CN1169955C - 编码具有抑制癌细胞生长功能的人蛋白的多核苷酸 - Google Patents
编码具有抑制癌细胞生长功能的人蛋白的多核苷酸 Download PDFInfo
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Abstract
本发明公开了一类新的具有抑癌功能的人蛋白,编码此多肽的多核苷酸和经重组技术产生该多肽的方法。本发明还公开了此多肽用于治疗多种疾病如癌症等的方法。本发明还公开了抗此多肽的拮抗剂及其治疗作用。本发明还公开了编码这类新的具有抑癌功能的人蛋白的多核苷酸的用途。
Description
本发明属于生物技术领域,具体地说,本发明涉及新的编码具有抑癌功能的人蛋白的多核苷酸,以及此多核苷酸编码的多肽。本发明还涉及此多核苷酸和多肽的用途和制备。
人基因组学研究目前是国际上的热点,除人染色体DNA大规模测序,表达序列测序(EST)的方法外,还缺少从功能开始的筛选具有功能基因的高通量的方法。
癌症是危害人类健康的主要疾病之一。为了有效地治疗和预防肿瘤,目前人们已越来越关注肿瘤的基因治疗。因此,本领域迫切需要开发研究具有抑癌功能的人蛋白及其激动剂/抑制剂。
本发明的目的是提供一类新的具有抑癌功能的人蛋白多肽以及其片段、类似物和衍生物。
本发明的另一目的是提供编码这些多肽的多核苷酸。
本发明的另一目的是提供生产这些多肽的方法以及该多肽和编码序列的用途。
在本发明的第一方面,提供新颖的分离出的具有抑癌功能的蛋白多肽,它包含具有选自下组的氨基酸序列的多肽:SEQ ID NO:2、SEQ ID NO:5、SEQ ID NO:8、SEQ IDNO:11、SEQ ID NO:14、SEQ ID NO:17、SEQ ID NO:20、SEQ ID NO:23、SEQ IDNO:26、SEQ ID NO:29、SEQ ID NO:32;或其保守性变异多肽、或其活性片段、或其活性衍生物。
较佳地,该多肽是具有选自下组的氨基酸序列的多肽:SEQ ID NO:2、SEQ IDNO:5、SEQ ID NO:8、SEQ ID NO:11、SEQ ID NO:14、SEQ ID NO:17、SEQ IDNO:20、SEQ ID NO:23、SEQ ID NO:26、SEQ ID NO:29、SEQ ID NO:32。
在本发明的第二方面,提供了一种分离的多核苷酸,它包含一核苷酸序列,该核苷酸序列与选自下组的一种核苷酸序列有至少85%相同性:(a)编码上述的具有抑癌功能的蛋白多肽的多核苷酸;(b)与多核苷酸(a)互补的多核苷酸。较佳地,该多核苷酸编码的多肽具有选自下组的氨基酸序列:SEQ ID NO:2、SEQ ID NO:5、SEQ ID NO:8、SEQ IDNO:11、SEQ ID NO:14、SEQ ID NO:17、SEQ ID NO:20、SEQ ID NO:23、SEQ IDNO:26、SEQ ID NO:29、SEQ ID NO:32。更佳地,该多核苷酸的序列选自下组:SEQ IDNO:3、SEQ ID NO:6、SEQ ID NO:9、SEQ ID NO:12、SEQ ID NO:15、SEQ ID NO:18、SEQ ID NO:21、SEQ ID NO:24、SEQ ID NO:27、SEQ ID NO:30、SEQ ID NO:33的编码区序列或全长序列。
在本发明的第三方面,提供了含有上述多核苷酸的载体,以及被该载体转化或转导的宿主细胞或者被上述多核苷酸直接转化或转导的宿主细胞。
在本发明的第四方面,提供了制备具有具有抑癌功能的蛋白活性的多肽的制备方法,该方法包含:(a)在适合表达具有抑癌功能的蛋白的条件下,培养上述被转化或转导的宿主细胞;(b)从培养物中分离出具有具有抑癌功能的蛋白活性的多肽。
在本发明的第五方面,提供了与上述的具有抑癌功能的蛋白多肽特异性结合的抗体。还提供了可用于检测的核酸分子,它含有上述的多核苷酸中连续的10-800个核苷酸。
在本发明的第六方面,提供了一种药物组合物,它含有安全有效量的本发明的具有抑癌功能的蛋白多肽以及药学上可接受的载体。这些药物组合物可治疗癌症以及细胞异常增殖等病症。
本发明的其它方面由于本文的技术的公开,对本领域的技术人员而言是显而易见的。
本发明采用大规模cDNA克隆转染癌细胞,在获得具有抑癌作用的基础上,经测序证明为新的基因,进一步得到全长cDNA克隆。DNA转染试验证明,本发明的具有抑癌功能的蛋白对癌细胞(肝癌细胞)具有抑制克隆形成的作用,其抑制率在50%或50%以上。
如本文所用,“分离的”是指物质从其原始环境中分离出来(如果是天然的物质,原始环境即是天然环境)。如活体细胞内的天然状态下的多聚核苷酸和多肽是没有分离纯化的,但同样的多聚核苷酸或多肽如从天然状态中同存在的其他物质中分开,则为分离纯化的。
如本文所用,“分离的具有抑癌功能的蛋白或多肽”是指具有抑癌功能的蛋白多肽基本上不含天然与其相关的其它蛋白、脂类、糖类或其它物质。本领域的技术人员能用标准的蛋白质纯化技术纯化具有抑癌功能的蛋白。基本上纯的多肽在非还原聚丙烯酰胺凝胶上能产生单一的主带。具有抑癌功能的蛋白多肽的纯度能用氨基酸序列分析。
本发明的多肽可以是重组多肽、天然多肽、合成多肽,优选重组多肽。本发明的多肽可以是天然纯化的产物,或是化学合成的产物,或使用重组技术从原核或真核宿主(例如,细菌、酵母、高等植物、昆虫和哺乳动物细胞)中产生。根据重组生产方案所用的宿主,本发明的多肽可以是糖基化的,或可以是非糖基化的。本发明的多肽还可包括或不包括起始的甲硫氨酸残基。
本发明还包括具有抑癌功能的人蛋白的片段、衍生物和类似物。如本文所用,术语“片段”、“衍生物”和“类似物”是指基本上保持本发明的天然具有抑癌功能的人蛋白相同的生物学功能或活性的多肽。本发明的多肽片段、衍生物或类似物可以是(i)有一个或多个保守或非保守性氨基酸残基(优选保守性氨基酸残基)被取代的多肽,而这样的取代的氨基酸残基可以是也可以不是由遗传密码编码的,或(ii)在一个或多个氨基酸残基中具有取代基团的多肽,或(iii)成熟多肽与另一个化合物(比如延长多肽半衰期的化合物,例如聚乙二醇)融合所形成的多肽,或(iv)附加的氨基酸序列融合到此多肽序列而形成的多肽(如前导序列或分泌序列或用来纯化此多肽的序列或蛋白原序列)。根据本文的教导,这些片段、衍生物和类似物属于本领域熟练技术人员公知的范围。
本发明的多核苷酸可以是DNA形式或RNA形式。DNA形式包括cDNA、基因组DNA或人工合成的DNA。DNA可以是单链的或是双链的。DNA可以是编码链或非编码链。以PP610蛋白(在本申请中,蛋白质的命名采用其克隆编号)为例,编码成熟多肽的编码区序列可以与SEQ ID NO:3所示的编码区序列相同或者是简并的变异体。如本文所用,“简并的变异体”在本发明中是指编码具有SEQ ID NO:2的蛋白质,但与SEQ ID NO:3所示的编码区序列有差别的核酸序列。以PP784蛋白为例,编码成熟多肽的编码区序列可以与SEQID NO:6所示的编码区序列相同或者是简并的变异体。如本文所用,“简并的变异体”在本发明中是指编码具有SEQ ID NO:5的蛋白质,但与SEQ ID NO:6所示的编码区序列有差别的核酸序列。对于其他具有抑癌功能的蛋白,可依此类推。对于其他具有抑癌功能的蛋白,可依此类推。
编码成熟多肽的多核苷酸包括:只编码成熟多肽的编码序列;成熟多肽的编码序列和各种附加编码序列;成熟多肽的编码序列(和任选的附加编码序列)以及非编码序列。
术语“编码多肽的多核苷酸”可以是包括编码此多肽的多核苷酸,也可以是还包括附加编码和/或非编码序列的多核苷酸。
本发明还涉及上述多核苷酸的变异体,其编码与本发明有相同的氨基酸序列的多肽或多肽的片段、类似物和衍生物。此多核苷酸的变异体可以是天然发生的等位变异体或非天然发生的变异体。这些核苷酸变异体包括取代变异体、缺失变异体和插入变异体。如本领域所知的,等位变异体是一个多核苷酸的替换形式,它可能是一个或多个核苷酸的取代、缺失或插入,但不会从实质上改变其编码的多肽的功能。
本发明还涉及与上述的序列杂交且两个序列之间具有至少50%,较佳地至少70%,更佳地至少80%相同性的多核苷酸。本发明特别涉及在严格条件下与本发明所述多核苷酸可杂交的多核苷酸。在本发明中,“严格条件”是指:(1)在较低离子强度和较高温度下的杂交和洗脱,如0.2×SSC,0.1%SDS,60℃;或(2)杂交时加有变性剂,如50%(v/v)甲酰胺,0.1%小牛血清/0.1%Ficoll,42℃等;或(3)仅在两条序列之间的相同性至少在95%以上,更好是97%以上时才发生杂交。并且,可杂交的多核苷酸编码的多肽与SEQ ID NO:2所示的成熟多肽有相同的生物学功能和活性。
本发明还涉及与上述的序列杂交的核酸片段。如本文所用,“核酸片段”的长度至少含15个核苷酸,较好是至少30个核苷酸,更好是至少50个核苷酸,最好是至少100个核苷酸以上。核酸片段可用于核酸的扩增技术(如PCR)以确定和/或分离编码具有抑癌功能的蛋白的多聚核苷酸。
本发明中的多肽和多核苷酸优选以分离的形式提供,更佳地被纯化至均质。
本发明的DNA序列能用几种方法获得。例如,用本领域熟知的杂交技术分离DNA。这些技术包括但不局限于:1)用探针与基因组或cDNA文库杂交以检出同源性核苷酸序列,和2)表达文库的抗体筛选以检出具有共同结构特征的克隆的DNA片段。
编码具有抑癌功能的蛋白的特异DNA片段序列产生也能用下列方法获得:1)从基因组DNA分离双链DNA序列;2)化学合成DNA序列以获得所需多肽的双链DNA。
上述提到的方法中,分离基因组DNA最不常用。当需要的多肽产物的整个氨基酸序列已知时,DNA序列的直接化学合成是经常选用的方法。如果所需的氨基酸的整个序列不清楚时,DNA序列的直接化学合成是不可能的,选用的方法是cDNA序列的分离。分离感兴趣的cDNA的标准方法是从高表达该基因的供体细胞分离mRNA并进行逆转录,形成质粒或噬菌体cDNA文库。提取mRNA的方法已有多种成熟的技术,试剂盒也可从商业途径获得(Qiagene)。而构建cDNA文库也是通常的方法(Sambrook,et al.,Molecular Cloning,A Laboratory Manual,Cold Spring Harbor Laboratory.New York,1989)。还可得到商业供应的cDNA文库,如Clontech公司的不同cDNA文库。当结合使用聚合酶反应技术时,即使极少的表达产物也能克隆。
可用常规方法从这些cDNA文库中筛选本发明的基因。这些方法包括(但不限于):(1)DNA-DNA或DNA-RNA杂交;(2)标志基因的功能出现或丧失;(3)测定具有抑癌功能的蛋白的转录本的水平;(4)通过免疫学技术或测定生物学活性,来检测基因表达的蛋白产物。上述方法可单用,也可多种方法联合应用。
在第(1)种方法中,杂交所用的探针是与本发明的多核苷酸的任何一部分同源,其长度至少15个核苷酸,较好是至少30个核苷酸,更好是至少50个核苷酸,最好是至少100个核苷酸。此外,探针的长度通常在2kb之内,较佳地为1kb之内。此处所用的探针通常是在本发明的基因DNA序列信息的基础上化学合成的DNA序列。本发明的基因本身或者片段当然可以用作探针。DNA探针的标记可用放射性同位素,荧光素或酶(如碱性磷酸酶)等。
在第(4)种方法中,检测具有抑癌功能的蛋白基因表达的蛋白产物可用免疫学技术如Western印迹法,放射免疫沉淀法,酶联免疫吸附法(ELISA)等。
应用PCR技术扩增DNA/RNA的方法(Saiki,et al.Science 1985;230:1350-1354)被优选用于获得本发明的基因。特别是很难从文库中得到全长的cDNA时,可优选使用RACE法(RACE-cDNA末端快速扩增法),用于PCR的引物可根据本文所公开的本发明的序列信息适当地选择,并可用常规方法合成。可用常规方法如通过凝胶电泳分离和纯化扩增的DNA/RNA片段。
如上所述得到的本发明的基因,或者各种DNA片段等的核苷酸序列的测定可用常规方法如双脱氧链终止法(Sanger et al.PNAS,1977,74:5463-5467)。这类核苷酸序列测定也可用商业测序试剂盒等。为了获得全长的cDNA序列,测序需反复进行。有时需要测定多个克隆的cDNA序列,才能拼接成全长的cDNA序列。
本发明也涉及包含本发明的多核苷酸的载体,以及用本发明的载体或具有抑癌功能的蛋白编码序列经基因工程产生的宿主细胞,以及经重组技术产生本发明所述多肽的方法。
通过常规的重组DNA技术,可利用本发明的多聚核苷酸序列可用来表达或生产重组的具有抑癌功能的蛋白多肽(Science,1984;224:1431)。一般来说有以下步骤:
(1).用本发明的编码具有抑癌功能的人蛋白的多核苷酸(或变异体),或用含有该多核苷酸的重组表达载体转化或转导合适的宿主细胞;
(2).在合适的培养基中培养的宿主细胞;
(3).从培养基或细胞中分离、纯化蛋白质。
本发明中,具有抑癌功能的人蛋白多核苷酸序列可插入到重组表达载体中。术语“重组表达载体”指本领域熟知的细菌质粒、噬菌体、酵母质粒、植物细胞病毒、哺乳动物细胞病毒如腺病毒、逆转录病毒或其他载体。在本发明中适用的载体包括但不限于:在细菌中表达的基于T7的表达载体(Rosenberg,et al.Gene,1987,56:125);在哺乳动物细胞中表达的pMSXND表达载体(Lee and Nathans,J Bio Chem.263:3521,1988)和在昆虫细胞中表达的来源于杆状病毒的载体。总之,只要能在宿主体内复制和稳定,任何质粒和载体都可以用。表达载体的一个重要特征是通常含有复制起点、启动子、标记基因和翻译控制元件。
本领域的技术人员熟知的方法能用于构建含具有抑癌功能的人蛋白编码DNA序列和合适的转录/翻译控制信号的表达载体。这些方法包括体外重组DNA技术、DNA合成技术、体内重组技术等(Sambroook,et al.Molecular Cloning,a Laboratory Manual,cold SpringHarbor Laboratory.New York,1989)。所述的DNA序列可有效连接到表达载体中的适当启动子上,以指导mRNA合成。这些启动子的代表性例子有:大肠杆菌的lac或trp启动子;λ噬菌体PL启动子;真核启动子包括CMV立即早期启动子、HSV胸苷激酶启动子、早期和晚期SV40启动子、反转录病毒的LTRs和其他一些已知的可控制基因在原核或真核细胞或其病毒中表达的启动子。表达载体还包括翻译起始用的核糖体结合位点和转录终止子。
此外,表达载体优选地包含一个或多个选择性标记基因,以提供用于选择转化的宿主细胞的表型性状,如真核细胞培养用的二氢叶酸还原酶、新霉素抗性以及绿色荧光蛋白(GFP),或用于大肠杆菌的四环素或氨苄青霉素抗性。
包含上述的适当DNA序列以及适当启动子或者控制序列的载体,可以用于转化适当的宿主细胞,以使其能够表达蛋白质。
宿主细胞可以是原核细胞,如细菌细胞;或是低等真核细胞,如酵母细胞;或是高等真核细胞,如哺乳动物细胞。代表性例子有:大肠杆菌,链霉菌属;鼠伤寒沙门氏菌的细菌细胞;真菌细胞如酵母;植物细胞;果蝇S2或Sf9的昆虫细胞;CHO、COS或Bowes黑素瘤细胞的动物细胞等。
本发明的多核苷酸在高等真核细胞中表达时,如果在载体中插入增强子序列时将会使转录得到增强。增强子是DNA的顺式作用因子,通常大约有10到300个碱基对,作用于启动子以增强基因的转录。可举的例子包括在复制起始点晚期一侧的100到270个碱基对的SV40增强子、在复制起始点晚期一侧的多瘤增强子以及腺病毒增强子等。
本领域一般技术人员都清楚如何选择适当的载体、启动子、增强子和宿主细胞。
用重组DNA转化宿主细胞可用本领域技术人员熟知的常规技术进行。当宿主为原核生物如大肠杆菌时,能吸收DNA的感受态细胞可在指数生长期后收获,用CaCl2法处理,所用的步骤在本领域众所周知。可供选择的是用MgCl2。如果需要,转化也可用电穿孔的方法进行。当宿主是真核生物,可选用如下的DNA转染方法:磷酸钙共沉淀法,常规机械方法如显微注射、电穿孔、脂质体包装等。
获得的转化子可以用常规方法培养,表达本发明的基因所编码的多肽。根据所用的宿主细胞,培养中所用的培养基可选自各种常规培养基。在适于宿主细胞生长的条件下进行培养。当宿主细胞生长到适当的细胞密度后,用合适的方法(如温度转换或化学诱导)诱导选择的启动子,将细胞再培养一段时间。
在上面的方法中的重组多肽可包被于细胞内、细胞外或在细胞膜上表达或分泌到细胞外。如果需要,可利用其物理的、化学的和其它特性通过各种分离方法分离和纯化重组的蛋白。这些方法是本领域技术人员所熟知的。这些方法的例子包括但并不限于:常规的复性处理、用蛋白沉淀剂处理(盐析方法)、离心、渗透破菌、超处理、超离心、分子筛层析(凝胶过滤)、吸附层析、离子交换层析、高效液相层析(HPLC)和其它各种液相层析技术及这些方法的结合。
重组的具有抑癌功能的人蛋白或多肽有多方面的用途。这些用途包括(但不限于):直接做为药物治疗具有抑癌功能的蛋白功能低下或丧失所致的疾病,和用于筛选促进或对抗具有抑癌功能的蛋白功能的抗体、多肽或其它配体。例如,抗体可用于激活或抑制具有抑癌功能的人蛋白的功能。用表达的重组具有抑癌功能的人蛋白筛选多肽库可用于寻找有治疗价值的能抑制或刺激具有抑癌功能的人蛋白功能的多肽分子。
本发明也提供了筛选药物以鉴定提高(激动剂)或阻遏(拮抗剂)具有抑癌功能的人蛋白的药剂的方法。激动剂提高具有抑癌功能的人蛋白刺激细胞增殖等生物功能,而拮抗剂阻止和治疗与细胞过度增殖有关的紊乱如各种癌症。例如,能在药物的存在下,将哺乳动物细胞或表达具有抑癌功能的人蛋白的膜制剂与标记的具有抑癌功能的人蛋白一起培养。然后测定药物提高或阻遏此相互作用的能力。
具有抑癌功能的人蛋白的拮抗剂包括筛选出的抗体、化合物、受体缺失物和类似物等。具有抑癌功能的人蛋白的拮抗剂可以与具有抑癌功能的人蛋白结合并消除其功能,或是抑制具有抑癌功能的人蛋白的产生,或是与多肽的活性位点结合使多肽不能发挥生物学功能。具有抑癌功能的人蛋白的拮抗剂可用于治疗用途。
在筛选作为拮抗剂的化合物时,可以将具有抑癌功能的蛋白加入生物分析测定中,通过测定化合物影响具有抑癌功能的蛋白和其受体之间的相互作用来确定化合物是否是拮抗剂。用上述筛选化合物的同样方法,可以筛选出起拮抗剂作用的受体缺失物和类似物。
本发明的多肽可直接用于疾病治疗,例如,各种恶性肿瘤、和细胞异常增殖等。
本发明的多肽,及其片段、衍生物、类似物或它们的细胞可以用来作为抗原以生产抗体。这些抗体可以是多克隆或单克隆抗体。多克隆抗体可以通过将此多肽直接注射动物的方法得到。制备单克隆抗体的技术包括杂交瘤技术,三瘤技术,人B-细胞杂交瘤技术,EBV-杂交瘤技术等。
可以将本发明的多肽和拮抗剂与合适的药物载体组合后使用。这些载体可以是水、葡萄糖、乙醇、盐类、缓冲液、甘油以及它们的组合。组合物包含安全有效量的多肽或拮抗剂以及不影响药物效果的载体和赋形剂。这些组合物可以作为药物用于疾病治疗。
本发明还提供含有一种或多种容器的药盒或试剂盒,容器中装有一种或多种本发明的药用组合物成分。与这些容器一起,可以有由制造、使用或销售药品或生物制品的政府管理机构所给出的指示性提示,该提示反映出生产、使用或销售的政府管理机构许可其在人体上施用。此外,本发明的多肽可以与其它的治疗化合物结合使用。
药物组合物可以以方便的方式给药,如通过局部、静脉内、腹膜内、肌内、皮下、鼻内或皮内的给药途径。具有抑癌功能的蛋白以有效地治疗和/或预防具体的适应症的量来给药。施用于患者的具有抑癌功能的蛋白的量和剂量范围将取决于许多因素,如给药方式、待治疗者的健康条件和诊断医生的判断。
具有抑癌功能的人蛋白的多聚核苷酸也可用于多种治疗目的。基因治疗技术可用于治疗由于具有抑癌功能的蛋白的无表达或异常/无活性的具有抑癌功能的蛋白的表达所致的细胞增殖、发育或代谢异常。重组的基因治疗载体(如病毒载体)可设计成表达变异的具有抑癌功能的蛋白,以抑制内源性的具有抑癌功能的蛋白活性。例如,一种变异的具有抑癌功能的蛋白可以是缩短的、缺失了信号传导功能域的具有抑癌功能的蛋白,虽可与下游的底物结合,但缺乏信号传导活性。因此重组的基因治疗载体可用于治疗具有抑癌功能的蛋白表达或活性异常所致的疾病。来源于病毒的表达载体如逆转录病毒、腺病毒、腺病毒相关病毒、单纯疱疹病毒、细小病毒等可用于将具有抑癌功能的蛋白基因转移至细胞内。构建携带具有抑癌功能的蛋白基因的重组病毒载体的方法可见于已有文献(Sambrook,etal.)。另外重组具有抑癌功能的人蛋白基因可包装到脂质体中转移至细胞内。
抑制具有抑癌功能的人蛋白mRNA的寡聚核苷酸(包括反义RNA和DNA)以及核酶也在本发明的范围之内。核酶是一种能特异性分解特定RNA的酶样RNA分子,其作用机制是核酶分子与互补的靶RNA特异性杂交后进行核酸内切作用。反义的RNA和DNA及核酶可用已有的任何RNA或DNA合成技术获得,如固相磷酸酰胺化学合成法合成寡核苷酸的技术已广泛应用。反义RNA分子可通过编码该RNA的DNA序列在体外或体内转录获得。这种DNA序列已整合到载体的RNA聚合酶启动子的下游。为了增加核酸分子的稳定性,可用多种方法对其进行修饰,如增加两侧的序列长度,核糖核苷之间的连接应用磷酸硫酯键或肽键而非磷酸二酯键。
多聚核苷酸导入组织或细胞内的方法包括:将多聚核苷酸直接注入到体内组织中;或在体外通过载体(如病毒、噬菌体或质粒等)先将多聚核苷酸导入细胞中,再将细胞移植到体内等。
本发明的多肽还可用作肽谱分析,例如,多肽可用物理的、化学或酶进行特异性切割,并进行一维或二维或三维的凝胶电泳分析。
本发明还提供了针对具有抑癌功能的人蛋白抗原决定簇的抗体。这些抗体包括(但不限于):多克隆抗体、单克隆抗体、嵌合抗体、单链抗体、Fab片段和Fab表达文库产生的片段。
抗具有抑癌功能的人蛋白的抗体可用于免疫组织化学技术中,检测活检标本中的具有抑癌功能的人蛋白。
与具有抑癌功能的人蛋白结合的单克隆抗体也可用放射性同位素标记,注入体内可跟踪其位置和分布。这种放射性标记的抗体可作为一种非创伤性诊断方法用于肿瘤细胞的定位和判断是否有转移。
本发明中的抗体可用于治疗或预防与具有抑癌功能的人蛋白相关的疾病。给予适当剂量的抗体可以刺激或阻断具有抑癌功能的人蛋白的产生或活性。
抗体也可用于设计针对体内某一特殊部位的免疫毒素。如具有抑癌功能的人蛋白高亲和性的单克隆抗体可与细菌或植物毒素(如白喉毒素,蓖麻蛋白,红豆碱等)共价结合。一种通常的方法是用巯基交联剂如SPDP,攻击抗体的氨基,通过二硫键的交换,将毒素结合于抗体上,这种杂交抗体可用于杀灭具有抑癌功能的人蛋白阳性的细胞。
多克隆抗体的生产可用具有抑癌功能的人蛋白或多肽免疫动物,如家兔,小鼠,大鼠等。多种佐剂可用于增强免疫反应,包括但不限于弗氏佐剂等。
具有抑癌功能的人蛋白单克隆抗体可用杂交瘤技术生产(Kohler and Milstein.Nature,1975,256:495-497)。将人恒定区和非人源的可变区结合的嵌合抗体可用已有的技术生产(Morrison et al,PNAS,1985,81:6851)。而已有的生产单链抗体的技术(U.S.PatNo.4946778)也可用于生产抗具有抑癌功能的人蛋白的单链抗体。
能与具有抑癌功能的人蛋白结合的多肽分子可通过筛选由各种可能组合的氨基酸结合于固相物组成的随机多肽库而获得。筛选时,必须对具有抑癌功能的人蛋白分子进行标记。
本发明还涉及定量和定位检测具有抑癌功能的人蛋白水平的诊断试验方法。这些试验是本领域所熟知的,且包括FISH测定和放射免疫测定。试验中所检测的具有抑癌功能的人蛋白水平,可以用作解释具有抑癌功能的人蛋白在各种疾病中的重要性和用于诊断具有抑癌功能的蛋白起作用的疾病。
具有抑癌功能的蛋白的多聚核苷酸可用于具有抑癌功能的蛋白相关疾病的诊断和治疗。在诊断方面,具有抑癌功能的蛋白的多聚核苷酸可用于检测具有抑癌功能的蛋白的表达与否或在疾病状态下具有抑癌功能的蛋白的异常表达。如具有抑癌功能的蛋白DNA序列可用于对活检标本的杂交以判断具有抑癌功能的蛋白的表达异常。杂交技术包括Southern印迹法,Northern印迹法、原位杂交等。这些技术方法都是公开的成熟技术,相关的试剂盒都可从商业途径得到。本发明的多核苷酸的一部分或全部可作为探针固定在微阵列(Microarray)或DNA芯片(又称为“基因芯片”)上,用于分析组织中基因的差异表达分析和基因诊断。用具有抑癌功能的蛋白特异的引物进行RNA-聚合酶链反应(RT-PCR)体外扩增也可检测具有抑癌功能的蛋白的转录产物。
