttcaaga aaatttttgt 1107accattattt gaatttatct ttctcttcca ggcttatcct gagtatttaa ttacttacca 1167gattatgagg cctgaaggta tggtcgatgg ataaatagtt attttaagaa actaattcca 1227ctgaacctaa aatcatcaaa gcagcagtgg cctctacgtt ttactccttt gctgaaaaaa 1287aatcatcttg cccacaggcc tgtggcaaaa ggataaaaat gtgaacgaag tttaacattc 1347tgacttgata aagctttaat aatgtacagt gttttctaaa tatttcctgt tttttcagca 1407ctttaacaga tgccattcca ggttaaactg ggttgtctgt actaaattat aaacagagtt 1467aacttgaacc ttttatatgt tatgcattga ttctaacaaa ctgtaatgcc ctcaacagaa 1527ctaattttac taatacaata ctgtgttctt taaaacacag catttacact gaatacaatt 1587tcatttgtaa aactgtaaat aagagctttt gtactagccc agtatttatt tacattgctt 1647tgtaatataa atctgtttta gaactgcaaa aaaaaaaaaa aaaaa 1692<210>44<211>119<212>PRT<213>人工序列<223>人工序列说明:非蛋白编码区的序列是载体序列<400>44Met Leu Phe His Gly Ser Pro Phe Val Asn Ala Ile Ile His Lys Gly1 5 10 15Phe Asp Glu Arg His Ala Tyr Ile Gly Gly Met Phe Gly Ala Gly Ile
20 25 30Tyr Phe Ala Glu Asn Ser Ser Lys Ser Asn Gln Tyr Val Tyr Gly Ile
35 40 45Gly Gly Gly Thr Gly Cys Pro Val His Lys Asp Arg Ser Cys Tyr Ile
50 55 60Cys His Arg Gln Leu Leu Phe Cys Arg Val Thr Leu Gly Lys Ser Phe65 70 75 80Leu Gln Phe Ser Ala Met Lys Met Ala His Ser Pro Pro Gly His His
85 90 95Ser Val Thr Gly Arg Pro Ser Val Asn Gly Leu Ala Leu Ala Glu Tyr
100 105 110Val Ile Tyr Arg Gly Glu Gln
115<210>45<211>582<212>DNA<213>人工序列<220><221>CDS<222>(1)..(480)<220><223>人工序列说明:Sequence not
specified as protein-coding is vector sequence<400>45gaa aga tac act cac cgg aga aaa gaa gtt tct gaa gaa aac cac aac 48Glu Arg Tyr Thr His Arg Arg Lys Glu Val Ser Glu Glu Asn His Asn1 5 10 15cat gcc aat gaa cga atg cra ttt cat ggg tct cct ttt gtg aat gca 96His Ala Asn Glu Arg Met Leu Phe His Gly Ser Pro Phe Val Asn Ala
20 25 30att atc cac aaa ggc ttt gat gaa agg cat gcg tac ata ggt ggt atg 144Ile Ile His Lys Gly Phe Asp Glu Arg His Ala Tyr Ile Gly Gly Met
35 40 45ttt gga gct ggc att tat ttt gct gaa aac tct tcc aaa agc aat caa 192Phe Gly Ala Gly Ile Tyr Phe Ala Glu Asn Ser Ser Lys Ser Asn Gln
50 55 60tat gta tat gga att gga gga ggt act ggg tgt cca gtt cac aaa gac 240Tyr Val Tyr Gly Ile Gly Gly Gly Thr Gly Cys Pro Val His Lys Asp65 70 75 80aga tct tgt tac att tgc cac agg cag ctg ctc ttt tgc cgg gta acc 288Arg Ser Cys Tyr Ile Cys His Arg Gln Leu Leu Phe Cys Arg Val Thr
85 90 95ttg gga aag tct ttc ctg cag ttc agt gca atg aaa atg gca cat tct 336Leu Gly Lys Ser Phe Leu Gln Phe Ser Ala Met Lys Met Ala His Ser
100 105 110cct cca ggt cat cac tca gtc act ggt agg ccc agt gta aat ggc cta 384Pro Pro Gly His His Ser Val Thr Gly Arg Pro Ser Val Asn Gly Leu
115 120 125gca tta gct gaa tat gtt att tac aga gga gaa cag gct tat cct gag 432Ala Leu Ala Glu Tyr Val Ile Tyr Arg Gly Glu Gln Ala Tyr Pro Glu
130 135 140tat tta att act tac cag att atg agg cct gaa ggt atg gtc gat gga 480Tyr Leu Ile Thr Tyr Gln Ile Met Arg Pro Glu Gly Met Val Asp Gly145 150 155 160taaatagtta ttttaagaaa ctaattccac tgaacctaaa atcatcaaag cagcagtggc 540ctctacgttt tactcctttg ctgaaaaaaa aaaaaaaaaa aa 582<210>46<211>160<212>PRT<213>人工序列<223>人工序列说明:Sequence not
specified as protein-coding is vector sequence<400>46Glu Arg Tyr Thr His Arg Arg Lys Glu Val Ser Glu Glu Asn His Asn1 5 10 15His Ala Asn Glu Arg Met Leu Phe His Gly Ser Pro Phe Val Asn Ala
20 25 30Ile Ile His Lys Gly Phe Asp Glu Arg His Ala Tyr Ile Gly Gly Met
35 40 45Phe Gly Ala Gly Ile Tyr Phe Ala Glu Asn Ser Ser Lys Ser Asn Gln
50 55 60Tyr Val Tyr Gly Ile Gly Gly Gly Thr Gly Cys Pro Val His Lys Asp65 70 75 80Arg Ser Cys Tyr Ile Cys His Arg Gln Leu Leu Phe Cys Arg Val Thr
85 90 95Leu Gly Lys Ser Phe Leu Gln Phe Ser Ala Met Lys Met Ala His Ser
100 105 110Pro Pro Gly His His Ser Val Thr Gly Arg Pro Ser Val Asn Gly Leu
115 120 125Ala Leu Ala Glu Tyr Val Ile Tyr Arg Gly Glu Gln Ala Tyr Pro Glu
130 135 140Tyr Leu Ile Thr Tyr Gln Ile Met Arg Pro Glu Gly Met Val Asp Gly145 150 155 160<210>47<211>23<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>47ctccggacaa caaggtctta acc 23<210>48<2ll>19<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>48ccacctatgt acgcatgcc 19<210>49<211>356<212>DNA<213>人(Homo sapiens)<400>49tccggacaac aaggtcttaa cccatattta actttgaaca cctctggtag tggaacaatt 60cttatagatc tgtctcctga tgataaagag tttcagtctg tggaggaaga gatgcaaagt 120acagttcgag agcacagaga tggaggtcat gcaggtggaa tcttcaacag atacaatatt 180ctcaagattc agaaggtttg taacaagaaa ctatgggaaa gatacactca ccggagaaaa 240gaagtttctg aagaaaacca caaccatgcc aatgaacgaa tgctatttca tgggtctcct 300tttgtgaatg caattatcca caaaggcttt gatgaaaggc atgcgtacat aggtgg 356<210>50<211>21<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>50atttaaccct cactaaaagg g 21<210>51<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>51aaaggctccc atcggcaaat 20<210>52<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>52gttgagggca ttacagtttg 20<210>53<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>53aaaacgtaga ggccactgct 20<210>54<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>54tggtgtagac tgacgccctt 20<210>55<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>55tccggtgagt gtatctttcc 20<210>56<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>56ctcctttgtc ttgggcattc 20<2l0>57<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>57atctgctctg ccctcttgtt 20<210>58<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>58gggtatcgcg gcaatttaca 20<210>59<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>59aacaagaggg cagagcagat 20<210>60<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>60tgccccatct caactaatac 20<210>61<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>61gtaatgccct caacagaact 20<210>62<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>62ggcgtcagtc tacaccactt 20<210>63<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>63taaattgccc gcgataccca 20<210>64<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>64cactcagtca ctggtaggcc 20<210>65<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>65atctgctctg ccctcttgtt 20<210>66<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>66tagttgagat ggggcacaag 20<210>67<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>67aaacgtagag gccactgctg 20<210>68<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>68cgggtaacct tgggaaagtc 20<210>69<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>69gggctttact gctttacaga 20<210>70<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>70gtaagggctg ctgacagtga 20<210>71<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>71ttactccagc agagggcact 20<210>72<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>72ctgacgccct tcaatgtctc 20<210>73<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>73ggtactaagg ccacaattca 20<210>74<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>74gggtatcgcg gcaatttaca 20<210>75<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>75gttgagggca ttacagtttg 20<210>76<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>76taacaagagg gcagagcaga 20<210>77<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>77agttctgttg agggcattac 20<210>78<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>78ggcctaccag tgactgagtg 20<210>79<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>79gggctagagg acctgaagag 20<210>80<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>80agtgccctct gctggagtaa 20<210>81<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>81ggcgtcagtc tacaccactt 20<210>82<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>82tgaattgtgg ccttagtacc 20<210>83<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>83atgcccaaga caaaggagga 20<210>84<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>84gtaatgccct caacagaact 20<210>85<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>85atctgctctg ccctcttgtt 20<210>86<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>86cgggtaacct tgggaaagtc 20<210>87<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>87ccggacaaca aggtcttaac 20<210>88<211>3353<212>DNA<213>人(Homo sapiens)<220><221>CDS<222>(1)..(2352)<400>88tgt gaa ctg ttg cta aga aaa gga gca aac atc aat gaa aag act aaa 48Cys Glu Leu Leu Leu Arg Lys Gly Ala Asn Ile Asn Glu Lys Thr Lys1 5 10 15gaa ttc ttg act cct ctg cac gtg gca tct gag aaa gct cat aat gat 96Gtu Phe Leu Thr Pro Leu His Val Ala Set Glu Lys Ala His Asn Asp
20 25 30gtt gtt gaa gta gtg gtg aaa cat gaa gca aag gtt aat gct ctg gat 144Val Val Glu Val Val Val Lys His Glu Ala Lys Val Asn Ala Leu Asp
35 40 45aat ctt ggt cag act tct cta cac aga gct gca tat tgt ggt cat cta 192Asn Leu Gly Gln Thr Ser Leu His Arg Ala Ala Tyr Cys Gly His Leu
50 55 60caa acc tgc cgc cta ctc ctg agc tat ggg tgt gat cct aac att ata 240Gln Thr Cys Arg Leu Leu Leu Ser Tyr Gly Cys Asp Pro Asn Ile Ile65 70 75 80tcc ctt cag ggc ttt act gct tta cag atg gga aat gaa aat gta cag 288Ser Leu Gln Gly Phe Thr Ala Leu Gln Met Gly Asn Glu Asn Val Gln
85 90 95caa ctc ctc caa gag ggt atc tca tta ggt aat tca gag gca gac aga 336Gln Leu Leu Gln Glu Gly Ile Ser Leu Gly Asn Ser Glu Ala Asp Arg
100 105 110caa ttg ctg gaa gct gca aag gct gga gat gtc gaa act gta aaa aaa 384Gln Leu Leu Glu Ala Ala Lys Ala Gly Asp Val Glu Thr Val Lys Lys
115 120 125ctg tgt act gtt cag agt gtc aac tgc aga gac att gaa ggg cgt cag 432Leu Cys Thr Val Gln Ser Val Asn Cys Arg Asp Ile Glu Gly Arg Gln
130 135 140tct aca cca ctt cat ttt gca gct ggg tat aac aga gtg tcc gtg gtg 480Ser Thr Pro Leu His Phe Ala Ala Gly Tyr Asn Arg Val Ser Val Val145 150 155 160gaa tat ctg cta cag cat gga gct gat gtg cat gct aaa gat aaa gga 528Glu Tyr Leu Leu Gln His Gly Ala Asp Val His Ala Lys Asp Lys Gly
165 170 175ggc ctt gta cct ttg cac aat gca tgt tct tat gga cat tat gaa gtt 576Gly Leu Val Pro Leu His Asn Ala Cys Ser Tyr Gly His Tyr Glu Val
180 185 190gca gaa ctt ctt gtt aaa cat gga gca gta gtt aat gta gct gat tta 624Ala Glu Leu Leu Val Lys His Gly Ala Val Val Asn Val Ala Asp Leu
195 200 205tgg aaa ttt aca cct tta cat gaa gca gca gca aaa gga aaa tat gaa 672Trp Lys Phe Thr Pro Leu His Glu Ala Ala Ala Lys Gly Lys Tyr Glu
210 215 220att tgc aaa ctt ctg ctc cag cat ggt gca gac cct aca aaa aaa aac 720Ile Cys Lys Leu Leu Leu Gln His Gly Ala Asp Pro Thr Lys Lys Asn225 230 235 240agg gat gga aat act cct ttg gat ctt gtt aaa gat gga gat aca gat 768Arg Asp Gly Asn Thr Pro Leu Asp Leu Val Lys Asp Gly Asp Thr Asp
245 250 255att caa gat ctg ctt agg gga gat gca gct ttg cta gat gct gcc aag 816Ile Gln Asp Leu Leu Arg Gly Asp Ala Ala Leu Leu Asp Ala Ala Lys
260 265 270aag ggt tgt tta gcc aga gtg aag aag ttg tct tct cct gat aat gta 864Lys Gly Cys Leu Ala Arg Val Lys Lys Leu Ser Ser Pro Asp Asn Val
275 280 285aat tgc cgc gat acc caa ggc aga cat tca aca cct tta cat tta gca 912Asn Cys Arg Asp Thr Gln Gly Arg His Ser Thr Pro Leu His Leu Ala
290 295 300gct ggt tat aat aat tta gaa gtt gca gag tat ttg tta caa cac gga 960Ala Gly Tyr Asn Asn Leu Glu Val Ala Glu Tyr Leu Leu Gln His Gly305 310 315 320gct gat gtg aat gcc caa gac aaa gga gga ctt att cct tta cat aat 1008Ala Asp Val Asn Ala Gln Asp Lys Gly Gly Leu Ile Pro Leu His Asn
325 330 335gca gca tct tac ggg cat gta gat gta gca gct cta cta ata aag tat 1056Ala Ala Ser Tyr Gly His Val Asp Val Ala Ala Leu Leu Ile Lys Tyr
340 345 350aat gca tgt gtc aat gcc acg gac aaa tgg gct ttc aca cct ttg cac 1104Asn Ala Cys Val Asn Ala Thr Asp Lys Trp Ala Phe Thr Pro Leu His
355 360 365gaa gca gcc caa aag gga cga aca cag ctt tgt gct ttg ttg cta gcc 1152Glu Ala Ala Gln Lys Gly Arg Thr Gln Leu Cys Ala Leu Leu Leu Ala
370 375 380cat gga gct gac ccg act ctt aaa aat cag gaa gga caa aca cct tta 1200His Gly Ala Asp Pro Thr Leu Lys Asn Gln Glu Gly Gln Thr Pro Leu385 390 395 400gat tta gtt tca gca gat gat gtc agc gct ctt ctg aca gca gcc atg 1248Asp Leu Val Ser Ala Asp Asp Val Ser Ala Leu Leu Thr Ala Ala Met
405 410 415ccc cca tct gct ctg ccc tct tgt tac aag cct caa gtg crc aat ggt 1296Pro Pro Ser Ala Leu Pro Ser Cys Tyr Lys Pro Gln Val Leu Asn Gly
420 425 430gtg aga agc cca gga gcc act gca gat gct ctc tct tca ggt cca tct 1344Val Arg Ser Pro Gly Ala Thr Ala Asp Ala Leu Ser Ser Gly Pro Ser
435 440 445agc cca tca agc ctt tct gca gcc agc agt ctt gac aac tta tct ggg 1392Ser Pro Ser Ser Leu Ser Ala Ala Ser Ser Leu Asp Asn Leu Ser Gly
450 455 460agt ttt tca gaa ctg tct tca gta gtt agt tca agt gga aca gag ggt 1440Ser Phe Ser Glu Leu Ser Ser Val Val Ser Ser Ser Gly Thr Glu Gly465 470 475 480gct tcc agt ttg gag aaa aag gag gtt cca gga gta gat ttt agc ata 1488Ala Ser Ser Leu Glu Lys Lys Glu Val Pro Gly Val Asp Phe Ser Ile
485 490 495act caa ttc gta agg aat ctt gga ctt gag cac cta atg gat ata ttt 1536Thr Gln Phe Val Arg Asn Leu Gly Leu Glu His Leu Met Asp Ile Phe
500 505 510gag aga gaa cag atc act ttg gat gta tta gtt gag atg ggg cac aag 1584Glu Arg Glu Gln Ile Thr Leu Asp Val Leu Val Glu Met Gly His Lys
515 520 525gag ctg aag gag att gga atc aat gct tat gga cat agg cac aaa cta 1632Glu Leu Lys Glu Ile Gly Ile Asn Ala Tyr Gly His Arg His Lys Leu
530 535 540att aaa gga gtc gag aga ctt atc tcc gga caa caa ggt ctt aac cca 1680Ile Lys Gly Val Glu Arg Leu Ile Ser Gly Gln Gln Gly Leu Asn Pro545 550 555 560tat tta act ttg aac acc tct ggt agt gga aca att ctt ata gat ctg 1728Tyr Leu Thr Leu Asn Thr Ser Gly Ser Gly Thr Ile Leu Ile Asp Leu
565 570 575tct cct gat gat aaa gag ttt cag tct gtg gag gaa gag atg caa agt 1776Ser Pro Asp Asp Lys Glu Phe Gln Ser Val Glu Glu Glu Met Gln Ser
580 585 590aca gtt cga gag cac aga gat gga ggt cat gca ggt gga atc ttc aac 1824Thr Val Arg Glu His Arg Asp Gly Gly His Ala Gly Gly Ile Phe Asn
595 600 605aga tac aat att ctc aag att cag aag gtt tgt aac aag aaa cta tgg 1872Arg Tyr Asn Ile Leu Lys Ile Gln Lys Val Cys Asn Lys Lys Leu Trp
610 615 620gaa aga tac act cac cgg aga aaa gaa gtt tct gaa gaa aac cac aac 1920Glu Arg Tyr Thr His Arg Arg Lys Glu Val Ser Glu Glu Asn His Asn625 630 635 640cat gcc aat gaa cga atg cta ttt cat ggg tct cct ttt gtg aat gca 1968His Ala Asn Glu Arg Met Leu Phe His Gly Ser Pro Phe Val Asn Ala
645 650 655att atc cac aaa ggc ttt gat gaa agg cat gcg tac ata ggt ggt atg 2016Ile Ile His Lys Gly Phe Asp Glu Arg His Ala Tyr Ile Gly Gly Met
660 665 670ttt gga gct ggc att tat ttt gct gaa aac tct tcc aaa agc aat caa 2064Phe Gly Ala Gly Ile Tyr Phe Ala Glu Asn Ser Ser Lys Ser Asn Gln
675 680 685tat gta tat gga att gga gga ggt act ggg tgt cca gtt cac aaa gac 2112Tyr Val Tyr Gly Ile Gly Gly Gly Thr Gly Cys Pro Val His Lys Asp
690 695 700aga tct tgt tac att tgc cac agg cag ctg ctc ttt tgc cgg gta acc 2160Arg Ser Cys Tyr Ile Cys His Arg Gln Leu Leu Phe Cys Arg Val Thr705 710 715 720ttg gga aag tct ttc ctg cag ttc agt gca atg aaa atg gca cat tct 2208Leu Gly Lys Ser Phe Leu Gln Phe Ser Ala Met Lys Met Ala His Ser
725 730 735cct cca ggt cat cac tca gtc act ggt agg ccc agt gta aat ggc cta 2256Pro Pro Gly His His Ser Val Thr Gly Arg Pro Ser Val Asn Gly Leu
740 745 750gca tta gct gaa tat gtt att tac aga gga gaa cag gct tat cct gag 2304Ala Leu Ala Glu Tyr Val Ile Tyr Arg Gly Glu Gln Ala Tyr Pro Glu
755 760 765tat tta att act tac cag att atg agg cct gaa ggt atg gtc gat gga 2352Tyr Leu Ile Thr Tyr Gln Ile Met Arg Pro Glu Gly Met Val Asp Gly
770 775 780taaatagtta ttttaagaaa ctaattccac tgaacctaaa atcatcaaag cagcagtggc 2412ctctacgttt tactcctttg ctgaaaaaaa atcatcttgc ccacaggcct gtggcaaaag 2472gataaaaatg tgaacgaagt ttaacattct gacttgataa agctttaata atgtacagtg 2532ttttctaaat atttcctgtt ttttcagcac tttaacagat gccattccag gttaaactgg 2592gttgtctgta ctaaattata aacagagtta acttgaacct tttatatgtt atgcattgat 2652tctaacaaac tgtaatgccc tcaacagaac taattttact aatacaatac tgtgttcttt 2712aaaacacagc atttacactg aatacaattt catttgtaaa actgtaaata agagcttttg 2772tactagccca gtatttattt acattgcttt gtaatataaa tctgttttag aactgcagcg 2832gtttacaaaa ttttttcata tgtattgttc atctatactt catcttacat cgtcatgatt 2892gagtgatctt tacatttgat tccagaggct atgttcagtt gttagttggg aaagattgag 2952ttatcagatt taatttgccg atgggagcct ttatctgtca ttagaaatct ttctcattta 3012agaacttatg aatatgctga agatttaatt tgtgatacct ttgtatgtat gagacacatt 3072ccaaagagct ctaactatga taggtcctga ttactaaaga agcttcttta ctggcctcaa 3132tttctagctt tcatgttgga aaattttctg cagtccttct gtgaaaatta gagcaaagtg 3192ctcctgtttt ttagagaaac taaatcttgc tgttgaacaa ttattgtgtt cttttcatgg 3252aacataagta ggatgttaca tttccagggt gggaagggta atcctaaatc atttcccaat 3312ctattctaat taccttaaat ctaaagggga aaaaaaaaat c 3353<210>89<211>784<212>PRT<213>人(Homo sapiens)<400>89Cys Glu Leu Leu Leu Arg Lys Gly Ala Asn Ile Asn Glu Lys Thr Lys1 5 10 15Glu Phe Leu Thr Pro Leu His Val Ala Ser Glu Lys Ala His Asn Asp
20 25 30Val Val Glu Val Val Val Lys His Glu Ala Lys Val Asn Ala Leu Asp
35 40 45Asn Leu Gly Gln Thr Ser Leu His Arg Ala Ala Tyr Cys Gly His Leu
50 55 60Gln Thr Cys Arg Leu Leu Leu Ser Tyr Gly Cys Asp Pro Asn Ile Ile65 70 75 80Ser Leu Gln Gly Phe Thr Ala Leu Gln Met Gly Asn Glu Asn Val Gln
85 90 95Gln Leu Leu Gln Glu Gly Ile Ser Leu Gly Asn Ser Glu Ala Asp Arg
100 105 110Gln Leu Leu Glu Ala Ala Lys Ala Gly Asp Val Glu Thr Val Lys Lys
115 120 125Leu Cys Thr Val Gln Ser Val Asn Cys Arg Asp Ile Glu Gly Arg Gln
130 135 140Ser Thr Pro Leu His Phe Ala Ala Gly Tyr Asn Arg Val Ser Val Val145 150 155 160Glu Tyr Leu Leu Gln His Gly Ala Asp Val His Ala Lys Asp Lys Gly
165 170 175Gly Leu Val Pro Leu His Asn Ala Cys Ser Tyr Gly His Tyr Glu Val
180 185 190Ala Glu Leu Leu Val Lys His Gly Ala Val Val Asn Val Ala Asp Leu
195 200 205Trp Lys Phe Thr Pro Leu His Glu Ala Ala Ala Lys Gly Lys Tyr Glu
210 215 220Ile Cys Lys Leu Leu Leu Gln His Gly Ala Asp Pro Thr Lys Lys Asn225 230 235 240Arg Asp Gly Asn Thr Pro Leu Asp Leu Val Lys Asp Gly Asp Thr Asp
245 250 255Ile Gln Asp Leu Leu Arg Gly Asp Ala Ala Leu Leu Asp Ala Ala Lys
260 265 270Lys Gly Cys Leu Ala Arg Val Lys Lys Leu Ser Ser Pro Asp Asn Val
275 280 285Asn Cys Arg Asp Thr Gln Gly Arg His Ser Thr Pro Leu His Leu Ala
290 295 300Ala Gly Tyr Asn Asn Leu Glu Val Ala Glu Tyr Leu Leu Gln His Gly305 310 315 320Ala Asp Val Asn Ala Gln Asp Lys Gly Gly Leu Ile Pro Leu His Asn
325 330 335Ala Ala Ser Tyr Gly His Val Asp Val Ala Ala Leu Leu Ile Lys Tyr
340 345 350Asn Ala Cys Val Asn Ala Thr Asp Lys Trp Ala Phe Thr Pro Leu His
355 360 365Glu Ala Ala Gln Lys Gly Arg Thr Gln Leu Cys Ala Leu Leu Leu Ala
370 375 380His Gly Ala Asp Pro Thr Leu Lys Asn Gln Glu Gly Gln Thr Pro Leu385 390 395 400Asp Leu Val Ser Ala Asp Asp Val Ser Ala Leu Leu Thr Ala Ala Met
405 410 415Pro Pro Ser Ala Leu Pro Ser Cys Tyr Lys Pro Gln Val Leu Asn Gly
420 425 430Val Arg Ser Pro Gly Ala Thr Ala Asp Ala Leu Ser Ser Gly Pro Ser
435 440 445Ser Pro Ser Ser Leu Ser Ala Ala Ser Ser Leu Asp Asn Leu Ser Gly
450 455 460Ser Phe Ser Glu Leu Ser Ser Val Val Ser Ser Ser Gly Thr Glu Gly465 470 475 480Ala Ser Ser Leu Glu Lys Lys Glu Val Pro Gly Val Asp Phe Ser Ile
485 490 495Thr Gln Phe Val Arg Asn Leu Gly Leu Glu His Leu Met Asp Ile Phe
500 505 510Glu Arg Glu Gln Ile Thr Leu Asp Val Leu Val Glu Met Gly His Lys
515 520 525Glu Leu Lys Glu Ile Gly Ile Asn Ala Tyr Gly His Arg His Lys Leu
530 535 540Ile Lys Gly Val Glu Arg Leu Ile Ser Gly Gln Gln Gly Leu Asn Pro545 550 555 560Tyr Leu Thr Leu Asn Thr Ser Gly Ser Gly Thr Ile Leu Ile Asp Leu
565 570 575Ser Pro Asp Asp Lys Glu Phe Gln Ser Val Glu Glu Glu Met Gln Ser
580 585 590Thr Val Arg Glu His Arg Asp Gly Gly His Ala Gly Gly Ile Phe Asn
595 600 605Arg Tyr Asn Ile Leu Lys Ile Gln Lys Val Cys Asn Lys Lys Leu Trp
610 615 620Glu Arg Tyr Thr His Arg Arg Lys Glu Val Ser Glu Glu Asn His Asn625 630 635 640His Ala Asn Glu Arg Met Leu Phe His Gly Ser Pro Phe Val Asn Ala
645 650 655Ile Ile His Lys Gly Phe Asp Glu Arg His Ala Tyr Ile Gly Gly Met
660 665 670Phe Gly Ala Gly Ile Tyr Phe Ala Glu Asn Ser Ser Lys Ser Asn Gln
675 680 685Tyr Val Tyr Gly Ile Gly Gly Gly Thr Gly Cys Pro Val His Lys Asp
690 695 700Arg Ser Cys Tyr Ile Cys His Arg Gln Leu Leu Phe Cys Arg Val Thr705 710 715 720Leu Gly Lys Ser Phe Leu Gln Phe Ser Ala Met Lys Met Ala His Ser
725 730 735Pro Pro Gly His His Ser Val Thr Gly Arg Pro Ser Val Asn Gly Leu
740 745 750Ala Leu Ala Glu Tyr Val Ile Tyr Arg Gly Glu Gln Ala Tyr Pro Glu
755 760 765Tyr Leu Ile Thr Tyr Gln Ile Met Arg Pro Glu Gly Met Val Asp Gly
770 775 780<210>90<211>3799<212>DNA<213>人(Homo sapiens)<220><221>CDS<222>(3)..(2270)<400>90aa gct cat aat gat gtt gtt gaa gta gtg gtg aaa cat gaa gca aag 47Ala His Asn Asp Val Val Glu Val Val Val Lys His Glu Ala Lys
1 5 10 15gtt aat gct ctg gat aat ctt ggt cag act tct cta cac aga gct gca 95Val Asn Ala Leu Asp Asn Leu Gly Gln Thr Ser Leu His Arg Ala Ala
20 25 30tat tgt ggt cat cta caa acc tgc cgc cta ctc ctg agc tat ggg tgt 143Tyr Cys Gly His Leu Gln Thr Cys Arg Leu Leu Leu Ser Tyr Gly Cys
35 40 45gat cct aac att ata tcc ctt cag ggc ttt act gct tta cag atg gga 191Asp Pro Asn Ile Ile Ser Leu Gln Gly Phe Thr Ala Leu Gln Met Gly
50 55 60aat gaa aat gta cag caa ctc ctc caa gag ggt atc tca tta ggt aat 239Asn Glu Asn Val Gln Gln Leu Leu Gln Glu Gly Ile Ser Leu Gly Asn
65 70 75tca gag gca gac aga caa ttg ctg gaa gct gca aag gct gga gat gtc 287Ser Glu Ala Asp Arg Gln Leu Leu Glu Ala Ala Lys Ala Gly Asp Val80 85 90 95gaa act gta aaa aaa ctg tgt act gtt cag agt gtc aac tgc aga gac 335Glu Thr Val Lys Lys Leu Cys Thr Val Gln Ser Val Asn Cys Arg Asp
100 105 110att gaa ggg cgt cag tct aca cca ctt cat ttt gca gct ggg tat aac 383Ile Glu Gly Arg Gln Ser Thr Pro Leu His Phe Ala Ala Gly Tyr Asn
115 120 125aga gtg tcc gtg gtg gaa tat ctg cta cag cat gga gct gat gtg cat 431Arg Val Ser Val Val Glu Tyr Leu Leu Gln His Gly Ala Asp Val His
130 135 140gct aaa gat aaa gga ggc ctt gta cct ttg cac aat gca tgt tct tat 479Ala Lys Asp Lys Gly Gly Leu Val Pro Leu His Asn Ala Cys Ser Tyr
145 150 155gga cat tat gaa gtt gca gaa ctt ctt gtt aaa cat gga gca gta gtt 527Gly His Tyr Glu Val Ala Glu Leu Leu Val Lys His Gly Ala Val Val160 165 170 175aat gta gct gat tta tgg aaa ttt aca cct tta cat gaa gca gca gca 575Asn Val Ala Asp Leu Trp Lys Phe Thr Pro Leu His Glu Ala Ala Ala
180 185 190aaa gga aaa tat gaa att tgc aaa ctt ctg ctc cag cat ggt gca gac 623Lys Gly Lys Tyr Glu Ile Cys Lys Leu Leu Leu Gln His Gly Ala Asp
195 200 205cct aca aaa aaa aac agg gat gga aat act cct ttg gat ctt gtt aaa 671Pro Thr Lys Lys Asn Arg Asp Gly Asn Thr Pro Leu Asp Leu Val Lys
210 215 220gat gga gat aca gat att caa gat ctg ctt agg gga gat gca gct ttg 719Asp Gly Asp Thr Asp Ile Gln Asp Leu Leu Arg Gly Asp Ala Ala Leu
225 230 235cta gat gct gcc aag aag ggt tgt tta gcc aga gtg aag aag ttg tct 767Leu Asp Ala Ala Lys Lys Gly Cys Leu Ala Arg Val Lys Lys Leu Ser240 245 250 255tct cct gat aat gta aat tgc cgc gat acc caa ggc aga cat tca aca 815Ser Pro Asp Asn Val Asn Cys Arg Asp Thr Gln Gly Arg His Ser Thr
260 265 270cct tta cat tta gca gct ggt tat aat aat tta gaa gtt gca gag tat 863Pro Leu His Leu Ala Ala Gly Tyr Asn Asn Leu Glu Val Ala Glu Tyr
275 280 285ttg tta caa cac gga gct gat gtg aat gcc caa gac aaa gga gga ctt 911Leu Leu Gln His Gly Ala Asp Val Asn Ala Gln Asp Lys Gly Gly Leu
290 295 300att cct tta cat aat gca gca tct tac ggg cat gta gat gta gca gct 959Ile Pro Leu His Asn Ala Ala Ser Tyr Gly His Val Asp Val Ala Ala
305 310 315cta cta ata aag tat aat gca tgt gtc aat gcc acg gac aaa tgg gct 1007Leu Leu Ile Lys Tyr Asn Ala Cys Val Asn Ala Thr Asp Lys Trp Ala320 325 330 335ttc aca cct ttg cac gaa gca gcc caa aag gga cga aca cag ctt tgt 1055Phe Thr Pro Leu His Glu Ala Ala Gln Lys Gly Arg Thr Gln Leu Cys
340 345 350gct ttg ttg cta gcc cat gga gct gac ccg act ctt aaa aat cag gaa 1103Ala Leu Leu Leu Ala His Gly Ala Asp Pro Thr Leu Lys Asn Gln Glu
355 360 365gga caa aca cct tta gat tta gtt tca gca gat gat gtc agc gct ctt 1151Gly Gln Thr Pro Leu Asp Leu Val Ser Ala Asp Asp Val Ser Ala Leu
370 375 380ctg aca gca gcc atg ccc cca tct gct ctg ccc tct tgt tac aag cct 1199Leu Thr Ala Ala Met Pro Pro Ser Ala Leu Pro Ser Cys Tyr Lys Pro
385 390 395caa gtg ctc aat ggt gtg aga agc cca gga gcc act gca gat gct ctc 1247Gln Val Leu Asn Gly Val Arg Ser Pro Gly Ala Thr Ala Asp Ala Leu400 405 410 415tct tca ggt cca tct agc cca tca agc ctt tct gca gcc agc agt ctt 1295Ser Ser Gly Pro Ser Ser Pro Ser Ser Leu Ser Ala Ala Ser Ser Leu
420 425 430gac aac tta tct ggg agt ttt tca gaa ctg tct tca gta gtt agt tca 1343Asp Asn Leu Ser Gly Ser Phe Ser Glu Leu Ser Ser Val Val Ser Ser
435 440 445agt gga aca gag ggt gct tcc agt ttg gag aaa aag gag gtt cca gga 1391Ser Gly Thr Glu Gly Ala Ser Ser Leu Glu Lys Lys Glu Val Pro Gly
450 455 460gta gat ttt agc ata act caa ttc gta agg aat ctt gga ctt gag cac 1439Val Asp Phe Ser Ile Thr Gln Phe Val Arg Asn Leu Gly Leu Glu His
465 470 475cta atg gat ata ttt gag aga gaa cag atc act ttg gat gta tta gtt 1487Leu Met Asp Ile Phe Glu Arg Glu Gln Ile Thr Leu Asp Val Leu Val480 485 490 495gag atg ggg cac aag gag ctg aag gag att gga atc aat gct tat gga 1535Glu Met Gly His Lys Glu Leu Lys Glu Ile Gly Ile Asn Ala Tyr Gly
500 505 510cat agg cac aaa cta att aaa gga gtc gag aga ctr atc rcc gga caa 1583His Arg His Lys Leu Ile Lys Gly Val Glu Arg Leu Ile Ser Gly Gln
515 520 525caa ggt ctt aac cca tat tta act ttg aac acc tct ggt agt gga aca 1631Gln Gly Leu Asn Pro Tyr Leu Thr Leu Asn Thr Ser Gly Ser Gly Thr
530 535 540att ctt ata gat ctg tct cct gat gat aaa gag ttt cag tct gtg gag 1679Ile Leu Ile Asp Leu Ser Pro Asp Asp Lys Glu Phe Gln Ser Val Glu
545 550 555gaa gag atg caa agt aca gtt cga gag cac aga gat gga ggt cat gca 1727Glu Glu Met Gln Ser Thr Val Arg Glu His Arg Asp Gly Gly His Ala560 565 570 575ggt gga atc ttc aac aga tac aat att ctc aag att cag aag gtt tgt 1775Gly Gly Ile Phe Asn Arg Tyr Asn Ile Leu Lys Ile Gln Lys Val Cys
580 585 590aac aag aaa cta tgg gaa aga tac act cac cgg aga aaa gaa gtt tct 1823Asn Lys Lys Leu Trp Glu Arg Tyr Thr His Arg Arg Lys Glu Val Ser
595 600 605gaa gaa aac cac aac cat gcc aat gaa cga atg cta ttt cat ggg tct 1871Glu Glu Asn His Asn His Ala Asn Glu Arg Met Leu Phe His Gly Ser
610 615 620cct ttt gtg aat gca att atc cac aaa ggc ttt gat gaa agg cat gcg 1919Pro Phe Val Asn Ala Ile Ile His Lys Gly Phe Asp Glu Arg His Ala
625 630 635tac ata ggt ggt atg ttt gga gct ggc att tat ttt gct gaa aac tct 1967Tyr Ile Gly Gly Met Phe Gly Ala Gly Ile Tyr Phe Ala Glu Asn Ser640 645 650 655tcc aaa agc aat caa tat gta tat gga att gga gga ggt act ggg tgt 2015Ser Lys Ser Asn Gln Tyr Val Tyr Gly Ile Gly Gly Gly Thr Gly Cys
660 665 670cca gtt cac aaa gac aga tct tgt tac att tgc cac agg cag ctg ctc 2063Pro Val His Lys Asp Arg Ser Cys Tyr Ile Cys His Arg Gln Leu Leu
675 680 685ttt tgc cgg gta acc ttg gga aag tct ttc ctg cag ttc agt gca atg 2111Phe Cys Arg Val Thr Leu Gly Lys Ser Phe Leu Gln Phe Ser Ala Met
690 695 700aaa atg gca cat tct cct cca ggt cat cac tca gtc act ggt agg ccc 2159Lys Met Ala His Ser Pro Pro Gly His His Ser Val Thr Gly Arg Pro
705 710 715agt gta aat ggc cta gca tta gct gaa tat gtt att tac aga gga gaa 2207Ser Val Asn Gly Leu Ala Leu Ala Glu Tyr Val Ile Tyr Arg Gly Glu720 725 730 735cag gct tat cct gag tat tta att act tac cag att atg agg cct gaa 2255Gln Ala Tyr Pro Glu Tyr Leu Ile Thr Tyr Gln Ile Met Arg Pro Glu
740 745 750ggt atg gtc gat gga taaatagtta ttttaagaaa ctaattccac tgaacctaaa 2310Gly Met Val Asp Gly
755atcatcaaag cagcagtggc ctctacgttt tactcctttg ctgaaaaaaa atcatcttgc 2370ccacaggcct gtggcaaaag gataaaaatg tgaacgaagt ttaacattct gacttgataa 2430agctttaata atgtacagtg ttttctaaat atttcctgtt ttttcagcac tttaacagat 2490gccattccag gttaaactgg gttgtctgta ctaaattata aacagagtta acttgaacct 2550tttatatgtt atgcattgat tctaacaaac tgtaatgccc tcaacagaac taattttact 2610aatacaatac tgtgttcttt aaaacacagc atttacactg aatacaattt catttgtaaa 2670actgtaaata agagcttttg tactagccca gtatttattt acattgcttt gtaatataaa 2730tctgttttag aactgcagcg gtttacaaaa ttttttcata tgtattgttc atctatactt 2790catcttacat cgtcatgatt gagtgatctt tacatttgat tccagaggct atgttcagtt 2850gttagttggg aaagattgag ttatcagatt taatttgcca ttaaacctta tggggttttc 2910tgttgcagac tgttgattga ccttactaaa tcccgaaatc taaaaaatga attgtggcct 2970tagtaccaca ccatctttaa agtctagtgt ttagtcccct tttccttcaa aactttccaa 3030caaatctagc gctttactga actcagaaca ttgttctctt tgagaatgtg aagattttaa 3090atagccaaag aattttcatg tataagagct agctaaatat agtatatcct gctctttcga 3150agaagataca aaactgttgc ctgtactaat gggtatagta gagcagttga agaactaaca 3210catacatgga cttttcggtc tgaatttgtg ttggcatcca tggtacttac tgttcagtag 3270gatgttattg caaggagcag agtgccctct gctggagtaa tcgcaattat tcttgcagca 3330gattaatttg acttgggtca tgaattcaac aaccagttac ttgcctttca tcatacaatt 3390tcttcggtag ttgagaattt ggtctacatt tatcaaatga ggaaagagtg tcacaaactc 3450taaaaagctg aaggagaccc cacacatctt ctcactgtca gcagccctta cttctgcaaa 3510atgttgaagg ataatgtttc tctgtttgca aagaagatgc ctctggctag aatgtttgtg 3570cagttataag caagggactg cttgtttttg taagttatct caactttatt cttgtgaaat 3630tgcaaaggaa gatcaataaa aagacttcat ttgaatgtaa atggtgtgaa atactgatgt 3690gttttgtaca tgtacataat atatttactt cctgctttca cattagtaat ctgagatggt 3750tctaccattt tataattaga aggagatgta ggggtgggag tggggaggg 3799<210>91<211>756<212>PRT<213>人(Homo sapiens)<400>91Ala His Asn Asp Val Val Glu Val Val Val Lys His Glu Ala Lys Val1 5 10 15Asn Ala Leu Asp Asn Leu Gly Gln Thr Ser Leu His Arg Ala Ala Tyr
20 25 30Cys Gly His Leu Gln Thr Cys Arg Leu Leu Leu Ser Tyr Gly Cys Asp
35 40 45Pro Asn Ile Ile Ser Leu Gln Gly Phe Thr Ala Leu Gln Met Gly Asn
50 55 60Glu Asn Val Gln Gln Leu Leu Gln Glu Gly Ile Ser Leu Gly Asn Ser65 70 75 80Glu Ala Asp Arg Gln Leu Leu Glu Ala Ala Lys Ala Gly Asp Val Glu
85 90 95Thr Val Lys Lys Leu Cys Thr Val Gln Ser Val Asn Cys Arg Asp Ile
100 105 110Glu Gly Arg Gln Ser Thr Pro Leu His Phe Ala Ala Gly Tyr Asn Arg
115 120 125Val Ser Val Val Glu Tyr Leu Leu Gln His Gly Ala Asp Val His Ala
130 135 140Lys Asp Lys Gly Gly Leu Val Pro Leu His Asn Ala Cys Ser Tyr Gly145 150 155 160His Tyr Glu Val Ala Glu Leu Leu Val Lys His Gly Ala Val Val Asn
165 170 175Val Ala Asp Leu Trp Lys Phe Thr Pro Leu His Glu Ala Ala Ala Lys
180 185 190Gly Lys Tyr Glu Ile Cys Lys Leu Leu Leu Gln His Gly Ala Asp Pro
195 200 205Thr Lys Lys Asn Arg Asp Gly Asn Thr Pro Leu Asp Leu Val Lys Asp
210 215 220Gly Asp Thr Asp Ile Gln Asp Leu Leu Arg Gly Asp Ala Ala Leu Leu225 230 235 240Asp Ala Ala Lys Lys Gly Cys Leu Ala Arg Val Lys Lys Leu Ser Ser
245 250 255Pro Asp Asn Val Asn Cys Arg Asp Thr Gln Gly Arg His Ser Thr Pro
260 265 270Leu His Leu Ala Ala Gly Tyr Asn Asn Leu Glu Val Ala Glu Tyr Leu
275 280 285Leu Gln His Gly Ala Asp Val Asn Ala Gln Asp Lys Gly Gly Leu Ile
290 295 300Pro Leu His Asn Ala Ala Ser Tyr Gly His Val Asp Val Ala Ala Leu305 310 315 320Leu Ile Lys Tyr Asn Ala Cys Val Asn Ala Thr Asp Lys Trp Ala Phe
325 330 335Thr Pro Leu His Glu Ala Ala Gln Lys Gly Arg Thr Gln Leu Cys Ala
340 345 350Leu Leu Leu Ala His Gly Ala Asp Pro Thr Leu Lys Asn Gln Glu Gly
355 360 365Gln Thr Pro Leu Asp Leu Val Ser Ala Asp Asp Val Ser Ala Leu Leu
370 375 380Thr Ala Ala Met Pro Pro Ser Ala Leu Pro Ser Cys Tyr Lys Pro Gln385 390 395 400Val Leu Asn Gly Val Arg Ser Pro Gly Ala Thr Ala Asp Ala Leu Ser
405 410 415Ser Gly Pro Ser Ser Pro Ser Ser Leu Ser Ala Ala Ser Ser Leu Asp
420 425 430Asn Leu Ser Gly Ser Phe Ser Glu Leu Ser Ser Val Val Ser Ser Ser
435 440 445Gly Thr Glu Gly Ala Ser Ser Leu Glu Lys Lys Glu Val Pro Gly Val
450 455 460Asp Phe Ser Ile Thr Gln Phe Val Arg Asn Leu Gly Leu Glu His Leu465 470 475 480Met Asp Ile Phe Glu Arg Glu Gln Ile Thr Leu Asp Val Leu Val Glu
485 490 495Met Gly His Lys Glu Leu Lys Glu Ile Gly Ile Asn Ala Tyr Gly His
500 505 510Arg His Lys Leu Ile Lys Gly Val Glu Arg Leu Ile Ser Gly Gln Gln
515 520 525Gly Leu Asn Pro Tyr Leu Thr Leu Asn Thr Ser Gly Ser Gly Thr Ile
530 535 540Leu Ile Asp Leu Ser Pro Asp Asp Lys Glu Phe Gln Ser Val Glu Glu545 550 555 560Glu Met Gln Ser Thr Val Arg Glu His Arg Asp Gly Gly His Ala Gly
565 570 575Gly Ile Phe Asn Arg Tyr Asn Ile Leu Lys Ile Gln Lys Val Cys Asn
580 585 590Lys Lys Leu Trp Glu Arg Tyr Thr His Arg Arg Lys Glu Val Ser Glu
595 600 605Glu Asn His Asn His Ala Asn Glu Arg Met Leu Phe His Gly Ser Pro
610 615 620Phe Val Asn Ala Ile Ile His Lys Gly Phe Asp Glu Arg His Ala Tyr625 630 635 640Ile Gly Gly Met Phe Gly Ala Gly Ile Tyr Phe Ala Glu Asn Ser Ser
645 650 655Lys Ser Asn Gln Tyr Val Tyr Gly Ile Gly Gly Gly Thr Gly Cys Pro
660 665 670Val His Lys Asp Arg Ser Cys Tyr Ile Cys His Arg Gln Leu Leu Phe
675 680 685Cys Arg Val Thr Leu Gly Lys Ser Phe Leu Gln Phe Ser Ala Met Lys
690 695 700Met Ala His Ser Pro Pro Gly His His Ser Val Thr Gly Arg Pro Ser705 710 715 720Val Asn Gly Leu Ala Leu Ala Glu Tyr Val Ile Tyr Arg Gly Glu Gln
725 730 735Ala Tyr Pro Glu Tyr Leu Ile Thr Tyr Gln Ile Met Arg Pro Glu Gly
740 745 750Met Val Asp Gly
755<210>92<211>2971<212>DNA<213>人(Homo sapiens)<400>92tgtgaactgt tgctaagaaa aggagcaaac atcaatgaaa agactaaaga attcttgact 60cctctgcacg tggcatctga gaaagctcat aatgatgttg ttgaagtagt ggtgaaacat 120gaagcaaagg ttaatgctct ggataatctt ggtcagactt ctctacacag agctgcatat 180tgtggtcatc tacaaacctg ccgcctactc ctgagctatg ggtgtgatcc taacattata 240tcccttcagg gctttactgc tttacagatg ggaaatgaaa atgtacagca actcctccaa 300gagggtatct cattaggtaa ttcagaggca gacagacaat tgctggaagc tgcaaaggct 360ggagatgtcg aaactgtaaa aaaactgtgt actgttcaga gtgtcaactg cagagacatt 420gaagggcgtc agtctacacc acttcatttt gcagctgggt ataacagagt gtccgtggtg 480gaatatctgc tacagcatgg agctgatgtg catgctaaag ataaaggagg ccttgtacct 540ttgcacaatg catgttctta tggacattat gaagttgcag aacttcttgt taaacatgga 600gcagtagtta atgtagctga tttatggaaa tttacacctt tacatgaagc agcagcaaaa 660ggaaaatatg aaatttgcaa acttctgctc cagcatggtg cagaccctac aaaaaaaaac 720agggatggaa atactccttt ggatcttgtt aaagatggag atacagatat tcaagatctg 780cttaggggag atgcagcttt gctagatgct gccaagaagg gttgtttagc cagagtgaag 840aagttgtctt ctcctgataa tgtaaattgc cgcgataccc aaggcagaca ttcaacacct 900ttacatttag cagctggtta taataattta gaagttgcag agtatttgtt acaacacgga 960gctgatgtga atgcccaaga caaaggagga cttattcctt tacataatgc agcatcttac 1020gggcatgtag atgtagcagc tctactaata aagtataatg catgtgtcaa tgccacggac 1080aaatgggctt tcacaccttt gcacgaagca gcccaaaagg gacgaacaca gctttgtgct 1140ttgttgctag cccatggagc tgacccgact cttaaaaatc aggaaggaca aacaccttta 1200gatttagttt cagcagatga tgtcagcgct cttctgacag cagccatgcc cccatctgct 1260ctgccctctt gttacaagcc tcaagtgctc aatggtgtga gaagcccagg agccactgca 1320gatgctctct cttcaggtcc atctagccca tcaagccttt ctgcagccag cagtcttgac 1380aacttatctg ggagtttttc agaactgtct tcagtagtta gttcaagtgg aacagagggt 1440gcttccagtt tggagaaaaa ggaggttcca ggagtagatt ttagcataac tcaattcgta 1500aggaatcttg gacttgagca cctaatggat atatttgaga gagaacagat cactttggat 1560gtattagttg agatggggca caaggagctg aaggagattg gaatcaatgc ttatggacat 1620aggcacaaac taattaaagg agtcgagaga cttatctccg gacaacaagg tcttaaccca 1680tatttaactt tgaacacctc tggtagtgga acaattctta tagatctgtc tcctgatgat 1740aaagagtttc agtctgtgga ggaagagatg caaagtacag ttcgagagca cagagatgga 1800ggtcatgcag gtggaatctt caacagatac aatattctca agattcagaa ggtttgtaac 1860aagaaactat gggaaagata cactcaccgg agaaaagaag tttctgaaga aaaccacaac 1920catgccaatg aacgaatgct atttcatggg tctccttttg tgaatgcaat tatccacaaa 1980ggctttgatg aaaggcatgc gtacataggt ggtatgtttg gagctggcat ttattttgct 2040gaaaactctt ccaaaagcaa tcaatatgta tatggaattg gaggaggtac tgggtgtcca 2100gttcacaaag acagatcttg ttacatttgc cacaggcagc tgctcttttg ccgggtaacc 2160ttgggaaagt ctttcctgca gttcagtgca atgaaaatgg cacattctcc tccaggtcat 2220cactcagtca ctggtaggcc cagtgtaaat ggcctagcat tagctgaata tgttatttac 2280agaggagaac aggcttatcc tgagtattta attacttacc agattatgag gcctgaaggt 2340atggtcgatg gataaatagt tattttaaga aactaattcc actgaaccta aaatcatcaa 2400agcagcagtg gcctctacgt tttactcctt tgctgaaaaa aaatcatctt gcccacaggc 2460ctgtggcaaa aggataaaaa tgtgaacgaa gtttaacatt ctgacttgat aaagctttaa 2520taatgtacag tgttttctaa atatttcctg ttttttcagc actttaacag atgccattcc 2580aggttaaact gggttgtctg tactaaatta taaacagagt taacttgaac cttttatatg 2640ttatgcattg attctaacaa actgtaatgc cctcaacaga actaatttta ctaatacaat 2700actgtgttct ttaaaacaca gcatttacac tgaatacaat ttcatttgta aaactgtaaa 2760taagagcttt tgtactagcc cagtatttat ttacattgct ttgtaatata aatctgtttt 2820agaactgcag cggtttacaa aattttttca tatgtattgt tcatctatac ttcatcttac 2880atcgtcatga ttgagtgatc tttacatttg attccagagg ctatgttcag ttgttagttg 2940ggaaagattg agttatcaga tttaatttgc c 2971<210>93<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>93gggcggaaag acgtagttga 20<210>94<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>94gcggctgttc accttctcag 20<210>95<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>95acgcaagtga tggcagaaag 20<210>96<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>96tcacttgcgt ggcagttgac 20<210>97<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>97gcggcaggtt tgtagatgac 20<210>98<211>1568<212>DNA<213>人(Homo sapiens)<220><221>CDS<222>(2)..(1567)<400>98g gcc agg atc atg tcg ggt cgc cgc tgc gcc ggc ggg gga gcg gcc tgc 49Ala Arg Ile Met Ser Gly Arg Arg Cys Ala Gly Gly Gly Ala Ala Cys
1 5 10 15gcg agc gcc gcg gcc gag gcc gtg gag ccg gcc gcc cga gag ctg ttc 97Ala Ser Ala Ala Ala Glu Ala Val Glu Pro Ala Ala Arg Glu Leu Phe
20 25 30gag gcg tgc cgc aac ggg gac gtg gaa cga gtc aag agg ctg gtg acg 145Glu Ala Cys Arg Asn Gly Asp Val Glu Arg Val Lys Arg Leu Val Thr
35 40 45cct gag aag gtg aac agc cgc gac acg gcg ggc agg aaa tcc acc ccg 193Pro Glu Lys Val Asn Ser Arg Asp Thr Ala Gly Arg Lys Ser Thr Pro
50 55 60ctg cac ttc gcc gca ggt ttt ggg cgg aaa gac gta gtt gaa tat ttg 241Leu His Phe Ala Ala Gly Phe Gly Arg Lys Asp Val Val Glu Tyr Leu65 70 75 80ctt cag aat ggt gca aat gtc caa gca cgt gat gat ggg ggc ctt att 289Leu Gln Asn Gly Ala Asn ValGln Ala Arg Asp Asp Gly Gly Leu Ile
85 90 95cct ctt cat aat gca tgc tct ttt ggt cat gct gaa gta gtc aat ctc 337Pro Leu His Asn Ala Cys Ser Phe Gly His Ala Glu Val Val Asn Leu
100 105 110ctt ttg cgg cat ggt gca gac ccc aat gct cga gat aat tgg aat tat 385Leu Leu Arg His Gly Ala Asp Pro Asn Ala Arg Asp Asn Trp Asn Tyr
115 120 125act cct ctc cat gaa gct gca att aaa gga aag att gat gtt tgc att 433Thr Pro Leu His Glu Ala Ala Ile Lys Gly Lys Ile Asp Val Cys Ile
130 135 140gtg ctg tta cag cat gga gct gag cca acc atc cga aat aca gat gga 481Val Leu Leu Gln His Gly Ala Glu Pro Thr Ile Arg Asn Thr Asp Gly145 150 155 160agg aca gca ttg gat tta gca gat cca tct gcc aaa gca gtg ctt act 529Arg Thr Ala Leu Asp Leu Ala Asp Pro Ser Ala Lys Ala Val Leu Thr
165 170 175ggt gaa tat aag aaa gat gaa ctc tta gaa agt gcc agg agt ggc aat 577Gly Glu Tyr Lys Lys Asp Glu Leu Leu Glu Ser Ala Arg Ser Gly Asn
180 185 190gaa gaa aaa atg atg gct cta ctc aca cca tta aat gtc aac tgc cac 625Glu Glu Lys Met Met Ala Leu Leu Thr Pro Leu Asn Val Asn Cys His
195 200 205gca agt gat ggc aga aag tca act cca tta cat ttg gca gca gga tat 673Ala Ser Asp Gly Arg Lys Ser Thr Pro Leu His Leu Ala Ala Gly Tyr
210 215 220aac aga gta aag att gta cag ctg tta ctg caa cat gga gct gat gtc 721Asn Arg Val Lys Ile Val Gln Leu Leu Leu Gln His Gly Ala Asp Val225 230 235 240cat gct aaa gat aaa ggt gat ctg gta cca tta cac aat gcc tgt tct 769His Ala Lys Asp Lys Gly Asp Leu Val Pro Leu His Asn Ala Cys Ser
245 250 255tat ggt cat tat gaa gta act gaa ctt ttg gtc aag cat ggt gcc tgt 817Tyr Gly His Tyr Glu Val Thr Glu Leu Leu Val Lys His Gly Ala Cys
260 265 270gta aat gca atg gac ttg tgg caa ttc act cct ctt cat gag gca gct 865ValAsn Ala Met Asp Leu Trp Gln Phe Thr Pro Leu His Glu Ala Ala
275 280 285tct aag aac agg gtt gaa gta tgt tct ctt ctc tta agt tat ggt gca 913Ser Lys Asn Arg Val Glu Val Cys Ser Leu Leu Leu Ser Tyr Gly Ala
290 295 300gac cca aca ctg ctc aat tgt cac aat aaa agt gct ata gac ttg gct 961Asp Pro Thr Leu Leu Asn Cys His Asn Lys Ser Ala Ile Asp Leu Ala305 310 315 320ccc aca cca cag tta aaa gaa aga tta gca tat gaa ttt aaa ggc cac 1009Pro Thr Pro Gln Leu Lys Glu Arg Leu Ala Tyr Glu Phe Lys Gly His
325 330 335tcg ttg ctg caa gct gca cga gaa gct gat gtt act cga atc aaa aaa 1057Ser Leu Leu Gln Ala Ala Arg Glu Ala Asp Val Thr Arg Ile Lys Lys
340 345 350cat ctc tct ctg gaa atg gtg aat ttc aag cat cct caa aca cat gaa 1105His Leu Ser Leu Glu Met Val Asn Phe Lys His Pro Gln Thr His Glu
355 360 365aca gca ttg cat tgt gct gct gca tct cca tat ccc aaa aga aag caa 1153Thr Ala Leu His Cys Ala Ala Ala Ser Pro Tyr Pro Lys Arg Lys Gln
370 375 380ata tgt gaa ctg ttg cta aga aaa gga gca aac atc aat gaa aag act 1201Ile Cys Glu Leu Leu Leu Arg Lys Gly Ala Asn Ile Asn Glu Lys Thr385 390 395 400aaa gaa ttc ttg act cct ctg cac gtg gca tct gag aaa gct cat aat 1249Lys Glu Phe Leu Thr Pro Leu His Val Ala Ser Glu Lys Ala His Asn
405 410 415gat gtt gtt gaa gta gtg gtg aaa cat gaa gca aag gtt aat gct ctg 1297Asp Val Val Glu Val Val Val Lys His Glu Ala Lys Val Asn Ala Leu
420 425 430gat aat ctt ggt cag act tct cta cac aga gct gca tat tgt ggt cat 1345Asp Asn Leu Gly Gln Thr Ser Leu His Arg Ala Ala Tyr Cys Gly His
435 440 445cta caa acc tgc cgc cta ctc ctg agc tat ggg tgt gat cct aac att 1393Leu Gln Thr Cys Arg Leu Leu Leu Ser Tyr Gly Cys Asp Pro Asn Ile
450 455 460ata tcc ctt cag ggc ttt act gct tta cag atg gga aat gaa aat gta 1441Ile Ser Leu Gln Gly Phe Thr Ala Leu Gln Met Gly Asn Glu Asn Val465 470 475 480cag caa ctc ctc caa gag ggt atc tca tta ggt aat tca gag gca gac 1489Gln Gln Leu Leu Gln Glu Gly Ile Ser Leu Gly Asn Ser Glu Ala Asp
485 490 495aga caa ttg ctg gaa gct gca aag gct gga gat gtc gaa act gta aaa 1537Arg Gln Leu Leu Glu Ala Ala Lys Ala Gly Asp Val Glu Thr Val Lys
500 505 510aaa ctg tgt act gtt cag agt gtc aac tgc a 1568Lys Leu Cys Thr Val Gln Ser Val Asn Cys
515 520<210>99<211>522<212>PRT<213>人(Homo sapiens)<400>99Ala Arg Ile Met Ser Gly Arg Arg Cys Ala Gly Gly Gly Ala Ala Cys1 5 10 15Ala Ser Ala Ala Ala Glu Ala Val Glu Pro Ala Ala Arg Glu Leu Phe
20 25 30Glu Ala Cys Arg Asn Gly Asp Val Glu Arg Val Lys Arg Leu Val Thr
35 40 45Pro Glu Lys Val Asn Ser Arg Asp Thr Ala Gly Arg Lys Ser Thr Pro
50 55 60Leu His Phe Ala Ala Gly Phe Gly Arg Lys Asp Val Val Glu Tyr Leu65 70 75 80Leu Gln Asn Gly Ala Asn ValGln Ala Arg Asp Asp Gly Gly Leu Ile
85 90 95Pro Leu His Asn Ala Cys Ser Phe Gly His Ala Glu Val Val Asn Leu
100 105 110Leu Leu Arg His Gly Ala Asp Pro Asn Ala Arg Asp Asn Trp Asn Tyr
115 120 125Thr Pro Leu His Glu Ala Ala Ile Lys Gly Lys Ile Asp Val Cys Ile
130 135 140Val Leu Leu Gln His Gly Ala Glu Pro Thr Ile Arg Asn Thr Asp Gly145 150 155 160Arg Thr Ala Leu Asp Leu Ala Asp Pro Ser Ala Lys Ala Val Leu Thr
165 170 175Gly Glu Tyr Lys Lys Asp Glu Leu Leu Glu Ser Ala Arg Ser Gly Asn
180 185 190Glu Glu Lys Met Met Ala Leu Leu Thr Pro Leu Asn Val Asn Cys His
195 200 205Ala Ser Asp Gly Arg Lys Ser Thr Pro Leu His Leu Ala Ala Gly Tyr
210 215 220Asn Arg Val Lys Ile Val Gln Leu Leu Leu Gln His Gly Ala Asp Val225 230 235 240His Ala Lys Asp Lys Gly Asp Leu Val Pro Leu His Asn Ala Cys Ser
245 250 255Tyr Gly His Tyr Glu Val Thr Glu Leu Leu Val Lys His Gly Ala Cys
260 265 270Val Asn Ala Met Asp Leu Trp Gln Phe Thr Pro Leu His Glu Ala Ala
275 280 285Ser Lys Asn Arg Val Glu Val Cys Ser Leu Leu Leu Ser Tyr Gly Ala
290 295 300Asp Pro Thr Leu Leu Asn Cys His Asn Lys Ser Ala Ile Asp Leu Ala305 310 315 320Pro Thr Pro Gln Leu Lys Glu Arg Leu Ala Tyr Glu Phe Lys Gly His
325 330 335Ser Leu Leu Gln Ala Ala Arg Glu Ala Asp Val Thr Arg Ile Lys Lys
340 345 350His Leu Ser Leu Glu Met Val Asn Phe Lys His Pro Gln Thr His Glu
355 360 365Thr Ala Leu His Cys Ala Ala Ala Ser Pro Tyr Pro Lys Arg Lys Gln
370 375 380Ile Cys Glu Leu Leu Leu Arg Lys Gly Ala Asn Ile Asn Glu Lys Thr385 390 395 400Lys Glu Phe Leu Thr Pro Leu His Val Ala Ser Glu Lys Ala His Asn
405 410 415Asp Val Val Glu Val Val Val Lys His Glu Ala Lys Val Asn Ala Leu
420 425 430Asp Asn Leu Gly Gln Thr Ser Leu His Arg Ala Ala Tyr Cys Gly His
435 440 445Leu Gln Thr Cys Arg Leu Leu Leu Ser Tyr Gly Cys Asp Pro Asn Ile
450 455 460Ile Ser Leu Gln Gly Phe Thr Ala Leu Gln Met Gly Asn Glu Asn Val465 470 475 480Gln Gln Leu Leu Gln Glu Gly Ile Ser Leu Gly Asn Ser Glu Ala Asp
485 490 495Arg Gln Leu Leu Glu Ala Ala Lys Ala Gly Asp Val Glu Thr Val Lys
500 505 510Lys Leu Cys Thr Val Gln Ser Val Asn Cys
515 520<210>100<211>4127<212>DNA<213>人(Homo sapiens)<220><221>CDS<222>(2)..(3508)<220><221>3’UTR<222>(3509)..(4127)<400>100g gcc agg atc atg tcg ggt cgc cgc tgc gcc ggc ggg gga gcg gcc tgc 49Ala Arg Ile Met Ser Gly Arg Arg Cys Ala Gly Gly Gly Ala Ala Cys
1 5 10 15gcg agc gcc gcg gcc gag gcc gtg gag ccg gcc gcc cga gag ctg ttc 97Ala Ser Ala Ala Ala Glu Ala Val Glu Pro Ala Ala Arg Glu Leu Phe
20 25 30gag gcg tgc cgc aac ggg gac gtg gaa cga gtc aag agg ctg gtg acg 145Glu Ala Cys Arg Asn Gly Asp Val Glu Arg Val Lys Arg Leu Val Thr
35 40 45cct gag aag gtg aac agc cgc gac acg gcg ggc agg aaa tcc acc ccg 193Pro Glu Lys Val Asn Ser Arg Asp Thr Ala Gly Arg Lys Ser Thr Pro
50 55 60ctg cac ttc gcc gca ggt ttt ggg cgg aaa gac gta gtt gaa tat ttg 241Leu His Phe Ala Ala Gly Phe Gly Arg Lys Asp Val Val Glu Tyr Leu65 70 75 80ctt cag aat ggt gca aat gtc caa gca cgt gat gat ggg ggc ctt att 289Leu Gln Asn Gly Ala Asn Val Gln Ala Arg Asp Asp Gly Gly Leu Ile
85 90 95cct ctt cat aat gca tgc tct ttt ggt cat gct gaa gta gtc aat ctc 337Pro Leu His Asn Ala Cys Ser Phe Gly His Ala Glu Val Val Asn Leu
100 105 110ctt ttg cga cat ggt gca gac ccc aat gct cga gat aat tgg aat tat 385Leu Leu Arg His Gly Ala Asp Pro Asn Ala Arg Asp Asn Trp Asn Tyr
115 120 125act cct ctc cat gaa gct gca att aaa gga aag att gat gtt tgc att 433Thr Pro Leu His Glu Ala Ala Ile Lys Gly Lys Ile Asp Val Cys Ile
130 135 140gtg ctg tta cag cat gga gct gag cca acc atc cga aat aca gat gga 481Val Leu Leu Gln His Gly Ala Glu Pro Thr Ile Arg Asn Thr Asp Gly145 150 155 160agg aca gca ttg gat tta gca gat cca tct gcc aaa gca gtg ctt act 529Arg Thr Ala Leu Asp Leu Ala Asp Pro Ser Ala Lys Ala Val Leu Thr
165 170 175ggt gaa tat aag aaa gat gaa ctc tta gaa agt gcc agg agt ggc aat 577Gly Glu Tyr Lys Lys Asp Glu Leu Leu Glu Ser Ala Arg Ser Gly Asn
180 185 190gaa gaa aaa atg atg gct cta ctc aca cca tta aat gtc aac tgc cac 625Glu Glu Lys Met Met Ala Leu Leu Thr Pro Leu Asn Val Asn Cys His
195 200 205gca agt gat ggc aga aag tca act cca tta cat ttg gca gca gga tat 673Ala Ser Asp Gly Arg Lys Ser Thr Pro Leu His Leu Ala Ala Gly Tyr
210 215 220aac aga gta aag att gta cag ctg tta ctg caa cat gga gct gat gtc 721Asn Arg Val Lys Ile Val Gln Leu Leu Leu Gln His Gly Ala Asp Val225 230 235 240cat gct aaa gat aaa ggt gat ctg gta cca tta cac aat gcc tgt tct 769His Ala Lys Asp Lys Gly Asp Leu Val Pro Leu His Asn Ala Cys Ser
245 250 255tat ggt cat tat gaa gta act gaa ctt ttg gtc aag cat ggt gcc tgt 817Tyr Gly His Tyr Glu Val Thr Glu Leu Leu Val Lys His Gly Ala Cys
260 265 270gta aat gca atg gac ttg tgg caa ttc act cct ctt cat gag gca gct 865Val Asn Ala Met Asp Leu Trp Gln Phe Thr Pro Leu His Glu Ala Ala
275 280 285tct aag aac agg gtt gaa gta tgt tct ctt ctc tta agt tat ggt gca 913Ser Lys Asn Arg Val Glu Val Cys Ser Leu Leu Leu Ser Tyr Gly Ala
290 295 300gac cca aca ctg ctc aat tgt cac aat aaa agt gct ata gac ttg gct 961Asp Pro Thr Leu Leu Asn Cys His Asn Lys Ser Ala Ile Asp Leu Ala305 310 315 320ccc aca cca cag tta aaa gaa aga tta gca tat gaa ttt aaa ggc cac 1009Pro Thr Pro Gln Leu Lys Glu Arg Leu Ala Tyr Glu Phe Lys Gly His
325 330 335tcg ttg ctg caa gct gca cga gaa gct gat gtt act cga atc aaa aaa 1057Ser Leu Leu Gln Ala Ala Arg Glu Ala Asp Val Thr Arg Ile Lys Lys
340 345 350cat ctc tct ctg gaa atg gtg aat ttc aag cat cct caa aca cat gaa 1105His Leu Ser Leu Glu Met Val Asn Phe Lys His Pro Gln Thr His Glu
355 360 365aca gca ttg cat tgt gct gct gca tct cca tat ccc aaa aga aag caa 1153Thr Ala Leu His Cys Ala Ala Ala Ser Pro Tyr Pro Lys Arg Lys Gln
370 375 380ata tgt gaa ctg ttg cta aga aaa gga gca aac atc aat gaa aag act 1201Ile Cys Glu Leu Leu Leu Arg Lys Gly Ala Asn Ile Asn Glu Lys Thr385 390 395 400aaa gaa ttc ttg act cct ctg cac gtg gca tct gag aaa gct cat aat 1249Lys Glu Phe Leu Thr Pro Leu His Val Ala Ser Glu Lys Ala His Asn
405 410 415gat gtt gtt gaa gta gtg gtg aaa cat gaa gca aag gtt aat gct ctg 1297Asp Val Val Glu Val Val Val Lys His Glu Ala Lys Val Asn Ala Leu
420 425 430gat aat ctt ggt cag act tct cta cac aga gct gca tat tgt ggt cat 1345Asp Asn Leu Gly Gln Thr Ser Leu His Arg Ala Ala Tyr Cys Gly His
435 440 445cta caa acc tgc cgc cta ctc ctg agc tat ggg tgt gat cct aac att 1393Leu Gln Thr Cys Arg Leu Leu Leu Ser Tyr Gly Cys Asp Pro Asn Ile
450 455 460ata tcc ctt cag ggc ttt act gct tta cag atg gga aat gaa aat gta 1441Ile Ser Leu Gln Gly Phe Thr Ala Leu Gln Met Gly Asn Glu Asn Val465 470 475 480cag caa ctc ctc caa gag ggt atc tca tta ggt aat tca gag gca gac 1489Gln Gln Leu Leu Gln Glu Gly Ile Ser Leu Gly Asn Ser Glu Ala Asp
485 490 495aga caa ttg ctg gaa gct gca aag gct gga gat gtc gaa act gta aaa 1537Arg Gln Leu Leu Glu Ala Ala Lys Ala Gly Asp Val Glu Thr Val Lys
500 505 510aaa ctg tgt act gtt cag agt gtc aac tgc aga gac att gaa ggg cgt 1585Lys Leu Cys Thr Val Gln Ser Val Asn Cys Arg Asp Ile Glu Gly Arg
515 520 525cag tct aca cca ctt cat ttt gca gct ggg tat aac aga gtg tcc gtg 1633Gln Ser Thr Pro Leu His Phe Ala Ala Gly Tyr Asn Arg Val Ser Val
530 535 540gtg gaa tat ctg cta cag cat gga gct gat gtg cat gct aaa gat aaa 1681Val Glu Tyr Leu Leu Gln His Gly Ala Asp Val His Ala Lys Asp Lys545 550 555 560gga ggc ctt gta cct ttg cac aat gca tgt tct tat gga cat tat gaa 1729Gly Gly Leu Val Pro Leu His Asn Ala Cys Ser Tyr Gly His Tyr Glu
565 570 575gtt gca gaa ctt ctt gtt aaa cat gga gca gta gtt aat gta gct gat 1777Val Ala Glu Leu Leu Val Lys His Gly Ala Val Val Asn Val Ala Asp
580 585 590tta tgg aaa ttt aca cct tta cat gaa gca gca gca aaa gga aaa tat 1825Leu Trp Lys Phe Thr Pro Leu His Glu Ala Ala Ala Lys Gly Lys Tyr
595 600 605gaa att tgc aaa ctt ctg ctc cag cat ggt gca gac cct aca aaa aaa 1873Glu Ile Cys Lys Leu Leu Leu Gln His Gly Ala Asp Pro Thr Lys Lys
610 615 620aac agg gat gga aat act cct ttg gat ctt gtt aaa gat gga gat aca 1921Asn Arg Asp Gly Asn Thr Pro Leu Asp Leu Val Lys Asp Gly Asp Thr625 630 635 640gat att caa gat ctg ctt agg gga gat gca gct ttg cta gat gct gcc 1969Asp Ile Gln Asp Leu Leu Arg Gly Asp Ala Ala Leu Leu Asp Ala Ala
645 650 655aag aag ggt tgt tta gcc aga gtg aag aag ttg tct tct cct gat aat 2017Lys Lys Gly Cys Leu Ala Arg Val Lys Lys Leu Ser Ser Pro Asp Asn
660 665 670gta aat tgc cgc gat acc caa ggc aga cat tca aca cct tta cat tta 2065Val Asn Cys Arg Asp Thr Gln Gly Arg His Ser Thr Pro Leu His Leu
675 680 685gca gct ggt tat aat aat tta gaa gtt gca gag tat ttg tta caa cac 2113Ala Ala Gly Tyr Asn Asn Leu Glu Val Ala Glu Tyr Leu Leu Gln His
690 695 700gga gct gat gtg aat gcc caa gac aaa gga gga ctt att cct tta cat 2161Gly Ala Asp Val Asn Ala Gln Asp Lys Gly Gly Leu Ile Pro Leu His705 710 715 720aat gca gca tct tac ggg cat gta gat gta gca gct cta cta ata aag 2209Asn Ala Ala Ser Tyr Gly His Val Asp Val Ala Ala Leu Leu Ile Lys
725 730 735tat aat gca tgt gtc aat gcc acg gac aaa tgg gct ttc aca cct ttg 2257Tyr Asn Ala Cys Val Asn Ala Thr Asp Lys Trp Ala Phe Thr Pro Leu
740 745 750cac gaa gca gcc caa aag gga cga aca cag ctt tgt gct ttg ttg cta 2305His Glu Ala Ala Gln Lys Gly Arg Thr Gln Leu Cys Ala Leu Leu Leu
755 760 765gcc cat gga gct gac ccg act ctt aaa aat cag gaa gga caa aca cct 2353Ala His Gly Ala Asp Pro Thr Leu Lys Asn Gln Glu Gly Gln Thr Pro
770 775 780tta gat tta gtt tca gca gat gat gtc agc gct ctt ctg aca gca gcc 2401Leu Asp Leu Val Ser Ala Asp Asp Val Ser Ala Leu Leu Thr Ala Ala785 790 795 800atg ccc cca tct gct ctg ccc tct tgt tac aag cct caa gtg ctc aat 2449Met Pro Pro Ser Ala Leu Pro Ser Cys Tyr Lys Pro Gln Val Leu Asn
805 810 815ggt gtg aga agc cca gga gcc act gca gat gct ctc tct tca ggt cca 2497Gly Val Arg Ser Pro Gly Ala Thr Ala Asp Ala Leu Ser Ser Gly Pro
820 825 830tct agc cca tca agc ctt tct gca gcc agc agt ctt gac aac tta tct 2545Ser Ser Pro Ser Ser Leu Ser Ala Ala Ser Ser Leu Asp Asn Leu Ser
835 840 845ggg agt ttt tca gaa ctg tct tca gta gtt agt tca agt gga aca gag 2593Gly Ser Phe Ser Glu Leu Ser Ser Val Val Ser Ser Ser Gly Thr Glu
850 855 860ggt gct tcc agt ttg gag aaa aag gag gtt cca gga gta gat ttt agc 2641Gly Ala Ser Ser Leu Glu Lys Lys Glu Val Pro Gly Val Asp Phe Ser865 870 875 880ata act caa ttc gta agg aat ctt gga ctt gag cac cta atg gat ata 2689Ile Thr Gln Phe Val Arg Asn Leu Gly Leu Glu His Leu Met Asp Ile
885 890 895ttt gag aga gaa cag atc act ttg gat gta tta gtt gag atg ggg cac 2737Phe Glu Arg Glu Gln Ile Thr Leu Asp Val Leu Val Glu Met Gly His
900 905 910aag gag ctg aag gag att gga atc aat gct tat gga cat agg cac aaa 2785Lys Glu Leu Lys Glu Ile Gly Ile Asn Ala Tyr Gly His Arg His Lys
915 920 925cta att aaa gga gtc gag aga ctt atc tcc gga caa caa ggt ctt aac 2833Leu Ile Lys Gly Val Glu Arg Leu Ile Ser Gly Gln Gln Gly Leu Asn
930 935 940cca tat tta act ttg aac acc tct ggt agt gga aca att ctt ata gat 2881Pro Tyr Leu Thr Leu Asn Thr Ser Gly Ser Gly Thr Ile Leu Ile Asp945 950 955 960ctg tct cct gat gat aaa gag ttt cag tct gtg gag gaa gag atg caa 2929Leu Ser Pro Asp Asp Lys Glu Phe Gln Ser Val Glu Glu Glu Met Gln
965 970 975agt aca gtt cga gag cac aga gat gga ggt cat gca ggt gga atc ttc 2977Ser Thr Val Arg Glu His Arg Asp Gly Gly His Ala Gly Gly Ile Phe
980 985 990aac aga tac aat att ctc aag att cag aag gtt tgt aac aag aaa cta 3025Asn Arg Tyr Asn Ile Leu Lys Ile Gln Lys Val Cys Asn Lys Lys Leu