检测具有抑癌功能的蛋白基因的突变也可用于诊断具有抑癌功能的蛋白相关的疾病。具有抑癌功能的蛋白突变的形式包括与正常野生型具有抑癌功能的蛋白DNA序列相比的点突变、易位、缺失、重组和其它任何异常等。可用已有的技术如Southern印迹法、DNA序列分析、PCR和原位杂交检测突变。另外,突变有可能影响蛋白的表达,因此用Northern印迹法、Western印迹法可间接判断基因有无突变。
本发明的序列对染色体鉴定也是有价值的。该序列会特异性地针对某条人染色体具体位置且并可以与其杂交。目前,需要鉴定染色体上的各基因的具体位点。现在,只有很少的基于实际序列数据(重复多态性)的染色体标记物可用于标记染色体位置。根据本发明,为了将这些序列与疾病相关基因相关联,其重要的第一步就是将这些DNA序列定位于染色体上。
简而言之,根据cDNA制备PCR引物(优选15-35bp),可以将序列定位于染色体上。然后,将这些引物用于PCR筛选含各条人染色体的体细胞杂合细胞。只有那些含有相应于引物的人基因的杂合细胞会产生扩增的片段。
体细胞杂合细胞的PCR定位法,是将DNA定位到具体染色体的快捷方法。使用本发明的的寡核苷酸引物,通过类似方法,可利用一组来自特定染色体的片段或大量基因组克隆而实现亚定位。可用于染色体定位的其它类似策略包括原位杂交、用标记的流式分选的染色体预筛选和杂交预选,从而构建染色体特异的cDNA库。
将cDNA克隆与中期染色体进行荧光原位杂交(FISH),可以在一个步骤中精确地进行染色体定位。此技术的综述,参见Verma等,Human Chromosomes:a Manual of BasicTechniques,Pergamon Press,New York(1988)。
一旦序列被定位到准确的染色体位置,此序列在染色体上的物理位置就可以与基因图数据相关联。这些数据可见于例如,V.Mckusick,Mendelian Inheritance in Man(可通过与Johns Hopkins University Welch Medical Library联机获得)。然后可通过连锁分析,确定基因与业已定位到染色体区域上的疾病之间的关系。
接着,需要测定患病和未患病个体间的cDNA或基因组序列差异。如果在一些或所有的患病个体中观察到某突变,而该突变在任何正常个体中未观察到,则该突变可能是疾病的病因。比较患病和未患病个体,通常涉及首先寻找染色体中结构的变化,如从染色体水平可见的或用基于cDNA序列的PCR可检测的缺失或易位。根据目前的物理作图和基因定位技术的分辨能力,被精确定位至与疾病有关的染色体区域的cDNA,可以是50至500个潜在致病基因间之一种(假定1兆碱基作图分辨能力和每20kb对应于一个基因)。
本发明的具有抑癌功能的蛋白核苷酸全长序列或其片段通常可以用PCR扩增法、重组法或人工合成的方法获得。对于PCR扩增法,可根据本发明所公开的有关核苷酸序列,尤其是开放阅读框序列来设计引物,并用市售的cDNA库或按本领域技术人员已知的常规方法所制备的cDNA库作为模板,扩增而得有关序列。当序列较长时,常常需要进行两次或多次PCR扩增,然后再将各次扩增出的片段按正确次序拼接在一起。
一旦获得了有关的序列,就可以用重组法来大批量地获得有关序列。这通常是将其克隆入载体,再转入细胞,然后通过常规方法从增殖后的宿主细胞中分离得到有关序列。
此外,还可用人工合成的方法来合成有关序列,尤其是片段长度较短时。通常,通过先合成多个小片段,然后再进行连接可获得序列很长的片段。
目前,已经可以完全通过化学合成来编码本发明蛋白(或其片段,或其衍生物)的DNA序列。然后可将该DNA序列引入本领域中的各种DNA分子(如载体)和细胞中。此外,还可通过化学合成将突变引入本发明蛋白序列中。
此外,由于本发明的具有抑癌功能的蛋白具有源自人的天然氨基酸序列,因此,与来源于其他物种的同族蛋白相比,预计在施用于人时将具有更高的活性和/或更低的副作用(例如在人体内的免疫原性更低或没有)。
下面结合具体实施例,进一步阐述本发明。应理解,这些实施例仅用于说明本发明而不用于限制本发明的范围。下列实施例中未注明具体条件的实验方法,通常按照常规条件如Sambrook等人,分子克隆:实验室手册(New York:Cold Spring Harbor LaboratoryPress,1989)中所述的条件,或按照制造厂商所建议的条件。
实施例1:cDNA基因的获得及对癌细胞克隆形成的抑制作用
PP610,PP784,PP827,PP1057,PP1551,PP1553,PP1665,PP2135,PP2281,PP2447,PP3111是通过用常规方法构建人胎盘cDNA文库获得的。取3、6、10月龄的胎盘组织,用Trizol试剂(GIBCO BRL公司)按厂方说明书提取总RNA,用mRNA提纯试剂盒(pharmacia公司)提取mRNA。用pCMV-script TMXR cDNA文库构建试剂盒(Seratagene公司)构建上述mRNA的cDNA文库。其中反转录酶改用MMLV-RT-Superscript II(GIBCO BRL),反转录反应在42℃进行。转化XL 10-Gold感受细胞,获得了1×106cfu/μg cDNA滴度的cDNA文库。第一轮随机挑取cDNA克隆,其后以高丰度cDNA克隆和已证明有抑癌细胞生长功能的cDNA克隆为探针,杂交筛选cDNA文库,挑取弱阳性及阴性克隆。用Qiagen 96孔板质粒抽提试剂盒,按厂家说明书进行质粒DNA的提取。质粒DNA和空载体同时转染肝癌细胞系7721。100ng DNA酒精沉淀干燥后,加6μlH2O溶解,待转染。每份DNA样品中加0.74μl脂质体及9.3μl无血清培液,混匀后,室温放置10分钟。每管中加150μl无血清培液,均分加入3孔生长于96孔板的7721细胞中,37℃放置2小时,每孔再加50μl无血清培液,37℃ 24小时。每孔换100μl全培液,37℃ 24小时,换含G418的全培液100μl,37℃ 24~48小时,边观察,边换G418浓度不等的培液。约2~3次后,直到镜检细胞有克隆形成,计数。发现以上克隆有抑制细胞克隆形成作用,结果如下表1所示。
表1cDNA克隆转染细胞(7721)克隆形成情况
cDNA克隆名称 | cDNA克隆数(三个重复) | 空载体克隆数(三个重复) |
PP610 | 0 0 0 | 21 30 42 |
PP784 | 1 3 2 | 18 24 25 |
PP827 | 5 3 1 | 31 38 32 |
PP1057 | 0 1 0 | 32 39 37 |
PP1551 | 0 0 0 | 27 25 26 |
PP1553 | 20 0 0 | 27 25 26 |
PP1665 | 17 18 18 | 22 25 13 |
PP2135 | 2 0 0 | 32 31 34 |
PP2281 | 0 0 0 | 41 36 33 |
PP2447 | 3 1 3 | 34 31 30 |
PP3111 | 0 1 0 | 12 18 20 |
对上述cDNA克隆采用双脱氧终止法,在ABI377 DNA自动测序仪上测定其一端近500bp的核苷酸序列。分析后,确定为新基因克隆,进行另一端测序,仍未获得全长cDNA序列,设计引物,再次进行测序,直到获得全长序列(SEQ ID NO:1、4、7、10、13、16、19、22、25、28、31)。
实施例2:从胎盘cDNA中PCR获得全长基因:
取3、6、10月龄的胎盘组织,用Trizol试剂(GIBCO BRL公司)按厂方说明书提取总RNA,用mRNA提纯试剂盒(pharmacia公司)提取mRNA。用MMLV-RT-Superscript II(GIBCO BRL),反转录酶在42℃进行反转录反应,获得胎盘cDNA。利用各个基因的特异引物(如下表所示),按97℃ 3’1个循环。94C 30″60℃ 30″72℃ 1’35个循环,72℃ 10’1个循环进行PCR扩增,获得含有完整开放阅读框序列的各蛋白基因的扩增产物。扩增产物经测序验证,与实施例1测得的序列相符,随后用常规技术将扩增产物转入宿主细胞,以获得重组蛋白。
基因特异引物
克隆名称 | 特异引物1(5′→3′) | 特异引物2(5′→3′) |
PP610 | GCTTCCTGCTCTCGCCGGAGTTT | GCAGGTGTGGACACCAGAGGGCT |
PP784 | CTTCCCGCAGAGTTGTGAAGCGA | ACGCCTAACAAAACCAGCACGCA |
PP827 | CCTCTGCACGCTAGGGTGGTCCT | CCATAAGGGAGACTGTTACATGGGGG |
PP1057 | GAGAGAAACGGGCTGGGGAGTGA | ACTCTGGCAAAGGGAGCCGTGAG |
PP1551 | GGAAGTGGCTGTTTGGCTGCTGA | TGCAGTGTGGATGCTCCTGGATG |
PP1553 | CAGGTGTGTGTGGGTCCTCCACC | TTGAAGGGCCCCAAAACAACAGC |
PP1665 | ACGCGTCCTGGAACCTGAGTGTG | ACACACCCACCCTGGCCATATCC |
PP2135 | CCTTCTGAGTCTGGGCGGCATCT | ATCTCCAGTGGGTGGGGCAACAT |
PP2281 | TCACTCTGGCTGCTGGGACATCA | CCACTTGTCCAAGAGCAGCTGGG |
PP2447 | CCTCTGCACGCTAGGGTGGTCCT | CAAGGGGTCAGTACCTGCTCGGG |
PP3111 | TGCGAGAGTAGCTGGCCAAGGTG | GGACATGGGCGTGTCTTTCCACA |
实施例3:cDNA克隆序列分析
1.PP610
A:核苷酸序列(SEQ ID NO:1)长度:1853bp
1 CTGAGCCTCT CCGTGCAATG ATTAACCCGG CGGGGCGGCC GGCGCGGGAC
51 CCGAGACGGA GGCGCGGGGC CGGGGCGGGA CCCCGCAGGA CCGCTCGGCT
101 TCCTGCTCTC GCCGGAGTTT CCGCGTAGAG GGCGCATCGC CGGCCCGGGG
151 CCCTTGGTGC GGCGTGGCGC AGGGCGCGGC GTGGGGCGCG CGTGGGCGCG
201 GCGCAGGCGG CCCGGGTCAC CATGAGGACT CTCCGCAGGT TGAAGTTCAT
251 GAGTTCGCCC AGCCTCAGTG ACCTGGGCAA GAGAGAGCCG GCCGCCGCCG
301 CGGACGAGCG GGGCACGCAG CAGCGCCGGG CCTGCGCCAA CGCCACCTGG
351 AACAGCATCC ACAACGGGGT GATCGCCGTC TTCCAGCGCA AGGGGCTGCC
401 CGACCAGGAG CTCTTCAGCC TCAACGAGGG CGTCCGGCAG CTGTTGAAGA
451 CAGAGCTGGG GTCCTTCTTC ACGGAGTACC TGCAGAACCA GCTGCTGACA
501 AAAGGCATGG TGATCCTrCG GGACAAGATT CGCTTCTATG AGGGACAGAA
551 GCTGCTGGAC TCACTGGCAG AGACCTGGGA CTTCTTCTTC AGTGACGTGC
601 TGCCCATGCT GCAGGCCATC TTCTACCCGG TGCAGGGCAA GGAGCCATCG
651 GTGCGCCAGC TGGCCCTGCT GCACTTCCGG AATGCCATCA CCCTCAGTGT
701 GAAGCTAGAG GATGCGCTGG CCCGGGCCCA TGCCCGTGTG CCCCCTGCCA
751 TCGTGCAGAT GCTGCTGGTG CTGCAGGGGG TACATGAGTC CAGGGGCGTG
801 ACTGAGGACT ACCTGCGCCT GGAGACGCTG GTCCAGAAGG TGGTGTCGCC
851 ATACCTGGGC ACCTACGGCC TCCACTCCAG CGAGGGGCCC TTCACCCATT
901 CCTGCATCCT GGAAAAGCGC CTCCTCCGCC GCTCCCGCTC GGGGGACGTG
951 CTGGCCAAGA ACCCTGTGGT GCGCTCCAAG AGCTACAACA CGCCTCTGCT
1001 GAACCCCGTG CAGGAGCACG AGGCGGAGGG CGCGGCGGCC GGCGGTACCA
1051 GCATCCGCAG GCACTCTGTG TCGGAGATGA CGTCCTGCCC CGAGCCTCAG
1101 GGCTTCTCCG ACCCGCCCGG CCAGGGCCCC ACCGGGACCT TCAGGTCCTC
1151 CCCGGCGCCC CACTCAGGGC CCTGCCCCAG CAGACTGTAC CCCACGACCC
1201 AGCCCCCTGA GCAGGGCTTG GATCCCACCC GCAGCTCCCT GCCCCGCTCC
1251 AGCCCGGAGA ACCTGGTGGA CCAGATCCTG GAGTCCGTGG ACTCGGATTC
1301 TGAAGGGATT TTCATTGACT TTGGCCGGGG CCGGGGCTCT GGCATGTCCG
1351 ACTTGGAGGG CTCTGGGGGC CGGCAGAGTG TCGTGTGAGG CCTCACAGCT
1401 GGCCTTGAGT TTTTACTGAC ACGTCCCTGT GTGCGGGGGT GTCCATGTGG
1451 CGTGTGTGTG AGTGAGACTT TTTTACTGCG TCCCGTCCCG CCAGCCCTAT
1501 CGGCCTCGTC ACTGGCCTTG GTCACTTTGT ATTTCTGTCT TGGTTGGAAA
1551 TACCATCAGC CTTCCTTGCT CGGCCCAGGT CTGTTTCAGG CATCTGAGTC
1601 GGCGTTTACC CAGGGGCCGG GCCAGAGACG GGGGTCGGCC GCTCGCTCCC
1651 ACGCTCCTCC TGCCCCAGCC CTCTGGTGTC CACACCTGCC CACAGAGAAT
1701 GTAAACCCAG TGGGCTCTGC CCACGCCGGG CCCCAAAGTG ACCAGACTCC
1751 AGCACACCTG TCTCCTCCTG CCTGGGGTGG CCATGGGGAT GGAAGGGGGT
1801 GGAATAAAAC CTGTCACCCT GGGAAAAAAA AAAAAAAAAA AAAAAAAAAA
1851 AAA
B:氨基酸序列(SEQ ID NO:2)长度:388个氨基酸
1 MRTLRRLKFM SSPSLSDLGK REPAAAADER GTQQRRACAN ATWNSIHNGV
51 IAVFQRKGLP DQELFSLNEG VRQLLKTELG SFFTEYLQNQ LLTKGMVILR
101 DKIRFYEGQK LLDSLAETWD FFFSDVLPML QAIFYPVQGK EPSVRQLALL
151 HFRNAITLSV KLEDALARAH ARVPPAIVQM LLVLQGVHES RGVTEDYLRL
201 ETLVQKVVSP YLGTYGLHSS EGPFTHSCIL EKRLLRRSRS GDVLAKNPVV
251 RSKSYNTPLL NPVQEHEAEG AAAGGTSIRR HSVSEMTSCP EPQGFSDPPG
301 QGPTGTFRSS PAPHSGPCPS RLYPTTQPPE QGLDPTRSSL PRSSPENLVD
351 QILESVDSDS EGIFIDFGRG RGSGMSDLEG SGGRQSVV
C.核苷酸及氨基酸组合序列(SEQ ID NO:3)
克隆号和蛋白名称:PP610
起始编码子: 222 ATG 终止编码子:1388 TGA
蛋白质分子量:42751.17
1 CT GAG CCT CTC CGT GCA ATG ATT AAC CCG GCG GGG CGG CCG GCG CGG 47
48 GAC CCG AGA CGG AGG CGC GGG GCC GGG GCG GGA CCC CGC AGG ACC GCT 95
96 CGG CTT CCT GCT CTC GCC GGA GTT TCC GCG TAG AGG GCG CAT CGC CGG 143
144 CCC GGG GCC CTT GGT GCG GCG TGG CGC AGG GCG CGG CGT GGG GCG CGC 191
192 GTG GGC GCG GCG CAG GCG GCC CGG GTC ACC ATG AGG ACT CTC CGC AGG 239
1 Met Arg Thr Leu Arg Arg 6
240 TTG AAG TTC ATG AGT TCG CCC AGC CTC AGT GAC CTG GGC AAG AGA GAG 287
7 Leu Lys Phe Met Ser Ser Pro Ser Leu Ser Asp Leu Gly Lys Arg Glu 22
288 CCG GCC GCC GCC GCG GAC GAG CGG GGC ACG CAG CAG CGC CGG GCC TGC 335
23 Pro Ala Ala Ala Ala Asp Glu Arg Gly Thr Gln Gln Arg Arg Ala Cys 38
336 GCC AAC GCC ACC TGG AAC AGC ATC CAC AAC GGG GTG ATC GCC GTC TTC 383
39 Ala Asn Ala Thr Trp Asn Ser Ile His Asn Gly Val Ile Ala Val Phe 54
384 CAG CGC AAG GGG CTG CCC GAC CAG GAG CTC TTC AGC CTC AAC GAG GGC 431
55 Gln Arg Lys Gly Leu Pro Asp Gln Glu Leu Phe Ser Leu Asn Glu Gly 70
432 GTC CGG CAG CTG TTG AAG ACA GAG CTG GGG TCC TTC TTC ACG GAG TAC 479
71 Val Arg Gln Leu Leu Lys Thr Glu Leu Gly Ser Phe Phe Thr Glu Tyr 86
480 CTG CAG AAC CAG CTG CTG ACA AAA GGC ATG GTG ATC CTT CGG GAC AAG 527
87 Leu Gln Asn Gln Leu Leu Thr Lys Gly Met Val Ile Leu Arg Asp Lys 102
528 ATT CGC TTC TAT GAG GGA CAG AAG CTG CTG GAC TCA CTG GCA GAG ACC 575
103 Ile Arg Phe Tyr Glu Gly Gln Lys Leu Leu Asp Ser Leu Ala Glu Thr 118
576 TGG GAC TTC TTC TTC AGT GAC GTG CTG CCC ATG CTG CAG GCC ATC TTC 623
119 Trp Asp Phe Phe Phe Ser Asp Val Leu Pro Met Leu Gln Ala Ile Phe 134
624 TAC CCG GTG CAG GGC AAG GAG CCA TCG GTG CGC CAG CTG GCC CTG CTG 671
135 Tyr Pro Val Gln Gly Lys Glu Pro Ser Val Arg Gln Leu Ala Leu Leu 150
672 CAC TTC CGG AAT GCC ATC ACC CTC AGT GTG AAG CTA GAG GAT GCG CTG 719
151 His Phe Arg Asn Ala Ile Thr Leu Ser Val Lys Leu Glu Asp Ala Leu 166
720 GCC CGG GCC CAT GCC CGT GTG CCC CCT GCC ATC GTG CAG ATG CTG CTG 767
167 Ala Arg Ala His Ala Arg Val Pro Pro Ala Ile Val Gln Met Leu Leu 182
768 GTG CTG CAG GGG GTA CAT GAG TCC AGG GGC GTG ACT GAG GAC TAC CTG 815
183 Val Leu Gln Gly Val His Glu Ser Arg Gly Val Thr Glu Asp Tyr Leu 198
816 CGC CTG GAG ACG CTG GTC CAG AAG GTG GTG TCG CCA TAC CTG GGC ACC 863
199 Arg Leu Glu Thr Leu Val Gln Lys Val Val Ser Pro Tyr Leu Gly Thr 214
864 TAC GGC CTC CAC TCC AGC GAG GGG CCC TTC ACC CAT TCC TGC ATC CTG 911
215 Tyr Gly Leu His Ser Ser Glu Gly Pro Phe Thr His Ser Cys Ile Leu 230
912 GAA AAG CGC CTC CTC CGC CGC TCC CGC TCG GGG GAC GTG CTG GCC AAG 959
231 Glu Lys Arg Leu Leu Arg Arg Ser Arg Ser Gly Asp Val Leu Ala Lys 246
960 AAC CCT GTG GTG CGC TCC AAG AGC TAC AAC ACG CCT CTG CTG AAC CCC 1007
247 Asn Pro Val Val Arg Ser Lys Ser Tyr Asn Thr Pro Leu Leu Asn Pro 262
1008 GTG CAG GAG CAC GAG GCG GAG GGC GCG GCG GCC GGC GGT ACC AGC ATC 1055
263 Val Gln Glu His Glu Ala Glu Gly Ala Ala Ala Gly Gly Thr Ser Ile 278
1056 CGC AGG CAC TCT GTG TCG GAG ATG ACG TCC TGC CCC GAG CCT CAG GGC 1103
279 Arg Arg His Ser Val Ser Glu Met Thr Ser Cys Pro Glu Pro Gln Gly 294
1104 TTC TCC GAC CCG CCC GGC CAG GGC CCC ACC GGG ACC TTC AGG TCC TCC 1151
295 Phe Ser Asp Pro Pro Gly Gln Gly Pro Thr Gly Thr Phe Arg Ser Ser 310
1152 CCG GCG CCC CAC TCA GGG CCC TGC CCC AGC AGA CTG TAC CCC ACG ACC 1199
311 Pro Ala Pro His Ser Gly Pro Cys Pro Ser Arg Leu Tyr Pro Thr Thr 326
1200 CAG CCC CCT GAG CAG GGC TTG GAT CCC ACC CGC AGC TCC CTG CCC CGC 1247
327 Gln Pro Pro Glu Gln Gly Leu Asp Pro Thr Arg Ser Ser Leu Pro Arg 342
1248 TCC AGC CCG GAG AAC CTG GTG GAC CAG ATC CTG GAG TCC GTG GAC TCG 1295
343 Ser Ser Pro Glu Asn Leu Val Asp Gln Ile Leu Glu Ser Val Asp Ser 358
1296 GAT TCT GAA GGG ATT TTC ATT GAC TTT GGC CGG GGC CGG GGC TCT GGC 1343
359 Asp Ser Glu Gly Ile Phe Ile Asp Phe Gly Arg Gly Arg Gly Ser Gly 374
1344 ATG TCC GAC TTG GAG GGC TCT GGG GGC CGG CAG AGT GTC GTG TGA GGC 1391
375 Met Ser Asp Leu Glu Gly Ser Gly Gly Arg Gln Ser Val Val *** 389
1392 CTC ACA GCT GGC CTT GAG TTT TTA CTG ACA CGT CCC TGT GTG CGG GGG 1439
1440 TGT CCA TGT GGC GTG TGT GTG AGT GAG ACT TTT TTA CTG CGT CCC GTC 1487
1488 CCG CCA GCC CTA TCG GCC TCG TCA CTG GCC TTG GTC ACT TTG TAT TTC 1535
1536 TGT CTT GGT TGG AAA TAC CAT CAG CCT TCC TTG CTC GGC CCA GGT CTG 1583
1584 TTT CAG GCA TCT GAG TCG GCG TTT ACC CAG GGG CCG GGC CAG AGA CGG 1631
1632 GGG TCG GCC GCT CGC TCC CAC GCT CCT CCT GCC CCA GCC CTC TGG TGT 1679
1680 CCA CAC CTG CCC ACA GAG AAT GTA AAC CCA GTG GGC TCT GCC CAC GCC 1727
1728 GGG CCC CAA AGT GAC CAG ACT CCA GCA CAC CTG TCT CCT CCT GCC TGG 1775
1776 GGT GGC CAT GGG GAT GGA AGG GGG TGG AAT AAA ACC TGT CAC CCT GGG 1823
1824 AAA AAA AAA AAA AAA AAA AAA AAA AAA AAA 1853
D:Blas+p同源比较结果
Query=pp610[gene=pp610](388字符)
>SP_HUM:O75982 O75982 homo sapiens(human).hypothetical 18.0kd
蛋白质(片段).11/1998 长度=158
分值=316bits(802),预期值=2e-85
相同性=158/158(100%),相似性=158/158(100%)
Query:73 QLLKTELGSFFTEYLQNQLLTKGMVILRDKIRFYEGQKLLDSLAETWDFFFSDVLPMLQA 132
QLLKTELGSFFTEYLQNQLLTKGMVILRDKIRFYEGQKLLDSLAETWDFFFSDVLPMLQA
Sbjct:1 QLLKTELGSFFTEYLQNQLLTKGMVILRDKIRFYEGQKLLDSLAETWDFFFSDVLPMLQA 60
Query:133 IFYPVQGKEPSVRQLALLHFRNAITLSVKLEDALARAHARVPPAIVQMLLVLQGVHESRG 192
IFYPVQGKEPSVRQLALLHFRNAITLSVKLEDALARAHARVPPAIVQMLLVLQGVHESRG
Sbjct:61 IFYPVQGKEPSVRQLALLHFRNAITLSVKLEDALARAHARVPPAIVQMLLVLQGVHESRG 120
Query:193 VTEDYLRLETLVQKVVSPYLGTYGLHSSEGPFTHSCIL 230
VTEDYLRLETLVQKVVSPYLGTYGLHSSEGPFTHSCIL
Sbjct:121 VTEDYLRLETLVQKVVSPYLGTYGLHSSEGPFTHSCIL 158
2.PP784
A:核苷酸序列(SEQ ID NO:4)长度:2145bp
1 GGCGATTCTT CCCGCAGAGT TGTGAAGCGA AAGGCTTACA ATTAAAAGGA
51 AGAAAAAAAA ATAAAGATAA TTCGGGAGTA CAATTGACAA AGCGTGTGGG
101 TCGCTCAGCC TCCAGCAGTA ACTGCTGATC TCCAGTTCTT GGAGGGTTCC
151 GGTGAGAAGA ACGCCCCTAC TGCGGTACTG AGGAAGCGGC AGGAGGAGAT
201 GCGGCCCCTG GACATAGACG AGGTGGAAGC GCCTGAGGAA GTGGAGGTGC
251 TGGAGCCCGA GGAGGATTTC GAGCAGTTCC TGCTCCCGGT CATCAACGAG
301 ATGCGCGAGG ACATCGCGTC TCTTATACGC GAGCACGGGC GGGCGTACCT
351 GCGGACCAGG AGCAAGCTGT GGGAGATGGA CAATATGCTT ATCCAGATCA
401 AAACGCAGGT GGAGGCCTCG GAGGAGAGCG CCCTCAATCA CGTGCAGCAC
451 CCGAGTGGCG AAGCCGACGA GAGAGTGTCG GAGTTGTGCG AGAAGGCTGA
501 GGAGAAAGCC AAGGAGATTG CGAAGATGGC AGAGATGCTG GTCGAGCTCG
551 TCTGGCGAAT AGAGAGAAGC GAGTCTTCTT GAAGGAGGAG ATCGGTGGTA
601 AGGTCGGAAA CTAGCTGGAG CTTGGATGAA ACTCAGTAGT CGCCCTAAGC
651 ATGGTTAAAC CGCGCCCTGT AAAAAGTTAT GTTCTAGGCC TGCATCCCCC
701 ATTTTATTGC ATCAAAAATT GAGCATTGGG AACAAAGTTG GGGTCAAGAG
751 GAAAGAATGC GTGCTGGTTT TGTTAGGCGT TAGTATACCG TTTTTTTTGT
801 GGCCTCTCCC TCCCACACTG GTAATTAGAG AAAGATAACA GTAACTTCGG
851 TTTAGTTTTT GTGAAACATA AAAGTCAATT CTAATAGGGC AGTCGCCAGA
901 AGTAGACCTG TCTAGGCACT AAGGGAGTTT GGGGAAAGCC AAAGAAGACC
951 TAGGCCATAG AGCACAGTGG AACGCAGGTG AGAACGCAGG GAAAGAGAAG
1001 TAAAGAGTAA AGCCAGAGGC CATTACCTGA AATTTCCAGA TTGTTCTATG
1051 AGACAGGTAT GTCAGAGGAC CGTGTCTCAA AGAAGTGGCA TTCTTCTGGG
1101 TGGTTCACAT TTATTTTTAA CATCAGGATA TGGAACTTTG AGAGTCAATG
1151 TCTTCATGGT AAATTTCTCC CAATTGTGCT TTAGGTTCAC AGCTGAGGAC
1201 TGTAGTCAAC TGGTAAGGAT GAACTGACAC CTTGAGGAAA CTGAAACAGC
1251 TTGTCATTTT CTCTGTTTTG TTTTTGTATG TTTTTTACCC CCATGTAACA
1301 GTCTCCCTTA TGGTCAGTGA TTGATGACCT CGATATGGAT TTAGATTATC
1351 AAATGTGTTT GGTTTTGGAA ATATAACTTT TGTCAAAGAA ATACTCTCAG
1401 AAGAGAAATG GGGCTTAATT AAGTTGTTTT TGTGGTCACG TTTATTCTTG
1451 TTACTTCGCT GTGTTTTTGA AATGTTGGGC ATGGCCTCGT ATTTTGCTGT
1501 TACCTTTGTG ACCTGATTGT TTTTTGGAAC ACGTCAAGAC GTGGGATCAG
1551 AATCTTCCAA CTTTAGAGGT GCAATGGAAG ACACTACGCT ACTTGGTTGA
1601 GCCTGGTGAA GAATGTATTA ATGAGACTGC TTTGCATAAA ACTGGGAAGA
1651 AAGAGAAGAC AGTTGGAGAT GGAAGATGGT TTTGTATATA TTTTGGAACT
1701 TTAGTTCCTC TGTGAGACGA AAGAGGAGAG CTATGTTTTG TGTCACATTG
1751 TCTGATATAT ATTGTGTAAC CTGTCAGGTG AGTTGATTTA GACAACATAG
1801 CTGACCTTTT ATGACAAGGC AGTTTGAATA GGGACTATTG TAATACCCTC
1851 ACACATTATA GGGGCATCAG AGAATGGCAT GGAAGAGACA GTCTACAGAG
1901 AGCTTTAAGA GGCCGGAGAA AGGAAAAGAC ATTATCAGGG CCTGGAAAGT
1951 CTCTTCCAGT TCATCAGGGT AGTAGACCTG TCAGATTGGA GGTCAAAGTC
2001 AAAAGTATAA TAAGTGTTAA TTTGCCTTGA AAAACAAAGA CCTAAAAAGC
2051 CTTTTTTAAG AAATAAATTT TTTGTTCACA TGACCAAAGC CCCTCCTGTG
2101 TGTGTTAATA AAAGAATCAT AAACAAAAAA AAAAAAAAAA AAAAA
B:氨基酸序列(SEQ ID NO:5)长度:127个氨基酸
1 MRPLDIDEVE APEEVEVLEP EEDFEQFLLP VINEMREDIA SLIREHGRAY
51 LRTRSKLWEM DNMLIQIKTQ VEASEESALN HVQHPSGEAD ERVSELCEKA
101 EEKAKEIAKM AEMLVELVWR IERSESS
C.核苷酸及氨基酸组合序列(SEQ ID NO:6)
克隆号和蛋白名称:PP784
起始编码子: 199 ATG 终止编码子:582 TGA
蛋白质分子量:14806.89
1 GGC GAT TCT TCC CGC AGA GTT GTG AAG CGA AAG GCT TAC AAT TAA AAG 48
49 GAA GAA AAA AAA ATA AAG ATA ATT CGG GAG TAC AAT TGA CAA AGC GTG 96
97 TGG GTC GCT CAG CCT CCA GCA GTA ACT GCT GAT CTC CAG TTC TTG GAG 144
145 GGT TCC GGT GAG AAG AAC GCC CCT ACT GCG GTA CTG AGG AAG CGG CAG 192
193 GAG GAG ATG CGG CCC CTG GAC ATA GAC GAG GTG GAA GCG CCT GAG GAA 240
1 Met Arg Pro Leu Asp Ile Asp Glu Val Glu Ala Pro Glu Glu 14
241 GTG GAG GTG CTG GAG CCC GAG GAG GAT TTC GAG CAG TTC CTG CTC CCG 288
15 Val Glu Val Leu Glu Pro Glu Glu Asp Phe Glu Gln Phe Leu Leu Pro 30
289 GTC ATC AAC GAG ATG CGC GAG GAC ATC GCG TCT CTT ATA CGC GAG CAC 336
31 Val Ile Asn Glu Met Arg Glu Asp Ile Ala Ser Leu Ile Arg Glu His 46
337 GGG CGG GCG TAC CTG CGG ACC AGG AGC AAG CTG TGG GAG ATG GAC AAT 384
47 Gly Arg Ala Tyr Leu Arg Thr Arg Ser Lys Leu Trp Glu Met Asp Asn 62
385 ATG CTT ATC CAG ATC AAA ACG CAG GTG GAG GCC TCG GAG GAG AGC GCC 432
63 Met Leu Ile Gln Ile Lys Thr Gln Val Glu Ala Ser Glu Glu Ser Ala 78
433 CTC AAT CAC GTG CAG CAC CCG AGT GGC GAA GCC GAC GAG AGA GTG TCG 480
79 Leu Asn His Val Gln His Pro Ser Gly Glu Ala Asp Glu Arg Val Ser 94
481 GAG TTG TGC GAG AAG GCT GAG GAG AAA GCC AAG GAG ATT GCG AAG ATG 528
95 Glu Leu Cys Glu Lys Ala Glu Glu Lys Ala Lys Glu Ile Ala Lys Met 110
529 GCA GAG ATG CTG GTC GAG CTC GTC TGG CGA ATA GAG AGA AGC GAG TCT 576
111 Ala Glu Met Leu Val Glu Leu Val Trp Arg Ile Glu Arg Ser Glu Ser 126
577 TCT TGA AGG AGG AGA TCG GTG GTA AGG TCG GAA ACT AGC TGG AGC TTG 624
127 Ser *** 128
625 GAT GAA ACT CAG TAG TCG CCC TAA GCA TGG TTA AAC CGC GCC CTG TAA 672
673 AAA GTT ATG TTC TAG GCC TGC ATC CCC CAT TTT ATT GCA TCA AAA ATT 720
721 GAG CAT TGG GAA CAA AGT TGG GGT CAA GAG GAA AGA ATG CGT GCT GGT 768
769 TTT GTT AGG CGT TAG TAT ACC GTT TTT TTT GTG GCC TCT CCC TCC CAC 816
817 ACT GGT AAT TAG AGA AAG ATA ACA GTA ACT TCG GTT TAG TTT TTG TGA 864
865 AAC ATA AAA GTC AAT TCT AAT AGG GCA GTC GCC AGA AGT AGA CCT GTC 912
913 TAG GCA CTA AGG GAG TTT GGG GAA AGC CAA AGA AGA CCT AGG CCA TAG 960
961 AGC ACA GTG GAA CGC AGG TGA GAA CGC AGG GAA AGA GAA GTA AAG AGT 1008
1009 AAA GCC AGA GGC CAT TAC CTG AAA TTT CCA GAT TGT TCT ATG AGA CAG 1056
1057 GTA TGT CAG AGG ACC GTG TCT CAA AGA AGT GGC ATT CTT CTG GGT GGT 1104
1105 TCA CAT TTA TTT TTA ACA TCA GGA TAT GGA ACT TTG AGA GTC AAT GTC 1152
1153 TTC ATG GTA AAT TTC TCC CAA TTG TGC TTT AGG TTC ACA GCT GAG GAC 1200
1201 TGT AGT CAA CTG GTA AGG ATG AAC TGA CAC CTT GAG GAA ACT GAA ACA 1248
1249 GCT TGT CAT TTT CTC TGT TTT GTT TTT GTA TGT TTT TTA CCC CCA TGT 1296
1297 AAC AGT CTC CCT TAT GGT CAG TGA TTG ATG ACC TCG ATA TGG ATT TAG 1344
1345 ATT ATC AAA TGT GTT TGG TTT TGG AAA TAT AAC TTT TGT CAA AGA AAT 1392
1393 ACT CTC AGA AGA GAA ATG GGG CTT AAT TAA GTT GTT TTT GTG GTC ACG 1440
1441 TTT ATT CTT GTT ACT TCG CTG TGT TTT TGA AAT GTT GGG CAT GGC CTC 1488
1489 GTA TTT TGC TGT TAC CTT TGT GAC CTG ATT GTT TTT TGG AAC ACG TCA 1536
1537 AGA CGT GGG ATC AGA ATC TTC CAA CTT TAG AGG TGC AAT GGA AGA CAC 1584
1585 TAC GCT ACT TGG TTG AGC CTG GTG AAG AAT GTA TTA ATG AGA CTG CTT 1632
1633 TGC ATA AAA CTG GGA AGA AAG AGA AGA CAG TTG GAG ATG GAA GAT GGT 1680
1681 TTT GTA TAT ATT TTG GAA CTT TAG TTC CTC TGT GAG ACG AAA GAG GAG 1728
1729 AGC TAT GTT TTG TGT CAC ATT GTC TGA TAT ATA TTG TGT AAC CTG TCA 1776
1777 GGT GAG TTG ATT TAG ACA ACA TAG CTG ACC TTT TAT GAC AAG GCA GTT 1824
1825 TGA ATA GGG ACT ATT GTA ATA CCC TCA CAC ATT ATA GGG GCA TCA GAG 1872
1873 AAT GGC ATG GAA GAG ACA GTC TAC AGA GAG CTT TAA GAG GCC GGA GAA 1920
1921 AGG AAA AGA CAT TAT CAG GGC CTG GAA AGT CTC TTC CAG TTC ATC AGG 1968
1969 GTA GTA GAC CTG TCA GAT TGG AGG TCA AAG TCA AAA GTA TAA TAA GTG 2016
2017 TTA ATT TGC CTT GAA AAA CAA AGA CCT AAA AAG CCT TTT TTA AGA AAT 2064
2065 AAA TTT TTT GTT CAC ATG ACC AAA GCC CCT CCT GTG TGT GTT AAT AAA 2112
2113 AGA ATC ATA AAC AAA AAA AAA AAA AAA AAA AAA 2145
3.PP827
A:核苷酸序列(SEQ ID NO:7)长度:1678bp
1 CCATGTTGGC CAGGCTGGTC TGAGAGAGAA CTCCTGACCT CGTGATCCGC
51 CTGCCTCAGC CTCCCAAAGT GCTGGGATGA CAGGCGTGCG CCACTGCACC
101 CGGCCCGTGA AGGTCTTTTC TGAAGGGAGC TACCTTTGAG AAGTGCCTGT
151 TGGGACAGAA CTCCCTGCTC TGGTCTGACC CTCTGCACGC TAGGGTGGTC
201 CTGCACCTGT CGGGGAGGGG GCCAAGGACA CAGCACTGGG CTGGGGCAGG
251 GCTCTGCGCT GGTGCTGCCT GCCATCCCAG TTCTGCCATT CCCCCAGCGC
301 ACCCCCGCAC CCGCCCCTCC AGCTGATGCC TGCTCCCTCT CTCTGCAGAA
351 CGGGCTCATG TCGGGGCTGA TGCAGATGCT GCTGCTGAAG GTGTCTGCAC
401 ACATCACCGA GCAGCTGGGC ATGGCCCCAG GTGGCGAGTT CAGGGAGGCC
451 TTCAAGGAGG TGGGCACAGG GTGAGGTAGG GGGTGGCCAC AGGGATGTGG
501 CTCACAGGGG TGGTCCGGGG TTCCCAGTGC TCCCCAACAC CCAGCCTCTG
551 CTCCAGGCCA GCAAGGTGCC TTTCTGCAAG TTCCACCTGG GTGACCGACC
601 CATCCCCGTC ACCTTCAAGA GGGCCATCGC AGCGCTCTCC TTCTGGCAGA
651 AGGTCAGGCT GGCTTGGGGC CTGTGCTTCC TGTCAGACCC CATCAGGTAG
701 GGCTGCCCCC GGGACCCTGG CCGGCCTGCA GGGTGGTCTG TGGGAGGCTC
751 CAGGCCCTCC TGTGCAGGTC CAAGCGCAGC CAATCCTCAC TCAAGGCCTT
801 CCCTGCCCTT TCCTTCCGCC ACAAATCCCA AACAAACGTG CTGTGGTCCC
851 TGCCCGGTGT CCACAGTGCC AGCCCCACCC CCCCAGCCCG TTGCCCATCC
901 CTGCGGGGCT GCAGCCATCC CTCTCCACAG CAAGGATGAC GTGGAACGCT
951 GCAAGCAGAA GGACCTACTG GAGCAGATGA TGGCCGAGAT GATTGGCGAG
1001 TTCCCAGACC TGCACCGCAC CATCGTCTCG GAGCGCGACG TCTACCTAAC
1051 CTACATGCTG CGCCAGGCCG CGCGGCGCCT CGAGCTGCCT CGGGCCTCTG
1101 ACGGTGACGG CCGCCCGCAG GCGTGGGACC CCCTGTGAGG CTGAGGCCCG
1151 AGCAGGTACT GACCCCTTGT CCTTCCCCAC AGCCGAGCCC AGGAAGTGCG
1201 TCCCCTCCGT GGTCGTGGGC GTCGTGGGCA TGGGCCACGT GCCTGGCATC
1251 GAGAAGAACT GGAGCACCGA CCTCAACATC CAGGAGATCA TGACCGTGCC
1301 CCCGCCGTCC GTCTCCGGCA GAGTGTCTCG GTTGGCCGTG AAGGCCGCCT
1351 TCTTCGGCCT GCTGGGCTAC AGCCTGTACT GGATGGGCCG CCGCACCGCG
1401 AGCCTGGTCC TGTCGCTGCC CGCCGCGCAG TACTGCCTGC AGAGGGTGAC