995 1000 1005tgg gaa aga tac act cac cgg aga aaa gaa gtt tct gaa gaa aac cac 3073Trp Glu Arg Tyr Thr His Arg Arg Lys Glu Val Ser Glu Glu Asn His1010 1015 1020aac cat gcc aat gaa cga atg cta ttt cat ggg tct cct ttt gtg aat 3121Asn His Ala Asn Glu Arg Met Leu Phe His Gly Ser Pro Phe Val Asn1025 1030 1035 1040gca att atc cac aaa ggc ttt gat gaa agg cat gcg tac ata ggt ggt 3169Ala Ile Ile His Lys Gly Phe Asp Glu Arg His Ala Tyr Ile Gly Gly
1045 1050 1055atg ttt gga gct ggc att tat ttt gct gaa aac tct tcc aaa agc aat 3217Met Phe Gly Ala Gly Ile Tyr Phe Ala Glu Asn Ser Ser Lys Ser Asn
1060 1065 1070caa tat gta tat gga att gga gga ggt act ggg tgt cca gtt cac aaa 3265Gln Tyr Val Tyr Gly Ile Gly Gly Gly Thr Gly Cys Pro Val His Lys
1075 1080 1085gac aga tct tgt tac att tgc cac agg cag ctg ctc ttt tgc cgg gta 3313Asp Arg Ser Cys Tyr Ile Cys His Arg Gln Leu Leu Phe Cys Arg Val1090 1095 1100acc ttg gga aag tct ttc ctg cag ttc agt gca atg aaa atg gca cat 3361Thr Leu Gly Lys Ser Phe Leu Gln Phe Ser Ala Met Lys Met Ala His1105 1110 1115 1120tct cct cca ggt cat cac tca gtc act ggt agg ccc agt gta aat ggc 3409Ser Pro Pro Gly His His Ser Val Thr Gly Arg Pro Ser Val Asn Gly
1125 1130 1135cta gca tta gct gaa tat gtt att tac aga gga gaa cag gct tat cct 3457Leu Ala Leu Ala Glu Tyr Val Ile Tyr Arg Gly Glu Gln Ala Tyr Pro
1140 1145 1150gag tat tta att act tac cag att atg agg cct gaa ggt atg gtc gat 3505Glu Tyr Leu Ile Thr Tyr Gln Ile Met Arg Pro Glu Gly Met Val Asp
1155 1160 1165gga taaatagtta ttttaagaaa ctaattccac tgaacctaaa atcatcaaag 3558Glycagcagtggc ctctacgttt tactcctttg ctgaaaaaaa atcatcttgc ccacaggcct 3618gtggcaaaag gataaaaatg tgaacgaagt ttaacattct gacttgataa agctttaata 3678atgtacagtg ttttctaaat atttcctgtt ttttcagcac tttaacagat gccattccag 3738gttaaactgg gttgtctgta ctaaattata aacagagtta acttgaacct tttatatgtt 3798atgcattgat tctaacaaac tgtaatgccc tcaacagaac taattttact aatacaatac 3858tgtgttcttt aaaacacagc atttacactg aatacaattt catttgtaaa actgtaaata 3918agagcttttg tactagccca gtatttattt acattgcttt gtaatataaa tctgttttag 3978aactgcagcg gtttacaaaa ttttttcata tgtattgttc atctatactt catcttacat 4038cgtcatgatt gagtgatctt tacatttgat tccagaggct atgttcagtt gttagttggg 4098aaagattgag ttatcagatt taatttgcc 4127<210>101<211>1169<212>PRT<213>人(Homo sapiens)<400>101Ala Arg Ile Met Ser Gly Arg Arg Cys Ala Gly Gly Gly Ala Ala Cys1 5 10 15Ala Ser Ala Ala Ala Glu Ala Val Glu Pro Ala Ala Arg Glu Leu Phe
20 25 30Glu Ala Cys Arg Asn Gly Asp Val Glu Arg Val Lys Arg Leu Val Thr
35 40 45Pro Glu Lys Val Asn Ser Arg Asp Thr Ala Gly Arg Lys Ser Thr Pro
50 55 60Leu His Phe Ala Ala Gly Phe Gly Arg Lys Asp Val Val Glu Tyr Leu65 70 75 80Leu Gln Asn Gly Ala Asn Val Gln Ala Arg Asp Asp Gly Gly Leu Ile
85 90 95Pro Leu His Asn Ala Cys Ser Phe Gly His Ala Glu Val Val Asn Leu
100 105 110Leu Leu Arg His Gly Ala Asp Pro Asn Ala Arg Asp Asn Trp Asn Tyr
115 120 125Thr Pro Leu His Glu Ala Ala Ile Lys Gly Lys Ile Asp Val Cys Ile
130 135 140Val Leu Leu Gln His Gly Ala Glu Pro Thr Ile Arg Asn Thr Asp Gly145 150 155 160Arg Thr Ala Leu Asp Leu Ala Asp Pro Ser Ala Lys Ala Val Leu Thr
165 170 175Gly Glu Tyr Lys Lys Asp Glu Leu Leu Glu Ser Ala Arg Ser Gly Asn
180 185 190Glu Glu Lys Met Met Ala Leu Leu Thr Pro Leu Asn Val Asn Cys His
195 200 205Ala Ser Asp Gly Arg Lys Ser Thr Pro Leu His Leu Ala Ala Gly Tyr
210 215 220Asn Arg Val Lys Ile Val Gln Leu Leu Leu Gln His Gly Ala Asp Val225 230 235 240His Ala Lys Asp Lys Gly Asp Leu Val Pro Leu His Asn Ala Cys Ser
245 250 255Tyr Gly His Tyr Glu Val Thr Glu Leu Leu Val Lys His Gly Ala Cys
260 265 270Val Asn Ala Met Asp Leu Trp Gln Phe Thr Pro Leu His Glu Ala Ala
275 280 285Ser Lys Asn Arg Val Glu Val Cys Ser Leu Leu Leu Ser Tyr Gly Ala
290 295 300Asp Pro Thr Leu Leu Asn Cys His Asn Lys Ser Ala Ile Asp Leu Ala305 310 315 320Pro Thr Pro Gln Leu Lys Glu Arg Leu Ala Tyr Glu Phe Lys Gly His
325 330 335Ser Leu Leu Gln Ala Ala Arg Glu Ala Asp Val Thr Arg Ile Lys Lys
340 345 350His Leu Ser Leu Glu Met Val Asn Phe Lys His Pro Gln Thr His Glu
355 360 365Thr Ala Leu His Cys Ala Ala Ala Ser Pro Tyr Pro Lys Arg Lys Gln
370 375 380Ile Cys Glu Leu Leu Leu Arg Lys Gly Ala Asn Ile Asn Glu Lys Thr385 390 395 400Lys Glu Phe Leu Thr Pro Leu His Val Ala Ser Glu Lys Ala His Asn
405 410 415Asp Val Val Glu Val Val Val Lys His Glu Ala Lys Val Asn Ala Leu
420 425 430Asp Asn Leu Gly Gln Thr Ser Leu His Arg Ala Ala Tyr Cys Gly His
435 440 445Leu Gln Thr Cys Arg Leu Leu Leu Ser Tyr Gly Cys Asp Pro Asn Ile
450 455 460Ile Ser Leu Gln Gly Phe Thr Ala Leu Gln Met Gly Asn Glu Asn Val465 470 475 480Gln Gln Leu Leu Gln Glu Gly Ile Ser Leu Gly Asn Ser Glu Ala Asp
485 490 495Arg Gln Leu Leu Glu Ala Ala Lys Ala Gly Asp Val Glu Thr Val Lys
500 505 510Lys Leu Cys Thr Val Gln Ser Val Asn Cys Arg Asp Ile Glu Gly Arg
515 520 525Gln Ser Thr Pro Leu His Phe Ala Ala Gly Tyr Asn Arg Val Ser Val
530 535 540Val Glu Tyr Leu Leu Gln His Gly Ala Asp Val His Ala Lys Asp Lys545 550 555 560Gly Gly Leu Val Pro Leu His Asn Ala Cys Ser Tyr Gly His Tyr Glu
565 570 575Val Ala Glu Leu Leu Val Lys His Gly Ala Val Val Asn Val Ala Asp
580 585 590Leu Trp Lys Phe Thr Pro Leu His Glu Ala Ala Ala Lys Gly Lys Tyr
595 600 605Glu Ile Cys Lys Leu Leu Leu Gln His Gly Ala Asp Pro Thr Lys Lys
610 615 620Asn Arg Asp Gly Asn Thr Pro Leu Asp Leu Val Lys Asp Gly Asp Thr625 630 635 640Asp Ile Gln Asp Leu Leu Arg Gly Asp Ala Ala Leu Leu Asp Ala Ala
645 650 655Lys Lys Gly Cys Leu Ala Arg Val Lys Lys Leu Ser Ser Pro Asp Asn
660 665 670Val Asn Cys Arg Asp Thr Gln Gly Arg His Ser Thr Pro Leu His Leu
675 680 685Ala Ala Gly Tyr Asn Asn Leu Glu Val Ala Glu Tyr Leu Leu Gln His
690 695 700Gly Ala Asp Val Asn Ala Gln Asp Lys Gly Gly Leu Ile Pro Leu His705 710 715 720Asn Ala Ala Ser Tyr Gly His Val Asp Val Ala Ala Leu Leu Ile Lys
725 730 735Tyr Asn Ala Cys Val Asn Ala Thr Asp Lys Trp Ala Phe Thr Pro Leu
740 745 750His Glu Ala Ala Gln Lys Gly Arg Thr Gln Leu Cys Ala Leu Leu Leu
755 760 765Ala His Gly Ala Asp Pro Thr Leu Lys Asn Gln Glu Gly Gln Thr Pro
770 775 780Leu Asp Leu Val Ser Ala Asp Asp Val Ser Ala Leu Leu Thr Ala Ala785 790 795 800Met Pro Pro Ser Ala Leu Pro Ser Cys Tyr Lys Pro Gln Val Leu Asn
805 810 815Gly Val Arg Ser Pro Gly Ala Thr Ala Asp Ala Leu Ser Ser Gly Pro
820 825 830Ser Ser Pro Ser Ser Leu Ser Ala Ala Ser Ser Leu Asp Asn Leu Ser
835 840 845Gly Ser Phe Ser Glu Leu Ser Ser Val Val Ser Ser Ser Gly Thr Glu
850 855 860Gly Ala Ser Ser Leu Glu Lys Lys Glu Val Pro Gly Val Asp Phe Ser865 870 875 880Ile Thr Gln Phe Val Arg Asn Leu Gly Leu Glu His Leu Met Asp Ile
885 890 895Phe Glu Arg Glu Gln Ile Thr Leu Asp Val Leu Val Glu Met Gly His
900 905 910Lys Glu Leu Lys Glu Ile Gly Ile Asn Ala Tyr Gly His Arg His Lys
915 920 925Leu Ile Lys Gly Val Glu Arg Leu Ile Ser Gly Gln Gln Gly Leu Asn
930 935 940Pro Tyr Leu Thr Leu Asn Thr Ser Gly Ser Gly Thr Ile Leu Ile Asp945 950 955 960Leu Ser Pro Asp Asp Lys Glu Phe Gln Ser Val Glu Glu Glu Met Gln
965 970 975Ser Thr Val Arg Glu His Arg Asp Gly Gly His Ala Gly Gly Ile Phe
980 985 990Asn Arg Tyr Asn Ile Leu Lys Ile Gln Lys Val Cys Asn Lys Lys Leu
995 1000 1005Trp Glu Arg Tyr Thr His Arg Arg Lys Glu Val Ser Glu Glu Asn His1010 1015 1020Asn His Ala Asn Glu Arg Met Leu Phe His Gly Ser Pro Phe Val Asn1025 1030 1035 1040Ala Ile Ile His Lys Gly Phe Asp Glu Arg His Ala Tyr Ile Gly Gly
1045 1050 1055Met Phe Gly Ala Gly Ile Tyr Phe Ala Glu Asn Ser Ser Lys Ser Asn
1060 1065 1070Gln Tyr Val Tyr Gly Ile Gly Gly Gly Thr Gly Cys Pro Val His Lys
1075 1080 1085Asp Arg Ser Cys Tyr Ile Cys His Arg Gln Leu Leu Phe Cys Arg Val1090 1095 1100Thr Leu Gly Lys Ser Phe Leu Gln Phe Ser Ala Met Lys Met Ala His1105 1110 1115 1120Ser Pro Pro Gly His His Ser Val Thr Gly Arg Pro Ser Val Asn Gly
1125 1130 1135Leu Ala Leu Ala Glu Tyr Val Ile Tyr Arg Gly Glu Gln Ala Tyr Pro
1140 1145 1150Glu Tyr Leu Ile Thr Tyr Gln Ile Met Arg Pro Glu Gly Met Val Asp
1155 1160 1165Gly<210>102<211>32<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>102gagcattggg gtctgcacca tgtcgcaaaa gg 32<210>103<211>27<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>103ccatcctaat acgactcact atagggc 27<210>104<211>647<212>DNA<213>人(Homo sapiens)<220><221>CDS<222>(2)..(646)<400>104g gag ctg gca gga ggg gcc ttg cca gct tcc gcc gcc gcg tcg ttt cag 49Glu Leu Ala Gly Gly Ala Leu Pro Ala Ser Ala Ala Ala Ser Phe Gln
1 5 10 15gac ccg gac ggc gga ttc gcg ctg cct ccg ccg ccg cgg ggc agc cgg 97Asp Pro Asp Gly Gly Phe Ala Leu Pro Pro Pro Pro Arg Gly Ser Arg
20 25 30ggg gca ggg agc cca gcg agg ggc gcg cgt ggg cgc ggc cat ggg act 145Gly Ala Gly Ser Pro Ala Arg Gly Ala Arg Gly Arg Gly His Gly Thr
35 40 45gcg ccg gat ccg gtg aca gca ggg agc caa gcg gcc cgg gcc ctg agc 193Ala Pro Asp Pro Val Thr Ala Gly Ser Gln Ala Ala Arg Ala Leu Ser
50 55 60gcg tct tct ccg ggg ggc ctc gcc ctc ctg ctc gcg ggg ccg ggg ctc 241Ala Ser Ser Pro Gly Gly Leu Ala Leu Leu Leu Ala Gly Pro Gly Leu65 70 75 80ctg ctc cgg ttg ctg gcg ctg ttg ctg gct gtg gcg gcg gcc agg atc 289Leu Leu Arg Leu Leu Ala Leu Leu Leu Ala Val Ala Ala Ala Arg Ile
85 90 95atg tcg ggt cgc cgc tgc gcc ggc ggg gga gcg gcc tgc gcg agc gcc 337Met Ser Gly Arg Arg Cys Ala Gly Gly Gly Ala Ala Cys Ala Ser Ala
100 105 110gcg gcc gag gcc gtg gag ccg gcc gcc cga gag ctg ttc gag gcg tgc 385Ala Ala Glu Ala Val Glu Pro Ala Ala Arg Glu Leu Phe Glu Ala Cys
115 120 125cgc aac ggg gac gtg gaa cga gtc aag agg ctg gtg acg cct gag aag 433Arg Asn Gly Asp Val Glu Arg Val Lys Arg Leu Val Thr Pro Glu Lys
130 135 140gtg aac agc cgc gac acg gcg ggc agg aaa tcc acc ccg ctg cac ttc 481Val Asn Ser Arg Asp Thr Ala Gly Arg Lys Ser Thr Pro Leu His Phe145 150 155 160gcc gca ggt ttt ggg cgg aaa gac gta gtt gaa tat ttg ctt cag aat 529Ala Ala Gly Phe Gly Arg Lys Asp Val Val Glu Tyr Leu Leu Gln Asn
165 170 175ggt gca aat gtc caa gca cgt gat gat ggg ggc ctt att cct ctt cat 577Gly Ala Asn Val Gln Ala Arg Asp Asp Gly Gly Leu Ile Pro Leu His
180 185 190aat gca tgc tct ttt ggt cat gct gaa gta gtc aat ctc ctt ttg cga 625Asn Ala Cys Ser Phe Gly His Ala Glu Val Val Asn Leu Leu Leu Arg
195 200 205cat ggt gca gac ccc aat gct c 647His Gly Ala Asp Pro Asn Ala
210 215<210>105<211>215<212>PRT<213>人(Homo sapiens)<400>105Glu Leu Ala Gly Gly Ala Leu Pro Ala Ser Ala Ala Ala Ser Phe Gln1 5 10 15Asp Pro Asp Gly Gly Phe Ala Leu Pro Pro Pro Pro Arg Gly Ser Arg
20 25 30Gly Ala Gly Ser Pro Ala Arg Gly Ala Arg Gly Arg Gly His Gly Thr
35 40 45Ala Pro Asp Pro Val Thr Ala Gly Ser Gln Ala Ala Arg Ala Leu Ser
50 55 60Ala Ser Ser Pro Gly Gly Leu Ala Leu Leu Leu Ala Gly Pro Gly Leu65 70 75 80Leu Leu Arg Leu Leu Ala Leu Leu Leu Ala Val Ala Ala Ala Arg Ile
85 90 95Met Ser Gly Arg Arg Cys Ala Gly Gly Gly Ala Ala Cys Ala Ser Ala
100 105 110Ala Ala Glu Ala Val Glu Pro Ala Ala Arg Glu Leu Phe Glu Ala Cys
115 120 125Arg Asn Gly Asp Val Glu Arg Val Lys Arg Leu Val Thr Pro Glu Lys
130 135 140Val Asn Ser Arg Asp Thr Ala Gly Arg Lys Ser Thr Pro Leu His Phe145 150 155 160Ala Ala Gly Phe Gly Arg Lys Asp Val Val Glu Tyr Leu Leu Gln Asn
165 170 175Gly Ala Asn Val Gln Ala Arg Asp Asp Gly Gly Leu Ile Pro Leu His
180 185 190Asn Ala Cys Ser Phe Gly His Ala Glu Val Val Asn Leu Leu Leu Arg
195 200 205His Gly Ala Asp Pro Asn Ala
210 215<210>106<211>4406<212>DNA<213>人(Homo sapiens)<220><221>CDS<222>(2)..(3787)<400>106g gag ctg gca gga ggg gcc ttg cca gct tcc gcc gcc gcg tcg ttt cag 49Glu Leu Ala Gly Gly Ala Leu Pro Ala Ser Ala Ala Ala Ser Phe Gln
1 5 10 15gac ccg gac ggc gga ttc gcg ctg cct ccg ccg ccg cgg ggc agc cgg 97Asp Pro Asp Gly Gly Phe Ala Leu Pro Pro Pro Pro Arg Gly Ser Arg
20 25 30ggg gca ggg agc cca gcg agg ggc gcg cgt ggg cgc ggc cat ggg act 145Gly Ala Gly Ser Pro Ala Arg Gly Ala Arg Gly Arg Gly His Gly Thr
35 40 45gcg ccg gat ccg gtg aca gca ggg agc caa gcg gcc cgg gcc ctg agc 193Ala Pro Asp Pro Val Thr Ala Gly Ser Gln Ala Ala Arg Ala Leu Ser
50 55 60gcg tct tct ccg ggg ggc ctc gcc ctc ctg ctc gcg ggg ccg ggg ctc 241Ala Ser Ser Pro Gly Gly Leu Ala Leu Leu Leu Ala Gly Pro Gly Leu65 70 75 80ctg ctc cgg ttg ctg gcg ctg ttg ctg gct gtg gcg gcg gcc agg atc 289Leu Leu Arg Leu Leu Ala Leu Leu Leu Ala Val Ala Ala Ala Arg Ile
85 90 95atg tcg ggt cgc cgc tgc gcc ggc ggg gga gcg gcc tgc gcg agc gcc 337Met Ser Gly Arg Arg Cys Ala Gly Gly Gly Ala Ala Cys Ala Ser Ala
100 105 110gcg gcc gag gcc gtg gag ccg gcc gcc cga gag ctg ttc gag gcg tgc 385Ala Ala Glu Ala Val Glu Pro Ala Ala Arg Glu Leu Phe Glu Ala Cys
115 120 125cgc aac ggg gac gtg gaa cga gtc aag agg ctg gtg acg cct gag aag 433Arg Asn Gly Asp Val Glu Arg Val Lys Arg Leu Val Thr Pro Glu Lys
130 135 140gtg aac agc cgc gac acg gcg ggc agg aaa tcc acc ccg ctg cac ttc 481Val Asn Ser Arg Asp Thr Ala Gly Arg Lys Ser Thr Pro Leu His Phe145 150 155 160gcc gca ggt ttt ggg cgg aaa gac gta gtt gaa tat ttg ctt cag aat 529Ala Ala Gly Phe Gly Arg Lys Asp Val Val Glu Tyr Leu Leu Gln Asn
165 170 175ggt gca aat gtc caa gca cgt gat gat ggg ggc ctt att cct ctt cat 577Gly Ala Asn Val Gln Ala Arg Asp Asp Gly Gly Leu Ile Pro Leu His
180 185 190aat gca tgc tct ttt ggt cat gct gaa gta gtc aat ctc ctt ttg cga 625Asn Ala Cys Ser Phe Gly His Ala Glu Val Val Asn Leu Leu Leu Arg
195 200 205cat ggt gca gac ccc aat gct cga gat aat tgg aat tat act cct ctc 673His Gly Ala Asp Pro Asn Ala Arg Asp Asn Trp Asn Tyr Thr Pro Leu
210 215 220cat gaa gct gca att aaa gga aag att gat gtt tgc att gtg ctg tta 721His Glu Ala Ala Ile Lys Gly Lys Ile Asp Val Cys Ile Val Leu Leu225 230 235 240cag cat gga gct gag cca acc atc cga aat aca gat gga agg aca gca 769Gln His Gly Ala Glu Pro Thr Ile Arg Asn Thr Asp Gly Arg Thr Ala
245 250 255ttg gat tta gca gat cca tct gcc aaa gca gtg ctt act ggt gaa tat 817Leu Asp Leu Ala Asp Pro Ser Ala Lys Ala Val Leu Thr Gly Glu Tyr
260 265 270aag aaa gat gaa ctc tta gaa agt gcc agg agt ggc aat gaa gaa aaa 865Lys Lys Asp Glu Leu Leu Glu Ser Ala Arg Ser Gly Asn Glu Glu Lys
275 280 285atg atg gct cta ctc aca cca tta aat gtc aac tgc cac gca agt gat 913Met Met Ala Leu Leu Thr Pro Leu Asn Val Asn Cys His Ala Ser Asp
290 295 300ggc aga aag tca act cca tta cat ttg gca gca gga tat aac aga gta 961Gly Arg Lys Ser Thr Pro Leu His Leu Ala Ala Gly Tyr Asn Arg Val305 310 315 320aag att gta cag ctg tta ctg caa cat gga get gat gtc cat gct aaa 1009Lys Ile Val Gln Leu Leu Leu Gln His Gly Ala Asp Val His Ala Lys
325 330 335gat aaa ggt gat ctg gta cca tta cac aat gcc tgt tct tat ggt cat 1057Asp Lys Gly Asp Leu Val Pro Leu His Asn Ala Cys Ser Tyr Gly His
340 345 350tat gaa gta act gaa ctt ttg gtc aag cat ggt gcc tgt gta aat gca 1105Tyr Glu Val Thr Glu Leu Leu Val Lys His Gly Ala Cys Val Asn Ala
355 360 365atg gac ttg tgg caa ttc act cct ctt cat gag gca gct tct aag aac 1153Met Asp Leu Trp Gln Phe Thr Pro Leu His Glu Ala Ala Ser Lys Asn
370 375 380agg gtt gaa gta tgt tct ctt ctc tta agt tat ggt gca gac cca aca 1201Arg Val Glu Val Cys Ser Leu Leu Leu Ser Tyr Gly Ala Asp Pro Thr385 390 395 400ctg ctc aat tgt cac aat aaa agt gct ata gac ttg gct ccc aca cca 1249Leu Leu Asn Cys His Asn Lys Ser Ala Ile Asp Leu Ala Pro Thr Pro
405 410 415cag tta aaa gaa aga tta gca tat gaa ttt aaa ggc cac tcg ttg ctg 1297Gln Leu Lys Glu Arg Leu Ala Tyr Glu Phe Lys Gly His Ser Leu Leu
420 425 430caa gct gca cga gaa gct gat gtt act cga atc aaa aaa cat ctc tct 1345Gln Ala Ala Arg Glu Ala Asp Val Thr Arg Ile Lys Lys His Leu Ser
435 440 445ctg gaa atg gtg aat ttc aag cat cct caa aca cat gaa aca gca ttg 1393Leu Glu Met Val Asn Phe Lys His Pro Gln Thr His Glu Thr Ala Leu
450 455 460cat tgt gct gct gca tct cca tat ccc aaa aga aag caa ata tgt gaa 1441His Cys Ala Ala Ala Ser Pro Tyr Pro Lys Arg Lys Gln Ile Cys Glu465 470 475 480ctg ttg cta aga aaa gga gca aac atc aat gaa aag act aaa gaa ttc 1489Leu Leu Leu Arg Lys Gly Ala Asn Ile Asn Glu Lys Thr Lys Glu Phe
485 490 495ttg act cct ctg cac gtg gca tct gag aaa gct cat aat gat gtt gtt 1537Leu Thr Pro Leu His Val Ala Ser Glu Lys Ala His Asn Asp Val Val
500 505 510gaa gta gtg gtg aaa cat gaa gca aag gtt aat gct ctg gat aat ctt 1585Glu Val Val Val Lys His Glu Ala Lys Val Asn Ala Leu Asp Asn Leu
515 520 525ggt cag act tct cta cac aga gct gca tat tgt ggt cat cta caa acc 1633Gly Gln Thr Ser Leu His Arg Ala Ala Tyr Cys Gly His Leu Gln Thr
530 535 540tgc cgc cta ctc ctg agc tat ggg tgt gat cct aac att ata tcc ctt 1681Cys Arg Leu Leu Leu Ser Tyr Gly Cys Asp Pro Asn Ile Ile Ser Leu545 550 555 560cag ggc ttt act gct tta cag atg gga aat gaa aat gta cag caa ctc 1729Gln Gly Phe Thr Ala Leu Gln Met Gly Asn Glu Asn Val Gln Gln Leu
565 570 575ctc caa gag ggt atc tca tta ggt aat tca gag gca gac aga caa ttg 1777Leu Gln Glu Gly Ile Ser Leu Gly Asn Ser Glu Ala Asp Arg Gln Leu
580 585 590ctg gaa gct gca aag gct gga gat gtc gaa act gta aaa aaa ctg tgt 1825Leu Glu Ala Ala Lys Ala Gly Asp Val Glu Thr Val Lys Lys Leu Cys
595 600 605act gtt cag agt gtc aac tgc aga gac att gaa ggg cgt cag tct aca 1873Thr Val Gln Set Val Asn Cys Arg Asp Ile Glu Gly Arg Gln Ser Thr
610 615 620cca ctt cat ttt gca gct ggg tat aac aga gtg tcc gtg gtg gaa tat 1921Pro Leu His Phe Ala Ala Gly Tyr Asn Arg Val Ser Val Val Glu Tyr625 630 635 640ctg cta cag cat gga gct gat gtg cat gct aaa gat aaa gga ggc ctt 1969Leu Leu Gln His Gly Ala Asp Val His Ala Lys Asp Lys Gly Gly Leu
645 650 655gta cct ttg cac aat gca tgt tct tat gga cat tat gaa gtt gca gaa 2017Val Pro Leu His Asn Ala Cys Ser Tyr Gly His Tyr Glu Val Ala Glu
660 665 670ctt ctt gtt aaa cat gga gca gta gtt aat gta gct gat tta tgg aaa 2065Leu Leu Val Lys His Gly Ala Val Val Asn Val Ala Asp Leu Trp Lys
675 680 685ttt aca cct tta cat gaa gca gca gca aaa gga aaa tat gaa att tgc 2113Phe Thr Pro Leu His Glu Ala Ala Ala Lys Gly Lys Tyr Glu Ile Cys
690 695 700aaa ctt ctg ctc cag cat ggt gca gac cct aca aaa aaa aac agg gat 2161Lys Leu Leu Leu Gln His Gly Ala Asp Pro Thr Lys Lys Asn Arg Asp705 710 715 720gga aat act cct ttg gat ctt gtt aaa gat gga gat aca gat att caa 2209Gly Asn Thr Pro Leu Asp Leu Val Lys Asp Gly Asp Thr Asp Ile Gln
725 730 735gat ctg ctt agg gga gat gca gct ttg cta gat gct gcc aag aag ggt 2257Asp Leu Leu Arg Gly Asp Ala Ala Leu Leu Asp Ala Ala Lys Lys Gly
740 745 750tgt tta gcc aga gtg aag aag ttg tct tct cct gat aat gta aat tgc 2305Cys Leu Ala Arg Val Lys Lys Leu Ser Ser Pro Asp Asn Val Asn Cys
755 760 765cgc gat acc caa ggc aga cat tca aca cct tta cat tta gca gct ggt 2353Arg Asp Thr Gln Gly Arg His Ser Thr Pro Leu His Leu Ala Ala Gly
770 775 780tat aat aat tta gaa gtt gca gag tat ttg tta caa cac gga gct gat 2401Tyr Asn Asn Leu Glu Val Ala Glu Tyr Leu Leu Gln His Gly Ala Asp785 790 795 800gtg aat gcc caa gac aaa gga gga ctt att cct tta cat aat gca gca 2449Val Asn Ala Gln Asp Lys Gly Gly Leu Ile Pro Leu His Asn Ala Ala
805 810 815tct tac ggg cat gta gat gta gca gct cta cta ata aag tat aat gca 2497Ser Tyr Gly His Val Asp Val Ala Ala Leu Leu Ile Lys Tyr Asn Ala
820 825 830tgt gtc aat gcc acg gac aaa tgg gct ttc aca cct ttg cac gaa gca 2545Cys Val Asn Ala Thr Asp Lys Trp Ala Phe Thr Pro Leu His Glu Ala
835 840 845gcc caa aag gga cga aca cag ctt tgt gct ttg ttg cta gcc cat gga 2593Ala Gln Lys Gly Arg Thr Gln Leu Cys Ala Leu Leu Leu Ala His Gly
850 855 860gct gac ccg act ctt aaa aat cag gaa gga caa aca cct tta gat tta 2641Ala Asp Pro Thr Leu Lys Asn Gln Glu Gly Gln Thr Pro Leu Asp Leu865 870 875 880gtt tca gca gat gat gtc agc gct ctt ctg aca gca gcc atg ccc cca 2689Val Ser Ala Asp Asp Val Ser Ala Leu Leu Thr Ala Ala Met Pro Pro
885 890 895tct gct ctg ccc tct tgt tac aag cct caa gtg ctc aat ggt gtg aga 2737Ser Ala Leu Pro Ser Cys Tyr Lys Pro Gln Val Leu Asn Gly Val Arg
900 905 910agc cca gga gcc act gca gat gct ctc tct tca ggt cca tct agc cca 2785Ser Pro Gly Ala Thr Ala Asp Ala Leu Ser Ser Gly Pro Ser Ser Pro
915 920 925tca agc ctt tct gca gcc agc agt ctt gac aac tta tct ggg agt ttt 2833Ser Ser Leu Ser Ala Ala Ser Ser Leu Asp Asn Leu Ser Gly Ser Phe
930 935 940tca gaa ctg tct tca gta gtt agt tca agt gga aca gag ggt gct tcc 2881Ser Glu Leu Ser Ser Val Val Ser Ser Ser Gly Thr Glu Gly Ala Ser945 950 955 960agt ttg gag aaa aag gag gtt cca gga gta gat ttt agc ata act caa 2929Ser Leu Glu Lys Lys Glu Val Pro Gly Val Asp Phe Ser Ile Thr Gln
965 970 975ttc gta agg aat ctt gga ctt gag cac cta atg gat ata ttt gag aga 2977Phe Val Arg Asn Leu Gly Leu Glu His Leu Met Asp Ile Phe Glu Arg
980 985 990gaa cag atc act ttg gat gta tta gtt gag atg ggg cac aag gag ctg 3025Glu Gln Ile Thr Leu Asp Val Leu Val Glu Met Gly His Lys Glu Leu
995 1000 1005aag gag att gga atc aat gct tat gga cat agg cac aaa cta att aaa 3073Lys Glu Ile Gly Ile Asn Ala Tyr Gly His Arg His Lys Leu Ile Lys1010 1015 1020gga gtc gag aga ctt atc tcc gga caa caa ggt ctt aac cca tat tta 3121Gly Val Glu Arg Leu Ile Ser Gly Gln Gln Gly Leu Asn Pro Tyr Leu1025 1030 1035 1040act ttg aac acc tct ggt agt gga aca att ctt ata gat ctg tct cct 3169Thr Leu Asn Thr Ser Gly Ser Gly Thr Ile Leu Ile Asp Leu Ser Pro
1045 1050 1055gat gat aaa gag ttt cag tct gtg gag gaa gag atg caa agt aca gtt 3217Asp Asp Lys Glu Phe Gln Ser Val Glu Glu Glu Met Gln Ser Thr Val
1060 1065 1070cga gag cac aga gat gga ggt cat gca ggt gga atc ttc aac aga tac 3265Arg Glu His Arg Asp Gly Gly His Ala Gly Gly Ile Phe Asn Arg Tyr
1075 1080 1085aat att ctc aag att cag aag gtt tgt aac aag aaa cta tgg gaa aga 3313Asn Ile Leu Lys Ile Gln Lys Val Cys Asn Lys Lys Leu Trp Glu Arg1090 1095 1100tac act cac cgg aga aaa gaa gtt tct gaa gaa aac cac aac cat gcc 3361Tyr Thr His Arg Arg Lys Glu Val Ser Glu Glu Asn His Asn His Ala1105 1110 1115 1120aat gaa cga atg cta ttt cat ggg tct cct ttt gtg aat gca att atc 3409Asn Glu Arg Met Leu Phe His Gly Ser Pro Phe Val Asn Ala Ile Ile
1125 1130 1135cac aaa ggc ttt gat gaa agg cat gcg tac ata ggt ggt atg ttt gga 3457His Lys Gly Phe Asp Glu Arg His Ala Tyr Ile Gly Gly Met Phe Gly
1140 1145 1150gct ggc att tat ttt gct gaa aac tct tcc aaa agc aat caa tat gta 3505Ala Gly Ile Tyr Phe Ala Glu Asn Ser Ser Lys Ser Asn Gln Tyr Val
1155 1160 1165tat gga att gga gga ggt act ggg tgt cca gtt cac aaa gac aga tct 3553Tyr Gly Ile Gly Gly Gly Thr Gly Cys Pro Val His Lys Asp Arg Ser1170 1175 1180tgt tac att tgc cac agg cag ctg ctc ttt tgc cgg gta acc ttg gga 3601Cys Tyr Ile Cys His Arg Gln Leu Leu Phe Cys Arg Val Thr Leu Gly1185 1190 1195 1200aag tct ttc ctg cag ttc agt gca atg aaa atg gca cat tct cct cca 3649Lys Ser Phe Leu Gln Phe Ser Ala Met Lys Met Ala His Ser Pro Pro
1205 1210 1215ggt cat cac tca gtc act ggt agg ccc agt gta aat ggc cta gca tta 3697Gly His His Ser Val Thr Gly Arg Pro Ser Val Asn Gly Leu Ala Leu
1220 1225 1230gct gaa tat gtt att tac aga gga gaa cag gct tat cct gag tat tta 3745Ala Glu Tyr Val Ile Tyr Arg Gly Glu Gln Ala Tyr Pro Glu Tyr Leu
1235 1240 1245att act tac cag att atg agg cct gaa ggt atg gtc gat gga 3787Ile Thr Tyr Gln Ile Met Arg Pro Glu Gly Met Val Asp Gly1250 1255 1260taaatagtta ttttaagaaa ctaattccac tgaacctaaa atcatcaaag cagcagtggc 3847ctctacgttt tactcctttg ctgaaaaaaa atcatcttgc ccacaggcct gtggcaaaag 3907gataaaaatg tgaacgaagt ttaacattct gacttgataa agctttaata atgtacagtg 3967ttttctaaat atttcctgtt ttttcagcac tttaacagat gccattccag gttaaactgg 4027gttgtctgta ctaaattata aacagagtta acttgaacct tttatatgtt atgcattgat 4087tctaacaaac tgtaatgccc tcaacagaac taattttact aatacaatac tgtgttcttt 4147aaaacacagc atttacactg aatacaattt catttgtaaa actgtaaata agagcttttg 4207tactagccca gtatttattt acattgcttt gtaatataaa tctgttttag aactgcagcg 4267gtttacaaaa ttttttcata tgtattgttc atctatactt catcttacat cgtcatgatt 4327gagtgatctt tacatttgat tccagaggct atgttcagtt gttagttggg aaagattgag 4387ttatcagatt taatttgcc 4406<210>107<211>1262<212>PRT<213>人(Homo sapiens)<400>107Glu Leu Ala Gly Gly Ala Leu Pro Ala Ser Ala Ala Ala Ser Phe Gln1 5 10 15Asp Pro Asp Gly Gly Phe Ala Leu Pro Pro Pro Pro Arg Gly Ser Arg
20 25 30Gly Ala Gly Ser Pro Ala Arg Gly Ala Arg Gly Arg Gly His Gly Thr
35 40 45Ala Pro Asp Pro Val Thr Ala Gly Ser Gln Ala Ala Arg Ala Leu Ser
50 55 60Ala Ser Ser Pro Gly Gly Leu Ala Leu Leu Leu Ala Gly Pro Gly Leu65 70 75 80Leu Leu Arg Leu Leu Ala Leu Leu Leu Ala Val Ala Ala Ala Arg Ile
85 90 95Met Ser Gly Arg Arg Cys Ala Gly Gly Gly Ala Ala Cys Ala Ser Ala
100 105 110Ala Ala Glu Ala Val Glu Pro Ala Ala Arg Glu Leu Phe Glu Ala Cys
115 120 125Arg Asn Gly Asp Val Glu Arg Val Lys Arg Leu Val Thr Pro Glu Lys
130 135 140Val Asn Ser Arg Asp Thr Ala Gly Arg Lys Ser Thr Pro Leu His Phe145 150 155 160Ala Ala Gly Phe Gly Arg Lys Asp Val Val Glu Tyr Leu Leu Gln Asn
165 170 175Gly Ala Asn Val Gln Ala Arg Asp Asp Gly Gly Leu Ile Pro Leu His
180 185 190Asn Ala Cys Ser Phe Gly His Ala Glu Val Val Asn Leu Leu Leu Arg
195 200 205His Gly Ala Asp Pro Asn Ala Arg Asp Asn Trp Asn Tyr Thr Pro Leu
210 215 220His Glu Ala Ala Ile Lys Gly Lys Ile Asp Val Cys Ile Val Leu Leu225 230 235 240Gln His Gly Ala Glu Pro Thr Ile Arg Asn Thr Asp Gly Arg Thr Ala
245 250 255Leu Asp Leu Ala Asp Pro Ser Ala Lys Ala Val Leu Thr Gly Glu Tyr
260 265 270Lys Lys Asp Glu Leu Leu Glu Ser Ala Arg Ser Gly Asn Glu Glu Lys
275 280 285Met Met Ala Leu Leu Thr Pro Leu Asn Val Asn Cys His Ala Ser Asp
290 295 300Gly Arg Lys Ser Thr Pro Leu His Leu Ala Ala Gly Tyr Asn Arg Val305 310 315 320Lys Ile Val Gln Leu Leu Leu Gln His Gly Ala Asp Val His Ala Lys
325 330 335Asp Lys Gly Asp Leu Val Pro Leu His Asn Ala Cys Ser Tyr Gly His
340 345 350Tyr Glu Val Thr Glu Leu Leu Val Lys His Gly Ala Cys Val Asn Ala
355 360 365Met Asp Leu Trp Gln Phe Thr Pro Leu His Glu Ala Ala Ser Lys Asn
370 375 380Arg Val Glu Val Cys Ser Leu Leu Leu Ser Tyr Gly Ala Asp Pro Thr385 390 395 400Leu Leu Asn Cys His Asn Lys Ser Ala Ile Asp Leu Ala Pro Thr Pro
405 410 415Gln Leu Lys Glu Arg Leu Ala Tyr Glu Phe Lys Gly His Ser Leu Leu
420 425 430Gln Ala Ala Arg Glu Ala Asp Val Thr Arg Ile Lys Lys His Leu Ser
435 440 445Leu Glu Mer Val Asn Phe Lys His Pro Gln Thr His Glu Thr Ala Leu
450 455 460His Cys Ala Ala Ala Ser Pro Tyr Pro Lys Arg Lys Gln Ile Cys Glu465 470 475 480Leu Leu Leu Arg Lys Gly Ala Asn Ile Asn Glu Lys Thr Lys Glu Phe
485 490 495Leu Thr Pro Leu His Val Ala Ser Glu Lys Ala His Asn Asp Val Val
500 505 510Glu Val Val Val Lys His Glu Ala Lys Val Asn Ala Leu Asp Asn Leu
515 520 525Gly Gln Thr Ser Leu His Arg Ala Ala Tyr Cys Gly His Leu Gln Thr
530 535 540Cys Arg Leu Leu Leu Ser Tyr Gly Cys Asp Pro Asn Ile Ile Ser Leu545 550 555 560Gln Gly Phe Thr Ala Leu Gln Met Gly Asn Glu Asn Val Gln Gln Leu
565 570 575Leu Gln Glu Gly Ile Ser Leu Gly Asn Ser Glu Ala Asp Arg Gln Leu
580 585 590Leu Glu Ala Ala Lys Ala Gly Asp Val Glu Thr Val Lys Lys Leu Cys
595 600 605Thr Val Gln Ser Val Asn Cys Arg Asp Ile Glu Gly Arg Gln Ser Thr
610 615 620Pro Leu His Phe Ala Ala Gly Tyr Asn Arg Val Ser Val Val Glu Tyr625 630 635 640Leu Leu Gln His Gly Ala Asp Val His Ala Lys Asp Lys Gly Gly Leu