1451 CGAGGCCCGG CACAAGTAGG AGACTGCTCC CCGCCCGCTC GGGCCCCTGA
1501 GGAGCCAGTG CCCCCGCGGC ACTTCTGGGT GCCAGGTGCA TCCTAGCCCG
1551 CCCGAGGCCC CTGCCACCCC CCATGGGGGT CTGGGCCCGG CCTCGCCTGC
1601 CCTCCTGGGC CAGTCACCCC TCCCCCAGCC CACCCAAATA AAGGATTATT
1651 TAACTGTCTG AAAAAAAAAA AAAAAAAA
B:氨基酸序列(SEQ ID NO:8)长度:161个氨基酸
1 MWLTGVVRGS QCSPTPSLCS RPARCLSASS TWVTDPSPSP SRGPSQRSPS
51 GRRSGWLGAC ASCQTPSGRA APGTLAGLQG GLWEAPGPPV QVQAQPILTQ
101 GLPCPFLPPQ IPNKRAVVPA RCPQCQPHPP SPLPIPAGLQ PSLSTARMTW
151 NAASRRTYWS R
C.核苷酸及氨基酸组合序列(SEQ ID NO:9)
克隆号和蛋白名称:PP827
起始编码子: 495 ATG 终止编码子:980 TGA
蛋白质分子量:16922.45
1 CC ATG TTG GCC AGG CTG GTC TGA GAG AGA ACT CCT GAC CTC GTG ATC 47
48 CGC CTG CCT CAG CCT CCC AAA GTG CTG GGA TGA CAG GCG TGC GCC ACT 95
96 GCA CCC GGC CCG TGA AGG TCT TTT CTG AAG GGA GCT ACC TTT GAG AAG 143
144 TGC CTG TTG GGA CAG AAC TCC CTG CTC TGG TCT GAC CCT CTG CAC GCT 191
192 AGG GTG GTC CTG CAC CTG TCG GGG AGG GGG CCA AGG ACA CAG CAC TGG 239
240 GCT GGG GCA GGG CTC TGC GCT GGT GCT GCC TGC CAT CCC AGT TCT GCC 287
288 ATT CCC CCA GCG CAC CCC CGC ACC CGC CCC TCC AGC TGA TGC CTG CTC 335
336 CCT CTC TCT GCA GAA CGG GCT CAT GTC GGG GCT GAT GCA GAT GCT GCT 383
384 GCT GAA GGT GTC TGC ACA CAT CAC CGA GCA GCT GGG CAT GGC CCC AGG 431
432 TGG CGA GTT CAG GGA GGC CTT CAA GGA GGT GGG CAC AGG GTG AGG TAG 479
480 GGG GTG GCC ACA GGG ATG TGG CTC ACA GGG GTG GTC CGG GGT TCC CAG 527
1 Met Trp Leu Thr Gly Val Val Arg Gly Ser Gln 11
528 TGC TCC CCA ACA CCC AGC CTC TGC TCC AGG CCA GCA AGG TGC CTT TCT 575
12 Cys Ser Pro Thr Pro Ser Leu Cys Ser Arg Pro Ala Arg Cys Leu Ser 27
576 GCA AGT TCC ACC TGG GTG ACC GAC CCA TCC CCG TCA CCT TCA AGA GGG 623
28 Ala Ser Ser Thr Trp Val Thr Asp Pro Ser Pro Ser Pro Ser Arg Gly 43
624 CCA TCG CAG CGC TCT CCT TCT GGC AGA AGG TCA GGC TGG CTT GGG GCC 671
44 Pro Ser Gln Arg Ser Pro Ser Gly Arg Arg Ser Gly Trp Leu Gly Ala 59
672 TGT GCT TCC TGT CAG ACC CCA TCA GGT AGG GCT GCC CCC GGG ACC CTG 719
60 Cys Ala Ser Cys Gln Thr Pro Ser Gly Arg Ala Ala Pro Gly Thr Leu 75
720 GCC GGC CTG CAG GGT GGT CTG TGG GAG GCT CCA GGC CCT CCT GTG CAG 767
76 Ala Gly Leu Gln Gly Gly Leu Trp Glu Ala Pro Gly Pro Pro Val Gln 91
768 GTC CAA GCG CAG CCA ATC CTC ACT CAA GGC CTT CCC TGC CCT TTC CTT 815
92 Val Gln Ala Gln Pro Ile Leu Thr Gln Gly Leu Pro Cys Pro Phe Leu 107
816 CCG CCA CAA ATC CCA AAC AAA CGT GCT GTG GTC CCT GCC CGG TGT CCA 863
108 Pro Pro Gln Ile Pro Asn Lys Arg Ala Val Val Pro Ala Arg Cys Pro 123
864 CAG TGC CAG CCC CAC CCC CCC AGC CCG TTG CCC ATC CCT GCG GGG CTG 911
124 Gln Cys Gln Pro His Pro Pro Ser Pro Leu Pro Ile Pro Ala Gly Leu 139
912 CAG CCA TCC CTC TCC ACA GCA AGG ATG ACG TGG AAC GCT GCA AGC AGA 959
140 Gln Pro Ser Leu Ser Thr Ala Arg Met Thr Trp Asn Ala Ala Ser Arg 155
960 AGG ACC TAC TGG AGC AGA TGA TGG CCG AGA TGA TTG GCG AGT TCC CAG 1007
156 Arg Thr Tyr Trp Ser Arg *** 162
1008 ACC TGC ACC GCA CCA TCG TCT CGG AGC GCG ACG TCT ACC TAA CCT ACA 1055
1056 TGC TGC GCC AGG CCG CGC GGC GCC TCG AGC TGC CTC GGG CCT CTG ACG 1103
1104 GTG ACG GCC GCC CGC AGG CGT GGG ACC CCC TGT GAG GCT GAG GCC CGA 1151
1152 GCA GGT ACT GAC CCC TTG TCC TTC CCC ACA GCC GAG CCC AGG AAG TGC 1199
1200 GTC CCC TCC GTG GTC GTG GGC GTC GTG GGC ATG GGC CAC GTG CCT GGC 1247
1248 ATC GAG AAG AAC TGG AGC ACC GAC CTC AAC ATC CAG GAG ATC ATG ACC 1295
1296 GTG CCC CCG CCG TCC GTC TCC GGC AGA GTG TCT CGG TTG GCC GTG AAG 1343
1344 GCC GCC TTC TTC GGC CTG CTG GGC TAC AGC CTG TAC TGG ATG GGC CGC 1391
1392 CGC ACC GCG AGC CTG GTC CTG TCG CTG CCC GCC GCG CAG TAC TGC CTG 1439
1440 CAG AGG GTG ACC GAG GCC CGG CAC AAG TAG GAG ACT GCT CCC CGC CCG 1487
1488 CTC GGG CCC CTG AGG AGC CAG TGC CCC CGC GGC ACT TCT GGG TGC CAG 1535
1536 GTG CAT CCT AGC CCG CCC GAG GCC CCT GCC ACC CCC CAT GGG GGT CTG 1583
1584 GGC CCG GCC TCG CCT GCC CTC CTG GGC CAG TCA CCC CTC CCC CAG CCC 1631
1632 ACC CAA ATA AAG GAT TAT TTA ACT GTC TGA AAA AAA AAA AAA AAA AA 1678
4.PP1057
A:核苷酸序列(SEQ ID NO:10)长度:2030bp
1 GGCACAGAGC TGGGCATGGG AGAGAAACGG GCTGGGGAGT GAGGCCAGGA
51 GTGGGATTCA GCTGCAGCAG GGCGCCCCCT CCAAACTGCA GCTGGTCTGG
101 CTTACTGTTT TGCCGTTCAA AAAGGTCGCG AATCCGTGGG ACTGAGCACG
151 GGGACCTCAC CCGCTAGCCA GCGTCTGCTG CACTTGATCA GGTGGGGCCT
201 TGGTGGGCGG CTGCCTTTCC TATACAGTTT GTCTTGTCAC CCTGGTTTCC
251 CACTGGGGCC AGGTCTCTTC TCCAGCCTCC ACCTGCCTGT CTGATCCAAG
301 AGCTGAGACA CGGCCACCCA GCACCAGTCA CTCCTCTGTT CACCTTAAGT
351 AACACACAAA TCGGGAACAG GAGGACAGAA CCGTTGGCAT TATCAGGATT
401 CGTGTTTTGT GGGGGTGGGA GTGGAGAGTA GGGTGGTCTT GTGAGTTGTG
451 CAGGGTGAAG ACCGCTTCCC TGAGACAGGG GCAGTGGTGC TGATGGAATG
501 TGGGGGAGGC CCACATTTGA GCAAAGCTGC CCTGCCCTTG TCCCCTGGCC
551 TGGCTTCCTG GTAAGGAGTT TCAGCCGCCT CCGCAGGAAC CCCCAAAGTG
601 CAGATTCCGG AGCAGACACA TCCGGGCGGA GAGACTCAGC AGACAAGTGC
651 TGCAGTTGCA CGGTGGGACC CGGGGCCTCG TGCGTTTTTT GCTGTGGGTG
701 GGGTGGGTGG GTTGGTTTAT GCCTATCAAT GCAATTTTTA ATTTTTGTTA
751 ATATCAACAG CAAAAGCCTA GTGCATTGGG AGATGTGCAA CCTCCCTGAA
801 AATCTTTTCT GTTTCTGGAG TACTTCAGGG GTGGCCTCTG GCCCCAGAGC
851 CTTTGCCACA GTGCTCCCAC CAGCCCCCAC CTCATCCGTC TGTTTGCAGA
901 GCCTCATCTA CAGGTCCCCA CGCTGCCTTC TTTACTCACT CTGCGCTTGG
951 CCGTTTTGTT ATTTGGCTTA GTCTACATTG GGCGGAAGTC TGTGTGCACA
1001 GAGTGGGTGT TCCTTCGAGC CCCTTCCACT CAGAGGGCCA CACCCAGCGA
1051 TGCCAGTGAA GGTGGCACAG CCTCTCTTCA GTTTCTCCTG ACTGTGATCT
1101 CACTGGGGTA GAATTCCCCT GAGAGAATTC CCTCACTCAC GGCTCCCTTT
1151 GCCAGAGTCA GTTCAATCAG GTCTGATGTG AGCAATTTAC ACACTTGTCT
1201 CAGAAAGTCC CTCAGGGTTT GTAGAGGACT GCAGGGGGGC ATCCGCTGCA
1251 GACTCAGCCT TTCTCTGCAG CCATCCTGCA GTGGGGGTGA GCGGGCACAG
1301 GCTGAGAACT GCTCTTGGGT GGTGGAAGCA GGTGTCACGG TGCAAGTCTC
1351 CCCCTGCACC CCTCCCCCAG CTTGAGCCGT GTCACCCCCC TCTCCCTCCA
1401 GCATGGGCCT GTGTCTCAGC TCTCTGGAAG GTGGCCCTGC CCGGACCCTC
1451 TTGCAGGTGT CCTGGTTTGA CTTGGAACTA GATGGCCATC TTTTCAGGCT
1501 TTGGTGGCCC AAGAGCAGTC TGGGTGGATG GAAGTGGCTG TCCCCTTCTC
1551 TTCAGCCCCT GCCCACCCAC TGGTGGAGGT GCTAACTAGC AGGACGTGGC
1601 ATAGGATGGG AGCTGGGCGT GAGGTGCTTT GGGGTCCATT CTTTGTCCCT
1651 CAGCTTCTCA GAGTCCGGCC AGCCCTTGTG TTCCCGTGCC CCACACTTTC
1701 CTCCTCCCCA CTGCAGTGAG TCAATAGTCC AGGGTGGGGC CTGGCCTCCC
1751 TGCCCTGATT GGGGACTCAG GAGGTAAGGC CTGGGGGGCT TCCTGCCCCC
1801 TCCTTGCCCA CCTGCCTGCC CCCGGGCAGC ACGGGAGGGA AAGCAGGGTG
1851 AGCACGCTTG TTGGTTTCAA ATGCACTTTC TGCTTGCATT GCCGTATCTG
1901 TGCGTTCCTT CATCCTGGTC CTGGCTTTAT GGAACACCAT GTTTTTAGCA
1951 TGTTTTTAAA TAAAAACGGA TAAAGTGTCA AAAGCCAAAA AAAAAAAAAA
2001 AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA
B:氨基酸序列(SEQ ID NO:11)长度:157个氨基酸
1 MWGRPTFEQS CPALVPWPGF LVRSFSRLRR NPQSADSGAD TSGRRDSADK
51 CCSCTVGPGA SCVFCCGWGG WVGLCLSMQF LIFVNINSKS LVHWEMCNLP
101 ENLFCFWSTS GVASGPRAFA TVLPPAPTSS VCLQSLIYRS PRCLLYSLCA
151 WPFCYLA
C.核苷酸及氨基酸组合序列(SEQ ID NO:12)
克隆号和蛋白名称:PP1057
起始编码子: 498 ATG 终止编码子:971 TAG
蛋白质分子量:17224.06
1 GG CAC AGA GCT GGG CAT GGG AGA GAA ACG GGC TGG GGA GTG AGG CCA 47
48 GGA GTG GGA TTC AGC TGC AGC AGG GCG CCC CCT CCA AAC TGC AGC TGG 95
96 TCT GGC TTA CTG TTT TGC CGT TCA AAA AGG TCG CGA ATC CGT GGG ACT 143
144 GAG CAC GGG GAC CTC ACC CGC TAG CCA GCG TCT GCT GCA CTT GAT CAG 191
192 GTG GGG CCT TGG TGG GCG GCT GCC TTT CCT ATA CAG TTT GTC TTG TCA 239
240 CCC TGG TTT CCC ACT GGG GCC AGG TCT CTT CTC CAG CCT CCA CCT GCC 287
288 TGT CTG ATC CAA GAG CTG AGA CAC GGC CAC CCA GCA CCA GTC ACT CCT 335
336 CTG TTC ACC TTA AGT AAC ACA CAA ATC GGG AAC AGG AGG ACA GAA CCG 383
384 TTG GCA TTA TCA GGA TTC GTG TTT TGT GGG GGT GGG AGT GGA GAG TAG 431
432 GGT GGT CTT GTG AGT TGT GCA GGG TGA AGA CCG CTT CCC TGA GAC AGG 479
480 GGC AGT GGT GCT GAT GGA ATG TGG GGG AGG CCC ACA TTT GAG CAA AGC 527
1 Met Trp Gly Arg Pro Thr Phe Glu Gln Ser 10
528 TGC CCT GCC CTT GTC CCC TGG CCT GGC TTC CTG GTA AGG AGT TTC AGC 575
11 Cys Pro Ala Leu Val Pro Trp Pro Gly Phe Leu Val Arg Ser Phe Ser 26
576 CGC CTC CGC AGG AAC CCC CAA AGT GCA GAT TCC GGA GCA GAC ACA TCC 623
27 Arg Leu Arg Arg Asn Pro Gln Ser Ala Asp Ser Gly Ala Asp Thr Ser 42
624 GGG CGG AGA GAC TCA GCA GAC AAG TGC TGC AGT TGC ACG GTG GGA CCC 671
43 Gly Arg Arg Asp Ser Ala Asp Lys Cys Cys Ser Cys Thr Val Gly Pro 58
672 GGG GCC TCG TGC GTT TTT TGC TGT GGG TGG GGT GGG TGG GTT GGT TTA 719
59 Gly Ala Ser Cys Val Phe Cys Cys Gly Trp Gly Gly Trp Val Gly Leu 74
720 TGC CTA TCA ATG CAA TTT TTA ATT TTT GTT AAT ATC AAC AGC AAA AGC 767
75 Cys Leu Ser Met Gln Phe Leu Ile Phe Val Asn Ile Asn Ser Lys Ser 90
768 CTA GTG CAT TGG GAG ATG TGC AAC CTC CCT GAA AAT CTT TTC TGT TTC 815
91 Leu Val His Trp Glu Met Cys Asn Leu Pro Glu Asn Leu Phe Cys Phe 106
816 TGG AGT ACT TCA GGG GTG GCC TCT GGC CCC AGA GCC TTT GCC ACA GTG 863
107 Trp Ser Thr Ser Gly Val Ala Ser Gly Pro Arg Ala Phe Ala Thr Val 122
864 CTC CCA CCA GCC CCC ACC TCA TCC GTC TGT TTG CAG AGC CTC ATC TAC 911
123 Leu Pro Pro Ala Pro Thr Ser Ser Val Cys Leu Gln Ser Leu Ile Tyr 138
912 AGG TCC CCA CGC TGC CTT CTT TAC TCA CTC TGC GCT TGG CCG TTT TGT 959
139 Arg Ser Pro Arg Cys Leu Leu Tyr Ser Leu Cys Ala Trp Pro Phe Cys 154
960 TAT TTG GCT TAG TCT ACA TTG GGC GGA AGT CTG TGT GCA CAG AGT GGG 1007
155 Tyr Leu Ala *** 158
1008 TGT TCC TTC GAG CCC CTT CCA CTC AGA GGG CCA CAC CCA GCG ATG CCA 1055
1056 GTG AAG GTG GCA CAG CCT CTC TTC AGT TTC TCC TGA CTG TGA TCT CAC 1103
1104 TGG GGT AGA ATT CCC CTG AGA GAA TTC CCT CAC TCA CGG CTC CCT TTG 1151
1152 CCA GAG TCA GTT CAA TCA GGT CTG ATG TGA GCA ATT TAC ACA CTT GTC 1199
1200 TCA GAA AGT CCC TCA GGG TTT GTA GAG GAC TGC AGG GGG GCA TCC GCT 1247
1248 GCA GAC TCA GCC TTT CTC TGC AGC CAT CCT GCA GTG GGG GTG AGC GGG 1295
1296 CAC AGG CTG AGA ACT GCT CTT GGG TGG TGG AAG CAG GTG TCA CGG TGC 1343
1344 AAG TCT CCC CCT GCA CCC CTC CCC CAG CTT GAG CCG TGT CAC CCC CCT 1391
1392 CTC CCT CCA GCA TGG GCC TGT GTC TCA GCT CTC TGG AAG GTG GCC CTG 1439
1440 CCC GGA CCC TCT TGC AGG TGT CCT GGT TTG ACT TGG AAC TAG ATG GCC 1487
1488 ATC TTT TCA GGC TTT GGT GGC CCA AGA GCA GTC TGG GTG GAT GGA AGT 1535
1536 GGC TGT CCC CTT CTC TTC AGC CCC TGC CCA CCC ACT GGT GGA GGT GCT 1583
1584 AAC TAG CAG GAC GTG GCA TAG GAT GGG AGC TGG GCG TGA GGT GCT TTG 1631
1632 GGG TCC ATT CTT TGT CCC TCA GCT TCT CAG AGT CCG GCC AGC CCT TGT 1679
1680 GTT CCC GTG CCC CAC ACT TTC CTC CTC CCC ACT GCA GTG AGT CAA TAG 1727
1728 TCC AGG GTG GGG CCT GGC CTC CCT GCC CTG ATT GGG GAC TCA GGA GGT 1775
1776 AAG GCC TGG GGG GCT TCC TGC CCC CTC CTT GCC CAC CTG CCT GCC CCC 1823
1824 GGG CAG CAC GGG AGG GAA AGC AGG GTG AGC ACG CTT GTT GGT TTC AAA 1871
1872 TGC ACT TTC TGC TTG CAT TGC CGT ATC TGT GCG TTC CTT CAT CCT GGT 1919
1920 CCT GGC TTT ATG GAA CAC CAT GTT TTT AGC ATG TTT TTA AAT AAA AAC 1967
1968 GGA TAA AGT GTC AAA AGC CAA AAA AAA AAA AAA AAA AAA AAA AAA AAA 2015
2016 AAA AAA AAA AAA AAA 2030
5.PP1551
A:核苷酸序列(SEQ ID NO:13)长度:2332bp
1 CACCTCTGGG GAAGGTGTGA GAGGAGAGAA GGAAGTGGCT GTTTGGCTGC
51 TGACAACATG AAGACTTCCT GCGATGAGAA CAGAGGCACA GGTGCCGGCC
101 CTGCAGCCCC CAGAACCCGG ACTGGAGGGG GCCATGGGGC GCCGGACCCT
151 GGCCCTGCCC TGGGTGCTGC TGACCCTGCG TGTCACTGCA GGGACCCCGG
201 AGGTGTGAGT ACAAGTTCGG ATGGAGGCCA CCGAGCTCTC GTCCTTCACC
251 ATCCGTTGTG GGTTCCTGGA GTCTGGCTCC ATCTCCCTGG TGACTGTGAG
301 CAGGGGGGGC CCCGATGGTG CTGGGGGGAC CACGCTGGCT GTGTTGCACC
351 CGGTTCTTGG CATCCAGCAA TGGGCCCCTG CTCGCCAGGC CCGCTGGGAA
401 ACCCAGAGCA GCATCTCTCT CATCCTGGAA GGCTCTGGGG CCAGCAGCCC
451 CTGCGCCAAC ACCACCTTCT GCTGCAAGTT TGCGTCCTTC CCTGAGGGCT
501 CCTGGGAGGC CTGTGGGAGC CTCCCGCCCA GCTCAGACCC AGGTGGGGCC
551 GGGGTGAGGG GTCCAGGAGG GCAGGGAGGG GCTCGGGAAC TGGCCATCCA
601 TCTGATTCTT GCCTCTGTGC CCAGGGCTCT CTGTCCCGCC GACTCCTGCC
651 CCCATTCTGC GGGCAGACCT GGCCGGGATC TTGGGGGTCT CAGGAGTCCT
701 TCTCTTTGAC TGTGGCTACC TCCTTCATCT GCTGTGCCGA CAGAAGCACC
751 GCCCTGCCCC TAGGCTCCAG CCATCCCACA CCAGCTCCTA GGCACTGAGA
801 GCACGAGCAT GGGCATAGCG CGCTGGCTAG GACTCCAGTA CCGTGAAGGG
851 AGGCAGTGAG AGCAGACATC TGTGCCTCAT TCCTGATCTC AAGGGGAAAG
901 CAAGAACAAG ACCTGTGAAG GGAAGGAGGG TGGATCGGCC TGACTGGCCC
951 TGGGAAGCAC AGAGGAGCCT ACTGGGACCA GGGAAGAGAC TGACAAGAGA
1001 AGGGAGGCCT GTCCCTGCCT GCTTCTGTCT CAAGAAGATT GTTATCACTT
1051 CCTCTGGTGA AATCCTGCCT TCCTGTCCCC ACGGCCCCTC AGCTGATGCC
1101 ACCTTACTCT GGAGAGGATG GGCAGAGCCT CCTATGGAGG GAGCCCTGTG
1151 GGTCACCAAA GCTCCAGTGA CCACCCCCGG GCAAAGACCT GCAGATCCCC
1201 TGTTAGAGAC GGGCCCAGGA CTTTGTGCGG GGTGCCCACT GTTGCTCTGA
1251 GCCTTCACTT CCTCAGGGAG GCTTCCTCAG GATCTCGAAC CTGCGGAAGG
1301 AGGACCAGTC TGTGTACTTC TGCCAAGTCC AGCTGGACAT ACAGATCAGG
1351 GAGGCTGTCG TGGCAGTCCA TCAAGGGGAC CCACCTCACC ATCACCCAGG
1401 CCCTCAGGCA GCCCCTCCAC AGGGCCCCTC TCCTGCCTGG ACAGCTCTGC
1451 TGGTCTCCCC GTCCCCTGGA GAAGAACAAG GCCATGGGTC GGCCCCTGCT
1501 GCTGCCCCTG CTGCTCCTGC TGCAGCCGCC AGCATTTCTG CAGCCTGGAT
1551 TATGTGAACC GGCTCTTTCT GAACTGGACA GAGGGTCAGG AGAGCGGCTT
1601 CCTCAGGATC TCAAACCTGC GGAAGGAGGA CCAGTCTGTG TATTTCTGCC
1651 GAGTCGAGCT GGACACCCGG AGATCAGGGA GGCAGCAGTT GCAGTCCATC
1701 AAGGGGACCA AACTCACCAT CACCCAGGCT GTCACAACCA CCACCACCTG
1751 GAGGCCCAGC AGCACAACCA CCATAGCCGG CCTCAGGGTC ACAGAAAGCA
1801 AAGGGCACTC AGAATCATGG CACCTAAGTC TGGACACTGC CATCAGGGTT
1851 GCATTGGCTG TCGCTGTGCT CAAAACTGTC ATTTTGGGAC TGCTGTGCCT
1901 CCTCCTCCTG TGGTGGAGGA GAAGGAAAGG TAGCAGGGCG CCAAGCAGTG
1951 ACTTCTGACC AACAGAGTGT GGGGAGAAGG GATGTGTATT AGCCCCGGAG
2001 GACGTGATGT GAGACCCGCT TGTGAGTCCT CCACACTCGT TCCCCATTGG
2051 CAAGATACAT GGAGAGCACC CTGAGGACCT TTAAAAGGCA AAGCCGCAAG
2101 GCAGAAGGAG GCTGGGTCCC TGAATCACCG ACTGGAGGAG AGTTACCTAC
2151 AAGAGCCTTC ATCCAGGAGC ATCCACACTG CAATGATATA GGAATGAGGT
2201 CTGAACTCCA CTGAATTAAA CCACTGGCAT TTGGGGGCTG TTTATTATAG
2251 CAGTGCAAAG AGTTCCTTTA TCCTCCCCAA GGATGGAAAA ATACAATTTA
2301 TTTTGCTTAC CATAAAAAAA AAAAAAAAAA AA
B:氨基酸序列(SEQ ID NO:14)长度:271个氨基酸
1 MGRASYGGSP VGHQSSSDHP RAKTCRSPVR DGPRTLCGVP TVALSLHFLR
51 EASSGSRTCG RRTSLCTSAK SSWTYRSGRL SWQSIKGTHL TITQALRQPL
101 HRAPLLPGQL CWSPRPLEKN KAMGRPLLLP LLLLLQPPAF LQPGLCEPAL
151 SELDRGSGER LPQDLKPAEG GPVCVFLPSR AGHPEIREAA VAVHQGDQTH
201 HHPGCHNHHH LEAQQHNHHS RPQGHRKQRA LRIMAPKSGH CHQGCIGCRC
251 AQNCHFGTAV PPPPVVEEKE R
C:核苷酸及氨基酸组合序列(SEQ ID NO:15)
克隆号和蛋白名称:PP1551
起始编码子: 1118 ATG 终止编码子:1933 TAG
蛋白质分子量:29628.42
1 C ACC TCT GGG GAA GGT GTG AGA GGA GAG AAG GAA GTG GCT GTT TGG 46
47 CTG CTG ACA ACA TGA AGA CTT CCT GCG ATG AGA ACA GAG GCA CAG GTG 94
95 CCG GCC CTC CAG CCC CCA GAA CCC GGA CTG GAG GGG GCC ATG GGG CGC 142
143 CGG ACC CTG GCC CTG CCC TGG GTG CTG CTG ACC CTG CGT GTC ACT GCA 190
191 GGG ACC CCG GAG GTG TGA GTA CAA GTT CGG ATG GAG GCC ACC GAG CTC 238
239 TCG TCC TTC ACC ATC CGT TGT GGG TTC CTG GAG TCT GGC TCC ATC TCC 286
287 CTG GTG ACT GTG AGC AGG GGG GGC CCC GAT GGT GCT GGG GGG ACC ACG 334
335 CTG GCT GTG TTG CAC CCG GAA CTT GGC ATC CAG CAA TGG GCC CCT GCT 382
383 CGC CAG GCC CGC TGG GAA ACC CAG AGC AGC ATC TCT CTC ATC CTG GAA 430
431 GGC TCT GGG GCC AGC AGC CCC TGC GCC AAC ACC ACC TTC TGC TGC AAG 478
479 TTT GCG TCC TTC CCT GAG GGC TCC TGG GAG GCC TGT GGG AGC CTC CCG 526
527 CCC AGC TCA GAC CCA GGT GGG GCC GGG GTG AGG GGT CCA GGA GGG CAG 574
575 GGA GGG GCT CGG GAA CTG GCC ATC CAT CTG ATT CTT GCC TCT GTG CCC 622
623 AGG GCT CTC TGT CCC GCC GAC TCC TGC CCC CAT TCT GCG GGC AGA CCT 670
671 GGC CGG GAT CTT GGG GGT CTC AGG AGT CCT TCT CTT TGA CTG TGG CTA 718
719 CCT CCT TCA TCT GCT GTG CCG ACA GAA GCA CCG CCC TGC CCC TAG GCT 766
767 CCA GCC ATC CCA CAC CAG CTC CTA GGC ACT GAG AGC ACG AGC ATG GGC 814
815 ATA GCG CGC TGG CTA GGA CTC CAG TAC CGT GAA GGG AGG CAG TGA GAG 862
863 CAG ACA TCT GTG CCT CAT TCC TGA TCT CAA GGG GAA AGC AAG AAC AAG 910
911 ACC TGT GAA GGG AAG GAG GGT GGA TCG GCC TGA CTG GCC CTG GGA AGC 958
959 ACA GAG GAG CCT ACT GGG ACC AGG GAA GAG ACT GAC AAG AGA AGG GAG 1006
1007 GCC TGT CCC TGC CTG CTT CTG TCT CAA GAA GAT TGT TAT CAC TTC CTC 1054
1055 TGG TGA AAT CCT GCC TTC CTG TCC CCA CGG CCC CTC AGC TGA TGC CAC 1102
1103 CTT ACT CTG GAG AGG ATG GGC AGA GCC TCC TAT GGA GGG AGC CCT GTG 1150
1 Met Gly Arg Ala Ser Tyr Gly Gly Ser Pro Val 11
1151 GGT CAC CAA AGC TCC AGT GAC CAC CCC CGG GCA AAG ACC TGC AGA TCC 1198
12 Gly His Gln Ser Ser Ser Asp His Pro Arg Ala Lys Thr Cys Arg Ser 27
1199 CCT GTT AGA GAC GGG CCC AGG ACT TTG TGC GGG GTG CCC ACT GTT GCT 1246
28 Pro Val Arg Asp Gly Pro Arg Thr Leu Cys Gly Val Pro Thr Val Ala 43
1247 CTG AGC CTT CAC TTC CTC AGG GAG GCT TCC TCA GGA TCT CGA ACC TGC 1294
44 Leu Ser Leu His Phe Leu Arg Glu Ala Ser Ser Gly Ser Arg Thr Cys 59
1295 GGA AGG AGG ACC AGT CTG TGT ACT TCT GCC AAG TCC AGC TGG ACA TAC 1342
60 Gly Arg Arg Thr Ser Leu Cys Thr Ser Ala Lys Ser Ser Trp Thr Tyr 75
1343 AGA TCA GGG AGG CTG TCG TGG CAG TCC ATC AAG GGG ACC CAC CTC ACC 1390
76 Arg Ser Gly Arg Leu Ser Trp Gln Ser Ile Lys Gly Thr His Leu Thr 91
1391 ATC ACC CAG GCC CTC AGG CAG CCC CTC CAC AGG GCC CCT CTC CTG CCT 1438
92 Ile Thr Gln Ala Leu Arg Gln Pro Leu His Arg Ala Pro Leu Leu Pro 107
1439 GGA CAG CTC TGC TGG TCT CCC CGT CCC CTG GAG AAG AAC AAG GCC ATG 1486
108 Gly Gln Leu Cys Trp Ser Pro Arg Pro Leu Glu Lys Asn Lys Ala Met 123
1487 GGT CGG CCC CTG CTG CTG CCC CTG CTG CTC CTG CTG CAG CCG CCA GCA 1534
124 Gly Arg Pro Leu Leu Leu Pro Leu Leu Leu Leu Leu Gln Pro Pro Ala 139
1535 TTT CTG CAG CCT GGA TTA TGT GAA CCG GCT CTT TCT GAA CTG GAC AGA 1582
140 Phe Leu Gln Pro Gly Leu Cys Glu Pro Ala Leu Ser Glu Leu Asp Arg 155
1583 GGG TCA GGA GAG CGG CTT CCT CAG GAT CTC AAA CCT GCG GAA GGA GGA 1630
156 Gly Ser Gly Glu Arg Leu Pro Gln Asp Leu Lys Pro Ala Glu Gly Gly 171
1631 CCA GTC TGT GTA TTT CTG CCG AGT CGA GCT GGA CAC CCG GAG ATC AGG 1678
172 Pro Val Cys Val Phe Leu Pro Ser Arg Ala Gly His Pro Glu Ile Arg 187
1679 GAG GCA GCA GTT GCA GTC CAT CAA GGG GAC CAA ACT CAC CAT CAC CCA 1726
188 Glu Ala Ala Val Ala Val His Gln Gly Asp Gln Thr His His His Pro 203
1727 GGC TGT CAC AAC CAC CAC CAC CTG GAG GCC CAG CAG CAC AAC CAC CAT 1774
204 Gly Cys His Asn His His His Leu Glu Ala Gln Gln His Asn His His 219
1775 AGC CGG CCT CAG GGT CAC AGA AAG CAA AGG GCA CTC AGA ATC ATG GCA 1822
220 Ser Arg Pro Gln Gly His Arg Lys Gln Arg Ala Leu Arg Ile Met Ala 235
1823 CCT AAG TCT GGA CAC TGC CAT CAG GGT TGC ATT GGC TGT CGC TGT GCT 1870
236 Pro Lys Ser Gly His Cys His Gln Gly Cys Ile Gly Cys Arg Cys Ala 251
1871 CAA AAC TGT CAT TTT GGG ACT GCT GTG CCT CCT CCT CCT GTG GTG GAG 1918
252 Gln Asn Cys His Phe Gly Thr Ala Val Pro Pro Pro Pro Val Val Glu 267
1919 GAG AAG GAA AGG TAG CAG GGC GCC AAG CAG TGA CTT CTG ACC AAC AGA 1966
268 Glu Lys Glu Arg *** 272
1967 GTG TGG GGA GAA GGG ATG TGT ATT AGC CCC GGA GGA CGT GAT GTG AGA 2014
2015 CCC GCT TGT GAG TCC TCC ACA CTC GTT CCC CAT TGG CAA GAT ACA TGG 2062
2063 AGA GCA CCC TGA GGA CCT TTA AAA GGC AAA GCC GCA AGG CAG AAG GAG 2110
2111 GCT GGG TCC CTG AAT CAC CGA CTG GAG GAG AGT TAC CTA CAA GAG CCT 2158
2159 TCA TCC AGG AGC ATC CAC ACT GCA ATG ATA TAG GAA TGA GGT CTG AAC 2206
2207 TCC ACT GAA TTA AAC CAC TGG CAT TTG GGG GCT GTT TAT TAT AGC AGT 2254
2255 GCA AAG AGT TCC TTT ATC CTC CCC AAG GAT GGA AAA ATA CAA TTT ATT 2302
2303 TTG CTT ACC ATA AAA AAA AAA AAA AAA AAA 2332
6.PP1553
A:核苷酸序列(SEQ ID NO:16)长度:2265bp
1 GGAAGGAGGA GGTCCAGGCC GGTAGTGGCC ATTCGCCGTG CCTCAGCGAG
51 CAGGTGTGTG TGGGTCCTCC ACCACTCACC TCTTGGTTAG CGGGAGTGTG
101 CTGCCCCCAC CCCCACCCCC GCACCCCCAT TCTACACAAG GCAGAAGAGG
151 CACGGGTTTT TCTGGGAGCG AATATCAAGT GCCTGAGAGC AACTACAGGA
201 CTAACTGTGT TTGGGTTGGG TGTAGTATAA ATAATAATAA TGGCTAATAT
251 TTCCTGAGCA TCTACTAAAT GCAAGGAATT GTGCTTGGTG TGTCATGTGG
301 ATTCTCTCTT GCATCTTCAT GATAAATGTT ATTGTCGCTG TTTTACCGAT
351 GAGGGTTGGA TTAGAGGGGT TAAACAACTT GTCTTAGGCT CCACAGCTGG
401 GAACAAGTGG GGCTGGGAAG CTGACTTCGT GCTCTTCACC ACCACAAAGG
451 ATGTGTGTGC ATCCTGGGGC ATGCCTGCCT CATGTGGGGG TGTCCTGGGC
501 TGAATTTCCT GGGCACTTCT CAGTGGAACT CTCTAGCCTC CTGGTTCGGA
551 ATGTCAACTA TGAGATCCCC TCACTGAAGA AGCAGATTGC CAAGTGCCAG
601 CAGCTGCAGC AAGAATACAG CCGCAAGGAG GAGGAGTGCC AGGCAGGGGC
651 TGCCGAGATG CGGGAGCAGT TCTACCACTC CTGCAAGCAG TATGGCATCA
701 CGGGCGAAAA TGTCCGAGGA GAACTGCTGG CCCTGGTGAA GGACCTGCCG
751 AGTCAGCTGG CTGAGATTGG GGCAGCGGCT CAGCAGTCCC TGGGGGAAGC
801 CATTGACGTG TACCAGGCGT CTGTGGGGTT TGTGTGTGAG AGGTAGAGAG
851 GCCTCAGCTT CTCCTGGTGG GGGTGCTTTG CCTGTGTTCC CCAGCTCATG
901 ACCCTTCTCC AGTTGTCTTG TTCCCATATA ACATTTGAAC TCTTTACACA
951 CCTGAACCTG TGGGGGCCTT GCCCATTTGA CCATGTGGCC CAGGCCAAAG
1001 CCCAGTGTTG GCCTTACGCA TGGTCGGCAG GAGAGTCAGT TGTGTGCTCT
1051 GTTGAAGCCC CACAGAGCAG GTGTTGCCAA TGCTGCGGTT CGTGCAGAAG
1101 CGGGGAAACT CAACGGTGTA CGAGTGGAGG ACAGGGACAG AGCCCTCTGT
1151 GGTGGAACGA CCCCACCTCG AGGAGCTTCC TGAGCAGGTG GCAGAAGATG
1201 CGATTGACTG GGGCGACTTT GGGGTAGAGG CAGTGTCTGA GGGGACTGAC
1251 TCTGGCATCT CTGCCGAGGC TGCTGGAATC GACTGGGGCA TCTTCCCGGA
1301 ATCAGATTCA AAGGATCCTG GAGGTGATGG GATAGACTGG GGAGACGATG
1351 CTGTTGCTTT GCAGATCACA GTGCTGGAAG CAGGAACCCA GGCTCCAGAA
1401 GGTGTTGCCA GGGGCCCAGA TGCCCTGACA CTGCTTGAAT ACACTGAGAC
1451 CCGGAATCAG TTCCTTGATG AGCTCATGGA GCTTGAGATC TTCTTAGCCC
1501 AGAGAGCAGT GGAGTTGAGT GAGGAGGCAG ATGTCCTGTC TGTGAGCCAG
1551 TTCCAGCTGG CTCCAGCCAT CCTGCAGGGC CAGACCAAAG AGAAGATGGT
1601 TACCATGGTG TCAGTGCTGG AGGATCTGAT TGGCAAGCTT ACCAGTCTTC
1651 AGCTGCAACA CCTGTTTATG ATCCTGGCCT CACCAAGGTA TGTGGACCGA
1701 GTGACTGAAT TCCTCCAGCA AAAGCTGAAG CAGTCCCAGC TGCTGGCTTT
1751 GAAGAAAGAG CTGATGGTGC AGAAGCAGCA GGAGGCACTT GAGGAGCAGG
1801 CGGCTCTGGA GCCTAAGCTG GACCTGCTAC TGGAGAAGAC CAAGGAGCTG
1851 CAGAAGCTGA TTGAAGCTGA CATCTCCAAG AGGTACAGCG GGCGCCCTGT
1901 GAACCTGATG GGAACCTCTC TGTGACACCC TCCGTGTTCT TGCCTGCCCA