645 650 655Val Pro Leu His Asn Ala Cys Ser Tyr Gly His Tyr Glu Val Ala Glu
660 665 670Leu Leu Val Lys His Gly Ala Val Val Asn Val Ala Asp Leu Trp Lys
675 680 685Phe Thr Pro Leu His Glu Ala Ala Ala Lys Gly Lys Tyr Glu Ile Cys
690 695 700Lys Leu Leu Leu Gln His Gly Ala Asp Pro Thr Lys Lys Asn Arg Asp705 710 715 720Gly Asn Thr Pro Leu Asp Leu Val Lys Asp Gly Asp Thr Asp Ile Gln
725 730 735Asp Leu Leu Arg Gly Asp Ala Ala Leu Leu Asp Ala Ala Lys Lys Gly
740 745 750Cys Leu Ala Arg Val Lys Lys Leu Ser Ser Pro Asp Asn Val Asn Cys
755 760 765Arg Asp Thr Gln Gly Arg His Ser Thr Pro Leu His Leu Ala Ala Gly
770 775 780Tyr Asn Asn Leu Glu Val Ala Glu Tyr Leu Leu Gln His Gly Ala Asp785 790 795 800Val Asn Ala Gln Asp Lys Gly Gly Leu Ile Pro Leu His Asn Ala Ala
805 810 815Ser Tyr Gly His Val Asp Val Ala Ala Leu Leu Ile Lys Tyr Asn Ala
820 825 830Cys Val Asn Ala Thr Asp Lys Trp Ala Phe Thr Pro Leu His Glu Ala
835 840 845Ala Gln Lys Gly Arg Thr Gln Leu Cys Ala Leu Leu Leu Ala His Gly
850 855 860Ala Asp Pro Thr Leu Lys Asn Gln Glu Gly Gln Thr Pro Leu Asp Leu865 870 875 880Val Ser Ala Asp Asp Val Ser Ala Leu Leu Thr Ala Ala Met Pro Pro
885 890 895Ser Ala Leu Pro Ser Cys Tyr Lys Pro Gln Val Leu Asn Gly Val Arg
900 905 910Ser Pro Gly Ala Thr Ala Asp Ala Leu Ser Ser Gly Pro Ser Ser Pro
915 920 925Ser Ser Leu Ser Ala Ala Ser Ser Leu Asp Asn Leu Ser Gly Ser Phe
930 935 940Ser Glu Leu Ser Ser Val Val Ser Ser Ser Gly Thr Glu Gly Ala Ser945 950 955 960Ser Leu Glu Lys Lys Glu Val Pro Gly Val Asp Phe Ser Ile Thr Gln
965 970 975Phe Val Arg Asn Leu Gly Leu Glu His Leu Met Asp Ile Phe Glu Arg
980 985 990Glu Gln Ile Thr Leu Asp Val Leu Val Glu Met Gly His Lys Glu Leu
995 1000 1005Lys Glu Ile Gly Ile Asn Ala Tyr Gly His Arg His Lys Leu Ile Lys1010 1015 1020Gly Val Glu Arg Leu Ile Ser Gly Gln Gln Gly Leu Asn Pro Tyr Leu1025 1030 1035 1040Thr Leu Asn Thr Ser Gly Ser Gly Thr Ile Leu Ile Asp Leu Ser Pro
1045 1050 1055Asp Asp Lys Glu Phe Gln Ser Val Glu Glu Glu Met Gln Ser Thr Val
1060 1065 1070Arg Glu His Arg Asp Gly Gly His Ala Gly Gly Ile Phe Asn Arg Tyr
1075 1080 1085Asn Ile Leu Lys Ile Gln Lys Val Cys Asn Lys Lys Leu Trp Glu Arg1090 1095 1100Tyr Thr His Arg Arg Lys Glu Val Ser Glu Glu Asn His Asn His Ala1105 1110 1115 1120Asn Glu Arg Met Leu Phe His Gly Ser Pro Phe Val Asn Ala Ile Ile
1125 1130 1135His Lys Gly Phe Asp Glu Arg His Ala Tyr Ile Gly Gly Met Phe Gly
1140 1145 1150Ala Gly Ile Tyr Phe Ala Glu Asn Ser Ser Lys Ser Asn Gln Tyr Val
1155 1160 1165Tyr Gly Ile Gly Gly Gly Thr Gly Cys Pro Val His Lys Asp Arg Ser1170 1175 1180Cys Tyr Ile Cys His Arg Gln Leu Leu Phe Cys Arg Val Thr Leu Gly1185 1190 1195 1200Lys Ser Phe Leu Gln Phe Ser Ala Met Lys Met Ala His Ser Pro Pro
1205 1210 1215Gly His His Ser Val Thr Gly Arg Pro Ser Val Asn Gly Leu Ala Leu
1220 1225 1230Ala Glu Tyr Val Ile Tyr Arg Gly Glu Gln Ala Tyr Pro Glu Tyr Leu
1235 1240 1245Ile Thr Tyr Gln Ile Met Arg Pro Glu Gly Met Val Asp Gly1250 1255 1260<210>108<211>436<212>DNA<213>人(Homo sapiens)<400>108ttttttttgc agttctaaaa cagatttata ttacaaagca atgtaaataa atactgggct 60agtacaaaag ctcttattta cagttttaca aatgaaattg tattcagtgt aaatgctgtg 120ttttaaagaa cacagtattg tattagtaaa attagttctg ttgagggcat tacagtttgt 180tagaatcaat gcataacata taaaaggttc aagttaactc tgtttataat ttagtacaga 240caacccagtt taacctggga tgggcatctg ttaaagtgct ggaaaaaaca gggaaatatt 300taggaaaaca ctggtacatt atttaaaggc tttntccaag gtcaggantg tttaaacttc 360gtttcacatt tttatccntt tggccacggc ctgtggggcn aggatggatt ttttttccgg 420ccaagggtgt taaacg 436<210>109<211>21<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>109cgcctgagaa ggtgaacagc c 21<210>110<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>110acgcctcgaa cagctctcgg 20<210>111<211>23<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>111gcgtgggcgc ggccatggga ctg 23<210>112<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>112cagcgcgaat ccgccgtccg 20<210>113<211>620<212>DNA<213>人(Homo sapiens)<220><221>CDS<222>(3)..(620)<400>113tt aaa aca aca aca aca aaa aac aca ata tgc agg atc gtt cgg ctt 47Lys Thr Thr Thr Thr Lys Asn Thr Ile Cys Arg Ile Val Arg Leu
l 5 10 15cag cag aac cca ccg caa aga tgg cgg tgg gac gaa gcc cct tct ccc 95Gln Gln Asn Pro Pro Gln Arg Trp Arg Trp Asp Glu Ala Pro Ser Pro
20 25 30gcc gcc gaa gcc tct cgc ctc aca ttt ccc aca aac cct tcg cgc cgc 143Ala Ala Glu Ala Ser Arg Leu Thr Phe Pro Thr Asn Pro Ser Arg Arg
35 40 45ctc gct agc cga aac ctg ccc agc cgg tgc ccg gcc act gcg cac gcg 191Leu Ala Ser Arg Asn Leu Pro Ser Arg Cys Pro Ala Thr Ala His Ala
50 55 60cgg gac gac gtc acg tgc gct ccc ggg gct gga cgg agc tgg cag gag 239Arg Asp Asp Val Thr Cys Ala Pro Gly Ala Gly Arg Ser Trp Gln Glu
65 70 75ctg gca gga ggg gcc ttg cca gct tcc gcc gcc gcg tcg ttt cag gac 287Leu Ala Gly Gly Ala Leu Pro Ala Ser Ala Ala Ala Ser Phe Gln Asp80 85 90 95ccg gac ggc gga ttc gcg ctg cct ccg ccg ccg cgg ggc agc cgg ggg 335Pro Asp Gly Gly Phe Ala Leu Pro Pro Pro Pro Arg Gly Ser Arg Gly
100 105 110gca ggg agc cca gcg agg ggc gcg cgt ggg cgc ggc cat ggg act gcg 383Ala Gly Ser Pro Ala Arg Gly Ala Arg Gly Arg Gly His Gly Thr Ala
115 120 125ccg gat ccg gtg aca gca ggg agc caa gcg gcc cgg gcc ctg agc gcg 431Pro Asp Pro Val Thr Ala Gly Ser Gln Ala Ala Arg Ala Leu Ser Ala
130 135 140tct tct ccg ggg ggc ctc gcc ctc ctg ctc gcg ggg ccg ggg ctc ctg 479Ser Ser Pro Gly Gly Leu Ala Leu Leu Leu Ala Gly Pro Gly Leu Leu
145 150 155ctc cgg ttg ctg gcg ctg ttg ctg gct gtg gcg gcg gcc agg atc atg 527Leu Arg Leu Leu Ala Leu Leu Leu Ala Val Ala Ala Ala Arg Ile Met160 165 170 175tcg ggt cgc cgc tgc gcc ggc ggg gga gcg gcc tgc gcg agc gcc gcg 575Ser Gly Arg Arg Cys Ala Gly Gly Gly Ala Ala Cys Ala Ser Ala Ala
180 185 190gcc gag gcc gtg gag ccg gcc gcc cga gag ctg ttc gag gcg tgc 620Ala Glu Ala Val Glu Pro Ala Ala Arg Glu Leu Phe Glu Ala Cys
195 200 205<210>114<211>206<212>PRT<213>人(Homo sapiens)<400>114Lys Thr Thr Thr Thr Lys Asn Thr Ile Cys Arg Ile Val Arg Leu Gln1 5 10 15Gln Asn Pro Pro Gln Arg Trp Arg Trp Asp Glu Ala Pro Ser Pro Ala
20 25 30Ala Glu Ala Ser Arg Leu Thr Phe Pro Thr Asn Pro Ser Arg Arg Leu
35 40 45Ala Ser Arg Asn Leu Pro Ser Arg Cys Pro Ala Thr Ala His Ala Arg
50 55 60Asp Asp Val Thr Cys Ala Pro Gly Ala Gly Arg Ser Trp Gln Glu Leu65 70 75 80Ala Gly Gly Ala Leu Pro Ala Ser Ala Ala Ala Ser Phe Gln Asp Pro
85 90 95Asp Gly Gly Phe Ala Leu Pro Pro Pro Pro Arg Gly Ser Arg Gly Ala
100 105 110Gly Ser Pro Ala Arg Gly Ala Arg Gly Arg Gly His Gly Thr Ala Pro
115 120 125Asp Pro Val Thr Ala Gly Ser Gln Ala Ala Arg Ala Leu Ser Ala Ser
130 135 140Ser Pro Gly Gly Leu Ala Leu Leu Leu Ala Gly Pro Gly Leu Leu Leu145 150 155 160Arg Leu Leu Ala Leu Leu Leu Ala Val Ala Ala Ala Arg Ile Met Ser
165 170 175Gly Arg Arg Cys Ala Gly Gly Gly Ala Ala Cys Ala Ser Ala Ala Ala
180 185 190Glu Ala Val Glu Pro Ala Ala Arg Glu Leu Phe Glu Ala Cys
195 200 205<210>115<211>1039<212>DNA<213>人(Homo sapiens)<220><221>CDS<222>(287)..(1039)<400>115gtacaatatt gatttacaaa aagttcctct aatcaatcct gagctaataa cttactgtgg 60aaagagtaat tgatcagagc catccctcca attggagtca actttcatga ctgttcggat 120ttcctttatt ttgggggcag ttcatccaaa cttctattaa acggcaacta gttcactttt 180gagaagtggt ttacaagaaa caacaacaac aacaacaaag cagttgcgga ggaaagaaaa 240gagacaaagt aaaaaaaacg gaaaagaaat ctcccaggag aaaggg atg tgg aag 295
Met Trp Lys
1ctg aaa aca cgg aca att tcc aca gta aga ctt cca aaa gaa tgt gca 343Leu Lys Thr Arg Thr Ile Ser Thr Val Arg Leu Pro Lys Glu Cys Ala
5 10 15aga tcc gag caa aac ttt caa ggg ctc ttt ttc agt gta atg gta gtg 391Arg Ser Glu Gln Asn Phe Gln Gly Leu Phe Phe Ser Val Met Val Val20 25 30 35aga aag ttc agc ctg gaa agc cca ggg ctt aaa aca aca aca aca aaa 439Arg Lys Phe Ser Leu Glu Ser Pro Gly Leu Lys Thr Thr Thr Thr Lys
40 45 50aac aca ata tgc agg atc gtt cgg ctt cag cag aac cca ccg caa aga 487Asn Thr Ile Cys Arg Ile Val Arg Leu Gln Gln Asn Pro Pro Gln Arg
55 60 65tgg cgg tgg gac gaa gcc cct tct ccc gcc gcc gaa gcc tct cgc ctc 535Trp Arg Trp Asp Glu Ala Pro Ser Pro Ala Ala Glu Ala Ser Arg Leu
70 75 80aca ttt ccc aca aac cct tcg cgc cgc ctc gct agc cga aac ctg ccc 583Thr Phe Pro Thr Asn Pro Ser Arg Arg Leu Ala Ser Arg Asn Leu Pro
85 90 95agc cgg tgc ccg gcc act gcg cac gcg cgg gac gac gtc acg tgc gct 631Ser Arg Cys Pro Ala Thr Ala His Ala Arg Asp Asp Val Thr Cys Ala100 105 110 115ccc ggg gct gga cgg agc tgg cag gag ctg gca gga ggg gcc ttg cca 679Pro Gly Ala Gly Arg Ser Trp Gln Glu Leu Ala Gly Gly Ala Leu Pro
120 125 130gct tcc gcc gcc gcg tcg ttt cag gac ccg gac ggc gga ttc gcg ctg 727Ala Ser Ala Ala Ala Ser Phe Gln Asp Pro Asp Gly Gly Phe Ala Leu
135 140 145cct ccg ccg ccg cgg ggc agc cgg ggg gca ggg agc cca gcg agg ggc 775Pro Pro Pro Pro Arg Gly Ser Arg Gly Ala Gly Ser Pro Ala Arg Gly
150 155 160gcg cgt ggg cgc ggc cat ggg act gcg ccg gat ccg gtg aca gca ggg 823Ala Arg Gly Arg Gly His Gly Thr Ala Pro Asp Pro Val Thr Ala Gly
165 170 175agc caa gcg gcc cgg gcc ctg agc gcg tct tct ccg ggg ggc ctc gcc 871Ser Gln Ala Ala Arg Ala Leu Ser Ala Ser Ser Pro Gly Gly Leu Ala180 185 190 195ctc ctg ctc gcg ggg ccg ggg ctc ctg ctc cgg ttg ctg gcg ctg ttg 919Leu Leu Leu Ala Gly Pro Gly Leu Leu Leu Arg Leu Leu Ala Leu Leu
200 205 210ctg gct gtg gcg gcg gcc agg atc atg tcg ggt cgc cgc tgc gcc ggc 967Leu Ala Val Ala Ala Ala Arg Ile Met Ser Gly Arg Arg Cys Ala Gly
215 220 225ggg gga gcg gcc tgc gcg agc gcc gcg gcc gag gcc gtg gag ccg gcc 1015Gly Gly Ala Ala Cys Ala Ser Ala Ala Ala Glu Ala Val Glu Pro Ala
230 235 240gcc cga gag ctg ttc gag gcg tgc 1039Ala Arg Glu Leu Phe Glu Ala Cys
245 250<210>116<211>251<212>PRT<213>人(Homo sapiens)<400>116Met Trp Lys Leu Lys Thr Arg Thr Ile Ser Thr ValArg Leu Pro Lys1 5 10 15Glu Cys Ala Arg Ser Glu Gln Asn Phe Gln Gly Leu Phe Phe Ser Val
20 25 30Met Val Val Arg Lys Phe Ser Leu Glu Ser Pro Gly Leu Lys Thr Thr
35 40 45Thr Thr Lys Asn Thr Ile Cys Arg Ile Val Arg Leu Gln Gln Asn Pro
50 55 60Pro Gln Arg Trp Arg Trp Asp Glu Ala Pro Ser Pro Ala Ala Glu Ala65 70 75 80Ser Arg Leu Thr Phe Pro Thr Asn Pro Ser Arg Arg Leu Ala Ser Arg
85 90 95Asn Leu Pro Ser Arg Cys Pro Ala Thr Ala His Ala Arg Asp Asp Val
100 105 110Thr Cys Ala Pro Gly Ala Gly Arg Ser Trp Gln Glu Leu Ala Gly Gly
115 120 125Ala Leu Pro Ala Ser Ala Ala Ala Ser Phe Gln Asp Pro Asp Gly Gly
130 135 140Phe Ala Leu Pro Pro Pro Pro Arg Gly Ser Arg Gly Ala Gly Ser Pro145 150 155 160Ala Arg Gly Ala Arg Gly Arg Gly His Gly Thr Ala Pro Asp Pro Val
165 170 175Thr Ala Gly Ser Gln Ala Ala Arg Ala Leu Ser Ala Ser Ser Pro Gly
180 185 190Gly Leu Ala Leu Leu Leu Ala Gly Pro Gly Leu Leu Leu Arg Leu Leu
195 200 205Ala Leu Leu Leu Ala Val Ala Ala Ala Arg Ile Met Ser Gly Arg Arg
210 215 220Cys Ala Gly Gly Gly Ala Ala Cys Ala Ser Ala Ala Ala Glu Ala Val225 230 235 240Glu Pro Ala Ala Arg Glu Leu Phe Glu Ala Cys
245 250<210>117<211>473<212>DNA<213>人(Homo sapiens)<220><221>CDS<222>(3)..(473)<400>117ct agc cga aac ctg ccc agc cgg tgc ccg gcc act gcg cac gcg cgg 47Ser Arg Asn Leu Pro Ser Arg Cys Pro Ala Thr Ala His Ala Arg
1 5 10 15gac gac gtc acg tgc gct ccc ggg gct gga cgg agc tgg cag gag ctg 95Asp Asp Val Thr Cys Ala Pro Gly Ala Gly Arg Ser Trp Gln Glu Leu
20 25 30gca gga ggg gcc ttg cca gct tcc gcc gcc gcg tcg ttt cag gac ccg 143Ala Gly Gly Ala Leu Pro Ala Ser Ala Ala Ala Ser Phe Gln Asp Pro
35 40 45gac ggc gga ttc gcg ctg cct ccg ccg ccg cgg ggc agc cgg ggg gca 191Asp Gly Gly Phe Ala Leu Pro Pro Pro Pro Arg Gly Ser Arg Gly Ala
50 55 60ggg agc cca gcg agg ggc gcg cgt ggg cgc ggc cat ggg act gcg ccg 239Gly Ser Pro Ala Arg Gly Ala Arg Gly Arg Gly His Gly Thr Ala Pro
65 70 75gat ccg gtg aca gca ggg agc caa gcg gcc cgg gcc ctg agc gcg tct 287Asp Pro Val Thr Ala Gly Ser Gln Ala Ala Arg Ala Leu Ser Ala Ser80 85 90 95tct ccg ggg ggc ctc gcc ctc ctg ctc gcg ggg ccg ggg ctc ctg ctc 335Ser Pro Gly Gly Leu Ala Leu Leu Leu Ala Gly Pro Gly Leu Leu Leu
100 105 110cgg ttg ctg gcg ctg ttg ctg gct gtg gcg gcg gcc agg atc atg tcg 383Arg Leu Leu Ala Leu Leu Leu Ala Val Ala Ala Ala Arg Ile Met Ser
115 120 125ggt cgc cgc tgc gcc ggc ggg gga gcg gcc tgc gcg agc gcc gcg gcc 431Gly Arg Arg Cys Ala Gly Gly Gly Ala Ala Cys Ala Ser Ala Ala Ala
130 135 140gag gcc gtg gag ccg gcc gcc cga gag ctg ttc gag gcg tgc 473Glu Ala Val Glu Pro Ala Ala Arg Glu Leu Phe Glu Ala Cys
145 150 155<210>118<211>157<212>PRT<213>人(Homo sapiens)<400>118Ser Arg Asn Leu Pro Ser Arg Cys Pro Ala Thr Ala His Ala Arg Asp1 5 10 15Asp Val Thr Cys Ala Pro Gly Ala Gly Arg Ser Trp Gln Glu Leu Ala
20 25 30Gly Gly Ala Leu Pro Ala Ser Ala Ala Ala Ser Phe Gln Asp Pro Asp
35 40 45Gly Gly Phe Ala Leu Pro Pro Pro Pro Arg Gly Ser Arg Gly Ala Gly
50 55 60Ser Pro Ala Arg Gly Ala Arg Gly Arg Gly His Gly Thr Ala Pro Asp65 70 75 80Pro Val Thr Ala Gly Ser Gln Ala Ala Arg Ala Leu Ser Ala Ser Ser
85 90 95Pro Gly Gly Leu Ala Leu Leu Leu Ala Gly Pro Gly Leu Leu Leu Arg
100 105 110Leu Leu Ala Leu Leu Leu Ala Val Ala Ala Ala Arg Ile Met Ser Gly
115 120 125Arg Arg Cys Ala Gly Gly Gly Ala Ala Cys Ala Ser Ala Ala Ala Glu
130 135 140Ala Val Glu Pro Ala Ala Arg Glu Leu Phe Glu Ala Cys145 150 155<210>119<211>22<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>119gttcctctaa tcaatcctga gc 22<210>120<211>26<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>120ggaaagagta attgatcaga gccatc 26<210>121<211>27<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>121cgccgaagcc tctcgcctca catttcc 27<210>122<21l>27<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>122ggaaatgtga ggcgagaggc ttcggcg 27<210>123<211>659<212>DNA<213>人(Homo sapiens)<400>123ggaaagagta attgatcaga gccatccctc caattggagt caacttccat gactgttcgg 60atttccttta ttttgggggc agttcatcca aacttctatt aaacggcaac tagttcactt 120ttgagaagtg gtttacaaga aacaacaaca acaacaacaa agcagttgcg gaggaaagaa 180aagagacaaa gtaaaaaaaa cggaaaagaa atctcccagg agaaagggat gtggaagctg 240aaaacacgga caatttccac agtaagactt ccaaaagaat gtgcaagatc cgagcaaaac 300tttcaagggc tctttttcag tgtaatggta gtgagaaagt tcagcctgga aagcccaggg 360cttaaaacaa caacaacaaa aaacacaata tgcaggatcg ttcggcttca gcagaaccca 420ccgcaaagat ggcggtggga cgaagcccct tctcccgccg ccgaagcctc tcgcctcaca 480tttcccacaa acccttcgcg ccgcctcgct agccgaaacc tgcccagccg gtgcccggcc 540actgcgcacg cgcgggacga cgtcacgtgc gctcccgggg ctggacggag ctggcaggag 600gggccttgcc agcttccgcc gccgcgtcgt ttcaggaccc ggacggcgga ttcgcgctg 659<210>124<211>669<212>DNA<213>人(Homo sapiens)<400>124ggaaagagta attgatcaga gccatccctc caattggagt caactttcat gactgttcgg 60atttccttta ttttgggggc agttcatcca aacttctatt aaacggcaac tagttcactt 120ttgagaagtg gtttacaaga aacaacaaca acaacaacaa agcagttgcg gaggaaagaa 180aagagacaaa gtaaaaaaaa cggaaaagaa atctcccagg agaaagggat gtggaagctg 240aaaacacgga caatttccac agtaagactt ccaaaagaat gtgcaagatc cgagcaaaac 300tttcaagggc tctttttcag tgtaatggta gtgagaaagt tcagcctgga aagcccaggg 360cttaaaacaa caacaacaaa aaacacaata tgcaggatcg ttcggcttca gcagaaccca 420ccgcaaagat ggcggtggga cgaagcccct tctcccgccg ccgaagcctc tcgcctcaca 480tttcccacaa acccttcgcg ccgcctcgct agccgaaacc tgcccagccg gtgcccggcc 540actgcgcacg cgcgggacga cgtcacgtgc gctcccgggg ctggacggag ctggcaggag 600gggccaggag gggccttgcc agcttccgcc gccgcgtcgt ttcaggaccc ggacggcgga 660ttcgcgctg 669<210>125<211>659<212>DNA<213>人(Homo sapiens)<400>125ggaaagagta attgatcaga gccatccctc caattggagt caacttccat gactgttcgg 60atttccttta ttttgggggc agttcatcca aacttctatt aaacggcaac tagttcactt 120ttgagaagtg gtttacaaga aacaacaaca acaacaacaa agcagttgcg gaggaaagaa 180aagagacaaa gtaaaaaaaa cggaaaagaa atctcccagg agaaagggat gtggaagctg 240aaaacacgga caatttccac agtaagactt ccaaaagaat gtgcaagatc cgagcaaaac 300tttcaagggc tctttttcag tgtaatggta gtgagaaagt tcagcctgga aagcccaggg 360cttaaaacaa caacaacaaa aaacacaata tgcaggatcg ttcggcttca gcagaaccca 420ccgcaaagat ggcggtggga cgaagcccct tctcccgccg ccgaagcctc tcgcctcaca 480tttcccacaa acccttcgcg ccgcctcgct agccgaaacc tgcccagccg gtgcccggcc 540actgcgcacg cgcgggacga cgtcacgtgc gctcccgggg ctggacggag ctggcaggag 600gggccttgcc agcttccgcc gccgcgtcgt ttcaggaccc ggacggcgga ttcgcgctg 659<210>126<211>659<212>DNA<213>人(Homo sapiens)<400>126ggaaagagta attgatcaga gccatccctc caattggagt caacttccat gactgttcgg 60atttccttta ttttgggggc agttcatcca aacttctatt aaacggcaac tagttcactt 120ttgagaagtg gtttacaaga aacaacaaca acaacaacaa agcagttgcg gaggaaagaa 180aagagacaaa gtaaaaaaaa cggaaaagaa atctcccagg agaaagggat gtggaagctg 240aaaacacgga caatttccac agtaagactt ccaaaagaat gtgcaagatc cgagcaaaac 300tttcaagggc tctttttcag tgtaatggta gtgagaaagt tcagcctgga aagcccaggg 360cttaaaacaa caacaacaaa aaacacaata tgcaggatcg ttcggcttca gcagaaccca 420ccgcaaagat ggcggtggga cgaagcccct tctcccgccg ccgaagcctc tcgcctcaca 480tttcccacaa acccttcgcg ccgcctcgct agccgaaacc tgcccagccg gtgcccggcc 540actgcgcacg cgcgggacga cgtcacgtgc gctcccgggg ctggacggag ctggcaggag 600gggccttgcc agcttccgcc gccgcgtcgt ttcaggaccc ggacggcgga ttcgcgctg 659<210>127<211>659<212>DNA<213>人(Homo sapiens)<400>127ggaaagagta attgatcaga gccatccctc caattggagt caacttccat gactgttcgg 60atttccttta ttttgggggc agttcatcca aacttctatt aaacggcaac tagttcactt 120ttgagaagtg gtttacaaga aacaacaaca acaacaacaa agcagttgcg gaggaaagaa 180aagagacaaa gtaaaaaaaa cggaaaagaa atctcccagg agaaagggat gtggaagctg 240aaaacacgga caatttccac agtaagactt ccaaaagaat gtgcaagatc cgagcaaaac 300tttcaagggc tctttttcag tgtaatggta gtgagaaagt tcagcctgga aagcccaggg 360cttaaaacaa caacaacaaa aaacacaata tgcaggatcg ttcggcttca gcagaaccca 420ccgcaaagat ggcggtggga cgaagcccct tctcccgccg ccgaagcctc tcgcctcaca 480tttcccacaa acccttcgcg ccgcctcgct agccgaaacc tgcccagccg gtgcccggcc 540actgcgcacg cgcgggacga cgtcacgtgc gctcccgggg ctggacggag ctggcaggag 600gggccttgcc agcttccgcc gccgcgtcgt ttcaggaccc ggacggcgga ttcgcgctg 659<210>128<211>669<212>DNA<213>人(Homo sapiens)<400>128ggaaagagta attgatcaga gccatccctc caattggagt caactttcat gactgttcgg 60atttccttta ttttgggggc agttcatcca aacttctatt aaacggcaac tagttcactt 120ttgagaagtg gtttacaaga aacaacaaca acaacaacaa agcagttgcg gaggaaagaa 180aagagacaaa gtaaaaaaaa cggaaaagaa atctcccagg agaaagggat gtggaagctg 240aaaacacgga caatttccac agtaagactt ccaaaagaat gtgcaagatc cgagcaaaac 300tttcaagggc tctttttcag tgtaatggta gtgagaaagt tcagcctgga aagcccaggg 360cttaaaacaa caacaacaaa aaacacaata tgcaggatcg ttcggcttca gcagaaccca 420ccgcaaagat ggcggtggga cgaagcccct tctcccgccg ccgaagcctc tcgcctcaca 480tttcccacaa acccttcgcg ccgcctcgct agccgaaacc tgcccagccg gtgcccggcc 540actgcgcatg cgcgggacga cgtcacgtgc gctcccgggg ctggacggag ctggcaggag 600ctggcaggag gggccttgcc agcttccgcc gccgcgtcgt ttcaggaccc ggacggcgga 660ttcgcgctg 669<210>129<211>597<212>DNA<213>人(Homo sapiens)<400>129ggaaagagta attgatcaga gccatccctc caattggagt caactttcat gactgttcgg 60atttccttta ttttgggggc agttcatcca aacttctatt aaacggcaac tagttcactt 120ttgagaagtg gtttacaaga aacaacaaca acaacaacaa agcagttgcg gaggaaagaa 180aagagacaaa gtaaaaaaaa cggaaaagaa atctcccagg agaaagggat gtggaagctg 240aaaacacgga caatttccac agtaagactt ccaaaagaat gtgcaagatc cgagcaaaac 300tttcaagggc tctttttcag tgtaatggta gtgagaaagt tcagcctgga aagcccaggg 360cttaaaacaa caacaacaaa aaacacaata tgcaggatcg ttcggcttca gcagaaccca 420ccgcaaagat ggcggtggga cgaagcccct tctcccgccg ccgaagcctc tcgcctcaca 480tttcccacaa acccttcgcg ccgcctcgct agccgaaacc tgcccagccg gtgcccggcc 540actgcgcacg cgcgggacga cgtcacgtgc gctcccgggg ctggacggag ctggcag 597<210>130<211>10<212>DNA<213>人(Homo sapiens)<400>130gagctggcag 10<210>131<211>30<212>DNA<213>人(Homo sapiens)<400>131gggctggacg gagctggcag gaggggcctt 30<210>132<211>5002<212>DNA<213>人(Homo sapiens)<220><221>CDS<222>(229)..(4383)<400>132ggaaagagta attgatcaga gccatccctc caattggagt caactttcat gactgttcgg 60atttccttta ttttgggggc agttcatcca aacttctatt aaacggcaac tagttcactt 120ttgagaagtg gtttacaaga aacaacaaca acaacaacaa agcagttgcg gaggaaagaa 180aagagacaaa gtaaaaaaaa cggaaaagaa atctcccagg agaaaggg atg tgg aag 237
Met Trp Lys
1ctg aaa aca cgg aca att tcc aca gta aga ctt cca aaa gaa tgt gca 285Leu Lys Thr Arg Thr Ile Ser Thr Val Arg Leu Pro Lys Glu Cys Ala
5 10 15aga tcc gag caa aac ttt caa ggg ctc ttt ttc agt gta atg gta gtg 333Arg Ser Glu Gln Asn Phe Gln Gly Leu Phe Phe Ser Val Met Val Val20 25 30 35aga aag ttc agc ctg gaa agc cca ggg ctt aaa aca aca aca aca aaa 381Arg Lys Phe Ser Leu Glu Ser Pro Gly Leu Lys Thr Thr Thr Thr Lys
40 45 50aac aca ata tgc agg atc gtt cgg ctt cag cag aac cca ccg caa aga 429Asn Thr Ile Cys Arg Ile Val Arg Leu Gln Gln Asn Pro Pro Gln Arg
55 60 65tgg cgg tgg gac gaa gcc cct tct ccc gcc gcc gaa gcc tct cgc ctc 477Trp Arg Trp Asp Glu Ala Pro Ser Pro Ala Ala Glu Ala Ser Arg Leu
70 75 80aca ttt ccc aca aac cct tcg cgc cgc ctc gct agc cga aac ctg ccc 525Thr Phe Pro Thr Asn Pro Ser Arg Arg Leu Ala Ser Arg Asn Leu Pro
85 90 95agc cgg tgc ccg gcc act gcg cac gcg cgg gac gac gtc acg tgc gct 573Ser Arg Cys Pro Ala Thr Ala His Ala Arg Asp Asp Val Thr Cys Ala100 105 110 115ccc ggg gct gga cgg agc tgg cag gag ctg gca gga ggg gcc ttg cca 621Pro Gly Ala Gly Arg Ser Trp Gln Glu Leu Ala Gly Gly Ala Leu Pro
120 125 130gct tcc gcc gcc gcg tcg ttt cag gac ccg gac ggc gga ttc gcg ctg 669Ala Ser Ala Ala Ala Ser Phe Gln Asp Pro Asp Gly Gly Phe Ala Leu
135 140 145cct ccg ccg ccg cgg ggc agc cgg ggg gca ggg agc cca gcg agg ggc 717Pro Pro Pro Pro Arg Gly Ser Arg Gly Ala Gly Ser Pro Ala Arg Gly
150 155 160gcg cgt ggg cgc ggc cat ggg act gcg ccg gat ccg gtg aca gca ggg 765Ala Arg Gly Arg Gly His Gly Thr Ala Pro Asp Pro Val Thr Ala Gly
165 170 175agc caa gcg gcc cgg gcc ctg agc gcg tct tct ccg ggg ggc ctc gcc 813Ser Gln Ala Ala Arg Ala Leu Ser Ala Ser Ser Pro Gly Gly Leu Ala180 185 190 195ctc ctg ctc gcg ggg ccg ggg ctc ctg ctc cgg ttg ctg gcg ctg ttg 861Leu Leu Leu Ala Gly Pro Gly Leu Leu Leu Arg Leu Leu Ala Leu Leu
200 205 210ctg gct gtg gcg gcg gcc agg atc atg tcg ggt cgc cgc tgc gcc ggc 909Leu Ala Val Ala Ala Ala Arg Ile Met Ser Gly Arg Arg Cys Ala Gly
215 220 225ggg gga gcg gcc tgc gcg agc gcc gcg gcc gag gcc gtg gag ccg gcc 957Gly Gly Ala Ala Cys Ala Ser Ala Ala Ala Glu Ala Val Glu Pro Ala
230 235 240gcc cga gag ctg ttc gag gcg tgc cgc aac ggg gac gtg gaa cga gtc 1005Ala Arg Glu Leu Phe Glu Ala Cys Arg Asn Gly Asp Val Glu Arg Val
245 250 255aag agg ctg gtg acg cct gag aag gtg aac agc cgc gac acg gcg ggc 1053Lys Arg Leu Val Thr Pro Glu Lys Val Asn Ser Arg Asp Thr Ala Gly260 265 270 275agg aaa tcc acc ccg ctg cac ttc gcc gca ggt ttt ggg cgg aaa gac 1101Arg Lys Ser Thr Pro Leu His Phe Ala Ala Gly Phe Gly Arg Lys Asp
280 285 290gta gtt gaa tat ttg ctt cag aat ggt gca aat gtc caa gca cgt gat 1149Val Val Glu Tyr Leu Leu Gln Asn Gly Ala Asn Val Gln Ala Arg Asp
295 300 305gat ggg ggc ctt att cct ctt cat aat gca tgc tct ttt ggt cat gct 1197Asp Gly Gly Leu Ile Pro Leu His Asn Ala Cys Ser Phe Gly His Ala
310 315 320gaa gta gtc aat ctc ctt ttg cga cat ggt gca gac ccc aat gct cga 1245Glu Val Val Asn Leu Leu Leu Arg His Gly Ala Asp Pro Asn Ala Arg
325 330 335gat aat tgg aat tat act cct ctc cat gaa gct gca att aaa gga aag 1293Asp Asn Trp Asn Tyr Thr Pro Leu His Glu Ala Ala Ile Lys Gly Lys340 345 350 355att gat gtt tgc att gtg ctg tta cag cat gga gct gag cca acc atc 1341Ile Asp Val Cys Ile Val Leu Leu Gln His Gly Ala Glu Pro Thr Ile
360 365 370cga aat aca gat gga agg aca gca ttg gat tta gca gat cca tct gcc 1389Arg Asn Thr Asp Gly Arg Thr Ala Leu Asp Leu Ala Asp Pro Ser Ala
375 380 385aaa gca gtg ctt act ggt gaa tat aag aaa gat gaa ctc tta gaa agt 1437Lys Ala Val Leu Thr Gly Glu Tyr Lys Lys Asp Glu Leu Leu Glu Ser
390 395 400gcc agg agt ggc aat gaa gaa aaa atg atg gct cta ctc aca cca tta 1485Ala Arg Ser Gly Asn Glu Glu Lys Met Met Ala Leu Leu Thr Pro Leu
405 410 415aat gtc aac tgc cac gca agt gat ggc aga aag tca act cca tta cat 1533Asn Val Asn Cys His Ala Ser Asp Gly Arg Lys Ser Thr Pro Leu His420 425 430 435ttg gca gca gga tat aac aga gta aag att gta cag ctg tta ctg caa 1581Leu Ala Ala Gly Tyr Asn Arg Val Lys Ile Val Gln Leu Leu Leu Gln
440 445 450cat gga gct gat gtc cat gct aaa gat aaa ggt gat ctg gta cca tta 1629His Gly Ala Asp Val His Ala Lys Asp Lys Gly Asp Leu Val Pro Leu
455 460 465cac aat gcc tgt tct tat ggt cat tat gaa gta act gaa ctt ttg gtc 1677His Asn Ala Cys Ser Tyr Gly His Tyr Glu Val Thr Glu Leu Leu Val
470 475 480aag cat ggt gcc tgt gta aat gca atg gac ttg tgg caa ttc act cct 1725Lys His Gly Ala Cys Val Asn Ala Met Asp Leu Trp Gln Phe Thr Pro
485 490 495ctt cat gag gca gct tct aag aac agg gtt gaa gta tgt tct ctt ctc 1773Leu His Glu Ala Ala Ser Lys Asn Arg Val Glu Val Cys Ser Leu Leu500 505 510 515tta agt tat ggt gca gac cca aca ctg ctc aat tgt cac aat aaa agt 1821Leu Ser Tyr Gly Ala Asp Pro Thr Leu Leu Asn Cys His Asn Lys Ser
520 525 530gct ata gac ttg gct ccc aca cca cag tta aaa gaa aga tta gca tat 1869Ala Ile Asp Leu Ala Pro Thr Pro Gln Leu Lys Glu Arg Leu Ala Tyr
535 540 545gaa ttt aaa ggc cac tcg ttg ctg caa gct gca cga gaa gct gat gtt 1917Glu Phe Lys Gly His Ser Leu Leu Gln Ala Ala Arg Glu Ala Asp Val
550 555 560act cga atc aaa aaa cat ctc tct ctg gaa atg gtg aat ttc aag cat 1965Thr Arg Ile Lys Lys His Leu Ser Leu Glu Met Val Asn Phe Lys His
565 570 575cct caa aca cat gaa aca gca ttg cat tgt gct gct gca tct cca tat 2013Pro Gln Thr His Glu Thr Ala Leu His Cys Ala Ala Ala Ser Pro Tyr580 585 590 595ccc aaa aga aag caa ata tgt gaa ctg ttg cta aga aaa gga gca aac 2061Pro Lys Arg Lys Gln Ile Cys Glu Leu Leu Leu Arg Lys Gly Ala Asn
600 605 610atc aat gaa aag act aaa gaa ttc ttg act cct ctg cac gtg gca tct 2109Ile Asn Glu Lys Thr Lys Glu Phe Leu Thr Pro Leu His Val Ala Ser
615 620 625gag aaa gct cat aat gat gtt gtt gaa gta gtg gtg aaa cat gaa gca 2157Glu Lys Ala His Asn Asp Val Val Glu Val Val Val Lys His Glu Ala
630 635 640aag gtt aat gct ctg gat aat ctt ggt cag act tct cta cac aga gct 2205Lys Val Asn Ala Leu Asp Asn Leu Gly Gln Thr Ser Leu His Arg Ala
645 650 655gca tat tgt ggt cat cta caa acc tgc cgc cta ctc ctg agc tat ggg 2253Ala Tyr Cys Gly His Leu Gln Thr Cys Arg Leu Leu Leu Ser Tyr Gly660 665 670 675tgt gat cct aac att ata tcc ctt cag ggc ttt act gct tta cag atg 2301Cys Asp Pro Asn Ile Ile Ser Leu Gln Gly Phe Thr Ala Leu Gln Met
680 685 690gga aat gaa aat gta cag caa ctc ctc caa gag ggt atc tca tta ggt 2349Gly Asn Glu Asn Val Gln Gln Leu Leu Gln Glu Gly Ile Ser Leu Gly
695 700 705aat tca gag gca gac aga caa ttg ctg gaa gct gca aag gct gga gat 2397Asn Ser Glu Ala Asp Arg Gln Leu Leu Glu Ala Ala Lys Ala Gly Asp
710 715 720gtc gaa act gta aaa aaa ctg tgt act gtt cag agt gtc aac tgc aga 2445Val Glu Thr Val Lys Lys Leu Cys Thr Val Gln Ser Val Asn Cys Arg
725 730 735gac att gaa ggg cgt cag tct aca cca ctt cat ttt gca gct ggg tat 2493Asp Ile Glu Gly Arg Gln Ser Thr Pro Leu His Phe Ala Ala Gly Tyr740 745 750 755aac aga gtg tcc gtg gtg gaa tat ctg cta cag cat gga gct gat gtg 2541Asn Arg Val Ser Val Val Glu Tyr Leu Leu Gln His Gly Ala Asp Val