1951 TCTTCTCCGC TTTTGGGATG AAGATGATAG CCAGGGCTGT TGTTTTGGGG
2001 CCCTTCAAGG CAAAAGACCA GGCTGACTGG AAGATGGAAA GCCACAGGAA
2051 GGAAGCGGCA CCTGATGGTG ATCTTGGCAC TCTCCATGTT CTCTACAAGA
2101 AGCTGTGGTG ATTGGCCCTG TGGTCTATCA GGCGAAAACC ACAGATTCTC
2151 CTTCTAGTTA GTATAGCGGA CTTAATAAAA GAGGAAAAAA CTCTTTAAAA
2201 AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA
2251 AAAAAAAAAA AAAAA
B:氨基酸序列(SEQ ID NO:17)长度:281个氨基酸
1 MLRFVQKRGN STVYEWRTGT EPSVVERPHL EELPEQVAED AIDWGDFGVE
51 AVSEGTDSGI SAEAAGIDWG IFPESDSKDP GGDGIDWGDD AVALQITVLE
101 AGTQAPEGVA RGPDALTLLE YTETRNQFLD ELMELEIFLA QRAVELSEEA
151 DVLSVSQFQL APAILQGQTK EKMVTMVSVL EDLIGKLTSL QLQHLFMILA
201 SPRYVDRVTE FLQQKLKQSQ LLALKKELMV QKQQEALEEQ AALEPKLDLL
251 LEKTKELQKL IEADISKRYS GRPVNLMGTS L
C:核苷酸及氨基酸序列(SEQ ID NO:18)
克隆号和蛋白名称:PP1553
起始编码子: 1080 ATG 终止编码子:1925 TGA
蛋白质分子量:31198.70
1 GG AAG GAG GAG GTC CAG GCC GGT AGT GGC CAT TCG CCG TGC CTC AGC 47
48 GAG CAG GTG TGT GTG GGT CCT CCA CCA CTC ACC TCT TGG TTA GCG GGA 95
96 GTG TGC TGC CCC CAC CCC CAC CCC CGC ACC CCC ATT CTA CAC AAG GCA 143
144 GAA GAG GCA CGG GTT TTT CTG GGA GCG AAT ATC AAG TGC CTG AGA GCA 191
192 ACT ACA GGA CTA ACT GTG TTT GGG TTG GGT GTA GTA TAA ATA ATA ATA 239
240 ATG GCT AAT ATT TCC TGA GCA TCT ACT AAA TGC AAG GAA TTG TGC TTG 287
288 GTG TGT CAT GTG GAT TCT CTC TTG CAT CTT CAT GAT AAA TGT TAT TGT 335
336 CGC TGT TTT ACC GAT GAG GGT TGG ATT AGA GGG GTT AAA CAA CTT GTC 383
384 TTA GGC TCC ACA GCT GGG AAC AAG TGG GGC TGG GAA GCT GAC TTC GTG 431
432 CTC TTC ACC ACC ACA AAG GAT GTG TGT GCA TCC TGG GGC ATG CCT GCC 479
480 TCA TGT GGG GGT GTC CTG GGC TGA ATT TCC TGG GCA CTT CTC AGT GGA 527
528 ACT CTC TAG CCT CCT GGT TCG GAA TGT CAA CTA TGA GAT CCC CTC ACT 575
576 GAA GAA GCA GAT TGC CAA GTG CCA GCA GCT GCA GCA AGA ATA CAG CCG 623
624 CAA GGA GGA GGA GTG CCA GGC AGG GGC TGC CGA GAT GCG GGA GCA GTT 671
672 CTA CCA CTC CTG CAA GCA GTA TGG CAT CAC GGG CGA AAA TGT CCG AGG 719
720 AGA ACT GCT GGC CCT GGT GAA GGA CCT GCC GAG TCA GCT GGC TGA GAT 767
768 TGG GGC AGC GGC TCA GCA GTC CCT GGG GGA AGC CAT TGA CGT GTA CCA 815
816 GGC GTC TGT GGG GTT TGT GTG TGA GAG GTA GAG AGG CCT CAG CTT CTC 863
864 CTG GTG GGG GTG CTT TGC CTG TGT TCC CCA GCT CAT GAC CCT TCT CCA 911
912 GTT GTC TTG TTC CCA TAT AAC ATT TGA ACT CTT TAC ACA CCT GAA CCT 959
960 GTG GGG GCC TTG CCC ATT TGA CCA TGT GGC CCA GGC CAA AGC CCA GTG 1007
1008 TTG GCC TTA CGC ATG GTC GGC AGG AGA GTC AGT TGT GTG CTC TGT TGA 1055
1056 AGC CCC ACA GAG CAG GTG TTG CCA ATG CTG CGG TTC GTG CAG AAG CGG 1103
1 Met Leu Arg Phe Val Gln Lys Arg 8
1104 GGA AAC TCA ACG GTG TAC GAG TGG AGG ACA GGG ACA GAG CCC TCT GTG 1151
9 Gly Asn Ser Thr Val Tyr Glu Trp Arg Thr Gly Thr Glu Pro Ser Val 24
1152 GTG GAA CGA CCC CAC CTC GAG GAG CTT CCT GAG CAG GTG GCA GAA GAT 1199
25 Val Glu Arg Pro His Leu Glu Glu Leu Pro Glu Gln Val Ala Glu Asp 40
1200 GCG ATT GAC TGG GGC GAC TTT GGG GTA GAG GCA GTG TCT GAG GGG ACT 1247
41 Ala Ile Asp Trp Gly Asp Phe Gly Val Glu Ala Val Ser Glu Gly Thr 56
1248 GAC TCT GGC ATC TCT GCC GAG GCT GCT GGA ATC GAC TGG GGC ATC TTC 1295
57 Asp Ser Gly Ile Ser Ala Glu Ala Ala Gly Ile Asp Trp Gly Ile Phe 72
1296 CCG GAA TCA GAT TCA AAG GAT CCT GGA GGT GAT GGG ATA GAC TGG GGA 1343
73 Pro Glu Ser Asp Ser Lys Asp Pro Gly Gly Asp Gly Ile Asp Trp Gly 88
1344 GAC GAT GCT GTT GCT TTG CAG ATC ACA GTG CTG GAA GCA GGA ACC CAG 1391
89 Asp Asp Ala Val Ala Leu Gln Ile Thr Val Leu Glu Ala Gly Thr Gln 104
1392 GCT CCA GAA GGT GTT GCC AGG GGC CCA GAT GCC CTG ACA CTG CTT GAA 1439
105 Ala Pro Glu Gly Val Ala Arg Gly Pro Asp Ala Leu Thr Leu Leu Glu 120
1440 TAC ACT GAG ACC CGG AAT CAG TTC CTT GAT GAG CTC ATG GAG CTT GAG 1487
121 Tyr Thr Glu Thr Arg Asn Gln Phe Leu Asp Glu Leu Met Glu Leu Glu 136
1488 ATC TTC TTA GCC CAG AGA GCA GTG GAG TTG AGT GAG GAG GCA GAT GTC 1535
137 Ile Phe Leu Ala Gln Arg Ala Val Glu Leu Ser Glu Glu Ala Asp Val 152
1536 CTG TCT GTG AGC CAG TTC CAG CTG GCT CCA GCC ATC CTG CAG GGC CAG 1583
153 Leu Ser Val Ser Gln Phe Gln Leu Ala Pro Ala Ile Leu Gln Gly Gln 168
1584 ACC AAA GAG AAG ATG GTT ACC ATG GTG TCA GTG CTG GAG GAT CTG ATT 1631
169 Thr Lys Glu Lys Met Val Thr Met Val Ser Val Leu Glu Asp Leu Ile 184
1632 GGC AAG CTT ACC AGT CTT CAG CTG CAA CAC CTG TTT ATG ATC CTG GCC 1679
185 Gly Lys Leu Thr Ser Leu Gln Leu Gln His Leu Phe Met Ile Leu Ala 200
1680 TCA CCA AGG TAT GTG GAC CGA GTG ACT GAA TTC CTC CAG CAA AAG CTG 1727
201 Ser Pro Arg Tyr Val Asp Arg Val Thr Glu Phe Leu Gln Gln Lys Leu 216
1728 AAG CAG TCC CAG CTG CTG GCT TTG AAG AAA GAG CTG ATG GTG CAG AAG 1775
217 Lys Gln Ser Gln Leu Leu Ala Leu Lys Lys Glu Leu Met Val Gln Lys 232
1776 CAG CAG GAG GCA CTT GAG GAG CAG GCG GCT CTG GAG CCT AAG CTG GAC 1823
233 Gln Gln Glu Ala Leu Glu Glu Gln Ala Ala Leu Glu Pro Lys Leu Asp 248
1824 CTG CTA CTG GAG AAG ACC AAG GAG CTG CAG AAG CTG ATT GAA GCT GAC 1871
249 Leu Leu Leu Glu Lys Thr Lys Glu Leu Gln Lys Leu Ile Glu Ala Asp 264
1872 ATC TCC AAG AGG TAC AGC GGG CGC CCT GTG AAC CTG ATG GGA ACC TCT 1919
265 Ile Ser Lys Arg Tyr Ser Gly Arg Pro Val Asn Leu Met Gly Thr Ser 280
1920 CTG TGA CAC CCT CCG TGT TCT TGC CTG CCC ATC TTC TCC GCT TTT GGG 1967
281 Leu *** 282
1968 ATG AAG ATG ATA GCC AGG GCT GTT GTT TTG GGG CCC TTC AAG GCA AAA 2015
2016 GAC CAG GCT GAC TGG AAG ATG GAA AGC CAC AGG AAG GAA GCG GCA CCT 2063
2064 GAT GGT GAT CTT GGC ACT CTC CAT GTT CTC TAC AAG AAG CTG TGG TGA 2111
2112 TTG GCC CTG TGG TCT ATC AGG CGA AAA CCA CAG ATT CTC CTT CTA GTT 2159
2160 AGT ATA GCG GAC TTA ATA AAA GAG GAA AAA ACT CTT TAA AAA AAA AAA 2207
2208 AAA AAA AAA AAA AAA AAA AAA AAA AAA AAA AAA AAA AAA AAA AAA AAA 2255
2256 AAA AAA AAA A 2265
7.PP1665
A:核苷酸序列(SEQ ID NO:19)长度:1559bp
1 CTGCCTAGCA GGCAGAGGCC CCTGGTGCGG AAGGTGGCTC CCGGCTTCCA
51 ACAGACATCA GGCTCCAAGG AGGCAGTCGC CAGCCTGCGG AGAGGCCACA
101 TCCAGCGGCT GAACCTGCGC TACACTCAGG TGTCCCGCCA GGAGCTCAGG
151 GACTACGCGT CCTGGAACCT GAGTGTGAAC CTCTACACAG TCAACGCACC
201 GTGGCTCTTC TCCCTGCTGT GGTGTGCGGG GGTCCCATCC GTCACCTCTG
251 ACAACTCCCA CACCCTGTCC CAGGTGCCTT CCCCCCTCTG GATCATGCCC
301 CCGGACGAGT ACTGTCTCAT GTGGGTCACT GCCGACCTGG TCTCCTTCAC
351 CCTCATCGTG GGCATCTTCG TGCTCCAGAA GTGGCGCCTG GGTGGCATAC
401 GGAGCTACAA CCCTGAGCAG ATCATGCTGA GTGCTGCGGT GCGCCGGACC
451 AGCCGGGACG TCAGCATCAT GAAGGAGAAG CTTATTTTCT CAGAGATCAG
501 CGATGGTGTA GAGGTCTCCG ATGTGCTCTC CGTATGTTCA GACAACAGTT
551 ATGACACATA TGCCAACAGC ACCGCCACCC CTGTGGGCCC CCGAGGGGGT
601 GGCAGCCACA CCAAGACCCT CATAGAGCGG AGTGGGCGTT AGCTGAAGAC
651 ATGTCTGTCC CACCTGTACC TGACACAGAA GCTGGGGAGC CTAGGAGAGC
701 TGGTGGAAGT GTGTCTGAAC TCGGAGTGCT CTGGGAGCGG GCTCCACAGC
751 CTCCTTGTGG GCTCCAGCCC CTTGTCAGCC GCAGCCTCTC TTGAGGGGGA
801 CTCCCTGTCT CCTGAGGCCC AGCTGGGCCA GGACTCCATC CTTTCAGATG
851 CCCCTGCAGG CCTGGGGCTC CTTCTGGGAA GTATGGGGCC TAGGGCTTGG
901 TCCCCCTCTT CTGAGGCCCT CTCCTGTATC CCGACCTGGA AGCTTTGATG
951 GGTCATGGGC CATGCCATAC CCCCTGTGGC AATGGAGTGT GTGGATGCTC
1001 ACCTGTGCCA TCTGTCCTCC TGTCTGTGCC AGGAGGCACC TGAGTTCTCT
1051 GCTGTTATCC TGCCCCAAGG GCCTGGGCCG AGCCTCTACC TGAAGCAACT
1101 CTGCTCTTCC TGTCAGTCTC AAAGCACAAG GAGGTTCAGC CCAGGAGGAA
1151 GCCAGCTGCA ATGTGGAGAC ACGTCCTCCT CCCCAACCCA CCTCATGCCA
1201 CCGCCAACCC CCTGCCCCAG GAGCGGGCCT GAGCCACGTC CCCTAGGAGC
1251 AGCTGGAGAT GGCCAAAAGA GTGAGCTCAG GACTACTGGA TCCCATGCCC
1301 AGGTGTCCAG CAGACCTCAA GGCAGAAGGG TCACCTAACC CAGGAGTCCA
1351 CAGACTGATG TGACCTCAGG TTCCCACATC AGTGGCCACA GGGCAGGGCC
1401 CACCTGGTAG AAGTGTTCTG GATATGGCCA GGGTGGGTGT GTGGCTAAGT
1451 GGGCCTGAAC AGAGGGAACC TAGGGCCCTT GGCCAATGTG ATTAAAGCTG
1501 CATCTTGAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA
1551 AAAAAAAAC
B:氨基酸序列(SEQ ID NO:20)长度:127个氨基酸
1 MGHAIPPVAM ECVDAHLCHL SSCLCQEAPE FSAVILPQGP GPSLYLKQLC
51 SSCQSQSTRR FSPGGSQLQC GDTSSSPTHL MPPPTPCPRS GPEPRPLGAA
101 GDGQKSELRT TGSHAQVSSR PQGRRVT
C.核苷酸及氨基酸组合序列(SEQ ID NO:21)
克隆号和蛋白名称:PP1665
起始编码子: 955 ATG 终止编码子:1338 TAA
蛋白质分子量:13303.38
1 CTG CCT AGC AGG CAG AGG CCC CTG GTG CGG AAG GTG GCT CCC GGC TTC 48
49 CAA CAG ACA TCA GGC TCC AAG GAG GCA GTC GCC AGC CTG CGG AGA GGC 96
97 CAC ATC CAG CGG CTG AAC CTG CGC TAC ACT CAG GTG TCC CGC CAG GAG 144
145 CTC AGG GAC TAC GCG TCC TGG AAC CTG AGT GTG AAC CTC TAC ACA GTC 192
193 AAC GCA CCG TGG CTC TTC TCC CTG CTG TGG TGT GCG GGG GTC CCA TCC 240
241 GTC ACC TCT GAC AAC TCC CAC ACC CTG TCC CAG GTG CCT TCC CCC CTC 288
289 TGG ATC ATG CCC CCG GAC GAG TAC TGT CTC ATG TGG GTC ACT GCC GAC 336
337 CTG GTC TCC TTC ACC CTC ATC GTG GGC ATC TTC GTG CTC CAG AAG TGG 384
385 CGC CTG GGT GGC ATA CGG AGC TAC AAC CCT GAG CAG ATC ATG CTG AGT 432
433 GCT GCG GTG CGC CGG ACC AGC CGG GAC GTC AGC ATC ATG AAG GAG AAG 480
481 CTT ATT TTC TCA GAG ATC AGC GAT GGT GTA GAG GTC TCC GAT GTG CTC 528
529 TCC GTA TGT TCA GAC AAC AGT TAT GAC ACA TAT GCC AAC AGC ACC GCC 576
577 ACC CCT GTG GGC CCC CGA GGG GGT GGC AGC CAC ACC AAG ACC CTC ATA 624
625 GAG CGG AGT GGG CGT TAG CTG AAG ACA TGT CTG TCC CAC CTG TAC CTG 672
673 ACA CAG AAG CTG GGG AGC CTA GGA GAG CTG GTG GAA GTG TGT CTG AAC 720
721 TCG GAG TGC TCT GGG AGC GGG CTC CAC AGC CTC CTT GTG GGC TCC AGC 768
769 CCC TTG TCA GCC GCA GCC TCT CTT GAG GGG GAC TCC CTG TCT CCT GAG 816
817 GCC CAG CTG GGC CAG GAC TCC ATC CTT TCA GAT GCC CCT GCA GGC CTG 864
865 GGG CTC CTT CTG GGA AGT ATG GGG CCT AGG GCT TGG TCC CCC TCT TCT 912
913 GAG GCC CTC TCC TGT ATC CCG ACC TGG AAG CTT TGA TGG GTC ATG GGC 960
1 Met Gly 2
961 CAT GCC ATA CCC CCT GTG GCA ATG GAG TGT GTG GAT GCT CAC CTG TGC 1008
3 His Ala Ile Pro Pro Val Ala Met Glu Cys Val Asp Ala His Leu Cys 18
1009 CAT CTG TCC TCC TGT CTG TGC CAG GAG GCA CCT GAG TTC TCT GCT GTT 1056
19 His Leu Ser Ser Cys Leu Cys Gln Glu Ala Pro Glu Phe Ser Ala Val 34
1057 ATC CTG CCC CAA GGG CCT GGG CCG AGC CTC TAC CTG AAG CAA CTC TGC 1104
35 Ile Leu Pro Gln Gly Pro Gly Pro Ser Leu Tyr Leu Lys Gln Leu Cys 50
1105 TCT TCC TGT CAG TCT CAA AGC ACA AGG AGG TTC AGC CCA GGA GGA AGC 1152
51 Ser Ser Cys Gln Ser Gln Ser Thr Arg Arg Phe Ser Pro Gly Gly Ser 66
1153 CAG CTG CAA TGT GGA GAC ACG TCC TCC TCC CCA ACC CAC CTC ATG CCA 1200
67 Gln Leu Gln Cys Gly Asp Thr Ser Ser Ser Pro Thr His Leu Met Pro 82
1201 CCG CCA ACC CCC TGC CCC AGG AGC GGG CCT GAG CCA CGT CCC CTA GGA 1248
83 Pro Pro Thr Pro Cys Pro Arg Ser Gly Pro Glu Pro Arg Pro Leu Gly 98
1249 GCA GCT GGA GAT GGC CAA AAG AGT GAG CTC AGG ACT ACT GGA TCC CAT 1296
99 Ala Ala Gly Asp Gly Gln Lys Ser Glu Leu Arg Thr Thr Gly Ser His 114
1297 GCC CAG GTG TCC AGC AGA CCT CAA GGC AGA AGG GTC ACC TAA CCC AGG 1344
115 Ala Gln Val Ser Ser Arg Pro Gln Gly Arg Arg Val Thr *** 128
1345 AGT CCA CAG ACT GAT GTG ACC TCA GGT TCC CAC ATC AGT GGC CAC AGG 1392
1393 GCA GGG CCC ACC TGG TAG AAG TGT TCT GGA TAT GGC CAG GGT GGG TGT 1440
1441 GTG GCT AAG TGG GCC TGA ACA GAG GGA ACC TAG GGC CCT TGG CCA ATG 1488
1489 TGA TTA AAG CTG CAT CTT GAA AAA AAA AAA AAA AAA AAA AAA AAA AAA 1536
1537 AAA AAA AAA AAA AAA AAA AAA AC 1559
8.PP2135
A:核苷酸序列(SEQ ID NO:22)长度:1188bp
1 GAAAACCCCG GCTTTGAAGC CTCACCACCT GCCCAGGGGA TACCCGAGGC
51 CAAAGTCAGG CACCCCCTGT CCTATGTGGC CCAGCGGCAG CCTTCTGAGT
101 CTGGGCGGCA TCTGCTTTCG GAGCCCAGCA CCCCCCTGTC TCCTCCAGGC
151 CCCGGAGACG TCTTCTTCCC ATCCCTGGAC CCTGTCCCTG ACTCTCCAAA
201 CTTTGAGGTC ATCTAGCCCA GCTGGGGGAC AGTGGGCTGT TGTGGCTGGG
251 TCTGGGGCAG GTGCATTTGA GCCAGGGCTG GCTCTGTGAG TGGCCTCCTT
301 GGCCTCGGCC CTGGTTCCCT CCCTCCTGCT CTGGGCTCAG ATACTGTGAC
351 ATCCCAGAAG CCCAGCCCCT CAACCCCTCT GGATGCTACA TGGGGATGCT
401 GGACGGCTCA GCCCCTGTTC CAAGGATTTT GGGGTGCTGA GATTCTCCCC
451 TAGAGACCTG AAATTCACCA GCTACAGATG CCAAATGACT TACATCTTAA
501 GAAGTCTCAG AACGTCCAGC CCTTCAGCAG CTCTCGTTCT GAGACATGAG
551 CCTTGGGATG TGGCAGCATC AGTGGGACAA GATGGACACT GGGCCACCCT
601 CCCAGGCACC AGACACAGGG CACGGTGGAG AGACTTCTCC CCCGTGGCCG
651 CCTTGGCTCC CCCGTTTTGC CCGAGGCTGC TCTTCTGTCA GACTTCCTCT
701 TTGTACCACA GTGGCTCTGG GGCCAGGCCT GCCTGCCCAC TGGCCATCGC
751 CACCTTCCCC AGCTGCCTCC TACCAGCAGT TTCTCTGAAG ATCTGTCAAC
801 AGGTTAAGTC AATCTGGGGC TTCCACTGCC TGCATTCCAG TCCCCAGAGC
851 TTGGTGGTCC CGAAACGGGA AGTACATATT GGGGCATGGT GGCCTCCGTG
901 AGCAAATGGT GTCTTGGGCA ATCTGAGGCC AGGACAGATG TTGCCCCACC
951 CACTGGAGAT GGTGCTGAGG GAGGTGGGTG GGGCCTTCTG GGAAGGTGAG
1001 TGGAAGAAGG CACCTGCCCC CCGCCCTCCC CATCCCCTAC TCCCACTGCT
1051 CAGCGCGGGC CATTGCAAGG GTGCCACACA ATGTCTTGTC CACCCTGGGA
1101 CACTTCTGAG TATGAAGCGG GATGCTATTA AAAACTACAT GGGGAAACAG
1151 GTGCAAACCC TGGAAAAAAA AAAAAAAAAA AAAAAAA
B:氨基酸序列(SEQ ID NO:23)长度:172个氨基酸
1 MLHGDAGRLS PCSKDFGVLR FSPRDLKFTS YRCQMTYILR SLRTSSPSAA
51 LVLRHEPWDV AASVGQDGHW ATLPGTRHRA RWRDFSPVAA LAPPFCPRLL
101 FCQTSSLYHS GSGARPACPL AIATFPSCLL PAVSLKICQQ VKSIWGFHCL
151 HSSPQSLVVP KREVHIGAWW PP
C:核苷酸及氨基酸组合序列(SEQ ID NO:24)
克隆号和蛋白名称:PP2135
起始编码子: 383 ATG 终止编码子:901 TGA
蛋白质分子量:19046.05
1 G AAA ACC CCG GCT TTG AAG CCT CAC CAC CTG CCC AGG GGA TAC CCG 46
47 AGG CCA AAG TCA GGC ACC CCC TGT CCT ATG TGG CCC AGC GGC AGC CTT 94
95 CTG AGT CTG GGC GGC ATC TGC TTT CGG AGC CCA GCA CCC CCC TGT CTC 142
143 CTC CAG GCC CCG GAG ACG TCT TCT TCC CAT CCC TGG ACC CTG TCC CTG 190
191 ACT CTC CAA ACT TTG AGG TCA TCT AGC CCA GCT GGG GGA CAG TGG GCT 238
239 GTT GTG GCT GGG TCT GGG GCA GGT GCA TTT GAG CCA GGG CTG GCT CTG 286
287 TGA GTG GCC TCC TTG GCC TCG GCC CTG GTT CCC TCC CTC CTG CTC TGG 334
335 GCT CAG ATA CTG TGA CAT CCC AGA AGC CCA GCC CCT CAA CCC CTC TGG 382
383 ATG CTA CAT GGG GAT GCT GGA CGG CTC AGC CCC TGT TCC AAG GAT TTT 430
1 Met Leu His Gly Asp Ala Gly Arg Leu Ser Pro Cys Ser Lys Asp Phe 16
431 GGG GTG CTG AGA TTC TCC CCT AGA GAC CTG AAA TTC ACC AGC TAC AGA 478
17 Gly Val Leu Arg Phe Ser Pro Arg Asp Leu Lys Phe Thr Ser Tyr Arg 32
479 TGC CAA ATG ACT TAC ATC TTA AGA AGT CTC AGA ACG TCC AGC CCT TCA 526
33 Cys Gln Met Thr Tyr Ile Leu Arg Ser Leu Arg Thr Ser Ser Pro Ser 48
527 GCA GCT CTC GTT CTG AGA CAT GAG CCT TGG GAT GTG GCA GCA TCA GTG 574
49 Ala Ala Leu Val Leu Arg His Glu Pro Trp Asp Val Ala Ala Ser Val 64
575 GGA CAA GAT GGA CAC TGG GCC ACC CTC CCA GGC ACC AGA CAC AGG GCA 622
65 Gly Gln Asp Gly His Trp Ala Thr Leu Pro Gly Thr Arg His Arg Ala 80
623 CGG TGG AGA GAC TTC TCC CCC GTG GCC GCC TTG GCT CCC CCG TTT TGC 670
81 Arg Trp Arg Asp Phe Ser Pro Val Ala Ala Leu Ala Pro Pro Phe Cys 96
671 CCG AGG CTG CTC TTC TGT CAG ACT TCC TCT TTG TAC CAC AGT GGC TCT 718
97 Pro Arg Leu Leu Phe Cys Gln Thr Ser Ser Leu Tyr His Ser Gly Ser 112
719 GGG GCC AGG CCT GCC TGC CCA CTG GCC ATC GCC ACC TTC CCC AGC TGC 766
113 Gly Ala Arg Pro Ala Cys Pro Leu Ala Ile Ala Thr Phe Pro Ser Cys 128
767 CTC CTA CCA GCA GTT TCT CTG AAG ATC TGT CAA CAG GTT AAG TCA ATC 814
129 Leu Leu Pro Ala Val Ser Leu Lys Ile Cys Gln Gln Val Lys Ser Ile 144
815 TGG GGC TTC CAC TGC CTG CAT TCC AGT CCC CAG AGC TTG GTG GTC CCG 862
145 Trp Gly Phe His Cys Leu His Ser Ser Pro Gln Ser Leu Val Val Pro 160
863 AAA CGG GAA GTA CAT ATT GGG GCA TGG TGG CCT CCG TGA GCA AAT GGT 910
161 Lys Arg Glu Val His Ile Gly Ala Trp Trp Pro Pro *** 173
911 GTC TTG GGC AAT CTG AGG CCA GGA CAG ATG TTG CCC CAC CCA CTG GAG 958
959 ATG GTG CTG AGG GAG GTG GGT GGG GCC TTC TGG GAA GGT GAG TGG AAG 1006
1007 AAG GCA CCT GCC CCC CGC CCT CCC CAT CCC CTA CTC CCA CTG CTC AGC 1054
1055 GCG GGC CAT IGC AAG GGT GCC ACA CAA TGT CTT GTC CAC CCT GGG ACA 1102
1103 CTT CTG AGT ATG AAG CGG GAT GCT ATT AAA AAC TAC ATG GGG AAA CAG 1150
1151 GTG CAA ACC CTG GAA AAA AAA AAA AAA AAA AAA AAA AA 1188
9.PP2281
A:核苷酸序列(SEQ ID NO:25)长度:1939bp
1 CAGACATTCA CTCTGGCTGC TGGGACATCA GAAAACAAAG TCTTCATCTC
51 TCTCTCCAGT TTCACCCACC CCACCCTTTG CTTTCATTTC AGGTGTGTTG
101 GTCTATATGA CAGGGAGGAG AGTAAAGGAG AGCAGGAGCA ATTGGCTGCC
151 TGCAAAGCCA GCTGGAGGTG AAGTGCAGGA AAGGAAAGGT CACCCCATTC
201 TACTCCATGG CCTCTCTGCT CCCAGCTGTG GTAGGCTCAC ATAGCCAGTG
251 TGATCGGTTT TTAAGAGGCA GTGCTTTTCA GCTTTTCTCC CTGATATATC
301 CATTTTGCTT CCCAGCACTT TTTAGGAGTA GTGAGAGCGC TTCCTGCCCT
351 TGTTGGAAGC CCCAGGGTGG ACTCTCAGCA CGAAGGTCTC TCCCTTAACT
401 GCTGCCCTTC CAAGACTTGC TCCCGAGATG GAGTGGGCGT GGTCTTCCAG
451 GCTGGCCCTT CCTTCTCCTC ACCGCCACCT TCCCTGCCCC AGCCCCAGCA
501 GCCATGGGTA CATGGGTCCC CAGCTCACCT ATGGATTCCC GCCAGTCTGC
551 CCAGCTGCAG TACTCACGCC CCATGGGGGA TCTTGGTCTG TTTTTCTTGT
601 GGGAGCCTAG TGGAGAGCAG ACGTGGCTTT TTATGTGTCT TGTTGGGGAG
651 GTGACTTGCA TGGTGGGGAC AAGGCTGTCG TGGCAACCTT GGGATCGAGT
701 TTGAGACTAA AGGATGTCAT GAGATCCCTG GCTTCTCCCC ATGTTGTTCC
751 CGGACAAGGG CAGAAGGGAG GCATGGCAAG GGACCTCTGC TGTCCTTACT
801 CAACAGTGGT CCTCATCCCT CCCCACCTCC CACTGCTTCC TGCAAGGGCA
851 CCAGTTGTAT GAGAAAGTTG GCCTTTGGAC TTAGGATTTC TTATTGTAGC
901 TAAGAGCCAT CTGAAGCAGC AGGTTGCAGG ACAAATGCTT CAGTCCACCG
951 AGAGCAGTAC CGTGTGGCCA AGAGGTGGAC TCAGAGCCTT CCTTGAGCTA
1001 AACTCGGCCA ACCAAGGCAC GCAGCATGTC CCCTCAGGTC TCCAGTCAGT
1051 CCAGGTTGAC CCTCAGTTCT GGACGTGTGT ATATAGCTGT ATTTAATACC
1101 TCAAGGTCAT TGTGGCTCTG GGGATGCCGG GGCAGGAGGA CGAGGGTGCG
1151 CTGTGGACAC AGCAGTCCGC GGAATTCCGT TCTGGGAAGC CAATGGTCGC
1201 CGGCACCCCT TGCTTCCTCC CTCTGTTGTC TGCCTGTGTG ACACACATCA
1251 ATGGCAATAA CTTCTTCCAA CTCCTCGCAG AAGTGGGAGA GGCCGGCAGC
1301 CTGCACCGAG AGGGACTTTC CTCTCTCTTG CTCCCCGCTT CGTTCTGTTT
1351 TGGCTGCAGA GAGTGGTTCA TCCATACTCT CATTCCCTCG CCTCCCCTTG
1401 TGGACGGGGG TCTTGCCTTT TCAATTCCTG TGTTTTGGTG TCTTCCCTTA
1451 TCTGCTACCC TGAATCACCT GTCCTGGTCT TGCTGTGTGA TGGGAACATG
1501 CTTGTAAACT GCGTAACAAA TCTACTTTGT GTATGTGTCT GTTTATGGGG
1551 GTGGTTTATT ATTTTTGCTG GTCCCTAGAC CACTTTGTAT GACCGTTTGC
1601 AGTCTGAGCA GGCCAGGGGC TGACAGCTAA TGTCAGGACC CTCAGCGGTG
1651 GAGCCTGCTG GGGGGACCCA GCTGCTCTTG GACAAGTGGC TGAGCTCCTA
1701 TCTGGCCTCC TCTTTTTTTT TTTTTCAAGT AATTTGTGTG TATTTCTAAC
1751 TGATGTATTG AAAAAATTCC TAGTATTTCA GTAAAAATGC CTGTTGTGAG
1801 ATGAACCTCC TGTAACTTCT ATCTGTTCTT TTTTGAGGCT CAGGGAGAAA
1851 CTAGCATTTT TTTTTTCCAA ACTACTTTTT GTCACTGTGA CAGTTGTAAA
1901 TAAAGTTTGA AAATGCTTTC CAAAAAAAAA AAAAAAAAA
B:氨基酸序列(SEQ ID NO:26)长度:127个氨基酸
1 MPGQEDEGAL WTQQSAEFRS GKPMVAGTPC FLPLLSACVT HINGNNFFQL
51 LAEVGEAGSL HREGLSSLLL PASFCFGCRE WFIHTLIPSP PLVDGGLAFS
101 IPVFWCLPLS ATLNHLSWSC CVMGTCL
C:核苷酸及氨基酸组合序列(SEQ ID NO:27)
克隆号和蛋白名称:PP2281
起始编码子: 1124 ATG 终止编码子:1507 TAA
蛋白质分子量:13698.16
1 C AGA CAT TCA CTC TGG CTG CTG GGA CAT CAG AAA ACA AAG TCT TCA 46
47 TCT CTC TCT CCA GTT TCA CCC ACC CCA CCC TTT GCT TTC ATT TCA GGT 94
95 GTG TTG GTC TAT ATG ACA GGG AGG AGA GTA AAG GAG AGC AGG AGC AAT 142
143 TGG CTG CCT GCA AAG CCA GCT GGA GGT GAA GTG CAG GAA AGG AAA GGT 190
191 CAC CCC ATT CTA CTC CAT GGC CTC TCT GCT CCC AGC TGT GGT AGG CTC 238
239 ACA TAG CCA GTG TGA TCG GTT TTT AAG AGG CAG TGC TTT TCA GCT TTT 286
287 CTC CCT GAT ATA TCC ATT TTG CTT CCC AGC ACT TTT TAG GAG TAG TGA 334
335 GAG CGC TTC CTG CCC TTG TTG GAA GCC CCA GGG TGG ACT CTC AGC ACG 382
383 AAG GTC TCT CCC TTA ACT GCT GCC CTT CCA AGA CTT GCT CCC GAG ATG 430
431 GAG TGG GCG TGG TCT TCC AGG CTG GCC CTT CCT TCT CCT CAC CGC CAC 478
479 CTT CCC TGC CCC AGC CCC AGC AGC CAT GGG TAC ATG GGT CCC CAG CTC 526
527 ACC TAT GGA TTC CCG CCA GTC TGC CCA GCT GCA GTA CTC ACG CCC CAT 574
575 GGG GGA TCT TGG TCT GTT TTT CTT GTG GGA GCC TAG TGG AGA GCA GAC 622
623 GTG GCT TTT TAT GTG TCT TGT TGG GGA GGT GAC TTG CAT GGT GGG GAC 670
671 AAG GCT GTC GTG GCA ACC TTG GGA TCG AGT TTG AGA CTA AAG GAT GTC 718
719 ATG AGA TCC CTG GCT TCT CCC CAT GTT GTT CCC GGA CAA GGG GAG AAG 766
767 GGA GGC ATG GCA AGG GAC CTC TGC TGT CCT TAC TCA ACA GTG GTC CTC 814
815 ATC CCT CCC CAC CTC CCA CTG CTT CCT CCA AGG GCA CCA GTT GTA TGA 862
863 GAA AGT TGG CCT TTG GAC TTA GGA TTT CTT ATT GTA GCT AAG AGC CAT 910
911 CTG AAG CAG CAG GTT GCA GGA CAA ATG CTT CAG TCC ACC GAG AGC AGT 958
959 ACC GTG TGG CCA AGA GGT GGA CTC AGA GCC TTC CTT GAG CTA AAC TCG 1006
1007 GCC AAC CAA GGC ACG CAG CAT GTC CCC TCA GGT CTC CAG TCA GTC CAG 1054
1055 GTT GAC CCT CAG TTC TGG ACG TGT GTA TAT AGC TGT ATT TAA TAC CTC 1102
1103 AAG GTC ATT GTG GCT CTG GGG ATG CCG GGG CAG GAG GAC GAG GGT GCG 1150
1 Met Pro Gly Gln Glu Asp Glu Gly Ala 9
1151 CTG TGG ACA CAG CAG TCC GCG GAA TTC CGT TCT GGG AAG CCA ATG GTC 1198
10 Leu Trp Thr Gln Gln Ser Ala Glu Phe Arg Ser Gly Lys Pro Met Val 25
1199 GCC GGC ACC CCT TGC TTC CTC CCT CTG TTG TCT GCC TGT GTG ACA CAC 1246
26 Ala Gly Thr Pro Cys Phe Leu Pro Leu Leu Ser Ala Cys Val Thr His 41
1247 ATC AAT GGC AAT AAC TTC TTC CAA CTC CTC GCA GAA GTG GGA GAG GCC 1294
42 Ile Asn Gly Asn Asn Phe Phe Gln Leu Leu Ala Glu Val Gly Glu Ala 57
1295 GGC AGC CTG CAC CGA GAG GGA CTT TCC TCT CTC TTG CTC CCC GCT TCG 1342
58 Gly Ser Leu His Arg Glu Gly Leu Ser Ser Leu Leu Leu Pro Ala Ser 73
1343 TTC TGT TTT GGC TGC AGA GAG TGG TTC ATC CAT ACT CTC ATT CCC TCG 1390
74 Phe Cys Phe Gly Cys Arg Glu Trp Phe Ile His Thr Leu Ile Pro Ser 89
1391 CCT CCC CTT GTG GAC GGG GGT CTT GCC TTT TCA ATT CCT GTG TTT TGG 1438
90 Pro Pro Leu Val Asp Gly Gly Leu Ala Phe Ser Ile Pro Val Phe Trp 105
1439 TGT CTT CCC TTA TCT GCT ACC CTG AAT CAC CTG TCC TGG TCT TGC TGT 1486
106 Cys Leu Pro Leu Ser Ala Thr Leu Asn His Leu Ser Trp Ser Cys Cys 121
1487 GTG ATG GGA ACA TGC TTG TAA ACT GCG TAA CAA ATC TAC TTT GTG TAT 1534
122 Val Met Gly Thr Cys Leu *** 128
1535 GTG TCT GTT TAT GGG GGT GGT TTA TTA TTT TTG CTG GTC CCT AGA CCA 1582
1583 CTT TGT ATG ACC GTT TGC AGT CTG AGC AGG CCA GGG GCT GAC AGC TAA 1630
1631 TGT CAG GAC CCT CAG CGG TGG AGC CTG CTG GGG GGA CCC AGC TGC TCT 1678
1679 TGG ACA AGT GGC TGA GCT CCT ATC TGG CCT CCT CTT TTT TTT TTT TTC 1726