760 765 770cat gct aaa gat aaa gga ggc ctt gta cct ttg cac aat gca tgt tct 2589His Ala Lys Asp Lys Gly Gly Leu Val Pro Leu His Asn Ala Cys Ser
775 780 785tat gga cat tat gaa gtt gca gaa ctt ctt gtt aaa cat gga gca gta 2637Tyr Gly His Tyr Glu Val Ala Glu Leu Leu Val Lys His Gly Ala Val
790 795 800gtt aat gta gct gat tta tgg aaa ttt aca cct tta cat gaa gca gca 2685Val Asn Val Ala Asp Leu Trp Lys Phe Thr Pro Leu His Glu Ala Ala
805 810 815gca aaa gga aaa tat gaa att tgc aaa ctt ctg ctc cag cat ggt gca 2733Ala Lys Gly Lys Tyr Glu Ile Cys Lys Leu Leu Leu Gln His Gly Ala820 825 830 835gac cct aca aaa aaa aac agg gat gga aat act cct ttg gat ctt gtt 2781Asp Pro Thr Lys Lys Asn Arg Asp Gly Asn Thr Pro Leu Asp Leu Val
840 845 850aaa gat gga gat aca gat att caa gat ctg ctt agg gga gat gca gct 2829Lys Asp Gly Asp Thr Asp Ile Gln Asp Leu Leu Arg Gly Asp Ala Ala
855 860 865ttg cta gat gct gcc aag aag ggt tgt tta gcc aga gtg aag aag ttg 2877Leu Leu Asp Ala Ala Lys Lys Gly Cys Leu Ala Arg Val Lys Lys Leu
870 875 880tct tct cct gat aat gta aat tgc cgc gat acc caa ggc aga cat tca 2925Ser Ser Pro Asp Asn Val Asn Cys Arg Asp Thr Gln Gly Arg His Ser
885 890 895aca cct tta cat tta gca gct ggt tat aat aat tta gaa gtt gca gag 2973Thr Pro Leu His Leu Ala Ala Gly Tyr Asn Asn Leu Glu Val Ala Glu900 905 910 915tat ttg tta caa cac gga gct gat gtg aat gcc caa gac aaa gga gga 3021Tyr Leu Leu Gln His Gly Ala Asp Val Asn Ala Gln Asp Lys Gly Gly
920 925 930ctt att cct tta cat aat gca gca tct tac ggg cat gta gat gta gca 3069Leu Ile Pro Leu His Asn Ala Ala Ser Tyr Gly His Val Asp Val Ala
935 940 945gct cta cta ata aag tat aat gca tgt gtc aat gcc acg gac aaa tgg 3117Ala Leu Leu Ile Lys Tyr Asn Ala Cys Val Asn Ala Thr Asp Lys Trp
950 955 960gct ttc aca cct ttg cac gaa gca gcc caa aag gga cga aca cag ctt 3165Ala Phe Thr Pro Leu His Glu Ala Ala Gln Lys Gly Arg Thr Gln Leu
965 970 975tgt gct ttg ttg cta gcc cat gga gct gac ccg act ctt aaa aat cag 3213Cys Ala Leu Leu Leu Ala His Gly Ala Asp Pro Thr Leu Lys Asn Gln980 985 990 995gaa gga caa aca cct tta gat tta gtt tca gca gat gat gtc agc gct 3261Glu Gly Gln Thr Pro Leu Asp Leu Val Ser Ala Asp Asp Val Ser Ala
1000 1005 1010ctt ctg aca gca gcc atg ccc cca tct gct ctg ccc tct tgt tac aag 3309Leu Leu Thr Ala Ala Met Pro Pro Ser Ala Leu Pro Ser Cys Tyr Lys
1015 1020 1025cct caa gtg ctc aat ggt gtg aga agc cca gga gcc act gca gat gct 3357Pro Gln Val Leu Asn Gly Val Arg Ser Pro Gly Ala Thr Ala Asp Ala
1030 1035 1040ctc tct tca ggt cca tct agc cca tca agc ctt tct gca gcc agc agt 3405Leu Ser Ser Gly Pro Ser Ser Pro Ser Ser Leu Ser Ala Ala Ser Ser1045 1050 1055ctt gac aac tta tct ggg agt ttt tca gaa ctg tct tca gta gtt agt 3453Leu Asp Asn Leu Ser Gly Ser Phe Ser Glu Leu Ser Ser Val Val Ser1060 1065 1070 1075tca agt gga aca gag ggt gct tcc agt ttg gag aaa aag gag gtt cca 3501Ser Ser Gly Thr Glu Gly Ala Ser Ser Leu Glu Lys Lys Glu Val Pro
1080 1085 1090gga gta gat ttt agc ata act caa ttc gta agg aat ctt gga ctt gag 3549Gly Val Asp Phe Ser Ile Thr Gln Phe Val Arg Asn Leu Gly Leu Glu
1095 1100 1105cac cta atg gat ata ttt gag aga gaa cag atc act ttg gat gta tta 3597His Leu Met Asp Ile Phe Glu Arg Glu Gln Ile Thr Leu Asp Val Leu
1110 1115 1120gtt gag atg ggg cac aag gag ctg aag gag att gga atc aat gct tat 3645Val Glu Met Gly His Lys Glu Leu Lys Glu Ile Gly Ile Asn Ala Tyr 1125 1130 1135gga cat agg cac aaa cta att aaa gga gtc gag aga ctt atc tcc gga 3693Gly His Arg His Lys Leu Ile Lys Gly Val Glu Arg Leu Ile Ser Gly1140 1145 1150 1155caa caa ggt ctt aac cca tat tta act ttg aac acc tct ggt agt gga 3741Gln Gln Gly Leu Asn Pro Tyr Leu Thr Leu Asn Thr Ser Gly Ser Gly
1160 1165 1170aca att ctt ata gat ctg tct cct gat gat aaa gag ttt cag tct gtg 3789Thr Ile Leu Ile Asp Leu Ser Pro Asp Asp Lys Glu Phe Gln Ser Val
1175 1180 1185gag gaa gag atg caa agt aca gtt cga gag cac aga gat gga ggt cat 3837Glu Glu Glu Met Gln Ser Thr Val Arg Glu His Arg Asp Gly Gly His
1190 1195 1200gca ggt gga atc ttc aac aga tac aat att ctc aag att cag aag gtt 3885Ala Gly Gly Ile Phe Asn Arg Tyr Asn Ile Leu Lys Ile Gln Lys Val1205 1210 1215tgt aac aag aaa cta tgg gaa aga tac act cac cgg aga aaa gaa gtt 3933Cys Asn Lys Lys Leu Trp Glu Arg Tyr Thr His Arg Arg Lys Glu Val1220 1225 1230 1235tct gaa gaa aac cac aac cat gcc aat gaa cga atg cta ttt cat ggg 3981Ser Glu Glu Asn His Asn His Ala Asn Glu Arg Met Leu Phe His Gly
1240 1245 1250tct cct ttt gtg aat gca att atc cac aaa ggc ttt gat gaa agg cat 4029Ser Pro Phe Val Asn Ala Ile Ile His Lys Gly Phe Asp Glu Arg His
1255 1260 1265gcg tac ata ggt ggt atg ttt gga gct ggc att tat ttt gct gaa aac 4077Ala Tyr Ile Gly Gly Met Phe Gly Ala Gly Ile Tyr Phe Ala Glu Asn
1270 1275 1280tct tcc aaa agc aat caa tat gta tat gga att gga gga ggt act ggg 4125Ser Ser Lys Ser Asn Gln Tyr Val Tyr Gly Ile Gly Gly Gly Thr Gly1285 1290 1295tgt cca gtt cac aaa gac aga tct tgt tac att tgc cac agg cag ctg 4173Cys Pro Val His Lys Asp Arg Ser Cys Tyr Ile Cys His Arg Gln Leu1300 1305 1310 1315ctc ttt tgc cgg gta acc ttg gga aag tct ttc ctg cag ttc agt gca 4221Leu Phe Cys Arg Val Thr Leu Gly Lys Ser Phe Leu Gln Phe Ser Ala
1320 1325 1330atg aaa atg gca cat tct cct cca ggt cat cac tca gtc act ggt agg 4269Met Lys Met Ala His Ser Pro Pro Gly His His Ser Val Thr Gly Arg
1335 1340 1345ccc agt gta aat ggc cta gca tta gct gaa tat gtt att tac aga gga 4317Pro Ser Val Asn Gly Leu Ala Leu Ala Glu Tyr Val Ile Tyr Arg Gly
1350 1355 1360gaa cag gct tat cct gag tat tta att act tac cag att atg agg cct 4365Glu Gln Ala Tyr Pro Glu Tyr Leu Ile Thr Tyr Gln Ile Met Arg Pro1365 1370 1375gaa ggt atg gtc gat gga taaatagtta ttttaagaaa ctaattccac 4413Glu Gly Met Val Asp Gly1380 1385tgaacctaaa atcatcaaag cagcagtggc ctctacgttt tactcctttg ctgaaaaaaa 4473atcatcttgc ccacaggcct gtggcaaaag gataaaaatg tgaacgaagt ttaacattct 4533gacttgataa agctttaata atgtacagtg ttttctaaat atttcctgtt ttttcagcac 4593tttaacagat gccattccag gttaaactgg gttgtctgta ctaaattata aacagagtta 4653acttgaacct tttatatgtt atgcattgat tctaacaaac tgtaatgccc tcaacagaac 4713taattttact aatacaatac tgtgttcttt aaaacacagc atttacactg aatacaattt 4773catttgtaaa actgtaaata agagcttttg tactagccca gtatttattt acattgcttt 4833gtaatataaa tctgttttag aactgcagcg gtttacaaaa ttttttcata tgtattgttc 4893atctatactt catcttacat cgtcatgatt gagtgatctt tacatttgat tccagaggct 4953atgttcagtt gttagttggg aaagattgag ttatcagatt taatttgcc 5002<210>133<211>1385<212>PRT<213>人(Homo sapiens)<400>133Met Trp Lys Leu Lys Thr Arg Thr Ile Ser Thr Val Arg Leu Pro Lys1 5 10 15Glu Cys Ala Arg Ser Glu Gln Asn Phe Gln Gly Leu Phe Phe Ser Val
20 25 30Met Val Val Arg Lys Phe Ser Leu Glu Ser Pro Gly Leu Lys Thr Thr
35 40 45Thr Thr Lys Asn Thr Ile Cys Arg Ile Val Arg Leu Gln Gln Asn Pro
50 55 60Pro Gln Arg Trp Arg Trp Asp Glu Ala Pro Ser Pro Ala Ala Glu Ala65 70 75 80Ser Arg Leu Thr Phe Pro Thr Asn Pro Ser Arg Arg Leu Ala Ser Arg
85 90 95Asn Leu Pro Ser Arg Cys Pro Ala Thr Ala His Ala Arg Asp Asp Val
100 105 110Thr Cys Ala Pro Gly Ala Gly Arg Ser Trp Gln Glu Leu Ala Gly Gly
115 120 125Ala Leu Pro Ala Ser Ala Ala Ala Ser Phe Gln Asp Pro Asp Gly Gly
130 135 140Phe Ala Leu Pro Pro Pro Pro Arg Gly Ser Arg Gly Ala Gly Ser Pro145 150 155 160Ala Arg Gly Ala Arg Gly Arg Gly His Gly Thr Ala Pro Asp Pro Val
165 170 175Thr Ala Gly Ser Gln Ala Ala Arg Ala Leu Ser Ala Ser Ser Pro Gly
180 185 190Gly Leu Ala Leu Leu Leu Ala Gly Pro Gly Leu Leu Leu Arg Leu Leu
195 200 205Ala Leu Leu Leu Ala Val Ala Ala Ala Arg Ile Met Ser Gly Arg Arg
210 215 220Cys Ala Gly Gly Gly Ala Ala Cys Ala Ser Ala Ala Ala Glu Ala Val225 230 235 240Glu Pro Ala Ala Arg Glu Leu Phe Glu Ala Cys Arg Asn Gly Asp Val
245 250 255Glu Arg Val Lys Arg Leu Val Thr Pro Glu Lys Val Asn Ser Arg Asp
260 265 270Thr Ala Gly Arg Lys Ser Thr Pro Leu His Phe Ala Ala Gly Phe Gly
275 280 285Arg Lys Asp Val Val Glu Tyr Leu Leu Gln Asn Gly Ala Asn Val Gln
290 295 300Ala Arg Asp Asp Gly Gly Leu Ile Pro Leu His Asn Ala Cys Ser Phe305 310 315 320Gly His Ala Glu Val Val Asn Leu Leu Leu Arg His Gly Ala Asp Pro
325 330 335Asn Ala Arg Asp Asn Trp Asn Tyr Thr Pro Leu His Glu Ala Ala Ile
340 345 350Lys Gly Lys Ile Asp Val Cys Ile Val Leu Leu Gln His Gly Ala Glu
355 360 365Pro Thr Ile Arg Asn Thr Asp Gly Arg Thr Ala Leu Asp Leu Ala Asp
370 375 380Pro Ser Ala Lys Ala Val Leu Thr Gly Glu Tyr Lys Lys Asp Glu Leu385 390 395 400Leu Glu Ser Ala Arg Ser Gly Asn Glu Glu Lys Met Met Ala Leu Leu
405 410 415Thr Pro Leu Asn Val Asn Cys His Ala Ser Asp Gly Arg Lys Ser Thr
420 425 430Pro Leu His Leu Ala Ala Gly Tyr Asn Arg Val Lys Ile Val Gln Leu
435 440 445Leu Leu Gln His Gly Ala Asp Val His Ala Lys Asp Lys Gly Asp Leu
450 455 460Val Pro Leu His Asn Ala Cys Ser Tyr Gly His Tyr Glu Val Thr Glu465 470 475 480Leu Leu Val Lys His Gly Ala Cys Val Asn Ala Met Asp Leu Trp Gln
485 490 495Phe Thr Pro Leu His Glu Ala Ala Ser Lys Asn Arg Val Glu Val Cys
500 505 510Ser Leu Leu Leu Ser Tyr Gly Ala Asp Pro Thr Leu Leu Asn Cys His
515 520 525Asn Lys Ser Ala Ile Asp Leu Ala Pro Thr Pro Gln Leu Lys Glu Arg
530 535 540Leu Ala Tyr Glu Phe Lys Gly His Ser Leu Leu Gln Ala Ala Arg Glu545 550 555 560Ala Asp Val Thr Arg Ile Lys Lys His Leu Ser Leu Glu Met Val Asn
565 570 575Phe Lys His Pro Gln Thr His Glu Thr Ala Leu His Cys Ala Ala Ala
580 585 590Ser Pro Tyr Pro Lys Arg Lys Gln Ile Cys Glu Leu Leu Leu Arg Lys
595 600 605Gly Ala Asn Ile Asn Glu Lys Thr Lys Glu Phe Leu Thr Pro Leu His
610 615 620Val Ala Ser Glu Lys Ala His Asn Asp Val Val Glu Val Val Val Lys625 630 635 640His Glu Ala Lys Val Asn Ala Leu Asp Asn Leu Gly Gln Thr Ser Leu
645 650 655His Arg Ala Ala Tyr Cys Gly His Leu Gln Thr Cys Arg Leu Leu Leu
660 665 670Ser Tyr Gly Cys Asp Pro Asn Ile Ile Ser Leu Gln Gly Phe Thr Ala
675 680 685Leu Gln Met Gly Asn Glu Asn Val Gln Gln Leu Leu Gln Glu Gly Ile
690 695 700Ser Leu Gly Asn Ser Glu Ala Asp Arg Gln Leu Leu Glu Ala Ala Lys705 710 715 720Ala Gly Asp Val Glu Thr Val Lys Lys Leu Cys Thr Val Gln Ser Val
725 730 735Asn Cys Arg Asp Ile Glu Gly Arg Gln Ser Thr Pro Leu His Phe Ala
740 745 750Ala Gly Tyr Asn Arg Val Ser Val Val Glu Tyr Leu Leu Gln His Gly
755 760 765Ala Asp Val His Ala Lys Asp Lys Gly Gly Leu Val Pro Leu His Asn
770 775 780Ala Cys Ser Tyr Gly His Tyr Glu Val Ala Glu Leu Leu Val Lys His785 790 795 800Gly Ala Val Val Asn Val Ala Asp Leu Trp Lys Phe Thr Pro Leu His
805 810 815Glu Ala Ala Ala Lys Gly Lys Tyr Glu Ile Cys Lys Leu Leu Leu Gln
820 825 830His Gly Ala Asp Pro Thr Lys Lys Asn Arg Asp Gly Asn Thr Pro Leu
835 840 845Asp Leu Val Lys Asp Gly Asp Thr Asp Ile Gln Asp Leu Leu Arg Gly
850 855 860Asp Ala Ala Leu Leu Asp Ala Ala Lys Lys Gly Cys Leu Ala Arg Val865 870 875 880Lys Lys Leu Ser Ser Pro Asp Asn Val Asn Cys Arg Asp Thr Gln Gly
885 890 895Arg His Ser Thr Pro Leu His Leu Ala Ala Gly Tyr Asn Asn Leu Glu
900 905 910Val Ala Glu Tyr Leu Leu Gln His Gly Ala Asp Val Asn Ala Gln Asp
915 920 925Lys Gly Gly Leu Ile Pro Leu His Asn Ala Ala Ser Tyr Gly His Val
930 935 940Asp Val Ala Ala Leu Leu Ile Lys Tyr Asn Ala Cys Val Asn Ala Thr945 950 955 960Asp Lys Trp Ala Phe Thr Pro Leu His Glu Ala Ala Gln Lys Gly Arg
965 970 975Thr Gln Leu Cys Ala Leu Leu Leu Ala His Gly Ala Asp Pro Thr Leu
980 985 990Lys Asn Gln Glu Gly Gln Thr Pro Leu Asp Leu Val Ser Ala Asp Asp
995 1000 1005Val Ser Ala Leu Leu Thr Ala Ala Met Pro Pro Ser Ala Leu Pro Ser1010 1015 1020Cys Tyr Lys Pro Gln Val Leu Asn Gly Val Arg Ser Pro Gly Ala Thr1025 1030 1035 1040Ala Asp Ala Leu Ser Ser Gly Pro Ser Ser Pro Ser Ser Leu Ser Ala
1045 1050 1055Ala Ser Ser Leu Asp Asn Leu Ser Gly Ser Phe Ser Glu Leu Ser Ser
1060 1065 1070Val Val Ser Ser Ser Gly Thr Glu Gly Ala Ser Ser Leu Glu Lys Lys
1075 1080 1085Glu Val Pro Gly Val Asp Phe Ser Ile Thr Gln Phe Val Arg Asn Leu1090 1095 1100Gly Leu Glu His Leu Met Asp Ile Phe Glu Arg Glu Gln Ile Thr Leu1105 1110 1115 1120Asp Val Leu Val Glu Met Gly His Lys Glu Leu Lys Glu Ile Gly Ile
1125 1130 1135Asn Ala Tyr Gly His Arg His Lys Leu Ile Lys Gly Val Glu Arg Leu
1140 1145 1150Ile Ser Gly Gln Gln Gly Leu Asn Pro Tyr Leu Thr Leu Asn Thr Ser
1155 1160 1165Gly Ser Gly Thr Ile Leu Ile Asp Leu Ser Pro Asp Asp Lys Glu Phe1170 1175 1180Gln Ser Val Glu Glu Glu Met Gln Ser Thr Val Arg Glu His Arg Asp1185 1190 1195 1200Gly Gly His Ala Gly Gly Ile Phe Asn Arg Tyr Asn Ile Leu Lys Ile
1205 1210 1215Gln Lys Val Cys Asn Lys Lys Leu Trp Glu Arg Tyr Thr His Arg Arg
1220 1225 1230Lys Glu Val Ser Glu Glu Asn His Asn His Ala Asn Glu Arg Met Leu
1235 1240 1245Phe His Gly Ser Pro Phe Val Asn Ala Ile Ile His Lys Gly Phe Asp1250 1255 1260Glu Arg His Ala Tyr Ile Gly Gly Met Phe Gly Ala Gly Ile Tyr Phe1265 1270 1275 1280Ala Glu Asn Ser Ser Lys Ser Asn Gln Tyr Val Tyr Gly Ile Gly Gly
1285 1290 1295Gly Thr Gly Cys Pro Val His Lys Asp Arg Ser Cys Tyr Ile Cys His
1300 1305 1310Arg Gln Leu Leu Phe Cys Arg Val Thr Leu Gly Lys Ser Phe Leu Gln
1315 1320 1325Phe Ser Ala Met Lys Met Ala His Ser Pro Pro Gly His His Ser Val1330 1335 1340Thr Gly Arg Pro Ser Val Asn Gly Leu Ala Leu Ala Glu Tyr Val Ile1345 1350 1355 1360Tyr Arg Gly Glu Gln Ala Tyr Pro Glu Tyr Leu Ile Thr Tyr Gln Ile
1365 1370 1375Met Arg Pro Glu Gly Met Val Asp Gly
1380 1385<210>134<211>4992<212>DNA<213>人(Homo sapiens)<220><221>CDS<222>(876)..(4373)<400>134ggaaagagta attgatcaga gccatccctc caattggagt caacttccat gactgttcgg 60atttccttta ttttgggggc agttcatcca aacttctatt aaacggcaac tagttcactt 120ttgagaagtg gtttacaaga aacaacaaca acaacaacaa agcagttgcg gaggaaagaa 180aagagacaaa gtaaaaaaaa cggaaaagaa atctcccagg agaaagggat gtggaagctg 240aaaacacgga caatttccac agtaagactt ccaaaagaat gtgcaagatc cgagcaaaac 300tttcaagggc tctttttcag tgtaatggta gtgagaaagt tcagcctgga aagcccaggg 360cttaaaacaa caacaacaaa aaacacaata tgcaggatcg ttcggcttca gcagaaccca 420ccgcaaagat ggcggtggga cgaagcccct tctcccgccg ccgaagcctc tcgcctcaca 480tttcccacaa acccttcgcg ccgcctcgct agccgaaacc tgcccagccg gtgcccggcc 540actgcgcacg cgcgggacga cgtcacgtgc gctcccgggg ctggacggag ctggcaggag 600gggccttgcc agcttccgcc gccgcgtcgt ttcaggaccc ggacggcgga ttcgcgctgc 660ctccgccgcc gcggggcagc cggggggcag ggagcccagc gaggggcgcg cgtgggcgcg 720gccatgggac tgcgccggat ccggtgacag cagggagcca agcggcccgg gccctgagcg 780cgtcttctcc ggggggcctc gccctcctgc tcgcggggcc ggggctcctg ctccggttgc 840tggcgctgtt gctggctgtg gcggcggcca ggatc atg tcg ggt cgc cgc tgc 893
Met Ser Gly Arg Arg Cys
1 5gcc ggc ggg gga gcg gcc tgc gcg agc gcc gcg gcc gag gcc gtg gag 941Ala Gly Gly Gly Ala Ala Cys Ala Ser Ala Ala Ala Glu Ala Val Glu
10 15 20ccg gcc gcc cga gag ctg ttc gag gcg tgc cgc aac ggg gac gtg gaa 989Pro Ala Ala Arg Glu Leu Phe Glu Ala Cys Arg Asn Gly Asp Val Glu
25 30 35cga gtc aag agg ctg gtg acg cct gag aag gtg aac agc cgc gac acg 1037Arg Val Lys Arg Leu Val Thr Pro Glu Lys Val Asn Ser Arg Asp Thr
40 45 50gcg ggc agg aaa tcc acc ccg ctg cac ttc gcc gca ggt ttt ggg cgg 1085Ala Gly Arg Lys Ser Thr Pro Leu His Phe Ala Ala Gly Phe Gly Arg55 60 65 70aaa gac gta gtt gaa tat ttg ctt cag aat ggt gca aat gtc caa gca 1133Lys Asp Val Val Glu Tyr Leu Leu Gln Asn Gly Ala Asn Val Gln Ala
75 80 85cgt gat gat ggg ggc ctt att cct ctt cat aat gca tgc tct ttt ggt 1181Arg Asp Asp Gly Gly Leu Ile Pro Leu His Asn Ala Cys Ser Phe Gly
90 95 100cat gct gaa gta gtc aat ctc ctt ttg cga cat ggt gca gac ccc aat 1229His Ala Glu Val Val Asn Leu Leu Leu Arg His Gly Ala Asp Pro Asn
105 110 115gct cga gat aat tgg aat tat act cct ctc cat gaa gct gca att aaa 1277Ala Arg Asp Asn Trp Asn Tyr Thr Pro Leu His Glu Ala Ala Ile Lys
120 125 130gga aag att gat gtt tgc att gtg ctg tta cag cat gga gct gag cca 1325Gly Lys Ile Asp Val Cys Ile Val Leu Leu Gln His Gly Ala Glu Pro135 140 145 150acc atc cga aat aca gat gga agg aca gca ttg gat tta gca gat cca 1373Thr Ile Arg Asn Thr Asp Gly Arg Thr Ala Leu Asp Leu Ala Asp Pro
155 160 165tct gcc aaa gca gtg ctt act ggt gaa tat aag aaa gat gaa ctc tta 1421Ser Ala Lys Ala Val Leu Thr Gly Glu Tyr Lys Lys Asp Glu Leu Leu
170 175 180gaa agt gcc agg agt ggc aat gaa gaa aaa atg atg gct cta ctc aca 1469Glu Ser Ala Arg Ser Gly Asn Glu Glu Lys Met Met Ala Leu Leu Thr
185 190 195cca tta aat gtc aac tgc cac gca agt gat ggc aga aag tca act cca 1517Pro Leu Asn Val Asn Cys His Ala Ser Asp Gly Arg Lys Ser Thr Pro
200 205 210tta cat ttg gca gca gga tat aac aga gta aag att gta cag ctg tta 1565Leu His Leu Ala Ala Gly Tyr Asn Arg Val Lys Ile Val Gln Leu Leu215 220 225 230ctg caa cat gga gct gat gtc cat gct aaa gat aaa ggt gat ctg gta 1613Leu Gln His Gly Ala Asp Val His Ala Lys Asp Lys Gly Asp Leu Val
235 240 245cca tta cac aat gcc tgt tct tat ggt cat tat gaa gta act gaa ctt 1661Pro Leu His Asn Ala Cys Ser Tyr Gly His Tyr Glu Val Thr Glu Leu
250 255 260ttg gtc aag cat ggt gcc tgt gta aat gca atg gac ttg tgg caa ttc 1709Leu Val Lys His Gly Ala Cys Val Asn Ala Met Asp Leu Trp Gln Phe
265 270 275act cct ctt cat gag gca gct tct aag aac agg gtt gaa gta tgt tct 1757Thr Pro Leu His Glu Ala Ala Ser Lys Asn Arg Val Glu Val Cys Ser
280 285 290ctt ctc tta agt tat ggt gca gac cca aca ctg ctc aat tgt cac aat 1805Leu Leu Leu Ser Tyr Gly Ala Asp Pro Thr Leu Leu Asn Cys His Asn295 300 305 310aaa agt gct ata gac ttg gct ccc aca cca cag tta aaa gaa aga tta 1853Lys Ser Ala Ile Asp Leu Ala Pro Thr Pro Gln Leu Lys Glu Arg Leu
315 320 325gca tat gaa ttt aaa ggc cac tcg ttg ctg caa gct gca cga gaa gct 1901Ala Tyr Glu Phe Lys Gly His Ser Leu Leu Gln Ala Ala Arg Glu Ala
330 335 340gat gtt act cga atc aaa aaa cat ctc tct ctg gaa atg gtg aat ttc 1949Asp Val Thr Arg Ile Lys Lys His Leu Ser Leu Glu Met Val Asn Phe
345 350 355aag cat cct caa aca cat gaa aca gca ttg cat tgt gct gct gca tct 1997Lys His Pro Gln Thr His Glu Thr Ala Leu His Cys Ala Ala Ala Ser
360 365 370cca tat ccc aaa aga aag caa ata tgt gaa ctg ttg cta aga aaa gga 2045Pro Tyr Pro Lys Arg Lys Gln Ile Cys Glu Leu Leu Leu Arg Lys Gly375 380 385 390gca aac atc aat gaa aag act aaa gaa ttc ttg act cct ctg cac gtg 2093Ala Asn Ile Asn Glu Lys Thr Lys Glu Phe Leu Thr Pro Leu His Val
395 400 405gca tct gag aaa gct cat aat gat gtt gtt gaa gta gtg gtg aaa cat 2141Ala Ser Glu Lys Ala His Asn Asp Val Val Glu Val Val Val Lys His
410 415 420gaa gca aag gtt aat gct ctg gat aat ctt ggt cag act tct cta cac 2189Glu Ala Lys Val Asn Ala Leu Asp Asn Leu Gly Gln Thr Ser Leu His
425 430 435aga gct gca tat tgt ggt cat cta caa acc tgc cgc cta ctc ctg agc 2237Arg Ala Ala Tyr Cys Gly His Leu Gln Thr Cys Arg Leu Leu Leu Ser
440 445 450tat ggg tgt gat cct aac att ata tcc ctt cag ggc ttt act gct tta 2285Tyr Gly Cys Asp Pro Asn Ile Ile Ser Leu Gln Gly Phe Thr Ala Leu455 460 465 470cag atg gga aat gaa aat gta cag caa ctc ctc caa gag ggt atc tca 2333Gln Met Gly Asn Glu Asn Val Gln Gln Leu Leu Gln Glu Gly Ile Ser
475 480 485tta ggt aat tca gag gca gac aga caa ttg ctg gaa gct gca aag gct 2381Leu Gly Asn Ser Glu Ala Asp Arg Gln Leu Leu Glu Ala Ala Lys Ala
490 495 500gga gat gtc gaa act gta aaa aaa ctg tgt act gtt cag agt gtc aac 2429Gly Asp Val Glu Thr Val Lys Lys Leu Cys Thr Val Gln Ser Val Asn
505 510 515tgc aga gac att gaa ggg cgt cag tct aca cca ctt cat ttt gca gct 2477Cys Arg Asp Ile Glu Gly Arg Gln Ser Thr Pro Leu His Phe Ala Ala
520 525 530ggg tat aac aga gtg tcc gtg gtg gaa tat ctg cta cag cat gga gct 2525Gly Tyr Asn Arg Val Ser Val Val Glu Tyr Leu Leu Gln His Gly Ala535 540 545 550gat gtg cat gct aaa gat aaa gga ggc ctt gta cct ttg cac aat gca 2573Asp Val His Ala Lys Asp Lys Gly Gly Leu Val Pro Leu His Asn Ala
555 560 565tgt tct tat gga cat tat gaa gtt gca gaa ctt ctt gtt aaa cat gga 2621Cys Ser Tyr Gly His Tyr Glu Val Ala Glu Leu Leu Val Lys His Gly
570 575 580gca gta gtt aat gta gct gat tta tgg aaa ttt aca cct tta cat gaa 2669Ala Val Val Asn Val Ala Asp Leu Trp Lys Phe Thr Pro Leu His Glu
585 590 595gca gca gca aaa gga aaa tat gaa att tgc aaa ctt ctg ctc cag cat 2717Ala Ala Ala Lys Gly Lys Tyr Glu Ile Cys Lys Leu Leu Leu Gln His
600 605 610ggt gca gac cct aca aaa aaa aac agg gat gga aat act cct ttg gat 2765Gly Ala Asp Pro Thr Lys Lys Asn Arg Asp Gly Asn Thr Pro Leu Asp615 620 625 630ctt gtt aaa gat gga gat aca gat att caa gat ctg ctt agg gga gat 2813Leu Val Lys Asp Gly Asp Thr Asp Ile Gln Asp Leu Leu Arg Gly Asp
635 640 645gca gct ttg cta gat gct gcc aag aag ggt tgt tta gcc aga gtg aag 2861Ala Ala Leu Leu Asp Ala Ala Lys Lys Gly Cys Leu Ala Arg Val Lys
650 655 660aag ttg tct tct cct gat aat gta aat tgc cgc gat acc caa ggc aga 2909Lys Leu Ser Ser Pro Asp Asn Val Asn Cys Arg Asp Thr Gln Gly Arg
665 670 675cat tca aca cct tta cat tta gca gct ggt tat aat aat tta gaa gtt 2957His Ser Thr Pro Leu His Leu Ala Ala Gly Tyr Asn Asn Leu Glu Val
680 685 690gca gag tat ttg tta caa cac gga gct gat gtg aat gcc caa gac aaa 3005Ala Glu Tyr Leu Leu Gln His Gly Ala Asp Val Asn Ala Gln Asp Lys695 700 705 710gga gga ctt att cct tta cat aat gca gca tct tac ggg cat gta gat 3053Gly Gly Leu Ile Pro Leu His Asn Ala Ala Ser Tyr Gly His Val Asp
715 720 725gta gca gct cta cta ata aag tat aat gca tgt gtc aat gcc acg gac 3101Val Ala Ala Leu Leu Ile Lys Tyr Asn Ala Cys Val Asn Ala Thr Asp
730 735 740aaa tgg gct ttc aca cct ttg cac gaa gca gcc caa aag gga cga aca 3149Lys Trp Ala Phe Thr Pro Leu His Glu Ala Ala Gln Lys Gly Arg Thr
745 750 755cag ctt tgt gct ttg ttg cta gcc cat gga gct gac ccg act ctt aaa 3197Gln Leu Cys Ala Leu Leu Leu Ala His Gly Ala Asp Pro Thr Leu Lys
760 765 770aat cag gaa gga caa aca cct tta gat tta gtt tca gca gat gat gtc 3245Asn Gln Glu Gly Gln Thr Pro Leu Asp Leu Val Ser Ala Asp Asp Val775 780 785 790agc gct ctt ctg aca gca gcc atg ccc cca tct gct ctg ccc tct tgt 3293Ser Ala Leu Leu Thr Ala Ala Met Pro Pro Ser Ala Leu Pro Ser Cys
795 800 805tac aag cct caa gtg ctc aat ggt gtg aga agc cca gga gcc act gca 3341Tyr Lys Pro Gln Val Leu Asn Gly Val Arg Ser Pro Gly Ala Thr Ala
810 815 820gat gct ctc tct tca ggt cca tct agc cca tca agc ctt tct gca gcc 3389Asp Ala Leu Ser Ser Gly Pro Ser Ser Pro Ser Ser Leu Ser Ala Ala
825 830 835agc agt ctt gac aac tta tct ggg agt ttt tca gaa ctg tct tca gta 3437Ser Ser Leu Asp Asn Leu Ser Gly Ser Phe Ser Glu Leu Ser Ser Val
840 845 850gtt agt tca agt gga aca gag ggt gct tcc agt ttg gag aaa aag gag 3485Val Ser Ser Ser Gly Thr Glu Gly Ala Ser Ser Leu Glu Lys Lys Glu855 860 865 870gtt cca gga gta gat ttt agc ata act caa ttc gta agg aat ctt gga 3533Val Pro Gly Val Asp Phe Ser Ile Thr Gln Phe Val Arg Asn Leu Gly
875 880 885ctt gag cac cta atg gat ata ttt gag aga gaa cag atc act ttg gat 3581Leu Glu His Leu Met Asp Ile Phe Glu Arg Glu Gln Ile Thr Leu Asp
890 895 900gta tta gtt gag atg ggg cac aag gag ctg aag gag att gga atc aat 3629Val Leu Val Glu Met Gly His Lys Glu Leu Lys Glu Ile Gly Ile Asn
905 910 915gct tat gga cat agg cac aaa cta att aaa gga gtc gag aga ctt atc 3677Ala Tyr Gly His Arg His Lys Leu Ile Lys Gly Val Glu Arg Leu Ile
920 925 930tcc gga caa caa ggt ctt aac cca tat tta act ttg aac acc tct ggt 3725Ser Gly Gln Gln Gly Leu Asn Pro Tyr Leu Thr Leu Asn Thr Ser Gly935 940 945 950agt gga aca att ctt ata gat ctg tct cct gat gat aaa gag ttt cag 3773Ser Gly Thr Ile Leu Ile Asp Leu Ser Pro Asp Asp Lys Glu Phe Gln
955 960 965tct gtg gag gaa gag atg caa agt aca gtt cga gag cac aga gat gga 3821Ser Val Glu Glu Glu Met Gln Ser Thr Val Arg Glu His Arg Asp Gly
970 975 980ggt cat gca ggt gga atc ttc aac aga tac aat att ctc aag att cag 3869Gly His Ala Gly Gly Ile Phe Asn Arg Tyr Asn Ile Leu Lys Ile Gln
985 990 995aag gtt tgt aac aag aaa cta tgg gaa aga tac act cac cgg aga aaa 3917Lys Val Cys Asn Lys Lys Leu Trp Glu Arg Tyr Thr His Arg Arg Lys1000 1005 1010gaa gtt tct gaa gaa aac cac aac cat gcc aat gaa cga atg cta ttt 3965Glu Val Ser Glu Glu Asn His Asn His Ala Asn Glu Arg Met Leu Phe1015 1020 1025 1030cat ggg tct cct ttt gtg aat gca att atc cac aaa ggc ttt gat gaa 4013His Gly Ser Pro Phe Val Asn Ala Ile Ile His Lys Gly Phe Asp Glu
1035 1040 1045agg cat gcg tac ata ggt ggt atg ttt gga gct ggc att tat ttt gct 4061Arg His Ala Tyr Ile Gly Gly Met Phe Gly Ala Gly Ile Tyr Phe Ala
1050 1055 1060gaa aac tct tcc aaa agc aat caa tat gta tat gga att gga gga ggt 4109Glu Asn Ser Ser Lys Ser Asn Gln Tyr Val Tyr Gly Ile Gly Gly Gly
1065 1070 1075act ggg tgt cca gtt cac aaa gac aga tct tgt tac att tgc cac agg 4157Thr Gly Cys Pro Val His Lys Asp Arg Ser Cys Tyr Ile Cys His Arg1080 1085 1090cag ctg ctc ttt tgc cgg gta acc ttg gga aag tct ttc ctg cag ttc 4205Gln Leu Leu Phe Cys Arg Val Thr Leu Gly Lys Ser Phe Leu Gln Phe1095 1100 1105 1110agt gca atg aaa atg gca cat tct cct cca ggt cat cac tca gtc act 4253Ser Ala Met Lys Met Ala His Ser Pro Pro Gly His His Ser Val Thr
1115 1120 1125ggt agg ccc agt gta aat ggc cta gca tta gct gaa tat gtt att tac 4301Gly Arg Pro Ser Val Asn Gly Leu Ala Leu Ala Glu Tyr Val Ile Tyr
1130 1135 1140aga gga gaa cag gct tat cct gag tat tta att act tac cag att atg 4349Arg Gly Glu Gln Ala Tyr Pro Glu Tyr Leu Ile Thr Tyr Gln Ile Met
1145 1150 1155agg cct gaa ggt atg gtc gat gga taaatagtta ttttaagaaa ctaattccac 4403Arg Pro Glu Gly Met Val Asp Gly1160 1165tgaacctaaa atcatcaaag cagcagtggc ctctacgttt tactcctttg ctgaaaaaaa 4463atcatcttgc ccacaggcct gtggcaaaag gataaaaatg tgaacgaagt ttaacattct 4523gacttgataa agctttaata atgtacagtg ttttctaaat atttcctgtt ttttcagcac 4583tttaacagat gccattccag gttaaactgg gttgtctgta ctaaattata aacagagtta 4643acttgaacct tttatatgtt atgcattgat tctaacaaac tgtaatgccc tcaacagaac 4703taattttact aatacaatac tgtgttcttt aaaacacagc atttacactg aatacaattt 4763catttgtaaa actgtaaata agagcttttg tactagccca gtatttattt acattgcttt 4823gtaatataaa tctgttttag aactgcagcg gtttacaaaa ttttttcata tgtattgttc 4883atctatactt catcttacat cgtcatgatt gagtgatctt tacatttgat tccagaggct 4943atgttcagtt gttagttggg aaagattgag ttatcagatt taatttgcc 4992<210>135<211>1166<212>PRT<213>人(Homo sapiens)<400>135Met Ser Gly Arg Arg Cys Ala Gly Gly Gly Ala Ala Cys Ala Ser Ala1 5 10 15Ala Ala Glu Ala Val Glu Pro Ala Ala Arg Glu Leu Phe Glu Ala Cys
20 25 30Arg Asn Gly Asp Val Glu Arg Val Lys Arg Leu Val Thr Pro Glu Lys
35 40 45Val Asn Ser Arg Asp Thr Ala Gly Arg Lys Ser Thr Pro Leu His Phe
50 55 60Ala Ala Gly Phe Gly Arg Lys Asp Val Val Glu Tyr Leu Leu Gln Asn65 70 75 80Gly Ala Asn Val Gln Ala Arg Asp Asp Gly Gly Leu Ile Pro Leu His
85 90 95Asn Ala Cys Ser Phe Gly His Ala Glu Val Val Asn Leu Leu Leu Arg
100 105 110His Gly Ala Asp Pro Asn Ala Arg Asp Asn Trp Asn Tyr Thr Pro Leu
115 120 125His Glu Ala Ala Ile Lys Gly Lys Ile Asp Val Cys Ile Val Leu Leu
130 135 140Gln His Gly Ala Glu Pro Thr Ile Arg Asn Thr Asp Gly Arg Thr Ala145 150 155 160Leu Asp Leu Ala Asp Pro Ser Ala Lys Ala Val Leu Thr Gly Glu Tyr
165 170 175Lys Lys Asp Glu Leu Leu Glu Ser Ala Arg Ser Gly Asn Glu Glu Lys
180 185 190Met Met Ala Leu Leu Thr Pro Leu Asn Val Asn Cys His Ala Ser Asp
195 200 205Gly Arg Lys Ser Thr Pro Leu His Leu Ala Ala Gly Tyr Asn Arg Val
210 215 220Lys Ile Val Gln Leu Leu Leu Gln His Gly Ala Asp Val His Ala Lys225 230 235 240Asp Lys Gly Asp Leu Val Pro Leu His Asn Ala Cys Ser Tyr Gly His
245 250 255Tyr Glu Val Thr Glu Leu Leu Val Lys His Gly Ala Cys Val Asn Ala
260 265 270Met Asp Leu Trp Gln Phe Thr Pro Leu His Glu Ala Ala Ser Lys Asn
275 280 285Arg Val Glu Val Cys Ser Leu Leu Leu Ser Tyr Gly Ala Asp Pro Thr
290 295 300Leu Leu Asn Cys His Asn Lys Ser Ala Ile Asp Leu Ala Pro Thr Pro305 310 315 320Gln Leu Lys Glu Arg Leu Ala Tyr Glu Phe Lys Gly His Ser Leu Leu
325 330 335Gln Ala Ala Arg Glu Ala Asp Val Thr Arg Ile Lys Lys His Leu Ser
340 345 350Leu Glu Met Val Asn Phe Lys His Pro Gln Thr His Glu Thr Ala Leu
355 360 365His Cys Ala Ala Ala Ser Pro Tyr Pro Lys Arg Lys Gln Ile Cys Glu
370 375 380Leu Leu Leu Arg Lys Gly Ala Asn Ile Asn Glu Lys Thr Lys Glu Phe385 390 395 400Leu Thr Pro Leu His Val Ala Ser Glu Lys Ala His Asn Asp Val Val
405 410 415Glu Val Val Val Lys His Glu Ala Lys Val Asn Ala Leu Asp Asn Leu
420 425 430Gly Gln Thr Ser Leu His Arg Ala Ala Tyr Cys Gly His Leu Gln Thr
435 440 445Cys Arg Leu Leu Leu Ser Tyr Gly Cys Asp Pro Asn Ile Ile Ser Leu
450 455 460Gln Gly Phe Thr Ala Leu Gln Met Gly Asn Glu Asn Val Gln Gln Leu465 470 475 480Leu Gln Glu Gly Ile Ser Leu Gly Asn Ser Glu Ala Asp Arg Gln Leu
485 490 495Leu Glu Ala Ala Lys Ala Gly Asp Val Glu Thr Val Lys Lys Leu Cys
500 505 510Thr Val Gln Ser Val Asn Cys Arg Asp Ile Glu Gly Arg Gln Ser Thr
515 520 525Pro Leu His Phe Ala Ala Gly Tyr Asn Arg Val Ser Val Val Glu Tyr
530 535 540Leu Leu Gln His Gly Ala Asp Val His Ala Lys Asp Lys Gly Gly Leu545 550 555 560Val Pro Leu His Asn Ala Cys Ser Tyr Gly His Tyr Glu Val Ala Glu
565 570 575Leu Leu Val Lys His Gly Ala Val Val Asn Val Ala Asp Leu Trp Lys
580 585 590Phe Thr Pro Leu His Glu Ala Ala Ala Lys Gly Lys Tyr Glu Ile Cys
595 600 605Lys Leu Leu Leu Gln His Gly Ala Asp Pro Thr Lys Lys Asn Arg Asp
610 615 620Gly Asn Thr Pro Leu Asp Leu Val Lys Asp Gly Asp Thr Asp Ile Gln625 630 635 640Asp Leu Leu Arg Gly Asp Ala Ala Leu Leu Asp Ala Ala Lys Lys Gly
645 650 655Cys Leu Ala Arg Val Lys Lys Leu Ser Ser Pro Asp Asn Val Asn Cys
660 665 670Arg Asp Thr Gln Gly Arg His Ser Thr Pro Leu His Leu Ala Ala Gly
675 680 685Tyr Asn Asn Leu Glu Val Ala Glu Tyr Leu Leu Gln His Gly Ala Asp
690 695 700Val Asn Ala Gln Asp Lys Gly Gly Leu Ile Pro Leu His Asn Ala Ala705 710 715 720Ser Tyr Gly His Val Asp Val Ala Ala Leu Leu Ile Lys Tyr Asn Ala
725 730 735Cys Val Asn Ala Thr Asp Lys Trp Ala Phe Thr Pro Leu His Glu Ala
740 745 750Ala Gln Lys Gly Arg Thr Gln Leu Cys Ala Leu Leu Leu Ala His Gly
755 760 765Ala Asp Pro Thr Leu Lys Asn Gln Glu Gly Gln Thr Pro Leu Asp Leu
770 775 780Val Ser Ala Asp Asp Val Ser Ala Leu Leu Thr Ala Ala Met Pro Pro785 790 795 800Ser Ala Leu Pro Ser Cys Tyr Lys Pro Gln Val Leu Asn Gly Val Arg
805 810 815Ser Pro Gly Ala Thr Ala Asp Ala Leu Ser Ser Gly Pro Ser Ser Pro
820 825 830Ser Ser Leu Ser Ala Ala Ser Ser Leu Asp Asn Leu Ser Gly Ser Phe
835 840 845Ser Glu Leu Ser Ser Val Val Ser Ser Ser Gly Thr Glu Gly Ala Ser
850 855 860Ser Leu Glu Lys Lys Glu Val Pro Gly Val Asp Phe Ser Ile Thr Gln865 870 875 880Phe Val Arg Asn Leu Gly Leu Glu His Leu Met Asp Ile Phe Glu Arg
885 890 895Glu Gln Ile Thr Leu Asp Val Leu Val Glu Met Gly His Lys Glu Leu
900 905 910Lys Glu Ile Gly Ile Asn Ala Tyr Gly His Arg His Lys Leu Ile Lys
915 920 925Gly Val Glu Arg Leu Ile Ser Gly Gln Gln Gly Leu Asn Pro Tyr Leu
930 935 940Thr Leu Asn Thr Ser Gly Ser Gly Thr Ile Leu Ile Asp Leu Ser Pro945 950 955 960Asp Asp Lys Glu Phe Gln Ser Val Glu Glu Glu Met Gln Ser Thr Val
965 970 975Arg Glu His Arg Asp Gly Gly His Ala Gly Gly Ile Phe Asn Arg Tyr
980 985 990Asn Ile Leu Lys Ile Gln Lys Val Cys Asn Lys Lys Leu Trp Glu Arg
995 1000 1005Tyr Thr His Arg Arg Lys Glu Val Ser Glu Glu Asn His Asn His Ala1010 1015 1020Asn Glu Arg Met Leu Phe His Gly Ser Pro Phe Val Asn Ala Ile Ile1025 1030 1035 1040His Lys Gly Phe Asp Glu Arg His Ala Tyr Ile Gly Gly Met Phe Gly
1045 1050 1055Ala Gly Ile Tyr Phe Ala Glu Asn Ser Ser Lys Ser Asn Gln Tyr Val
1060 1065 1070Tyr Gly Ile Gly Gly Gly Thr Gly Cys Pro Val His Lys Asp Arg Ser
1075 1080 1085Cys Tyr Ile Cys His Arg Gln Leu Leu Phe Cys Arg Val Thr Leu Gly1090 1095 1100Lys Ser Phe Leu Gln Phe Ser Ala Met Lys Met Ala His Ser Pro Pro1105 1110 1115 1120Gly His His Ser Val Thr Gly Arg Pro Ser Val Asn Gly Leu Ala Leu
1125 1130 1135Ala Glu Tyr Val Ile Tyr Arg Gly Glu Gln Ala Tyr Pro Glu Tyr Leu
1140 1145 1150Ile Thr Tyr Gln Ile Met Arg Pro Glu Gly Met Val Asp Gly
1155 1160 1165<210>136<211>3045<212>DNA<213>人(Homo sapiens)<220><221>CDS<222>(1)..(3042)<400>136atg gcg gag tct tcg gat aag ctc tat cga gtc gag tac gcc aag agc 48Met Ala Glu Ser Ser Asp Lys Leu Tyr Arg Val Glu Tyr Ala Lys Ser1 5 10 15ggg cgc gcc tct tgc aag aaa tgc agc gag agc atc ccc aag gac tcg 96Gly Arg Ala Ser Cys Lys Lys Cys Ser Glu Ser Ile Pro Lys Asp Ser
20 25 30ctc cgg atg gcc atc atg gtg cag tcg ccc atg ttt gat gga aaa gtc 144Leu Arg Met Ala Ile Met Val Gln Ser Pro Met Phe Asp Gly Lys Val
35 40 45cca cac tgg tac cac ttc tcc tgc ttc tgg aag gtg ggc cac tcc atc 192Pro His Trp Tyr His Phe Ser Cys Phe Trp Lys Val Gly His Ser Ile
50 55 60cgg cac cct gac gtt gag gtg gat ggg ttc tct gag ctt cgg tgg gat 240Arg His Pro Asp Val Glu Val Asp Gly Phe Ser Glu Leu Arg Trp Asp65 70 75 80gac cag cag aaa gtc aag aag aca gcg gaa gct gga gga gtg aca ggc 288Asp Gln Gln Lys Val Lys Lys Thr Ala Glu Ala Gly Gly Val Thr Gly
85 90 95aaa ggc cag gat gga att ggt agc aag gca gag aag act ctg ggt gac 336Lys Gly Gln Asp Gly Ile Gly Ser Lys Ala Glu Lys Thr Leu Gly Asp
100 105 110ttt gca gca gag tat gcc aag tcc aac aga agt acg tgc aag ggg tgt 384Phe Ala Ala Glu Tyr Ala Lys Ser Asn Arg Ser Thr Cys Lys Gly Cys
115 120 125atg gag aag ata gaa aag ggc cag gtg cgc ctg tcc aag aag atg gtg 432Met Glu Lys Ile Glu Lys Gly Gln Val Arg Leu Ser Lys Lys Met Val
130 135 140gac ccg gag aag cca cag cta ggc atg att gac cgc tgg tac cat cca 480Asp Pro Glu Lys Pro Gln Leu Gly Met Ile Asp Arg Trp Tyr His Pro145 150 155 160ggc tgc ttt gtc aag aac agg gag gag ctg ggt ttc cgg ccc gag tac 528Gly Cys Phe Val Lys Asn Arg Glu Glu Leu Gly Phe Arg Pro Glu Tyr
165 170 175agt gcg agt cag ctc aag ggc ttc agc ctc ctt gct aca gag gat aaa 576Ser Ala Ser Gln Leu Lys Gly Phe Ser Leu Leu Ala Thr Glu Asp Lys
180 185 190gaa gcc ctg aag aag cag ctc cca gga gtc aag agt gaa gga aag aga 624Glu Ala Leu Lys Lys Gln Leu Pro Gly Val Lys Ser Glu Gly Lys Arg
195 200 205aaa ggc gat gag gtg gat gga gtg gat gaa gtg gcg aag aag aaa tct 672Lys Gly Asp Glu Val Asp Gly Val Asp Glu Val Ala Lys Lys Lys Ser
210 215 220aaa aaa gaa aaa gac aag gat agt aag ctt gaa aaa gcc cta aag gct 720Lys Lys Glu Lys Asp Lys Asp Ser Lys Leu Glu Lys Ala Leu Lys Ala225 230 235 240cag aac gac ctg atc tgg aac atc aag gac gag cta aag aaa gtg tgt 768Gln Asn Asp Leu Ile Trp Asn Ile Lys Asp Glu Leu Lys Lys Val Cys
245 250 255tca act aat gac ctg aag gag cta ctc atc ttc aac aag cag caa gtg 816Ser Thr Asn Asp Leu Lys Glu Leu Leu Ile Phe Asn Lys Gln Gln Val
260 265 270cct tct ggg gag tcg gcg atc ttg gac cga gta gct gat ggc atg gtg 864Pro Ser Gly Glu Ser Ala Ile Leu Asp Arg Val Ala Asp Gly Met Val
275 280 285ttc ggt gcc ctc ctt ccc tgc gag gaa tgc tcg ggt cag ctg gtc ttc 912Phe Gly Ala Leu Leu Pro Cys Glu Glu Cys Ser Gly Gln Leu Val Phe
290 295 300aag agc gat gcc tat tac tgc act ggg gac gtc act gcc tgg acc aag 960Lys Ser Asp Ala Tyr Tyr Cys Thr Gly Asp Val Thr Ala Trp Thr Lys305 310 315 320tgt atg gtc aag aca cag aca ccc aac cgg aag gag tgg gta acc cca 1008Cys Met Val Lys Thr Gln Thr Pro Asn Arg Lys Glu Trp Val Thr Pro
325 330 335aag gaa ttc cga gaa atc tct tac ctc aag aaa ttg aag gtt aaa aag 1056Lys Glu Phe Arg Glu Ile Ser Tyr Leu Lys Lys Leu Lys Val Lys Lys
340 345 350cag gac cgt ata ttc ccc cca gaa acc agc gcc tcc gtg gcg gcc acg 1104Gln Asp Arg Ile Phe Pro Pro Glu Thr Ser Ala Ser Val Ala Ala Thr
355 360 365cct ccg ccc tcc aca gcc tcg gct cct gct gct gtg aac tcc tct gct 1152Pro Pro Pro Ser Thr Ala Ser Ala Pro Ala Ala Val Asn Ser Ser Ala
370 375 380tca gca gat aag cca tta tcc aac atg aag atc ctg act ctc ggg aag 1200Ser Ala Asp Lys Pro Leu Ser Asn Met Lys Ile Leu Thr Leu Gly Lys385 390 395 400ctg tcc cgg aac aag gat gaa gtg aag gcc atg att gag aaa ctc ggg 1248Leu Ser Arg Asn Lys Asp Glu Val Lys Ala Met Ile Glu Lys Leu Gly
405 410 415ggg aag ttg acg ggg acg gcc aac aag gct tcc ctg tgc atc agc acc 1296Gly Lys Leu Thr Gly Thr Ala Asn Lys Ala Ser Leu Cys Ile Ser Thr
420 425 430aaa aag gag gtg gaa aag atg aat aag aag atg gag gaa gta aag gaa 1344Lys Lys Glu Val Glu Lys Met Asn Lys Lys Met Glu Glu Val Lys Glu
435 440 445gcc aac atc cga gtt gtg tct gag gac ttc ctc cag gac gtc tcc gcc 1392Ala Asn Ile Arg Val Val Ser Glu Asp Phe Leu Gln Asp Val Ser Ala
450 455 460tcc acc aag agc ctt cag gag ttg ttc tta gcg cac atc ttg tcc cct 1440Ser Thr Lys Ser Leu Gln Glu Leu Phe Leu Ala His Ile Leu Ser Pro465 470 475 480tgg ggg gca gag gtg aag gca gag cct gtt gaa gtt gtg gcc cca aga 1488Trp Gly Ala Glu Val Lys Ala Glu Pro Val Glu Val Val Ala Pro Arg
485 490 495ggg aag tca ggg gct gcg ctc tcc aaa aaa agc aag ggc cag gtc aag 1536Gly Lys Ser Gly Ala Ala Leu Ser Lys Lys Ser Lys Gly Gln Val Lys
500 505 510gag gaa ggt atc aac aaa tct gaa aag aga atg aaa tta act ctt aaa 1584Glu Glu Gly Ile Asn Lys Ser Glu Lys Arg Met Lys Leu Thr Leu Lys
515 520 525gga gga gca gct gtg gat cct gat tct gga ctg gaa cac tct gcg cat 1632Gly Gly Ala Ala Val Asp Pro Asp Ser Gly Leu Glu His Ser Ala His
530 535 540gtc ctg gag aaa ggt ggg aag gtc ttc agt gcc acc ctt ggc ctg gtg 1680Val Leu Glu Lys Gly Gly Lys Val Phe Ser Ala Thr Leu Gly Leu Val545 550 555 560gac atc gtt aaa gga acc aac tcc tac tac aag ctg cag ctt ctg gag 1728Asp Ile Val Lys Gly Thr Asn Ser Tyr Tyr Lys Leu Gln Leu Leu Glu
565 570 575gac gac aag gaa aac agg tat tgg ata ttc agg tcc tgg ggc cgt gtg 1776Asp Asp Lys Glu Asn Arg Tyr Trp Ile Phe Arg Ser Trp Gly Arg Val
580 585 590ggt acg gtg atc ggt agc aac aaa ctg gaa cag atg ccg tcc aag gag 1824Gly Thr Val Ile Gly Ser Asn Lys Leu Glu Gln Met Pro Ser Lys Glu
595 600 605gat gcc att gag cag ttc atg aaa tta tat gaa gaa aaa acc ggg aac 1872Asp Ala Ile Glu Gln Phe Met Lys Leu Tyr Glu Glu Lys Thr Gly Asn
610 615 620gct tgg cac tcc aaa aat ttc acg aag tat ccc aaa aag ttt tac ccc 1920Ala Trp His Ser Lys Asn Phe Thr Lys Tyr Pro Lys Lys Phe Tyr Pro625 630 635 640ctg gag att gac tat ggc cag gat gaa gag gca gtg aag aag ctc aca 1968Leu Glu Ile Asp Tyr Gly Gln Asp Glu Glu Ala Val Lys Lys Leu Thr
645 650 655gta aat cct ggc acc aag tcc aag ctc ccc aag cca gtt cag gac ctc 2016Val Asn Pro Gly Thr Lys Ser Lys Leu Pro Lys Pro Val Gln Asp Leu
660 665 670atc aag atg atc ttt gat gtg gaa agt atg aag aaa gcc atg gtg gag 2064Ile Lys Met Ile Phe Asp Val Glu Ser Met Lys Lys Ala Met ValGlu
675 680 685tat gag atc gac ctt cag aag atg ccc ttg ggg aag ctg agc aaa agg 2112Tyr Glu Ile Asp Leu Gln Lys Met Pro Leu Gly Lys Leu Ser Lys Arg
690 695 700cag atc cag gcc gca tac tcc atc ctc agt gag gtc cag cag gcg gtg 2160Gln Ile Gln Ala Ala Tyr Ser Ile Leu Ser Glu Val Gln Gln Ala Val705 710 715 720tct cag ggc agc agc gac tct cag atc ctg gat ctc tca aat cgc ttt 2208Ser Gln Gly Ser Ser Asp Ser Gln Ile Leu Asp Leu Ser Asn Arg Phe
725 730 735tac acc ctg atc ccc cac gac ttt ggg atg aag aag cct ccg ctc ctg 2256Tyr Thr Leu Ile Pro His Asp Phe Gly Met Lys Lys Pro Pro Leu Leu
740 745 750aac aat gca gac agt gtg cag gcc aag gtg gaa atg ctt gac aac ctg 2304Asn Asn Ala Asp Ser Val Gln Ala Lys Val Glu Met Leu Asp Asn Leu
755 760 765ctg gac atc gag gtg gcc tac agt ctg ctc agg gga ggg tct gat gat 2352Leu Asp Ile Glu Val Ala Tyr Ser Leu Leu Arg Gly Gly Ser Asp Asp
770 775 780agc agc aag gat ccc atc gat gtc aac tat gag aag ctc aaa act gac 2400Ser Ser Lys Asp Pro Ile Asp Val Asn Tyr Glu Lys Leu Lys Thr Asp785 790 795 800att aag gtg gtt gac aga gat tct gaa gaa gcc gag atc atc agg aag 2448Ile Lys Val Val Asp Arg Asp Ser Glu Glu Ala Glu Ile Ile Arg Lys
805 810 815tat gtt aag aac act cat gca acc aca cac agt gcg tat gac ttg gaa 2496Tyr Val Lys Asn Thr His Ala Thr Thr His Ser Ala Tyr Asp Leu Glu
820 825 830gtc atc gat atc ttt aag ata gag cgt gaa ggc gaa tgc cag cgt tac 2544Val Ile Asp Ile Phe Lys Ile Glu Arg Glu Gly Glu Cys Gln Arg Tyr
835 840 845aag ccc ttt aag cag ctt cat aac cga aga ttg ctg tgg cac ggg tcc 2592Lys Pro Phe Lys Gln Leu His Asn Arg Arg Leu Leu Trp His Gly Ser
850 855 860agg acc acc aac ttt gct ggg atc ctg tcc cag ggt ctt cgg ata gcc 2640Arg Thr Thr Asn Phe Ala Gly Ile Leu Ser Gln Gly Leu Arg Ile Ala865 870 875 880ccg cct gaa gcg ccc gtg aca ggc tac atg ttt ggt aaa ggg atc tat 2688Pro Pro Glu Ala Pro Val Thr Gly Tyr Met Phe Gly Lys Gly Ile Tyr
885 890 895ttc gct gac atg gtc tcc aag agt gcc aac tac tac cat acg tct cag 2736Phe Ala Asp Met Val Ser Lys Ser Ala Asn Tyr Tyr His Thr Ser Gln
900 905 910gga gac cca ata ggc tta atc ctg ttg gga gaa gtt gcc ctt gga aac 2784Gly Asp Pro Ile Gly Leu Ile Leu Leu Gly Glu Val Ala Leu Gly Asn
915 920 925atg tat gaa ctg aag cac gct tca cat atc agc agg tta ccc aag ggc 2832Met Tyr Glu Leu Lys His Ala Ser His Ile Ser Arg Leu Pro Lys Gly
930 935 940aag cac agt gtc aaa ggt ttg ggc aaa act acc cct gat cct tca gct 2880Lys His Ser Val Lys Gly Leu Gly Lys Thr Thr Pro Asp Pro Ser Ala945 950 955 960aac att agt ctg gat ggt gta gac gtt cct ctt ggg acc ggg att tca 2928Asn Ile Ser Leu Asp Gly Val Asp Val Pro Leu Gly Thr Gly Ile Ser
965 970 975tct ggt gtg ata gac acc tct cta cta tat aac gag tac att gtc tat 2976Ser Gly Val Ile Asp Thr Ser Leu Leu Tyr Asn Glu Tyr Ile Val Tyr
980 985 990gat att gct cag gta aat ctg aag tat ctg ctg aaa ctg aaa ttc aat 3024Asp Ile Ala Gln Val Asn Leu Lys Tyr Leu Leu Lys Leu Lys Phe Asn
995 1000 1005ttt aag acc tcc ctg tgg taa 3045Phe Lys Thr Ser Leu Trp1010<210>137<211>1014<212>PRT<213>人(Homo sapiens)<400>137Met Ala Glu Ser Ser Asp Lys Leu Tyr Arg Val Glu Tyr Ala Lys Ser1 5 10 15Gly Arg Ala Ser Cys Lys Lys Cys Ser Glu Ser Ile Pro Lys Asp Ser
20 25 30Leu Arg Met Ala Ile Met Val Gln Ser Pro Met Phe Asp Gly Lys Val
35 40 45Pro His Trp Tyr His Phe Ser Cys Phe Trp Lys Val Gly His Ser Ile
50 55 60Arg His Pro Asp Val Glu Val Asp Gly Phe Ser Glu Leu Arg Trp Asp65 70 75 80Asp Gln Gln Lys Val Lys Lys Thr Ala Glu Ala Gly Gly Val Thr Gly
85 90 95Lys Gly Gln Asp Gly Ile Gly Ser Lys Ala Glu Lys Thr Leu Gly Asp
100 105 110Phe Ala Ala Glu Tyr Ala Lys Ser Asn Arg Ser Thr Cys Lys Gly Cys
115 120 125Met Glu Lys Ile Glu Lys Gly Gln Val Arg Leu Ser Lys Lys Met Val
130 135 140Asp Pro Glu Lys Pro Gln Leu Gly Met Ile Asp Arg Trp Tyr His Pro145 150 155 160Gly Cys Phe Val Lys Asn Arg Glu Glu Leu Gly Phe Arg Pro Glu Tyr
165 170 175Ser Ala Ser Gln Leu Lys Gly Phe Ser Leu Leu Ala Thr Glu Asp Lys
180 185 190Glu Ala Leu Lys Lys Gln Leu Pro Gly Val Lys Ser Glu Gly Lys Arg
195 200 205Lys Gly Asp Glu Val Asp Gly Val Asp Glu Val Ala Lys Lys Lys Ser
210 215 220Lys Lys Glu Lys Asp Lys Asp Ser Lys Leu Glu Lys Ala Leu Lys Ala225 230 235 240Gln Asn Asp Leu Ile Trp Asn Ile Lys Asp Glu Leu Lys Lys Val Cys
245 250 255Ser Thr Asn Asp Leu Lys Glu Leu Leu Ile Phe Asn Lys Gln Gln Val
260 265 270Pro Ser Gly Glu Ser Ala Ile Leu Asp Arg Val Ala Asp Gly Met Val
275 280 285Phe Gly Ala Leu Leu Pro Cys Glu Glu Cys Ser Gly Gln Leu Val Phe
290 295 300Lys Ser Asp Ala Tyr Tyr Cys Thr Gly Asp Val Thr Ala Trp Thr Lys305 310 315 320Cys Met Val Lys Thr Gln Thr Pro Asn Arg Lys Glu Trp Val Thr Pro
325 330 335Lys Glu Phe Arg Glu Ile Ser Tyr Leu Lys Lys Leu Lys Val Lys Lys
340 345 350Gln Asp Arg Ile Phe Pro Pro Glu Thr Ser Ala Ser Val Ala Ala Thr
355 360 365Pro Pro Pro Ser Thr Ala Ser Ala Pro Ala Ala Val Asn Ser Ser Ala
370 375 380Ser Ala Asp Lys Pro Leu Ser Asn Met Lys Ile Leu Thr Leu Gly Lys385 390 395 400Leu Ser Arg Asn Lys Asp Glu Val Lys Ala Met Ile Glu Lys Leu Gly
405 410 415Gly Lys Leu Thr Gly Thr Ala Asn Lys Ala Ser Leu Cys Ile Ser Thr
420 425 430Lys Lys Glu Val Glu Lys Met Asn Lys Lys Met Glu Glu Val Lys Glu
435 440 445Ala Asn Ile Arg Val Val Ser Glu Asp Phe Leu Gln Asp Val Ser Ala
450 455 460Ser Thr Lys Ser Leu Gln Glu Leu Phe Leu Ala His Ile Leu Ser Pro465 470 475 480Trp Gly Ala Glu Val Lys Ala Glu Pro Val Glu Val Val Ala Pro Arg
485 490 495Gly Lys Ser Gly Ala Ala Leu Ser Lys Lys Ser Lys Gly Gln Val Lys
500 505 510Glu Glu Gly Ile Asn Lys Ser Glu Lys Arg Met Lys Leu Thr Leu Lys
515 520 525Gly Gly Ala Ala Val Asp Pro Asp Ser Gly Leu Glu His Ser Ala His
530 535 540Val Leu Glu Lys Gly Gly Lys Val Phe Ser Ala Thr Leu Gly Leu Val545 550 555 560Asp Ile Val Lys Gly Thr Asn Ser Tyr Tyr Lys Leu Gln Leu Leu Glu
565 570 575Asp Asp Lys Glu Asn Arg Tyr Trp Ile Phe Arg Ser Trp Gly Arg Val
580 585 590Gly Thr Val Ile Gly Ser Asn Lys Leu Glu Gln Met Pro Ser Lys Glu
595 600 605Asp Ala Ile Glu Gln Phe Met Lys Leu Tyr Glu Glu Lys Thr Gly Asn
610 615 620Ala Trp His Ser Lys Asn Phe Thr Lys Tyr Pro Lys Lys Phe Tyr Pro625 630 635 640Leu Glu Ile Asp Tyr Gly Gln Asp Glu Glu Ala Val Lys Lys Leu Thr
645 650 655Val Asn Pro Gly Thr Lys Ser Lys Leu Pro Lys Pro Val Gln Asp Leu
660 665 670Ile Lys Met Ile Phe Asp Val Glu Ser Met Lys Lys Ala Met Val Glu
675 680 685Tyr Glu Ile Asp Leu Gln Lys Met Pro Leu Gly Lys Leu Ser Lys Arg
690 695 700Gln Ile Gln Ala Ala Tyr Ser Ile Leu Ser Glu Val Gln Gln Ala Val705 710 715 720Ser Gln Gly Ser Ser Asp Ser Gln Ile Leu Asp Leu Ser Asn Arg Phe
725 730 735Tyr Thr Leu Ile Pro His Asp Phe Gly Met Lys Lys Pro Pro Leu Leu
740 745 750Asn Asn Ala Asp Ser Val Gln Ala Lys Val Glu Met Leu Asp Asn Leu
755 760 765Leu Asp Ile Glu Val Ala Tyr Ser Leu Leu Arg Gly Gly Ser Asp Asp
770 775 780Ser Ser Lys Asp Pro Ile Asp Val Asn Tyr Glu Lys Leu Lys Thr Asp785 790 795 800Ile Lys Val Val Asp Arg Asp Ser Glu Glu Ala Glu Ile Ile Arg Lys
805 810 815Tyr Val Lys Asn Thr His Ala Thr Thr His Ser Ala Tyr Asp Leu Glu
820 825 830Val Ile Asp Ile Phe Lys Ile Glu Arg Glu Gly Glu Cys Gln Arg Tyr
835 840 845Lys Pro Phe Lys Gln Leu His Asn Arg Arg Leu Leu Trp His Gly Ser
850 855 860Arg Thr Thr Asn Phe Ala Gly Ile Leu Ser Gln Gly Leu Arg Ile Ala865 870 875 880Pro Pro Glu Ala Pro Val Thr Gly Tyr Met Phe Gly Lys Gly Ile Tyr
885 890 895Phe Ala Asp Met Val Ser Lys Ser Ala Asn Tyr Tyr His Thr Ser Gln
900 905 910Gly Asp Pro Ile Gly Leu Ile Leu Leu Gly Glu Val Ala Leu Gly Asn
915 920 925Met Tyr Glu Leu Lys His Ala Ser His Ile Ser Arg Leu Pro Lys Gly
930 935 940Lys His Ser Val Lys Gly Leu Gly Lys Thr Thr Pro Asp Pro Ser Ala945 950 955 960Asn Ile Ser Leu Asp Gly Val Asp Val Pro Leu Gly Thr Gly Ile Ser
965 970 975Ser Gly Val Ile Asp Thr Ser Leu Leu Tyr Asn Glu Tyr Ile Val Tyr
980 985 990Asp Ile Ala Gln Val Asn Leu Lys Tyr Leu Leu Lys Leu Lys Phe Asn
995 1000 1005Phe Lys Thr Ser Leu Trp1010<210>138<211>5482<212>DNA<213>黄猩猩果蝇(Drosophila melanogaster)<220><221>CDS<222>(474)..(4016)<400>138aaacaatgca atatttcgcg gagcagtgaa ttaatccgga aataatcgtc cgtgcccaga 60gctttggagg ccaagtacca gggagctagt cccagagttg gtcgcagtct cagtaaaaac 120gaatcgtagc caaccgcgat ccttgcaacc gtgctgtgtc gaaccaaaga aatcctattg 180attttgggtc tgcaattgtg cattaaatat taagcaaaaa cgagggctgg tcgcgtggca 240gccagtggca aattgttgct cctgcggcat aggcaggaca cctggataca ggatgcgggc 300aagccagcga cggacaacgg cgaggcttgt gtaggacggg cagagcaact gctggaggag 360agaactggac tgggagtgga aaacccgaaa gcccactgaa tattgcgctt gttttttgtt 420gcctattttt ttcggggcgt gtgtgtgcca aagcgtagca aacaagcaca aca atg 476
Met
1gcc aac agc agc cga agt cgg gcc att ttg agc gtt aat ctc gat gcg 524Ala Asn Ser Ser Arg Ser Arg Ala Ile Leu Ser Val Asn Leu Asp Ala
5 10 15gtc atg gcc aac gat ccg ctg agg gag ctc tcc gag gcc tgc aaa acg 572Val Met Ala Asn Asp Pro Leu Arg Glu Leu Ser Glu Ala Cys Lys Thr
20 25 30ggc gag atc gcc aag gtg aag aag cta ata acg cct cag acc gtg aac 620Gly Glu Ile Ala Lys Val Lys Lys Leu Ile Thr Pro Gln Thr ValAsn
35 40 45gcc agg gat acg gcg gga cgc aaa tcc aca cca ttg cat ttc gca gcg 668Ala Arg Asp Thr Ala Gly Arg Lys Ser Thr Pro Leu His Phe Ala Ala50 55 60 65ggt tat gga cgc cgg gaa gtg gtt gaa ttc ctg ctg aac agc ggc gcc 716Gly Tyr Gly Arg Arg Glu Val Val Glu Phe Leu Leu Asn Ser Gly Ala
70 75 80tcc ata cag gcg tgt gac gag ggt ggg ctg cac ccg ctg cac aac tgt 764Ser Ile Gln Ala Cys Asp Glu Gly Gly Leu His Pro Leu His Asn Cys
85 90 95tgc tcc ttt ggc cac gcc gag gta gtt cga ttg ttg ctg aag gca ggt 812Cys Ser Phe Gly His Ala Glu Val Val Arg Leu Leu Leu Lys Ala Gly
100 105 110gcc agt cca aac acc acc gac aac tgg aac tac acg cca ttg cac gag 860Ala Ser Pro Asn Thr Thr Asp Asn Trp Asn Tyr Thr Pro Leu His Glu
115 120 125gcg gcc agc aag ggc aag gtg gat gtg tgc ctg gct ctg ttg cag cat 908Ala Ala Ser Lys Gly Lys Val Asp Val Cys Leu Ala Leu Leu Gln His130 135 140 145ggc gca aac cat acg atc cgc aac tcg gag cag aag aca cca ctg gag 956Gly Ala Asn His Thr Ile Arg Asn Ser Glu Gln Lys Thr Pro Leu Glu
150 155 160ctg gcg gac gag gcg acg cgt ccc gta ttg acc ggc gaa tat cga aag 1004Leu Ala Asp Glu Ala Thr Arg Pro Va1 Leu Thr Gly Glu Tyr Arg Lys
165 170 175gat gag ctg ctt gaa gcc gca cgc tcg ggg gcc gag gat cgc ctg ctg 1052Asp Glu Leu Leu Glu Ala Ala Arg Ser Gly Ala Glu Asp Arg Leu Leu
180 185 190gcc cta ctc acg cca ctc aat gtc aac tgt cat gcc agc gat gga cga 1100Ala Leu Leu Thr Pro Leu Asn Val Asn Cys His Ala Ser Asp Gly Arg
195 200 205cgc tca acg ccg ctc cat ctg gca gcg ggc tac aat cgg atc ggc atc 1148Arg Ser Thr Pro Leu His Leu Ala Ala Gly Tyr Asn Arg Ile Gly Ile210 215 220 225gtg gaa att ctg ctg gcc aac gga gcg gat gta cat gct aag gac aag 1196Val Glu Ile Leu Leu Ala Asn Gly Ala Asp Val His Ala Lys Asp Lys
230 235 240ggc ggt ctg gtg ccg ctg cac aat gcc tgc tcc tac gga cacttc gat 1244Gly Gly Leu Va1 Pro Leu His Asn Ala Cys Ser Tyr Gly His Phe Asp
245 250 255gtg acc aag ctg ctt atc cag gcg ggc gcc aat gtc aac gcc aac gat 1292Val Thr Lys Leu Leu Ile Gln Ala Gly Ala Asn Val Asn Ala Asn Asp
260 265 270ctg tgg gcc ttt acg ccg ctc cac gag gcc gcc tcc aaa agt cgc gtc 1340Leu Trp Ala Phe Thr Pro Leu His Glu Ala Ala Ser Lys Ser Arg Val
275 280 285gag gtc tgc agc ctg ctg ctc agt cgt gga gcg gat ccc acc ctc cta 1388Glu Val Cys Ser Leu Leu Leu Ser Arg Gly Ala Asp Pro Thr Leu Leu290 295 300 305aac tgc cac agc aag tcg gcc atc gat gcg gcg ccc acc agg gag ctg 1436Asn Cys His Ser Lys Ser Ala Ile Asp Ala Ala Pro Thr Arg Glu Leu
310 315 320aga gag cgg att gcc ttt gaa tac aag ggt cac tgc ctg ctg gac gcc 1484Arg Glu Arg Ile Ala Phe Glu Tyr Lys Gly His Cys Leu Leu Asp Ala
325 330 335tgt cga aag tgt gat gtg tcc cgt gcc aag aag ctg gta tgc gca gag 1532Cys Arg Lys Cys Asp Val Ser Arg Ala Lys Lys Leu Val Cys Ala Glu
340 345 350att gtt aac ttc gtg cat cca tat aca gga gac act ccg ctc cac ctg 1580Ile Val Asn Phe Val His Pro Tyr Thr Gly Asp Thr Pro Leu His Leu
355 360 365gcc gtt gtc agt ccg gat ggg aag cgc aag cag ctg atg gaa ctg ctg 1628Ala Val Val Ser Pro Asp Gly Lys Arg Lys Gln Leu Met Glu Leu Leu370 375 380 385acc aga aag gga tcc ttg ctg aac gag aaa aac aag gct ttc ctc acg 1676Thr Arg Lys Gly Ser Leu Leu Asn Glu Lys Asn Lys Ala Phe Leu Thr
390 395 400ccc ctg cat ttg gct gcc gag ctg ctt cac tac gat gcc atg gag gtg 1724Pro Leu His Leu Ala Ala Glu Leu Leu His Tyr Asp Ala Met Glu Val
405 410 415ctg cta aag cag ggc gcc aag gtt aat gca ttg gac agt ctt gga caa 1772Leu Leu Lys Gln Gly Ala Lys Val Asn Ala Leu Asp Ser Leu Gly Gln
420 425 430acg cca ctg cat cgg tgc gcc cgt gat gag caa gcg gtg cga ctg ctg 1820Thr Pro Leu His Arg Cys Ala Arg Asp Glu Gln Ala Val Arg Leu Leu
435 440 445ctc tcg tac gca gcg gac acg aat atc gtt tcc ctt gag gga ctt acg 1868Leu Ser Tyr Ala Ala Asp Thr Asn Ile Val Ser Leu Glu Gly Leu Thr450 455 460 465gcc gct caa ttg gcc tcg gac agc gtg ctg aag ctg ctc aag aat cct 1916Ala Ala Gln Leu Ala Ser Asp Ser Val Leu Lys Leu Leu Lys Asn Pro
470 475 480ccg gac agt gag aca cat tta ctg gag gca gcc aag gcg gga gat ctg 1964Pro Asp Ser Glu Thr His Leu Leu Glu Ala Ala Lys Ala Gly Asp Leu
485 490 495gac act gtg cgc cgt ata gtg ctc aac aat ccg att tcg gtc aat tgc 2012Asp Thr Val Arg Arg Ile Val Leu Asn Asn Pro Ile Ser Val Asn Cys
500 505 510cgg gat ttg gac gga cga cat tcc aca cct ttg cac ttt gct gct ggg 2060Arg Asp Leu Asp Gly Arg His Ser Thr Pro Leu His Phe Ala Ala Gly
515 520 525ttt aat aga gtg cca gtg gtt cag ttt ctt ttg gaa cac ggc gcc gag 2108Phe Asn Arg Val Pro Val Val Gln Phe Leu Leu Glu His Gly Ala Glu530 535 540 545gtt tat gcg gct gac aag ggc gga ctg gtg ccc ctg cac aat gcc tgc 2156Val Tyr Ala Ala Asp Lys Gly Gly Leu Val Pro Leu His Asn Ala Cys
550 555 560tct tat ggg cac tat gag gta acc gaa ctg ctg gtc aag cac gga gcc 2204Ser Tyr Gly His Tyr Glu Val Thr Glu Leu Leu Val Lys His Gly Ala
565 570 575aat gta aat gta tcg gat ttg tgg aag ttt act cct ctt cat gaa gct 2252Asn Val Asn Val Ser Asp Leu Trp Lys Phe Thr Pro Leu His Glu Ala
580 585 590gcc gcc aag gga aag tat gat att tgc aag ctg ctc ttg aaa cat ggc 2300Ala Ala Lys Gly Lys Tyr Asp Ile Cys Lys Leu Leu Leu Lys His Gly
595 600 605gct gat cca atg aag aag aat cgg gat ggc gcg aca cca gcg gat ttg 2348Ala Asp Pro Met Lys Lys Asn Arg Asp Gly Ala Thr Pro Ala Asp Leu610 615 620 625gtt aag gaa tct gat cac gat gtt gca gag ctg ctg aga gga ccg tcc 2396Val Lys Glu Ser Asp His Asp Val Ala Glu Leu Leu Arg Gly Pro Ser
630 635 640gct ctg cta gac gca gca aag aaa gga aac ttg gca cgg gta cag cga 2444Ala Leu Leu Asp Ala Ala Lys Lys Gly Asn Leu Ala Arg Val Gln Arg
645 650 655ttg gtt aca ccg gaa tcc att aat tgc cgg gac gcg cag ggc agg aat 2492Leu Val Thr Pro Glu Ser Ile Asn Cys Arg Asp Ala Gln Gly Arg Asn
660 665 670tcc aca cca ctt cac ctg gcc gcc gga tat aac aac ttt gag tgt gcc 2540Ser Thr Pro Leu His Leu Ala Ala Gly Tyr Asn Asn Phe Glu Cys Ala
675 680 685gag tac ctt ctg gag aat gga gcc gat gtt aat gca cag gac aag ggg 2588Glu Tyr Leu Leu Glu Asn Gly Ala Asp Val Asn Ala Gln Asp Lys Gly690 695 700 705gga cta ata cct ctg cac aat gcc agc agc tat ggg cat ttg gat att 2636Gly Leu Ile Pro Leu His Asn Ala Ser Ser Tyr Gly His Leu Asp Ile
710 715 720gcg gca ctg cta att aag cac aag acg gtt gtc aat gcg aca gat aaa 2684Ala Ala Leu Leu Ile Lys His Lys Thr Val Val Asn Ala Thr Asp Lys
725 730 735tgg gga ttc aca ccg ctc cac gag gct gca cag aag ggg cgc act caa 2732Trp Gly Phe Thr Pro Leu His Glu Ala Ala Gln Lys Gly Arg Thr Gln
740 745 750ttg tgc tcg ctc ttg ttg gcc cac ggt gcc gat gcc tat atg aaa aac 2780Leu Cys Ser Leu Leu Leu Ala His Gly Ala Asp Ala Tyr Met Lys Asn
755 760 765cag gag ggg cag acg ccc att gag ttg gcc acg gca gat gat gtt aag 2828Gln Glu Gly Gln Thr Pro Ile Glu Leu Ala Thr Ala Asp Asp Val Lys770 775 780 785tgc ttg ctc cag gac gcg atg gcc acc tcg ttg agt caa cag gcg ttg 2876Cys Leu Leu Gln Asp Ala Met Ala Thr Ser Leu Ser Gln Gln Ala Leu
790 795 800agt gct tcc acg caa tcg ctg aca agc agt tcc ccg gca cca gat gca 2924Ser Ala Ser Thr Gln Ser Leu Thr Ser Ser Ser Pro Ala Pro Asp Ala
805 810 815act gct gct gcg gct ccg ggc aca tct tca tcg tcc tca tcc gca atc 2972Thr Ala Ala Ala Ala Pro Gly Thr Ser Ser Ser Ser Ser Ser Ala Ile
820 825 830cta tcg ccc acc acg gaa acg gtg ttg ctg ccc acc ggt gcc tcc atg 3020Leu Ser Pro Thr Thr Glu Thr Val Leu Leu Pro Thr Gly Ala Ser Met
835 840 845att ctg agt gtt cct gtt cca ctt cca ctg tcc agt agc acg cgc atc 3068Ile Leu Ser Val Pro Val Pro Leu Pro Leu Ser Ser Ser Thr Arg Ile850 855 860 865agt ccc gcc caa gga gca gag gcc aat ggg gct gag ggc tcc tct tcg 3116Ser Pro Ala Gln Gly Ala Glu Ala Asn Gly Ala Glu Gly Ser Ser Ser
870 875 880gat gat cta ctg ccg gat gcg gat acc ata aca aat gtg tcc gga ttc 3164Asp Asp Leu Leu Pro Asp Ala Asp Thr Ile Thr Asn Val Ser Gly Phe
885 890 895cta agc agc cag cag ctg cat cat cta atc gaa ctg ttc gag cgc gaa 3212Leu Ser Ser Gln Gln Leu His His Leu Ile Glu Leu Phe Glu Arg Glu
900 905 910caa atc acc ttg gac att cta gcc gag atg ggc cac gac gat ctc aag 3260Gln Ile Thr Leu Asp Ile Leu Ala Glu Met Gly His Asp Asp Leu Lys
915 920 925cag gtg ggc gtc tcc gcc tac ggc ttc cgc cac aag ata ctc aag gga 3308Gln Val Gly Val Ser Ala Tyr Gly Phe Arg His Lys Ile Leu Lys Gly930 935 940 945atc gcc cag ctg agg tcc acc aca ggc att ggt aac aac gtg aat cta 3356Ile Ala Gln Leu Arg Ser Thr Thr Gly Ile Gly Asn Asn Val Asn Leu
950 955 960tgc aca ttg ttg gtg gac ttg ctg ccg gac gat aag gag ttt gtg gcc 3404Cys Thr Leu Leu Val Asp Leu Leu Pro Asp Asp Lys Glu Phe Val Ala
965 970 975gtc gag gag gag atg cag gcc acg att cgt gaa cat cgt gat aat gga 3452Val Glu Glu Glu Met Gln Ala Thr Ile Arg Glu His Arg Asp Asn Gly
980 985 990cag gct gga ggt tat ttc act cga tat aac atc att cgg gtg caa aag 3500Gln Ala Gly Gly Tyr Phe Thr Arg Tyr Asn Ile Ile Arg Val Gln Lys
995 1000 1005gta caa aat cga aag ctg tgg gag cgt tat gct cat cga cgg caa gag 3548Val Gln Asn Arg Lys Leu Trp Glu Arg Tyr Ala His Arg Arg Gln Glu1010 1015 1020 1025atc gcc gag gag aat ttc ctg cag tcc aac gag cgt atg ctc ttc cac 3596Ile Ala Glu Glu Asn Phe Leu Gln Ser Asn Glu Arg Met Leu Phe His
1030 1035 1040ggt agt ccc ttc atc aac gca att gtg caa cgc gga ttc gac gag cgc 3644Gly Ser Pro Phe Ile Asn Ala Ile Val Gln Arg Gly Phe Asp Glu Arg
1045 1050 1055cac gcc tac att ggc ggc atg ttt ggg gct ggc att tat ttc gcc gag 3692His Ala Tyr Ile Gly Gly Met Phe Gly Ala Gly Ile Tyr Phe Ala Glu
1060 1065 1070cat agc tcg aaa agc aac cag tat gtg tac gga att ggc ggc ggc att 3740His Ser Ser Lys Ser Asn Gln Tyr Val Tyr Gly Ile Gly Gly Gly Ile1075 1080 1085ggc tgt ccc tcg cac aag gat aag tcc tgc tac gtg tgt cct aga caa 3788Gly Cys Pro Ser His Lys Asp Lys Ser Cys Tyr Val Cys Pro Arg Gln1090 1095 1100 1105ttg ctg ctg tgc cga gtg gcg tta ggc aaa tcc ttc ttg caa tac agt 3836Leu Leu Leu Cys Arg Val Ala Leu Gly Lys Ser Phe Leu Gln Tyr Ser
1110 1115 1120gca atg aag atg gcc cat gca ccg ccg gga cac cac tcg gtg gtg ggc 3884Ala Met Lys Met Ala His Ala Pro Pro Gly His His Ser Val Val Gly
1125 1130 1135aga ccc tcg gcg ggt ggc ttg cat ttc gcc gaa tac gtt gtc tat cgg 3932Arg Pro Ser Ala Gly Gly Leu His Phe Ala Glu Tyr Val Val Tyr Arg
1140 1145 1150ggc gaa cag tct tat ccg gag tac ttg ata acc tac caa atc gtc aag 3980Gly Glu Gln Ser Tyr Pro Glu Tyr Leu Ile Thr Tyr Gln Ile Val Lys1155 1160 1165ccc gat gac agc agt agt gga acg gag gat aca aga tgatggatgc 4026Pro Asp Asp Ser Ser Ser Gly Thr Glu Asp Thr Arg1170 1175 1180cctctgtcgg gtccacgccc acaaccacgt cgcccgcgct gcaccagccg caaacgcaac 4086aacaaccgca gcagcaacag cagcagcagc cgcaaccaca acaacagcag aaggcaccac 4146tgccgttgcc accgccacaa cagcagacct cagctccagt tgccaagagg cggccgaaac 4206atgccaaacc atcgctgcag ttgcagtatc agccctatca gccccagcac cacccggttg 4266ttgcaaccgc cgctgctgtg accaccaccc aaccttcgcc cgctggcgtt tttgcgcaca 4326gcaataacaa caataatacg agcagcggaa atgtgaataa taacaacaat gacatgtcgc 4386cggtgtcgaa cagcaatagc tactcctcgg tggacaccaa ccagacgctg ctcaactcgc 4446tggccaacca gcagcgcaac catcgacagc cacagaatca tcatcatcag cagcagcagc 4506aggcgaatcg cagccaaaag tatagtcaat ttatgatcat cacacccgcc gtttccatag 4566atcgcgactt cgagtacgag tcgcatttgg actttgagga tttcgccaat gcggcccaca 4626acaatggcaa tctgtttcga cttggattgc ggcggagtga tagcagcagc gacgacagcg 4686gccacagcag cgatagcagc agtttccgct cgaattacaa tccctacttg catcacagtc 4746gccagcattt gctatcgaaa ggtggtaatg gtggtggcgg cggtgctagt cgtcacttct 4806acgccttcac ctcgtcgtgg cgctggtgca gtcttctgtg cgccgccatg cgctgctttg 4866gggccggggg agccgggcac gggaatgctc cgtacagcag ctcgctgcaa catcatcgac 4926taagacgctg ttcgtcgtac aatgcggaga atgcctacga gcactttgcg gcccccttca 4986aggcgcgcaa gacgcgcgac cacatgaata acatcaccta cgagttgtga cggtggtacg 5046acatgctggt ggttattgat ctctatgtct ccgttggcct cccgtctatt ttattacaat 5106tactagctat agatgtcgtg tcctgtgtgt ctctctctct ctcgttgttt attatattac 5166tctataatat atcacgtaag ggcgagctag cgagatggat tcgttcggtt tggatcgaat 5226tggattggat tcgattgggt taatcaaaag tgaagcacag tttttgagtg attttaattc 5286gaaatacgga aaatgcgatt cgattatacg aggttacaag ttctttgccg atgaatgcat 5346tacattacat tacattacgc tcgcgcgttt atttaagtgt ttaagcttag ttaatttaaa 5406caataattaa aactccaatt aaatttaaat atacaaatac atatacatca atcgaaaaaa 5466aaaaaaaaaa aaaaaa 5482<210>139<211>1181<212>PRT<213>黄猩猩果蝇(Drosophila melanogaster)<400>139Met Ala Asn Ser Ser Arg Ser Arg Ala Ile Leu Ser Val Asn Leu Asp1 5 10 15Ala Val Met Ala Asn Asp Pro Leu Arg Glu Leu Ser Glu Ala Cys Lys
20 25 30Thr Gly Glu Ile Ala Lys Val Lys Lys Leu Ile Thr Pro Gln Thr Val
35 40 45Asn Ala Arg Asp Thr Ala Gly Arg Lys Ser Thr Pro Leu His Phe Ala
50 55 60Ala Gly Tyr Gly Arg Arg Glu Val Val Glu Phe Leu Leu Asn Ser Gly65 70 75 80Ala Ser Ile Gln Ala Cys Asp Glu Gly Gly Leu His Pro Leu His Asn
85 90 95Cys Cys Ser Phe Gly His Ala Glu Val Val Arg Leu Leu Leu Lys Ala
100 105 110Gly Ala Ser Pro Asn Thr Thr Asp Asn Trp Asn Tyr Thr Pro Leu His
115 120 125Glu Ala Ala Ser Lys Gly Lys Val Asp Val Cys Leu Ala Leu Leu Gln
130 135 140His Gly Ala Asn His Thr Ile Arg Asn Ser Glu Gln Lys Thr Pro Leu145 150 155 160Glu Leu Ala Asp Glu Ala Thr Arg Pro Val Leu Thr Gly Glu Tyr Arg
165 170 175Lys Asp Glu Leu Leu Glu Ala Ala Arg Ser Gly Ala Glu Asp Arg Leu
180 185 190Leu Ala Leu Leu Thr Pro Leu Asn Val Asn Cys His Ala Ser Asp Gly
195 200 205Arg Arg Ser Thr Pro Leu His Leu Ala Ala Gly Tyr Asn Arg Ile Gly
210 215 220Ile Val Glu Ile Leu Leu Ala Asn Gly Ala Asp Val His Ala Lys Asp225 230 235 240Lys Gly Gly Leu Val Pro Leu His Asn Ala Cys Ser Tyr Gly His Phe
245 250 255Asp Val Thr Lys Leu Leu Ile Gln Ala Gly Ala Asn Val Asn Ala Asn
260 265 270Asp Leu Trp Ala Phe Thr Pro Leu His Glu Ala Ala Ser Lys Ser Arg
275 280 285Val Glu Val Cys Ser Leu Leu Leu Ser Arg Gly Ala Asp Pro Thr Leu
290 295 300Leu Asn Cys His Ser Lys Ser Ala Ile Asp Ala Ala Pro Thr Arg Glu305 310 315 320Leu Arg Glu Arg Ile Ala Phe Glu Tyr Lys Gly His Cys Leu Leu Asp
325 330 335Ala Cys Arg Lys Cys Asp Val Ser Arg Ala Lys Lys Leu Val Cys Ala
340 345 350Glu Ile Val Asn Phe Val His Pro Tyr Thr Gly Asp Thr Pro Leu His
355 360 365Leu Ala Val Val Ser Pro Asp Gly Lys Arg Lys Gln Leu Met Glu Leu
370 375 380Leu Thr Arg Lys Gly Ser Leu Leu Asn Glu Lys Asn Lys Ala Phe Leu385 390 395 400Thr Pro Leu His Leu Ala Ala Glu Leu Leu His Tyr Asp Ala Met Glu
405 410 415Val Leu Leu Lys Gln Gly Ala Lys Val Asn Ala Leu Asp Ser Leu Gly
420 425 430Gln Thr Pro Leu His Arg Cys Ala Arg Asp Glu Gln Ala Val Arg Leu
435 440 445Leu Leu Ser Tyr Ala Ala Asp Thr Asn Ile Val Ser Leu Glu Gly Leu
450 455 460Thr Ala Ala Gln Leu Ala Ser Asp Ser Val Leu Lys Leu Leu Lys Asn465 470 475 480Pro Pro Asp Ser Glu Thr His Leu Leu Glu Ala Ala Lys Ala Gly Asp
485 490 495Leu Asp Thr Val Arg Arg Ile Val Leu Asn Asn Pro Ile Ser Val Asn
500 505 510Cys Arg Asp Leu Asp Gly Arg His Ser Thr Pro Leu His Phe Ala Ala
515 520 525Gly Phe Asn Arg Val Pro Val Val Gln Phe Leu Leu Glu His Gly Ala
530 535 540Glu Val Tyr Ala Ala Asp Lys Gly Gly Leu Val Pro Leu His Asn Ala545 550 555 560Cys Ser Tyr Gly His Tyr Glu Val Thr Glu Leu Leu Val Lys His Gly
565 570 575Ala Asn Val Asn Val Ser Asp Leu Trp Lys Phe Thr Pro Leu His Glu
580 585 590Ala Ala Ala Lys Gly Lys Tyr Asp Ile Cys Lys Leu Leu Leu Lys His
595 600 605Gly Ala Asp Pro Met Lys Lys Asn Arg Asp Gly Ala Thr Pro Ala Asp
610 615 620Leu Val Lys Glu Ser Asp His Asp Val Ala Glu Leu Leu Arg Gly Pro625 630 635 640Ser Ala Leu Leu Asp Ala Ala Lys Lys Gly Asn Leu Ala Arg Val Gln
645 650 655Arg Leu Val Thr Pro Glu Ser Ile Asn Cys Arg Asp Ala Gln Gly Arg
660 665 670Asn Ser Thr Pro Leu His Leu Ala Ala Gly Tyr Asn Asn Phe Glu Cys
675 680 685Ala Glu Tyr Leu Leu Glu Asn Gly Ala Asp Val Asn Ala Gln Asp Lys
690 695 700Gly Gly Leu Ile Pro Leu His Asn Ala Ser Ser Tyr Gly His Leu Asp705 710 715 720Ile Ala Ala Leu Leu Ile Lys His Lys Thr Val Val Asn Ala Thr Asp
725 730 735Lys Trp Gly Phe Thr Pro Leu His Glu Ala Ala Gln Lys Gly Arg Thr
740 745 750Gln Leu Cys Ser Leu Leu Leu Ala His Gly Ala Asp Ala Tyr Met Lys
755 760 765Asn Gln Glu Gly Gln Thr Pro Ile Glu Leu Ala Thr Ala Asp Asp Val
770 775 780Lys Cys Leu Leu Gln Asp Ala Met Ala Thr Ser Leu Ser Gln Gln Ala785 790 795 800Leu Ser Ala Ser Thr Gln Ser Leu Thr Ser Ser Ser Pro Ala Pro Asp
805 810 815Ala Thr Ala Ala Ala Ala Pro Gly Thr Ser Ser Ser Ser Ser Ser Ala
820 825 830Ile Leu Ser Pro Thr Thr Glu Thr Val Leu Leu Pro Thr Gly Ala Ser
835 840 845Met Ile Leu Ser Val Pro Val Pro Leu Pro Leu Ser Ser Ser Thr Arg
850 855 860Ile Ser Pro Ala Gln Gly Ala Glu Ala Asn Gly Ala Glu Gly Ser Ser865 870 875 880Ser Asp Asp Leu Leu Pro Asp Ala Asp Thr Ile Thr Asn Val Ser Gly
885 890 895Phe Leu Ser Ser Gln Gln Leu His His Leu Ile Glu Leu Phe Glu Arg
900 905 910Glu Gln Ile Thr Leu Asp Ile Leu Ala Glu Met Gly His Asp Asp Leu
915 920 925Lys Gln Val Gly Val Ser Ala Tyr Gly Phe Arg His Lys Ile Leu Lys
930 935 940Gly Ile Ala Gln Leu Arg Ser Thr Thr Gly Ile Gly Asn Asn Val Asn945 950 955 960Leu Cys Thr Leu Leu Val Asp Leu Leu Pro Asp Asp Lys Glu Phe Val
965 970 975Ala Val Glu Glu Glu Met Gln Ala Thr Ile Arg Glu His Arg Asp Asn
980 985 990Gly Gln Ala Gly Gly Tyr Phe Thr Arg Tyr Asn Ile Ile Arg Val Gln
995 1000 1005Lys Val Gln Asn Arg Lys Leu Trp Glu Arg Tyr Ala His Arg Arg Gln1010 1015 1020Glu Ile Ala Glu Glu Asn Phe Leu Gln Ser Asn Glu Arg Met Leu Phe1025 1030 1035 1040His Gly Ser Pro Phe Ile Asn Ala Ile Val Gln Arg Gly Phe Asp Glu
1045 1050 1055Arg His Ala Tyr Ile Gly Gly Met Phe Gly Ala Gly Ile Tyr Phe Ala
1060 1065 1070Glu His Ser Ser Lys Ser Asn Gln Tyr Val Tyr Gly Ile Gly Gly Gly
1075 1080 1085Ile Gly Cys Pro Ser His Lys Asp Lys Ser Cys Tyr Val Cys Pro Arg1090 1095 1100Gln Leu Leu Leu Cys Arg Val Ala Leu Gly Lys Ser Phe Leu Gln Tyr1105 1110 1115 1120Ser Ala Met Lys Met Ala His Ala Pro Pro Gly His His Ser Val Val
1125 1130 1135Gly Arg Pro Ser Ala Gly Gly Leu His Phe Ala Glu Tyr Val Val Tyr
1140 1145 1150Arg Gly Glu Gln Ser Tyr Pro Glu Tyr Leu Ile Thr Tyr Gln Ile Val
1155 1160 1165Lys Pro Asp Asp Ser Ser Ser Gly Thr Glu Asp Thr Arg1170 1175 1180<210>140<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>140ggcctgaagg tatggtcgat 20<210>141<21l>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>141tgagggcatt acagtttgtt 20<210>142<211>346<212>DNA<213>人(Homo sapiens)<400>142ggcctgaagg tatggtcgat ggataaatag ttattttaag aaactaattc cactgaacct 60aaaatcatca aagcagcagt ggcctctacg ttttactcct ttgctgaaaa aaaatcatct 120tgcccacagg cctgtggcaa aaggataaaa atgtgaacga agtttaacat tctgacttga 180taaagcttta ataatgtaca gtgttttcta aatatttcct gttttttcag cactttaaca 240gatgccattc caggttaaac tgggttgtct gtactaaatt ataaacagag ttaacttgaa 300ccttttatat gttatgcatt gattctaaca aactgtaatg ccctca 346<210>143<211>29<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>143gccgaattcg gcctgaaggt atggtcgat 29<210>144<211>33<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>144gccgaattct agatgagggc attacagttt gtt 33<210>145<211>362<212>DNA<213>人(Homo sapiens)<400>145gaattcggcc tgaaggtatg gtcgatggat aaatagttat tttaagaaac taattccact 60gaacctaaaa tcatcaaagc agcagtggcc tctacgtttt actcctttgc tgaaaaaaaa 120tcatcttgcc cacaggcctg tggcaaaagg ataaaaatgt gaacgaagtt taacattctg 180acttgataaa gctttaataa tgtacagtgt tttctaaata tttcctgttt tttcagcact 240ttaacagatg ccattccagg ttaaactggg ttgtctgtac taaattataa acagagttaa 300cttgaacctt ttatatgtta tgcattgatt ctaacaaact gtaatgccct catctagaat 360tc 362<210>146<211>5616<212>DNA<213>人(Homo sapiens)<400>146tgaatgcctg ctggtgaagg ccagatcaga tttcaacctg ggactggatt acagaggatt 60gtttctaata acaacatcaa tattctagaa gtccctgaca gcctagaaat aagctgtttg 120tcttctataa agcattgcta tagtgatgaa tagtatgagt aactgataca tactcaactg 180ctactgttcc ctttgaggaa atgtttacag gggcggcctt ttaacatatc tcaggctcat 240tttcattgca attatccatt tctaaaacaa gattgcttcg atctagactt ggaaatggaa 300aataagaaaa ccaatgcttt ttcaaatgtt cacaattcac acactacatt tgttttgtta 360tgcatgacgt gtctataaca aatatacaca tacgacaggc aacaagcttg tttttgattt 420gccagacatg catcattggc tattgtttgt ttgttttttg tttttttgtg ttttttgggt 480tactttgaaa atgagccaga gccttcttga ggatattttg cacaaagtca cgctgacaaa 540atcattagca gtgcaaccca agcttctggc tgagcaagat tcagtttcca ctttttaaaa 600tttttttatt ttgctctgta gctgcacttc tcgttatcat aaattgagat gaaaaggaaa 660aaacatcaag ttttagtacc tttttatgaa ttggcctatc ttacaagaga agggcacaaa 720caccaacctg acttaggaac gcctaaattc agagaagtca aagccggtga aggccacttg 780ctctttccaa cacaagcctg ccacagaggt cttcgggaca gtactggaga tgcaggttga 840cacgggcttg agttccaagg tgaaaaaact ggggaggctg tgaaggaaga gctgcattaa 900ggagggtgag gagcgtgtgg ttctgtatca tggcagcccc aatggatcca ggggatgcct 960ccaaaaaata catgcttccc ttcccttaat ctgtactgtt gggattgtta cccctccaaa 1020ttagctgcct tatttcaaaa gtcagtgaaa ttactgcact tgatgagggt cacaaaaata 1080ccacttgatt gtttctttag ttgagaatgc tgggattcag actcgaatag tggatagata 1140cacacaaatg caaggacttt tttgtttact ccagatttgg ggtttatttt gagtggcatg 1200cttcaaatag ttcataaaga tccttgcatt aaatttctga accatttctt caaacttctt 1260agtgtgttta gacaaggaga acaaaaattg aaaccaaagc cctttctgtt attttttcaa 1320tgaaggtgag aaagaaatac catacaattt tctttgtgaa attactgttt attttcatca 1380acatttacca agtgccattg acatttataa aaaaaatgat cctttatagt tcttacactt 1440gcccttttca ccttaactga atatgaattg agtgcactaa cttatttact tgatatactg 1500tgcatctact ctgctttgaa gcgaaagaaa tataaacacg aggaggaata ggaaagacag 1560tgtgacacaa acttgccatt gcaattcaaa gccctgaaaa cgatgggttt aatgcaaggt 1620gattaagctg tgacctcctt taatctcctg aagcaaaata aaatggttac atgcaaaact 1680tctagaaata cactcttaaa atatatacat tttgctttga ttttggcttc aacccagtgc 1740tggaactagg catccagact agtttgaatg tttgtagctg aatttttatg ggtcctcaaa 1800attaaatcga gaattagcct cagttgttgc ttcttttgaa gtttcagtga cccaagctgg 1860gtgtttgtgt cttggctact tgtttaatag cactagaatt ccaggtgaag ctttgagagt 1920tgatattcat taagagggct ttttttcccc ttctttcctt ctcttttgct gtaacaaagg 1980gttgaagaaa ttgccatctg tgtagttttc agtagctgtc aagtgtgtct tacttacctt 2040cccccagacg tagtttaaaa tggtaaacac agctgtgatt tttagttaag taaaagagtt 2100aatatgatat agatatggaa agctttatgg cttcattaaa aagataaacc actacctaac 2160tgtggttgta tgttgtttcc atcatactaa ctagatgaat ggatgcgcca gttttcatct 2220tggtccttac acttgagaag ttaaactgtg gttcagtatt taaactgcca gtgttatacg 2280tctcatgctc tgtgtgccag gtgaaggtac tgtgtaagga agacatttgc ggtgcttctt 2340gtcctataat gattcaagta tatagtagtt cttgaaagag tgtgcatata ttactcatct 2400gcttaagaga gtgggttaat ggatatatca gaggagccaa atacattttt ttcagaactt 2460gaaaaccaaa ggtcatcatg agtgcactca aaagttagga caagtttatt acatttggga 2520ttttcatctg tagccgtatg aagaaccctt tccaatataa aagcatggca ttaaattagg 2580ctgaagtctt ttattttttg tatatgtact atatagaaat actagcaagt taggatcatc 2640caatatggcc taccccgaaa tggcccctct gtttccctaa ccacatggaa gaaagaatct 2700gaacgtctcc accggctcta cccgagttcc aaaactaaag ggcttctcca gacctgatgg 2760ttccagttta cctgctgttg gcctgctgga tacttgactc aggcataaat taagtgccct 2820ggtcccgaac tttctcccag tatttgacct ccttccctct ttcctaaatt actagtctgg 2880aattaaaatt agctccagca atgacctttg actccattca ttttctcctc atcttgggtc 2940ttaaaaaagg agaccagata cctcctagct tttgtatcac aaccaggaat gggtattagg 3000cctcatgcgc tttgctcaga acactgccgc tttgttaaca aatgacagca tggaacccag 3060agttttgatt cgatgcaaaa taacagcagt gcaaccagga ttcttgtttt ccttttcctt 3120cttggagttt ggaatttcta gcttttcaag cagcataagt agaatcaaca ttaggatgtt 3180ttcatgaaat agcatcctta tacttctttg agcttgatgt tagtggctag actgatttcc 3240ctttgctctc aaaatacaaa gtgcattgaa gtatacagag aaatgcctga atatggcaag 3300caaataatgt agattaacat tctattattg tatccgtttt acaaaaaata aaattttgat 3360atatgccgga gaacggcatt agaatgcaat aagttgtcta ggtttttctg tttcagtgtc 3420tctcccaatg gcacgaaggg ttattgggca ttgtccccac ccccgccttt ttaacatgtg 3480cactatctgg attcctgtaa atggccttgc aaacagaagt ggtgtgtatt ttcaagcacc 3540tttcccccat tgtatccgaa tccctcttgt gtgatatctg tgacaaatac cattcttctt 3600gtgttttctg ttgggactaa ttgtctcacg taaagctata gaccttacta atttggcagg 3660tattcaaaac tgccattaag ataggatttc atgtcagata cgtatttaaa gagtaaagtc 3720aaatttgttt aatgtcagat cagtgacaga agtgaaaaga aagtaattgt gaaagtgatg 3780tttgagctat tgtacacatc tagcatatgg aaagcaaatg cactcgaaaa ctactattct 3840agaacatgag gcttcttcag caacttgtgc actctgccat taataaatta aatttttccc 3900ctctagaaag ccttaactat ggcggaaact ttttaacctt ttatatttta ataaataaaa 3960cattgtagtc ccatttctta gtgtttgaaa ggtgtgtcag tgagtcggcc atgtctccat 4020gtgtttcaga cctgttcatc ttattttatg atggtatatt tcataagtaa tattccctta 4080catgcaatgg agctgattaa aattaatcca tttcaatttc tccatattgg aacttcctca 4140gctaccagat ttctggtttg gagaagtgct ggaaagattt caaagcctat tcagttgtgt 4200atgtggggat acgacagcaa ctgtgatacc ttgtagaata tgagtgatat gcaagctgtg 4260ttttttaatt gttttaaaat gtaaattatg gttatgctaa agtgaaaacc tagaggaagc 4320taatgatttt atatactttg cacgaccaaa tatggtcgta gtatgacgag ttttatacat 4380tgccagagag ttctgcctcc tctgaaataa cattcgcact gtagattgca tttcggcttt 4440tcctcctttc acattctttt ttgctttaca cttcacgtct tcgcacctgc cctacctccc 4500atcctttcaa agaggtttct ttcacgttcc agaattcaga ttgttctgtg atttctttta 4560catcagtcta cccatttctg caggcagccc tgaaagccct tgtgttgatt cagagtgttt 4620gcagagaaat gcagttgaac cctggtagtg gggtgtccct cacacacccg cgcacccctc 4680ccaaagttca ggatgaaagg ctagaaaacc cattcaaagt taggaaagaa cacagatctt 4740tgaggccgat agcctagacc tagaagatga ccttgagtat gtaaacattg tctccgtgac 4800acaaaacact gaaactcttc atgtgcatat aacacctgct tctgctccca ttgtttcaag 4860ctcatcttat ctttgtagta gtaatgtttg tctttgatac ctacaaacta aaaaggtact 4920tttatcaagg tttctcaaaa catttacaaa accagctttg agaaaatgtt atgttgcctg 4980gcaacagcac tcggagtagt aattgtgttt tctcattgtg atgttggtct gtgtgagcaa 5040ccagtgtagt gactctttgg ttcattattc gtgttgtttt tatttttagt ctctgtgtga 5100cccaacagtg gcaggggtta caaccccctc tcctttcttt tttgtattta tctatttgta 5160ggattgtcag atcaagtaca agatgcccag ttaagtttga atttcagaga aacaatttca 5220cgttaagaat gtttcatgca atatttggca tatatttaca gtaaaagcat tcattatttg 5280tctgaaattc aaatttaact gagcatgctg gtttttctca ttgtttggtt tttctaaatc 5340tggcaatcct acagctgtgg tcatgggaaa tcacctacag catgttaaag tcctctagtc 5400atcatctcgt cacctgaaat ggaagtcctt tttccctcac cctccacttc tttccaaagg 5460agggcatcaa ggaacttaac ctgcctgcct ggtgggtttc tatttaagac atctttgtga 5520ttatatttaa cctgcaattg tgctttggct taatgtctag ctcactgtac ttgtaaatga 5580ttaatattca ataaaaccat ttttaaagta aaaaaa 5616<210>147<211>29<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>147gccgaattcc ttgtttttga tttgccaga 29<210>148<211>34<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>148gccgaattcc ggctttgact tctctgaatt tagg 34<210>149<211>372<212>DNA<213>人(Homo sapiens)<400>149gaattccttg tttttgattt gccagacatg catcattggc tattgtttgt ttgttttttg 60tttttttgtg ttttttgggt tactttgaaa atgagccaga gccttcttga ggatattttg 120cacaaagtca cgctgacaaa atcattagca gtgcaaccca agcttctggc tgagcaagat 180tcagtttcca ctttttaaaa tttttttatt ttgctctgta gctgcacttc tcgttatcat 240aaattgagat gaaaaggaaa aaacatcaag ttttagtacc tttttatgaa ttggcctatc 300ttacaagaga agggcacaaa caccaacctg acttaggaac gcctaaattc agagaagtca 360aagccggaat tc 372<210>150<211>1320<212>DNA<213>人(Homo sapiens)<220><221>CDS<222>(1)..(1317)<400>150atg gcg gag gat gtt tcc tca gcg gcc ccg agc ccg cgg cgg tgt gcg 48Met Ala Glu Asp Val Ser Ser Ala Ala Pro Ser Pro Arg Arg Cys Ala1 5 10 15gat ggt agg gat gcc gac cct act gag gag cag atg gca gaa aca gag 96Asp Gly Arg Asp Ala Asp Pro Thr Glu Glu Gln Met Ala Glu Thr Glu
20 25 30aga aac gac gag gag cag ttc gaa tgc cag gaa ctg ctc gag tgc cag 144Arg Asn Asp Glu Glu Gln Phe Glu Cys Gln Glu Leu Leu Glu Cys Gln
35 40 45gtg cag gtg ggg gcc ccc gag gag gag gag gag gag gag gag gac gcg 192Val Gln Val Gly Ala Pro Glu Glu Glu Glu Glu Glu Glu Glu Asp Ala
50 55 60ggc ctg gtg gcc gag gcc gag gcc gtg gct gcc ggc tgg atg ctc gat 240Gly Leu Val Ala Glu Ala Glu Ala Val Ala Ala Gly Trp Met Leu Asp65 70 75 80ttc ctc tgc ctc tct ctt tgc cga gct ttc cgc gac ggc cgc tcc gag 288Phe Leu Cys Leu Ser Leu Cys Arg Ala Phe Arg Asp Gly Arg Ser Glu
85 90 95gac ttc cgc agg acc cgc aac agc gca gag gct att att cat gga cta 336Asp Phe Arg Arg Thr Arg Asn Ser Ala Glu Ala Ile Ile His Gly Leu
100 105 110tcc agt cta aca gct tgc cag ttg aga acg ata tac ata tgt cag ttt 384Ser Ser Leu Thr Ala Cys Gln Leu Arg Thr Ile Tyr Ile Cys Gln Phe
115 120 125ttg aca aga att gca gca gga aaa acc ctt gat gca cag ttt gaa aat 432Leu Thr Arg Ile Ala Ala Gly Lys Thr Leu Asp Ala Gln Phe Glu Asn
130 135 140gat gaa cga att aca ccc ttg gaa tca gcc ctg atg att tgg ggt tca 480Asp Glu Arg Ile Thr Pro Leu Glu Ser Ala Leu Met Ile Trp Gly Ser145 150 155 160att gaa aag gaa cat gac aaa ctt cat gaa gaa ata cag aat tta att 528Ile Glu Lys Glu His Asp Lys Leu His Glu Glu Ile Gln Asn Leu Ile
165 170 175aaa att cag gct ata gct gtt tgt atg gaa aat ggc aac ttt aaa gaa 576Lys Ile Gln Ala Ile Ala Val Cys Met Glu Asn Gly Asn Phe Lys Glu
180 185 190gca gaa gaa gtc ttt gaa aga ata ttt ggt gat cca aat tct cat atg 624Ala Glu Glu Val Phe Glu Arg Ile Phe Gly Asp Pro Asn Ser His Met
195 200 205cct ttc aaa agc aaa ttg ctt atg ata atc tct cag aaa gat aca ttt 672Pro Phe Lys Ser Lys Leu Leu Met Ile Ile Ser Gln Lys Asp Thr Phe
210 215 220cat tcc ttt ttt caa cac ttc agc tac aac cac atg atg gag aaa att 720His Ser Phe Phe Gln His Phe Ser Tyr Asn His Met Met Glu Lys Ile225 230 235 240aag agt tat gtg aat tat gtg cta agt gaa aaa tca tca acc ttt cta 768Lys Ser Tyr Val Asn Tyr Val Leu Ser Glu Lys Ser Ser Thr Phe Leu
245 250 255atg aag gca gcg gca aaa gta gta gaa agc aaa agg aca aga aca ata 816Met Lys Ala Ala Ala Lys Val Val Glu Ser Lys Arg Thr Arg Thr Ile
260 265 270act tct caa gat aaa cct agt ggt aat gat gtt gaa atg gaa act gaa 864Thr Ser Gln Asp Lys Pro Ser Gly Asn Asp Val Glu Met Glu Thr Glu
275 280 285gct aat ttg gat aca aga aaa agt gtt agt gac aaa cag tct gcg gta 912Ala Asn Leu Asp Thr Arg Lys Ser Val Ser Asp Lys Gln Ser Ala Val
290 295 300act gaa tcc tca gag ggt aca gta tcc tta ttg agg tct cac aag aat 960Thr Glu Ser Ser Glu Gly Thr Val Ser Leu Leu Arg Ser His Lys Asn305 310 315 320ctt ttc tta tct aag ttg caa cat gga acc cag caa caa gac ctt aat 1008Leu Phe Leu Ser Lys Leu Gln His Gly Thr Gln Gln Gln Asp Leu Asn
325 330 335aag aaa gaa aga aga gta gga act cct caa agt aca aaa aag aaa aaa 1056Lys Lys Glu Arg Arg Val Gly Thr Pro Gln Ser Thr Lys Lys Lys Lys
340 345 350gaa agc aga aga gcc act gaa agc aga ata cct gtt tca aag agt cag 1104Glu Ser Arg Arg Ala Thr Glu Ser Arg Ile Pro Val Ser Lys Ser Gln
355 360 365ccg gta act cct gaa aaa cat cga gct aga aaa aga cag gca tgg ctt 1152Pro Val Thr Pro Glu Lys His Arg Ala Arg Lys Arg Gln Ala Trp Leu
370 375 380tgg gaa gaa gac aag aat ttg aga tct ggc gtg agg aaa tat gga gag 1200Trp Glu Glu Asp Lys Asn Leu Arg Ser Gly Val Arg Lys Tyr Gly Glu385 390 395 400gga aac tgg tct aaa ata ctg ttg cat tat aaa ttc aac aac cgg aca 1248Gly Asn Trp Ser Lys Ile Leu Leu His Tyr Lys Phe Asn Asn Arg Thr
405 410 415agt gtc atg tta aaa gac aga tgg agg acc atg aag aaa cta aaa ctg 1296Ser Val Met Leu Lys Asp Arg Trp Arg Thr Met Lys Lys Leu Lys Leu
420 425 430att tcc tca gac agc gaa gac tga 1320Ile Ser Ser Asp Ser Glu Asp
435<210>151<211>439<212>PRT<213>人(Homo sapiens)<400>151Met Ala Glu Asp Val Ser Ser Ala Ala Pro Ser Pro Arg Arg Cys Ala1 5 10 15Asp Gly Arg Asp Ala Asp Pro Thr Glu Glu Gln Met Ala Glu Thr Glu
20 25 30Arg Asn Asp Glu Glu Gln Phe Glu Cys Gln Glu Leu Leu Glu Cys Gln
35 40 45Val Gln Val Gly Ala Pro Glu Glu Glu Glu Glu Glu Glu Glu Asp Ala
50 55 60Gly Leu Val Ala Glu Ala Glu Ala Val Ala Ala Gly Trp Met Leu Asp65 70 75 80Phe Leu Cys Leu Ser Leu Cys Arg Ala Phe Arg Asp Gly Arg Ser Glu
85 90 95Asp Phe Arg Arg Thr Arg Asn Ser Ala Glu Ala Ile Ile His Gly Leu
100 105 110Ser Ser Leu Thr Ala Cys Gln Leu Arg Thr Ile Tyr Ile Cys Gln Phe
115 120 125Leu Thr Arg Ile Ala Ala Gly Lys Thr Leu Asp Ala Gln Phe Glu Asn
130 135 140Asp Glu Arg Ile Thr Pro Leu Glu Ser Ala Leu Met Ile Trp Gly Ser145 150 155 160Ile Glu Lys Glu His Asp Lys Leu His Glu Glu Ile Gln Asn Leu Ile
165 170 175Lys Ile Gln Ala Ile Ala Val Cys Met Glu Asn Gly Asn Phe Lys Glu
180 185 190Ala Glu Glu Val Phe Glu Arg Ile Phe Gly Asp Pro Asn Ser His Met
195 200 205Pro Phe Lys Ser Lys Leu Leu Met Ile Ile Ser Gln Lys Asp Thr Phe
210 215 220His Ser Phe Phe Gln His Phe Ser Tyr Asn His Met Met Glu Lys Ile225 230 235 240Lys Ser Tyr Val Asn Tyr Val Leu Ser Glu Lys Ser Ser Thr Phe Leu
245 250 255Met Lys Ala Ala Ala Lys Val Val Glu Ser Lys Arg Thr Arg Thr Ile
260 265 270Thr Ser Gln Asp Lys Pro Ser Gly Asn Asp Val Glu Met Glu Thr Glu
275 280 285Ala Asn Leu Asp Thr Arg Lys Ser Val Ser Asp Lys Gln Ser Ala Val
290 295 300Thr Glu Ser Ser Glu Gly Thr Val Ser Leu Leu Arg Ser His Lys Asn305 310 315 320Leu Phe Leu Ser Lys Leu Gln His Gly Thr Gln Gln Gln Asp Leu Asn
325 330 335Lys Lys Glu Arg Arg Val Gly Thr Pro Gln Ser Thr Lys Lys Lys Lys
340 345 350Glu Ser Arg Arg Ala Thr Glu Ser Arg Ile Pro Val Ser Lys Ser Gln
355 360 365Pro Val Thr Pro Glu Lys His Arg Ala Arg Lys Arg Gln Ala Trp Leu
370 375 380Trp Glu Glu Asp Lys Asn Leu Arg Ser Gly Val Arg Lys Tyr Gly Glu385 390 395 400Gly Asn Trp Ser Lys Ile Leu Leu His Tyr Lys Phe Asn Asn Arg Thr
405 410 415Ser Val Met Leu Lys Asp Arg Trp Arg Thr Met Lys Lys Leu Lys Leu
420 425 430Ile Ser Ser Asp Ser Glu Asp
435<210>152<211>39<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>152gccccgggga tcctcatggc ggaggatgtt tcctcagcg 39<210>153<211>33<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>153tcccggggat cctcacacca ggcccgcgtc ctc 33<210>154<211>201<212>DNA<213>人(Homo sapiens)<220><221>CDS<222>(1)..(201)<400>154atg gcg gag gat gtt tcc tca gcg gcc ccg agc ccg cgg ggc tgt gcg 48Met Ala Glu Asp Val Ser Ser Ala Ala Pro Ser Pro Arg Gly Cys Ala1 5 10 15gat ggt agg gat gcc gac cct act gag gag cag atg gca gaa aca gag 96Asp Gly Arg Asp Ala Asp Pro Thr Glu Glu Gln Met Ala Glu Thr Glu
20 25 30aga aac gac gag gag cag ttc gaa tgc cag gaa ctg ctc gag tgc cag 144Arg Asn Asp Glu Glu Gln Phe Glu Cys Gln Glu Leu Leu Glu Cys Gln
35 40 45gtg cag gtg ggg gcc ccc gag gag gag gag gag gag gag gag gac gcg 192Val Gln Val Gly Ala Pro Glu Glu Glu Glu Glu Glu Glu Glu Asp Ala
50 55 60ggc ctg gtg 201Gly Leu Val65<210>155<211>67<212>PRT<213>人(Homo sapiens)<400>155Met Ala Glu Asp Val Ser Ser Ala Ala Pro Ser Pro Arg Gly Cys Ala1 5 10 15Asp Gly Arg Asp Ala Asp Pro Thr Glu Glu Gln Met Ala Glu Thr Glu
20 25 30Arg Asn Asp Glu Glu Gln Phe Glu Cys Gln Glu Leu Leu Glu Cys Gln
35 40 45Val Gln Val Gly Ala Pro Glu Glu Glu Glu Glu Glu Glu Glu Asp Ala
50 55 60Gly Leu Val65<210>156<211>38<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>156cgcaggatcc ccttcactcc tcttcatgag gcagcttc 38<210>157<211>48<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>157ggatccgcta aatatctgta tctccatctt taacaagatc caaaggag 48<210>158<211>21<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>158gccgacttcg agtttgagca g 21<210>159<211>1103<212>DNA<213>人(Homo sapiens)<220><221>CDS<222>(9)..(1094)<400>159ggatcccc ttc act cct ctt cat gag gca gct tct aag aac agg gtt gaa 50
Phe Thr Pro Leu His Glu Ala Ala Ser Lys Asn Arg Val Glu
1 5 10gta tgt tct ctt ctc tta agt tat ggt gca gac cca aca ctg ctc aat 98Val Cys Ser Leu Leu Leu Ser Tyr Gly Ala Asp Pro Thr Leu Leu Asn15 20 25 30tgt cac aat aaa agt gct ata gac ttg gct ccc aca cca cag tta aaa 146Cys His Asn Lys Ser Ala Ile Asp Leu Ala Pro Thr Pro Gln Leu Lys
35 40 45gaa aga tta gca tat gaa ttt aaa ggc cac tcg ttg ctg caa gct gca 194Glu Arg Leu Ala Tyr Glu Phe Lys Gly His Ser Leu Leu Gln Ala Ala
50 55 60cga gaa gct gat gtt act cga atc aaa aaa cat ctc tct ctg gaa atg 242Arg Glu Ala Asp Val Thr Arg Ile Lys Lys His Leu Ser Leu Glu Met
65 70 75gtg aat ttc aag cat cct caa aca cat gaa aca gca ttg cat tgt gct 290Val Asn Phe Lys His Pro Gln Thr His Glu Thr Ala Leu His Cys Ala
80 85 90gct gca tct cca tat ccc aaa aga aag caa ata tgt gaa ctg ttg cta 338Ala Ala Ser Pro Tyr Pro Lys Arg Lys Gln Ile Cys Glu Leu Leu Leu95 100 105 110aga aaa gga gca aac atc aat gaa aag act aaa gaa ttc ttg act cct 386Arg Lys Gly Ala Asn Ile Asn Glu Lys Thr Lys Glu Phe Leu Thr Pro
115 120 125ctg cac gtg gca tct gag aaa gct cat aat gat gtt gtt gaa gta gtg 434Leu His Val Ala Ser Glu Lys Ala His Asn Asp Val Val Glu Val Val
130 135 140gtg aaa cat gaa gca aag gtt aat gct ctg gat aat ctt ggt cag act 482Val Lys His Glu Ala Lys Val Asn Ala Leu Asp Asn Leu Gly Gln Thr
145 150 155tct cta cac aga gct gca tat tgt ggt cat cta caa acc tgc cgc cta 530Ser Leu His Arg Ala Ala Tyr Cys Gly His Leu Gln Thr Cys Arg Leu
160 165 170ctc ctg agc tat ggg tgt gat cct aac att ata tcc ctt cag ggc ttt 578Leu Leu Ser Tyr Gly Cys Asp Pro Asn Ile Ile Ser Leu Gln Gly Phe175 180 185 190act gct tta cag atg gga aat gaa aat gta cag caa ctc ctc caa gag 626Thr Ala Leu Gln Met Gly Asn Glu Asn Val Gln Gln Leu Leu Gln Glu
195 200 205ggt atc tca tta ggt aat tca gag gca gac aga caa ttg ctg gaa gct 674Gly Ile Ser Leu Gly Asn Ser Glu Ala Asp Arg Gln Leu Leu Glu Ala
210 215 220gca aag gct gga gat gtc gaa act gta aaa aaa ctg tgt act gtt cag 722Ala Lys Ala Gly Asp Val Glu Thr Val Lys Lys Leu Cys Thr Val Gln
225 230 235agt gtc aac tgc aga gac att gaa ggg cgt cag tct aca cca ctt cat 770Ser Val Asn Cys Arg Asp Ile Glu Gly Arg Gln Ser Thr Pro Leu His
240 245 250ttt gca gct ggg tat aac aga gtg tcc gtg gtg gaa tat ctg cta cag 818Phe Ala Ala Gly Tyr Asn Arg Val Ser Val Val Glu Tyr Leu Leu Gln255 260 265 270cat gga gct gat gtg cat gct aaa gat aaa gga ggc ctt gta cct ttg 866His Gly Ala Asp Val His Ala Lys Asp Lys Gly Gly Leu Val Pro Leu
275 280 285cac aat gca tgt tct tat gga cat tat gaa gtt gca gaa ctt ctt gtt 914His Asn Ala Cys Ser Tyr Gly His Tyr Glu Val Ala Glu Leu Leu Val
290 295 300aaa cat gga gca gta gtt aat gta gct gat tta tgg aaa ttt aca cct 962Lys His Gly Ala Val Val Asn Val Ala Asp Leu Trp Lys Phe Thr Pro
305 310 315tta cat gaa gca gca gca aaa gga aaa tat gaa att tgc aaa ctt ctg 1010Leu His Glu Ala Ala Ala Lys Gly Lys Tyr Glu Ile Cys Lys Leu Leu
320 325 330ctc cag cat ggt gca gac cct aca aaa aaa aac agg gat gga aat act 1058Leu Gln His Gly Ala Asp Pro Thr Lys Lys Asn Arg Asp Gly Asn Thr335 340 345 350cct ttg gat ctt gtt aaa gat gga gat aca gat att tagcggatc 1103Pro Leu Asp Leu Val Lys Asp Gly Asp Thr Asp Ile
355 360<210>160<211>362<212>PRT<213>人(Homo sapiens)<400>160Phe Thr Pro Leu His Glu Ala Ala Ser Lys Asn Arg Val Glu Val Cys1 5 10 15Ser Leu Leu Leu Ser Tyr Gly Ala Asp Pro Thr Leu Leu Asn Cys His
20 25 30Asn Lys Ser Ala Ile Asp Leu Ala Pro Thr Pro Gln Leu Lys Glu Arg
35 40 45Leu Ala Tyr Glu Phe Lys Gly His Ser Leu Leu Gln Ala Ala Arg Glu
50 55 60Ala Asp Val Thr Arg Ile Lys Lys His Leu Ser Leu Glu Met Val Asn65 70 75 80Phe Lys His Pro Gln Thr His Glu Thr Ala Leu His Cys Ala Ala Ala
85 90 95Ser Pro Tyr Pro Lys Arg Lys Gln Ile Cys Glu Leu Leu Leu Arg Lys
100 105 110Gly Ala Asn Ile Asn Glu Lys Thr Lys Glu Phe Leu Thr Pro Leu His
115 120 125Val Ala Ser Glu Lys Ala His Asn Asp Val Val Glu Val Val Val Lys
130 135 140His Glu Ala Lys Val Asn Ala Leu Asp Asn Leu Gly Gln Thr Ser Leu145 150 155 160His Arg Ala Ala Tyr Cys Gly His Leu Gln Thr Cys Arg Leu Leu Leu
165 170 175Ser Tyr Gly Cys Asp Pro Asn Ile Ile Ser Leu Gln Gly Phe Thr Ala
180 185 190Leu Gln Met Gly Asn Glu Asn Val Gln Gln Leu Leu Gln Glu Gly Ile
195 200 205Ser Leu Gly Asn Ser Glu Ala Asp Arg Gln Leu Leu Glu Ala Ala Lys
210 215 220Ala Gly Asp Val Glu Thr Val Lys Lys Leu Cys Thr Val Gln Ser Val225 230 235 240Asn Cys Arg Asp Ile Glu Gly Arg Gln Ser Thr Pro Leu His Phe Ala
245 250 255Ala Gly Tyr Asn Arg Val Ser Val Val Glu Tyr Leu Leu Gln His Gly
260 265 270Ala Asp Val His Ala Lys Asp Lys Gly Gly Leu Val Pro Leu His Asn
275 280 285Ala Cys Ser Tyr Gly His Tyr Glu Val Ala Glu Leu Leu Val Lys His
290 295 300Gly Ala Val Val Asn Val Ala Asp Leu Trp Lys Phe Thr Pro Leu His305 310 315 320Glu Ala Ala Ala Lys Gly Lys Tyr Glu Ile Cys Lys Leu Leu Leu Gln
325 330 335His Gly Ala Asp Pro Thr Lys Lys Asn Arg Asp Gly Asn Thr Pro Leu
340 345 350Asp Leu Val Lys Asp Gly Asp Thr Asp Ile
355 360<210>161<211>39<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>161cgtcgaccca tggcggagtc ttcggataag ctctatcga 39<210>162<211>39<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>162ggaaacgcgt ttggtgccag gatttactgt cagcttctt 39<210>163<211>39<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>163cttaaacgcg ttgaaggaca aacaccttta gatttagtt 39<210>164<211>79<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>164gtcgaaagcg gccgcttagc ctccgaactg tggatgcctc cacgctccat cgaccatacc 60ttcaggcctc ataatctgg 79<210>165<211>17<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>165tttgttcgcc cagactc 17<210>166<211>22<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>166tatgtttcag gttcaggggg ag 22<210>167<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>167gcggaagctg gaggagtgac 20<210>168<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>168gtcactcctc cagcttccgc 20<210>169<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>169aagccctgaa gaagcagctc 20<210>170<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>170gagctgcttc ttcagggctt 20<210>171<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>171cagacaccca accggaagga 20<210>172<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>172tccttccggt tgggtgtctg 20<210>173<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>173tccgcctcca ccaagagcct 20<210>174<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>174aggctcttgg tggaggcgga 20<210>175<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>175tggcctggtg gacatcgtta 20<210>176<211>20<212>DNA<213>人工序列<220><223>人工序列说明:引物<400>176taacgatgtc caccaggcca 20<210>177<211>3308<212>DNA<213>人工序列<220><223>人工序列说明:Parp1a-Tank2b
融合物<220><221>CDS<222>(1)..(3297)<400>177atg aga ggc tcc cat cac cat cac cat cac gat tac gat atc cca acg 48Met Arg Gly Ser His His His His His His Asp Tyr Asp Ile Pro Thr1 5 10 15acc gaa aac ctg tat ttt cag ggc gcc atg gat ccg gaa ttc aaa ggc 96Thr Glu Asn Leu Tyr Phe Gln Gly Ala Met Asp Pro Glu Phe Lys Gly
20 25 30cta cgt cga ccc atg gcg gag tct tcg gat aag ctc tat cga gtc gag 144Leu Arg Arg Pro Met Ala Glu Ser Ser Asp Lys Leu Tyr Arg Val Glu
35 40 45tac gcc aag agc ggg cgc gcc tct tgc aag aaa tgt agc gag agc atc 192Tyr Ala Lys Ser Gly Arg Ala Ser Cys Lys Lys Cys Ser Glu Ser Ile
50 55 60ccc aag gac tcg ctc cgg atg gcc atc atg gtg cag tcg ccc atg ttt 240Pro Lys Asp Ser Leu Arg Met Ala Ile Met Val Gln Ser Pro Met Phe65 70 75 80gat gga aaa gtc cca cac tgg tac cac ttc tcc tgc ttc tgg aag gtg 288Asp Gly Lys Val Pro His Trp Tyr His Phe Ser Cys Phe Trp Lys Val
85 90 95ggc cac tcc atc cgg cac cct gac gtt gag gtg gat ggg ttc tct gag 336Gly His Ser Ile Arg His Pro Asp Val Glu Val Asp Gly Phe Ser Glu
100 105 110ctt cgg tgg gat gac cag cag aaa gtc aag aag aca gcg gaa gct gga 384Leu Arg Trp Asp Asp Gln Gln Lys Val Lys Lys Thr Ala Glu Ala Gly
115 120 125gga gtg aca ggc aaa ggc cag gat gga att ggt agc aag gca gag aag 432Gly Val Thr Gly Lys Gly Gln Asp Gly Ile Gly Ser Lys Ala Glu Lys
130 135 140act ctg ggt gac ttt gca gca gag tat gtc aag tcc aac aga agt acg 480Thr Leu Gly Asp Phe Ala Ala Glu Tyr Val Lys Ser Asn Arg Ser Thr145 150 155 160tgc aag ggg tgt atg gag aag ata gaa aag ggc cag gtg cgc ctg tcc 528Cys Lys Gly Cys Met Glu Lys Ile Glu Lys Gly Gln Val Arg Leu Ser
165 170 175aag aag atg gtg gac ccg gag aag cca cag cta ggc atg att gac cgc 576Lys Lys Met Val Asp Pro Glu Lys Pro Gln Leu Gly Met Ile Asp Arg
180 185 190tgg tac cat cca ggc tgc ttt gtc aag aac agg gag gag ctg ggt ttc 624Trp Tyr His Pro Gly Cys Phe Val Lys Asn Arg Glu Glu Leu Gly Phe
195 200 205cgg ccc gag tac agt gcg agt cag ctc aag ggc ttc agc ctc ctt gct 672Arg Pro Glu Tyr Ser Ala Ser Gln Leu Lys Gly Phe Ser Leu Leu Ala
210 215 220aca gag gat aaa gaa gcc ctg aag aag cag ctc cca gga gtc aag agt 720Thr Glu Asp Lys Glu Ala Leu Lys Lys Gln Leu Pro Gly Val Lys Ser225 230 235 240gaa gga aag aga aaa ggc gat gag gtg gat gga gtg gat gaa gtg gcg 768Glu Gly Lys Arg Lys Gly Asp Glu Val Asp Gly Val Asp Glu Val Ala
245 250 255aag aag aaa tct aaa aaa gaa aaa gac aag gat agt aag ctt gaa aaa 816Lys Lys Lys Ser Lys Lys Glu Lys Asp Lys Asp Ser Lys Leu Glu Lys
260 265 270gcc cta aag gct cag aac gac ctg atc tgg aac atc aag gac gag cta 864Ala Leu Lys Ala Gln Asn Asp Leu Ile Trp Asn Ile Lys Asp Glu Leu
275 280 285aag aaa gtg tgt tca act aat gac ctg aag gag cta ctc atc ttc aac 912Lys Lys Val Cys Ser Thr Asn Asp Leu Lys Glu Leu Leu Ile Phe Asn
290 295 300aag cag caa gtg cct tct ggg gag tcg gcg atc ttg gac cga gta gct 960Lys Gln Gln Val Pro Ser Gly Glu Ser Ala Ile Leu Asp Arg Val Ala305 310 315 320gat ggc atg gtg ttc ggt gcc ctc ctt ccc tgc gag gaa tgc tcg ggt 1008Asp Gly Met Val Phe Gly Ala Leu Leu Pro Cys Glu Glu Cys Ser Gly
325 330 335cag ctg gtc ttc aag agc gat gcc tat tac tgc act ggg gac gtc act 1056Gln Leu Val Phe Lys Ser Asp Ala Tyr Tyr Cys Thr Gly Asp Val Thr
340 345 350gcc tgg acc aag tgt atg gtc aag aca cag aca ccc aac cgg aag gag 1104Ala Trp Thr Lys Cys Met Val Lys Thr Gln Thr Pro Asn Arg Lys Glu
355 360 365tgg gta acc cca aag gaa ttc cga gaa atc tct tac ctc aag aaa ttg 1152Trp Val Thr Pro Lys Glu Phe Arg Glu Ile Ser Tyr Leu Lys Lys Leu
370 375 380aag gtt aaa aag cag gac cgt ata ttc ccc cca gaa acc agc gcc tcc 1200Lys Val Lys Lys Gln Asp Arg Ile Phe Pro Pro Glu Thr Ser Ala Ser385 390 395 400gtg gcg gcc acg cct ccg ccc tcc aca gcc tcg gct cct gct gct gtg 1248Val Ala Ala Thr Pro Pro Pro Ser Thr Ala Ser Ala Pro Ala Ala Val
405 410 415aac tcc tct gct tca gca gat aag cca tta tcc aac atg aag atc ctg 1296Asn Ser Ser Ala Ser Ala Asp Lys Pro Leu Ser Asn Met Lys Ile Leu
420 425 430act ctc ggg aag ctg tcc cgg aac aag gat gaa gtg aag gcc atg att 1344Thr Leu Gly Lys Leu Ser Arg Asn Lys Asp Glu Val Lys Ala Met Ile
435 440 445gag aaa ctc ggg ggg aag ttg acg ggg acg gcc aac aag gct tcc ctg 1392Glu Lys Leu Gly Gly Lys Leu Thr Gly Thr Ala Asn Lys Ala Ser Leu
450 455 460tgc atc agc acc aaa aag gag gtg gaa aag atg aat aag aag atg gag 1440Cys Ile Ser Thr Lys Lys Glu Val Glu Lys Met Asn Lys Lys Met Glu465 470 475 480gaa gta aag gaa gcc aac atc cga gtt gtg tct gag gac ttc ctc cag 1488Glu Val Lys Glu Ala Asn Ile Arg Val Val Ser Glu Asp Phe Leu Gln
485 490 495gac gtc tcc gcc tcc acc aag agc ctt cag gag ttg ttc tta gcg cac 1536Asp Val Ser Ala Ser Thr Lys Ser Leu Gln Glu Leu Phe Leu Ala His
500 505 510atc ttg tcc cct tgg ggg gca gag gtg aag gca gag cct gtt gaa gtt 1584Ile Leu Ser Pro Trp Gly Ala Glu Val Lys Ala Glu Pro Val Glu Val
515 520 525gtg gcc cca aga ggg aag tca ggg gct gcg ctc tcc aaa aaa agc aag 1632Val Ala Pro Arg Gly Lys Ser Gly Ala Ala Leu Ser Lys Lys Ser Lys
530 535 540ggc cag gtc aag gag gaa ggt atc aac aaa tct gaa aag aga atg aaa 1680Gly Gln Val Lys Glu Glu Gly Ile Asn Lys Ser Glu Lys Arg Met Lys545 550 555 560tta act ctt aaa gga gga gca gct gtg gat cct gat tct gga ctg gaa 1728Leu Thr Leu Lys Gly Gly Ala Ala Val Asp Pro Asp Ser Gly Leu Glu
565 570 575cac tct gcg cat gtc ctg gag aaa ggt ggg aag gtc ttc agt gcc acc 1776His Ser Ala His Val Leu Glu Lys Gly Gly Lys Val Phe Ser Ala Thr
580 585 590ctt ggc ctg gtg gac atc gtt aaa gga acc aac tcc tac tac aag ctg 1824Leu Gly Leu Val Asp Ile Val Lys Gly Thr Asn Ser Tyr Tyr Lys Leu
595 600 605cag ctt ctg gag gac gac aag gaa aac agg tat tgg ata ttc agg tcc 1872Gln Leu Leu Glu Asp Asp Lys Glu Asn Arg Tyr Trp Ile Phe Arg Ser
610 615 620tgg ggc cgt gtg ggt acg gtg atc ggt agc aac aaa ctg gaa cag atg 1920Trp Gly Arg Val Gly Thr Val Ile Gly Ser Asn Lys Leu Glu Gln Met625 630 635 640ccg tcc aag gag gat gcc att gag cac ttc atg aaa tta tat gaa gaa 1968Pro Ser Lys Glu Asp Ala Ile Glu His Phe Met Lys Leu Tyr Glu Glu
645 650 655aaa acc ggg aac gct tgg cac tcc aaa aat ttc acg aag tat ccc aaa 2016Lys Thr Gly Asn Ala Trp His Ser Lys Asn Phe Thr Lys Tyr Pro Lys
660 665 670aag ttc tac ccc ctg gag att gac tat ggc cag gat gaa gag gca gtg 2064Lys Phe Tyr Pro Leu Glu Ile Asp Tyr Gly Gln Asp Glu Glu Ala Val
675 680 685aag aag ctg aca gta aat cct ggc acc aaa cgc gtt gaa gga caa aca 2112Lys Lys Leu Thr Val Asn Pro Gly Thr Lys Arg Val Glu Gly Gln Thr
690 695 700cct tta gat tta gtt tca gca gat gat gtc agc gct ctt ctg aca gca 2160Pro Leu Asp Leu Val Ser Ala Asp Asp Val Ser Ala Leu Leu Thr Ala705 710 715 720gcc atg ccc cca tct gct ctg ccc tct tgt tac aag cct caa gtg ctc 2208Ala Met Pro Pro Ser Ala Leu Pro Ser Cys Tyr Lys Pro Gln Val Leu
725 730 735aat ggt gtg aga agc cca gga gcc act gca gat gct ctc tct tca ggt 2256Asn Gly Val Arg Ser Pro Gly Ala Thr Ala Asp Ala Leu Ser Ser Gly
740 745 750cca tct agc cca tca agc ctt tct gca gcc agc agt ctt gac aac tta 2304Pro Ser Ser Pro Ser Ser Leu Ser Ala Ala Ser Ser Leu Asp Asn Leu
755 760 765tct ggg agt ttt tca gaa ctg tct tca gta gtt agt tca agt gga aca 2352Ser Gly Ser Phe Ser Glu Leu Ser Ser Val Val Ser Ser Ser Gly Thr
770 775 780gag ggt gct tcc agt ttg gag aaa aag gag gtt cca gga gta gat ttt 2400Glu Gly Ala Ser Ser Leu Glu Lys Lys Glu Val Pro Gly Val Asp Phe785 790 795 800agc ata act caa ttc gta agg aat ctt gga ctt gag cac cta atg gat 2448Ser Ile Thr Gln Phe Val Arg Asn Leu Gly Leu Glu His Leu Met Asp
805 810 815ata ttt gag aga gaa cag atc act ttg gat gta tta gtt gag atg ggg 2496Ile Phe Glu Arg Glu Gln Ile Thr Leu Asp Val Leu Val Glu Met Gly
820 825 830cac aag gag ctg aag gag att gga atc aat gct tat gga cat agg cac 2544His Lys Glu Leu Lys Glu Ile Gly Ile Asn Ala Tyr Gly His Arg His
835 840 845aaa cta att aaa gga gtc gag aga ctt atc tcc gga caa caa ggt ctt 2592Lys Leu Ile Lys Gly Val Glu Arg Leu Ile Ser Gly Gln Gln Gly Leu
850 855 860aac cca tat tta act ttg aac acc tct ggt agt gga aca att ctt ata 2640Asn Pro Tyr Leu Thr Leu Asn Thr Ser Gly Ser Gly Thr Ile Leu Ile865 870 875 880gat ctg tct cct gat gat aaa gag ttt cag tct gtg gag gaa gag atg 2688Asp Leu Ser Pro Asp Asp Lys Glu Phe Gln Ser Val Glu Glu Glu Met
885 890 895caa agt aca gtt cga gag cac aga gat gga ggt cat gca ggt gga atc 2736Gln Ser Thr Val Arg Glu His Arg Asp Gly Gly His Ala Gly Gly Ile
900 905 910ttc aac aga tac aat att ctc aag att cag aag gtt tgt aac aag aaa 2784Phe Asn Arg Tyr Asn Ile Leu Lys Ile Gln Lys Val Cys Asn Lys Lys
915 920 925cta tgg gaa aga tac act cac cgg aga aaa gaa gtt tct gaa gaa aac 2832Leu Trp Glu Arg Tyr Thr His Arg Arg Lys Glu Val Ser Glu Glu Asn
930 935 940cac aac cat gcc aat gaa cga atg cta ttt cat ggg tct cct ttt gtg 2880His Asn His Ala Asn Glu Arg Met Leu Phe His Gly Ser Pro Phe Val945 950 955 960aat gca att atc cac aaa ggc ttt gat gaa agg cat gcg tac ata ggt 2928Asn Ala Ile Ile His Lys Gly Phe Asp Glu Arg His Ala Tyr Ile Gly
965 970 975ggt atg ttt gga gct ggc att tat ttt gct gaa aac tct tcc aaa agc 2976Gly Met Phe Gly Ala Gly Ile Tyr Phe Ala Glu Asn Ser Ser Lys Ser
980 985 990aat caa tat gta tat gga att gga gga ggt act ggg tgt cca gtt cac 3024Asn Gln Tyr Val Tyr Gly Ile Gly Gly Gly Thr Gly Cys Pro Val His
995 1000 1005aaa gac aga tct tgt tac att tgc cac agg cag ctg ctc ttt tgc cgg 3072Lys Asp Arg Ser Cys Tyr Ile Cys His Arg Gln Leu Leu Phe Cys Arg1010 1015 1020gta acc ttg gga aag tct ttc ctg cag ttc agt gca atg aaa atg gca 3120Val Thr Leu Gly Lys Ser Phe Leu Gln Phe Ser Ala Met Lys Met Ala1025 1030 1035 1040cat tct cct cca ggt cat cac tca gtc act ggt agg ccc agt gta aat 3168His Ser Pro Pro Gly His His Ser Val Thr Gly Arg Pro Ser Val Asn
1045 1050 1055ggc cta gca tta gct gaa tat gtt att tac aga gga gaa cag gct tat 3216Gly Leu Ala Leu Ala Glu Tyr Val Ile Tyr Arg Gly Glu Gln Ala Tyr
1060 1065 1070cct gag tat tta att act tac cag att atg agg cct gaa ggt atg gtc 3264Pro Glu Tyr Leu Ile Thr Tyr Gln Ile Met Arg Pro Glu Gly Met Val
1075 1080 1085gat gga gcg tgg agg cat cca cag ttc gga ggc taagcggccg c 3308Asp Gly Ala Trp Arg His Pro Gln Phe Gly Gly1090 1095<210>178<211>1099<212>PRT<213>人工序列<223>人工序列说明:Parp1a-Tank2b
融合物<400>178Met Arg Gly Ser His His His His His His Asp Tyr Asp Ile Pro Thr1 5 10 15Thr Glu Asn Leu Tyr Phe Gln Gly Ala Met Asp Pro Glu Phe Lys Gly
20 25 30Leu Arg Arg Pro Met Ala Glu Ser Ser Asp Lys Leu Tyr Arg Val Glu
35 40 45Tyr Ala Lys Ser Gly Arg Ala Ser Cys Lys Lys Cys Ser Glu Ser Ile
50 55 60Pro Lys Asp Ser Leu Arg Met Ala Ile Met Val Gln Ser Pro Met Phe65 70 75 80Asp Gly Lys Val Pro His Trp Tyr His Phe Ser Cys Phe Trp Lys Val
85 90 95Gly His Ser Ile Arg His Pro Asp Val Glu Val Asp Gly Phe Ser Glu
100 105 110Leu Arg Trp Asp Asp Gln Gln Lys Val Lys Lys Thr Ala Glu Ala Gly
115 120 125Gly Val Thr Gly Lys Gly Gln Asp Gly Ile Gly Ser Lys Ala Glu Lys
130 135 140Thr Leu Gly Asp Phe Ala Ala Glu Tyr Val Lys Ser Asn Arg Ser Thr145 150 155 160Cys Lys Gly Cys Met Glu Lys Ile Glu Lys Gly Gln Val Arg Leu Ser
165 170 175Lys Lys Met Val Asp Pro Glu Lys Pro Gln Leu Gly Met Ile Asp Arg
180 185 190Trp Tyr His Pro Gly Cys Phe Val Lys Asn Arg Glu Glu Leu Gly Phe
195 200 205Arg Pro Glu Tyr Ser Ala Ser Gln Leu Lys Gly Phe Ser Leu Leu Ala
210 215 220Thr Glu Asp Lys Glu Ala Leu Lys Lys Gln Leu Pro Gly Val Lys Ser225 230 235 240Glu Gly Lys Arg Lys Gly Asp Glu Val Asp Gly Val Asp Glu Val Ala
245 250 255Lys Lys Lys Ser Lys Lys Glu Lys Asp Lys Asp Ser Lys Leu Glu Lys
260 265 270Ala Leu Lys Ala Gln Asn Asp Leu Ile Trp Asn Ile Lys Asp Glu Leu
275 280 285Lys Lys Val Cys Ser Thr Asn Asp Leu Lys Glu Leu Leu Ile Phe Asn
290 295 300Lys Gln Gln Val Pro Ser Gly Glu Ser Ala Ile Leu Asp Arg Val Ala305 310 315 320Asp Gly Met Val Phe Gly Ala Leu Leu Pro Cys Glu Glu Cys Ser Gly
325 330 335Gln Leu Val Phe Lys Ser Asp Ala Tyr Tyr Cys Thr Gly Asp Val Thr
340 345 350Ala Trp Thr Lys Cys Met Val Lys Thr Gln Thr Pro Asn Arg Lys Glu
355 360 365Trp Val Thr Pro Lys Glu Phe Arg Glu Ile Ser Tyr Leu Lys Lys Leu
370 375 380Lys Val Lys Lys Gln Asp Arg Ile Phe Pro Pro Glu Thr Ser Ala Ser385 390 395 400Val Ala Ala Thr Pro Pro Pro Ser Thr Ala Ser Ala Pro Ala Ala Val
405 410 415Asn Ser Ser Ala Ser Ala Asp Lys Pro Leu Ser Asn Met Lys Ile Leu
420 425 430Thr Leu Gly Lys Leu Ser Arg Asn Lys Asp Glu Val Lys Ala Met Ile
435 440 445Glu Lys Leu Gly Gly Lys Leu Thr Gly Thr Ala Asn Lys Ala Ser Leu
450 455 460Cys Ile Ser Thr Lys Lys Glu Val Glu Lys Met Asn Lys Lys Met Glu465 470 475 480Glu Val Lys Glu Ala Asn Ile Arg Val Val Ser Glu Asp Phe Leu Gln
485 490 495Asp Val Ser Ala Ser Thr Lys Ser Leu Gln Glu Leu Phe Leu Ala His
500 505 510Ile Leu Ser Pro Trp Gly Ala Glu Val Lys Ala Glu Pro Val Glu Val
515 520 525Val Ala Pro Arg Gly Lys Ser Gly Ala Ala Leu Ser Lys Lys Ser Lys
530 535 540Gly Gln Val Lys Glu Glu Gly Ile Asn Lys Ser Glu Lys Arg Met Lys545 550 555 560Leu Thr Leu Lys Gly Gly Ala Ala Val Asp Pro Asp Ser Gly Leu Glu
565 570 575His Ser Ala His Val Leu Glu Lys Gly Gly Lys Val Phe Ser Ala Thr
580 585 590Leu Gly Leu Val Asp Ile Val Lys Gly Thr Asn Ser Tyr Tyr Lys Leu
595 600 605Gln Leu Leu Glu Asp Asp Lys Glu Asn Arg Tyr Trp Ile Phe Arg Ser
610 615 620Trp Gly Arg Val Gly Thr Val Ile Gly Ser Asn Lys Leu Glu Gln Met625 630 635 640Pro Ser Lys Glu Asp Ala Ile Glu His Phe Met Lys Leu Tyr Glu Glu
645 650 655Lys Thr Gly Asn Ala Trp His Ser Lys Asn Phe Thr Lys Tyr Pro Lys
660 665 670Lys Phe Tyr Pro Leu Glu Ile Asp Tyr Gly Gln Asp Glu Glu Ala Val
675 680 685Lys Lys Leu Thr Val Asn Pro Gly Thr Lys Arg Val Glu Gly Gln Thr
690 695 700Pro Leu Asp Leu Val Ser Ala Asp Asp Val Ser Ala Leu Leu Thr Ala705 710 715 720Ala Met Pro Pro Ser Ala Leu Pro Ser Cys Tyr Lys Pro Gln Val Leu
725 730 735Asn Gly Val Arg Ser Pro Gly Ala Thr Ala Asp Ala Leu Ser Ser Gly
740 745 750Pro Ser Ser Pro Ser Ser Leu Ser Ala Ala Ser Ser Leu Asp Asn Leu
755 760 765Ser Gly Ser Phe Ser Glu Leu Ser Ser Val Val Ser Ser Ser Gly Thr
770 775 780Glu Gly Ala Ser Ser Leu Glu Lys Lys Glu Val Pro Gly Val Asp Phe785 790 795 800Ser Ile Thr Gln Phe Val Arg Asn Leu Gly Leu Glu His Leu Met Asp
805 810 815Ile Phe Glu Arg Glu Gln Ile Thr Leu Asp Val Leu Val Glu Met Gly
820 825 830His Lys Glu Leu Lys Glu Ile Gly Ile Asn Ala Tyr Gly His Arg His
835 840 845Lys Leu Ile Lys Gly Val Glu Arg Leu Ile Ser Gly Gln Gln Gly Leu
850 855 860Asn Pro Tyr Leu Thr Leu Asn Thr Ser Gly Ser Gly Thr Ile Leu Ile865 870 875 880Asp Leu Ser Pro Asp Asp Lys Glu Phe Gln Ser Val Glu Glu Glu Met
885 890 895Gln Ser Thr Val Arg Glu His Arg Asp Gly Gly His Ala Gly Gly Ile
900 905 910Phe Asn Arg Tyr Asn Ile Leu Lys Ile Gln Lys Val Cys Asn Lys Lys
915 920 925Leu Trp Glu Arg Tyr Thr His Arg Arg Lys Glu Val Ser Glu Glu Asn
930 935 940His Asn His Ala Asn Glu Arg Met Leu Phe His Gly Ser Pro Phe Val945 950 955 960Asn Ala Ile Ile His Lys Gly Phe Asp Glu Arg His Ala Tyr Ile Gly
965 970 975Gly Met Phe Gly Ala Gly Ile Tyr Phe Ala Glu Asn Ser Ser Lys Ser
980 985 990Asn Gln Tyr Val Tyr Gly Ile Gly Gly Gly Thr Gly Cys Pro Val His
995 1000 1005Lys Asp Arg Ser Cys Tyr Ile Cys His Arg Gln Leu Leu Phe Cys Arg1010 1015 1020Val Thr Leu Gly Lys Ser Phe Leu Gln Phe Ser Ala Met Lys Met Ala1025 1030 1035 1040His Ser Pro Pro Gly His His Ser Val Thr Gly Arg Pro Ser Val Asn
1045 1050 1055Gly Leu Ala Leu Ala Glu Tyr Val Ile Tyr Arg Gly Glu Gln Ala Tyr
1060 1065 1070Pro Glu Tyr Leu Ile Thr Tyr Gln Ile Met Arg Pro Glu Gly Met Val
1075 1080 1085Asp Gly Ala Trp Arg His Pro Gln Phe Gly Gly1090 1095
Claims (26)
1.纯化的和分离的坦科聚合酶2多肽。
2.权利要求1的多肽,包括SEQ ID NO:133所示氨基酸序列。
3.权利要求1的多肽,包括SEQ ID NO:135所示氨基酸序列。
4.一种编码权利要求1的多肽的多核苷酸。
5.权利要求4的多核苷酸,包括SEQ ID NO:132所示核苷酸序列的编码区。
6.权利要求4的多核苷酸,包括SEQ ID NO:134所示核苷酸序列的编码区。
7.一种选自下列一组的多核苷酸:
(a)权利要求4的多核苷酸,
(b)互补于(a)的多核苷酸的多核苷酸,
(c)能在中等严格杂交条件下与(a)或(b)的多核苷酸杂交的多核苷酸。
8.权利要求7的多核苷酸,其中,该多核苷酸是DNA分子或RNA分子。
9.权利要求8的多核苷酸,还包括一个可检测的标记部分。
10.一种表达结构,包括权利要求4的多核苷酸。
11.一种用权利要求10的表达结构所转化或转染过的宿主细胞。
12.权利要求4的多核苷酸,其中,该多核苷酸可操作地连接于异源启动子上。
13.一种宿主细胞,包括权利要求12的多核苷酸。
14.一种用于生产坦科聚合酶2多肽的方法,包括以下步骤:
a)在适合表达该多肽的条件下生长权利要求11或13的宿主细胞;
b)从宿主细胞中或宿主细胞所生长的培养基中分离所述多肽。
15.一种能够与权利要求1的多肽发生专一性免疫反应的抗体。
16.权利要求15的抗体,其中,该抗体选自下列一组:单克隆抗体、多克隆抗体、单链抗体(scFv抗体)、嵌合抗体、双功能/双专一性抗体、人源化抗体、人抗体、CDR-移植抗体、Fab片段、Fab’片段、F(ab’)2片段、和Fv片段。
17.一种能产生权利要求15的抗体的细胞系。
18.一种能与权利要求15的抗体发生专一性免疫反应的抗独特型抗体。
19.一种用于鉴定坦科聚合酶2多肽的结合配偶体的方法,包括:
a)在容许坦科聚合酶2多肽和测试化合物结合的条件下,让坦科聚合酶2多肽与一种测试化合物接触;
b)检测测试化合物和坦科聚合酶2多肽的结合;和
c)鉴定作为坦科聚合酶2多肽结合配偶体的测试化合物。
20.权利要求19的方法,其中,所述专一性结合配偶体能选择性地或专一性地调节坦科聚合酶2多肽的生物活性。
21.一种用于鉴定坦科聚合酶2多核苷酸的结合配偶体的方法,包括:
a)在容许坦科聚合酶2多核苷酸和测试化合物结合的条件下,让坦科聚合酶2多核苷酸与一种测试化合物接触;
b)检测测试化合物和坦科聚合酶2多核苷酸的结合;和
c)鉴定作为坦科聚合酶2多核苷酸专一性结合配偶体的测试化合物。
22.权利要求21的方法,其中,所述结合配偶体能选择性地或专一性地调节坦科聚合酶2多核苷酸的活性。
23.一种治疗具有由聚(ADP-核糖)聚合酶活性介导的医学症状的动物的方法,包括给所述动物服用能有效抑制该动物体内的坦科聚合酶2活性的量的坦科聚合酶2抑制化合物。
24.权利要求23的方法,其中,所述医学症状与肿瘤组织生长相关。
25.权利要求24的方法,其中,所述肿瘤组织是选自下列一组的癌:癌症、肉瘤、白血病和淋巴瘤。
26.权利要求25的方法,其中,所述癌选自下列一组:ACTH生成肿瘤,急性淋巴细胞性白血病、急性非淋巴细胞性白血病、肾上腺皮质癌、膀胱癌、脑癌、乳腺癌、宫颈癌、慢性淋巴细胞性白血病、慢性髓细胞性白血病、结肠直肠癌、皮肤T细胞淋巴瘤,子宫内膜癌、食道癌、Ewing’s肉瘤、胆囊癌、毛细胞白血病、头颈癌、Hodgkin’s淋巴瘤、Kaposi’s肉瘤、肾癌、肝癌、肺癌(小细胞和非小细胞),恶性腹膜渗出、恶性胸膜渗出、黑素瘤、间皮瘤、多发性骨髓瘤、成神经细胞瘤、神经胶质瘤、非Hodgkin’s淋巴瘤、骨肉瘤、卵巢癌、卵巢(生殖细胞)癌、胰腺癌、阴茎癌、前列腺癌、成视网膜细胞瘤、皮肤癌、软组织肉瘤、鳞状细胞癌、胃癌、睾丸癌、甲状腺癌、滋养层肿瘤、子宫癌、阴道癌、外阴癌和Wilm’s肿瘤。
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US14158299P | 1999-06-29 | 1999-06-29 | |
US60/141,582 | 1999-06-29 |
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US (1) | US20030190739A1 (zh) |
EP (1) | EP1192259A1 (zh) |
JP (1) | JP2003503062A (zh) |
KR (1) | KR20020025093A (zh) |
CN (1) | CN1371421A (zh) |
AU (1) | AU5775300A (zh) |
CA (1) | CA2370568A1 (zh) |
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CA2360318A1 (en) | 1999-04-09 | 2000-10-19 | Geron Corporation | A second mammalian tankyrase |
US20050074825A1 (en) * | 1999-10-25 | 2005-04-07 | Ying Luo | Tankyrase H, compositions involved in the cell cycle and methods of use |
US6589725B1 (en) | 1999-10-25 | 2003-07-08 | Rigel Pharmaceuticals, Inc. | Tankyrase H, compositions involved in the cell cycle and methods of use |
US6887675B1 (en) * | 1999-10-25 | 2005-05-03 | Rigel Pharmaceuticals, Inc. | Tankyrase H, compositions involved in the cell cycle and methods of use |
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US6277613B1 (en) * | 1998-06-10 | 2001-08-21 | The Rockefeller University | TRF1 binding protein, methods of use thereof |
CA2360318A1 (en) * | 1999-04-09 | 2000-10-19 | Geron Corporation | A second mammalian tankyrase |
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CA2370568A1 (en) | 2001-01-04 |
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KR20020025093A (ko) | 2002-04-03 |
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