1727 AAG TTT TTT GTG TGT ATT TCT AAC TGA TGT ATT GAA AAA ATT CCT AGT 1774
1775 ATT TCA GTA AAA ATG CCT GTT GTG AGA TGA ACC TCC TGT AAC TTC TAT 1822
1823 CTG TTC TTT TTT GAG GCT CAG GGA GAA ACT AGC ATT TTT TTT TTC CAA 1870
1871 ACT ACT TTT TGT CAC TGT GAC AGT TGT AAA TAA AGT TTG AAA ATG CTT 1918
1919 TCC AAA AAA AAA AAA AAA AAA 1939
10.PP2447
A:核苷酸序列(SEQ ID NO:28)长度:1678bp
1 CCATGTTGGC CAGGCTGGTC TGAGAGAGAA CTCCTGACCT CGTGATCCGC
51 CTGCCTCAGC CTCCCAAAGT GCTGGGATGA CAGGCGTGCG CCACTGCACC
101 CGGCCCGTGA AGGTCTTTTC TGAAGGGAGC TACCTTTGAG AAGTGCCTGT
151 TGGGACAGAA CTCCCTGCTC TGGTCTGACC CTCTGCACGC TAGGGTGGTC
201 CTGCACCTGT CGGGGAGGGG GCCAAGGACA CAGCACTGGG CTGGGGCAGG
251 GCTCTGCGCT GGTGCTGCCT GCCATCCCAG TTCTGCCATT CCCCCAGCGC
301 ACCCCCGCAC CCGCCCCTCC AGCTGATGCC TGCTCCCTCT CTCTGCAGAA
351 CGGGCTCATG TCGGGGCTGA TGCAGATGCT GCTGCTGAAG GTGTCTGCAC
401 ACATCACCGA GCAGCTGGGC ATGGCCCCAG GTGGCGAGTT CAGGGAGGCC
451 TTCAAGGAGG TGGGCACAGG GTGAGGTAGG GGGTGGCCAC AGGGATGTGG
501 CTCACAGGGG TGGTCCGGGG TTCCCAGTGC TCCCCAACAC CCAGCCTCTG
551 CTCCAGGCCA GCAAGGTGCC TTTCTGCAAG TTCCACCTGG GTGACCGACC
601 CATCCCCGTC ACCTTCAAGA GGGCCATCGC AGCGCTCTCC TTCTGGCAGA
651 AGGTCAGGCT GGCTTGGGGC CTGTGCTTCC TGTCAGACCC CATCAGGTAG
701 GGCTGCCCCC GGGACCCTGG CCGGCCTGCA GGGTGGTCTG TGGGAGGCTC
751 CAGGCCCTCC TGTGCAGGTC CAAGCGCAGC CAATCCTCAC TCAAGGCCTT
801 CCCTGCCCTT TCCTTCCGCC ACAAATCCCA AACAAACGTG CTGTGGTCCC
851 TGCCCGGTGT CCACAGTGCC AGCCCCACCC CCCCAGCCCG TTGCCCATCC
901 CTGCGGGGCT GCAGCCATCC CTCTCCACAG CAAGGATGAC GTGGAACGCT
951 GCAAGCAGAA GGACCTACTG GAGCAGATGA TGGCCGAGAT GATTGGCGAG
1001 TTCCCAGACC TGCACCGCAC CATCGTCTCG GAGCGCGACG TCTACCTAAC
1051 CTACATGCTG CGCCAGGCCG CGCGGCGCCT CGAGCTGCCT CGGGCCTCTG
1101 ACGGTGACGG CCGCCCGCAG GCGTGGGACC CCCTGTGAGG CTGAGGCCCG
1151 AGCAGGTACT GACCCCTTGT CCTTCCCCAC AGCCGAGCCC AGGAAGTGCG
1201 TCCCCTCCGT GGTCGTGGGC GTCGTGGGCA TGGGCCACGT GCCTGGCATC
1251 GAGAAGAACT GGAGCACCGA CCTCAACATC CAGGAGATCA TGACCGTGCC
1301 CCCGCCGTCC GTCTCCGGCA GAGTGTCTCG GTTGGCCGTG AAGGCCGCCT
1351 TCTTCGGCCT GCTGGGCTAC AGCCTGTACT GGATGGGCCG CCGCACCGCG
1401 AGCCTGGTCC TGTCGCTGCC CGCCGCGCAG TACTGCCTGC AGAGGGTGAC
1451 CGAGGCCCGG CACAAGTAGG AGACTGCTCC CCGCCCGCTC GGGCCCCTGA
1501 GGAGCCAGTG CCCCCGCGGC ACTTCTGGCT GCCAGGTGCA TCCTAGCCCG
1551 CCCGAGGCCC CTGCCACCCC CCATGGGGGT CTGGGCCCGG CCTCGCCTGC
1601 CCTCCTGGGC CAGTCACCCC TCCCCCAGCC CACCCAAATA AAGGATTATT
1651 TAACTGTCTG AAAAAAAAAA AAAAAAAA
B:氨基酸序列(SEQ ID NO:29)长度:161个氨基酸
1 MWLTGVVRGS QCSPTPSLCS RPARCLSASS TWVTDPSPSP SRGPSQRSPS
51 GRRSGWLGAC ASCQTPSGRA APGTLAGLQG GLWEAPGPPV QVQAQPILTQ
101 GLPCPFLPPQ IPNKRAVVPA RCPQCQPHPP SPLPIPAGLQ PSLSTARMTW
151 NAASRRTYWS R
C:核苷酸及氨基酸组合序列(SEQ ID NO:30)
克隆号和蛋白名称:PP2447
起始编码子: 495 ATG 终止编码子:980 TGA
蛋白质分子量:16922.45
1 CC ATG TTG GCC AGG CTG GTC TGA GAG AGA ACT CCT GAC CTC GTG ATC 47
48 CGC CTG CCT CAG CCT CCC AAA GTG CTG GGA TGA CAG GCG TGC GCC ACT 95
96 GCA CCC GGC CCG TGA AGG TCT TTT CTG AAG GGA GCT ACC TTT GAG AAG 143
144 TGC CTG TTG GGA CAG AAC TCC CTG CTC TGG TCT GAC CCT CTG CAC GCT 191
192 AGG GTG GTC CTG CAC CTG TCG GGG AGG GGG CCA AGG ACA CAG CAC TGG 239
240 GCT GGG GCA GGG CTC TGC GCT GGT GCT GCC TGC CAT CCC AGT TCT GCC 287
288 ATT CCC CCA GCG CAC CCC CGC ACC CGC CCC TCC AGC TGA TGC CTG CTC 335
336 CCT CTC TCT GCA GAA CGG GCT CAT GTC GGG GCT GAT GCA GAT GCT GCT 383
384 GCT GAA GGT GTC TGC ACA CAT CAC CGA GCA GCT GGG CAT GGC CCC AGG 431
432 TGG CGA GTT CAG GGA GGC CTT CAA GGA GGT GGG CAC AGG GTG AGG TAG 479
480 GGG GTG GCC ACA GGG ATG TGG CTC ACA GGG GTG GTC CGG GGT TCC CAG 527
1 Met Trp Leu Thr Gly Val Val Arg Gly Ser Gln 11
528 TGC TCC CCA ACA CCC AGC CTC TGC TCC AGG CCA GCA AGG TGC CTT TCT 575
12 Cys Ser Pro Thr Pro Ser Leu Cys Ser Arg Pro Ala Arg Cys Leu Ser 27
576 GCA AGT TCC ACC TGG GTG ACC GAC CCA TCC CCG TCA CCT TCA AGA GGG 623
28 Ala Ser Ser Thr Trp Val Thr Asp Pro Ser Pro Ser Pro Ser Arg Gly 43
624 CCA TCG CAG CGC TCT CCT TCT GGC AGA AGG TCA GGC TGG CTT GGG GCC 671
44 Pro Ser Gln Arg Ser Pro Ser Gly Arg Arg Ser Gly Trp Leu Gly Ala 59
672 TGT GCT TCC TGT CAG ACC CCA TCA GGT AGG GCT GCC CCC GGG ACC CTG 719
60 Cys Ala Ser Cys Gln Thr Pro Ser Gly Arg Ala Ala Pro Gly Thr Leu 75
720 GCC GGC CTG CAG GGT GGT CTG TGG GAG GCT CCA GGC CCT CCT GTG CAG 767
76 Ala Gly Leu Gln Gly Gly Leu Trp Glu Ala Pro Gly Pro Pro Val Gln 91
768 GTC CAA GCG CAG CCA ATC CTC ACT CAA GGC CTT CCC TGC CCT TTC CTT 815
92 Val Gln Ala Gln Pro Ile Leu Thr Gln Gly Leu Pro Cys Pro Phe Leu 107
816 CCG CCA CAA ATC CCA AAC AAA CGT GCT GTG GTC CCT GCC CGG TGT CCA 863
108 Pro Pro Gln Ile Pro Asn Lys Arg Ala Val Val Pro Ala Arg Cys Pro 123
864 CAG TGC CAG CCC CAC CCC CCC AGC CCG TTG CCC ATC CCT GCG GGG CTG 911
124 Gln Cys Gln Pro His Pro Pro Ser Pro Leu Pro Ile Pro Ala Gly Leu 139
912 CAG CCA TCC CTC TCC ACA GCA AGG ATG ACG TGG AAC GCT GCA AGC AGA 959
140 Gln Pro Ser Leu Ser Thr Ala Arg Met Thr Trp Asn Ala Ala Ser Arg 155
960 AGG ACC TAC TGG AGC AGA TGA TGG CCG AGA TGA TTG GCG AGT TCC CAG 1007
156 Arg Thr Tyr Trp Ser Arg *** 162
1008 ACC TGC ACC GCA CCA TCG TCT CGG AGC GCG ACG TCT ACC TAA CCT ACA 1055
1056 TGC TGC GCC AGG CCG CGC GGC GCC TCG AGC TGC CTC GGG CCT CTG ACG 1103
1104 GTG ACG GCC GCC CGC AGG CGT GGG ACC CCC TGT GAG GCT GAG GCC CGA 1151
1152 GCA GGT ACT GAC CCC TTG TCC TTC CCC ACA GCC GAG CCC AGG AAG TGC 1199
1200 GTC CCC TCC GTG GTC GTG GGC GTC GTG GGC ATG GGC CAC GTG CCT GGC 1247
1248 ATC GAG AAG AAC TGG AGC ACC GAC CTC AAC ATC CAG GAG ATC ATG ACC 1295
1296 GTG CCC CCG CCG TCC GTC TCC GGC AGA GTG TCT CGG TTG GCC GTG AAG 1343
1344 GCC GCC TTC TTC GGC CTG CTG GGC TAC AGC CTG TAC TGG ATG GGC CGC 1391
1392 CGC ACC GCG AGC CTG GTC CTG TCG CTG CCC GCC GCG CAG TAC TGC CTG 1439
1440 CAG AGG GTG ACC GAG GCC CGG CAC AAG TAG GAG ACT GCT CCC CGC CCG 1487
1488 CTC GGG CCC CTG AGG AGC CAG TGC CCC CGC GGC ACT TCT GGG TGC CAG 1535
1536 GTG CAT CCT AGC CCG CCC GAG GCC CCT GCC ACC CCC CAT GGG GGT CTG 1583
1584 GGC CCG GCC TCG CCT GCC CTC CTG GGC CAG TCA CCC CTC CCC CAG CCC 1631
1632 ACC CAA ATA AAG GAT TAT TTA ACT GTC TGA AAA AAA AAA AAA AAA AA 1678
11.PP3111
A:核苷酸序列(SEQ ID NO:31)长度:1034bp
1 AGGACGTGGA GCGCTGCCTC CGGGACACGG GTGTGCAGGG CGTCATGAGC
51 GCAGAGGGCA ACCTGCACAA CCCCGCCCTG TTCGAGGGCC GGAGCCCTGC
101 CGTGTGGGAG CTGGCCGAGG AGTATCTGGA CATCGTGCGG GAGCACCCCT
151 GCCCCCTGTC CTACGTCCGG GCCCACCTCT TCAAGCTGTG GCACCACACG
201 CTGCAGGTGC ACCAGGAGCT GCGAGAGTAG CTGGCCAAGG TGAAGACCCT
251 GGAGGGCATC GCTGCTGTGA GCCAGGAGCT GAAGCTGCGG TGTCAGGAGG
301 AGATATCCAG GCAGGAGGGA GCGAAGCCCA CCGGCGACTT GCCCTTCCAC
351 TGGATCTGCC AGCCCTACAT CCGGCCGGGG CCCAGGGAGG GGAGCAAGGA
401 GAAGGCAGGT GCGCGCAGCA AGCGGGCCCT GGAGGAAGAG GAGGGTGGCA
451 CGGAGGTCCT GTCCAAGAAC AAGCAAAAGA AGCAGCTGAG GAACCCCCAC
501 AAGACCTTCG ACCCCTCTCT GAAGCCAAAA TATGCAAAGT GTGACCAGTG
551 TGGAAACCCA AAGGGCAACA GATGTGTGTT CAGCCTGTGC CGCGGCTGCT
601 GCAAGAAGCG AGCCTCCAAA GAGACTGCAG ACTGCCCAGG TCACGGATTG
651 CTTTTTAAAA CCAAATTGGA GAAGTCTCTG GCCTGGAAAG AGGCCCAGCC
701 TGAGCTGCAG GAGCCTCAGC CAGCAGCACC TGGAACACCA GGTGGCTTCT
751 CCGAAGTCAT GGGCAGTGCC CTGGCCTGAA GGCCCACAAC CCCCACCCCC
801 AGGACTGCTG CTGGAGCCTG GACACGTCCT ACTTAAGAAA ATGCCTTTTA
851 CTCAGGGAAT CTCCTGCTAC TTAATGTGGA AAGACACGCC CATGTCCCCC
901 TTCGGCCCAC TCTGGGGGCC TGGAAATGCT GCAGTGGGGA GCAGGCCCCA
951 GGCTGGACCT GCCCTGTCCT CAGCACGCGT GTGCAAAAGT GAACAATAAA
1001 TCATTTCAAA GATGCGAAAA AAAAAAAAAA AAAA
B:氨基酸序列(SEQ ID NO:32)长度:155个氨基酸
1 MQSVTSVETQ RATDVCSACA AAAARSEPPK RLQTAQVTDC FLKPNWRSLW
51 PGKRPSLSCR SLSQQHLEHQ VASPKSWAVP WPEGPQPPPP GLLLEPGHVL
101 LKKMPFTQGI SCYLMWKDTP MSPFGPLWGP GNAAVGSRPQ AGPALSSARV
151 CKSEQ
C:核苷酸及氨基酸组合序列(SEQ ID NO:33)
克隆号和蛋白名称:PP3111
起始编码子:532 ATG 终止编码子:999 TAA
蛋白质分子量:16780.42
1 AGG ACG TGG AGC GCT GCC TCC GGG ACA CGG GTG TGC AGG GCG TCA TGA 48
49 GCG CAG AGG GCA ACC TGC ACA ACC CCG CCC TGT TCG AGG GCC GGA GCC 96
97 CTG CCG TGT GGG AGC TGG CCG AGG AGT ATC TGG ACA TCG TGC GGG AGC 144
145 ACC CCT GCC CCC TGT CCT ACG TCC GGG CCC ACC TCT TCA AGC TGT GGC 192
193 ACC ACA CGC TGC AGG TGC ACC AGG AGC TGC GAG AGT AGC TGG CCA AGG 240
241 TGA AGA CCC TGG AGG GCA TCG CTG CTG TGA GCC AGG AGC TGA AGC TGC 288
289 GGT GTC AGG AGG AGA TAT CCA GGC AGG AGG GAG CGA AGC CCA CCG GCG 336
337 ACT TGC CCT TCC ACT GGA TCT GCC AGC CCT ACA TCC GGC CGG GGC CCA 384
385 GGG AGG GGA GCA AGG AGA AGG CAG GTG CGC GCA GCA AGC GGG CCC TGG 432
433 AGG AAG AGG AGG GTG GCA CGG AGG TCC TGT CCA AGA ACA AGC AAA AGA 480
481 AGC AGC TGA GGA ACC CCC ACA AGA CCT TCG ACC CCT CTC TGA AGC CAA 528
529 AAT ATG CAA AGT GTG ACC AGT GTG GAA ACC CAA AGG GCA ACA GAT GTG 576
1 Met Gln Ser Val Thr Ser Val Glu Thr Gln Arg Ala Thr Asp Val 15
577 TGT TCA GCC TGT GCC GCG GCT GCT GCA AGA AGC GAG CCT CCA AAG AGA 624
16 Cys Ser Ala Cys Ala Ala Ala Ala Ala Arg Ser Glu Pro Pro Lys Arg 31
625 CTG CAG ACT GCC CAG GTC ACG GAT TGC TTT TTA AAA CCA AAT TGG AGA 672
32 Leu Gln Thr Ala Gln Val Thr Asp Cys Phe Leu Lys Pro Asn Trp Arg 47
673 AGT CTC TGG CCT GGA AAG AGG CCC AGC CTG AGC TGC AGG AGC CTC AGC 720
48 Ser Leu Trp Pro Gly Lys Arg Pro Ser Leu Ser Cys Arg Ser Leu Ser 63
721 CAG CAG CAC CTG GAA CAC CAG GTG GCT TCT CCG AAG TCA TGG GCA GTG 768
64 Gln Gln His Leu Glu His Gln Val Ala Ser Pro Lys Ser Trp Ala Val 79
769 CCC TGG CCT GAA GGC CCA CAA CCC CCA CCC CCA GGA CTG CTG CTG GAG 816
80 Pro Trp Pro Glu Gly Pro Gln Pro Pro Pro Pro Gly Leu Leu Leu Glu 95
817 CCT GGA CAC GTC CTA CTT AAG AAA ATG CCT TTT ACT CAG GGA ATC TCC 864
96 Pro Gly His Val Leu Leu Lys Lys Met Pro Phe Thr Gln Gly Ile Ser 111
865 TGC TAC TTA ATG TGG AAA GAC ACG CCC ATG TCC CCC TTC GGC CCA CTC 912
112 Cys Tyr Leu Met Trp Lys Asp Thr Pro Met Ser Pro Phe Gly Pro Leu 127
913 TGG GGG CCT GGA AAT GCT GCA GTG GGG AGC AGG CCC CAG GCT GGA CCT 960
128 Trp Gly Pro Gly Asn Ala Ala Val Gly Ser Arg Pro Gln Ala Gly Pro 143
961 GCC CTG TCC TCA GCA CGC GTG TGC AAA AGT GAA CAA TAA ATC ATT TCA 1008
144 Ala Leu Ser Ser Ala Arg Val Cys Lys Ser Glu Gln *** 156
1009 AAG ATG CGA AAA AAA AAA AAA AAA AA 1034
Claims (5)
1.一种分离的多核苷酸,其特征在于,它包含一核苷酸序列,该核苷酸序列选自下组:
(a)编码具有抑癌功能的人蛋白的多核苷酸,所述蛋白具有选自下组的氨基酸序列:
SEQ ID NO:2、SEQ ID NO:5、SEQ ID NO:8、SEQ ID NO:11、SEQ ID NO:14、SEQ IDNO:17、SEQ ID NO:20、SEQ ID NO:23、SEQ ID NO:26、SEQ ID NO:29、SEQ IDNO:32;
(b)与多核苷酸(a)互补的多核苷酸。
2.如权利要求1所述的多核苷酸,其特征在于,该多核苷酸编码的多肽具有选自下组的氨基酸序列:SEQ ID NO:2、SEQ ID NO:5、SEQ ID NO:8、SEQ ID NO:11、SEQ IDNO:14、SEQ ID NO:17、SEQ ID NO:20、SEQ ID NO:23、SEQ ID NO:26、SEQ IDNO:29、SEQ ID NO:32。
3.如权利要求1所述的多核苷酸,其特征在于,该多核苷酸的序列选自下组:
SEQ ID NO:3、SEQ ID NO:6、SEQ ID NO:9、SEQ ID NO:12、SEQ ID NO:15、SEQ ID NO:18、SEQ ID NO:21、SEQ ID NO:24、SEQ ID NO:27、SEQ ID NO:30、SEQID NO:33的编码区序列或全长序列。
4.一种载体,其特征在于,它含有权利要求1所述的多核苷酸。
5.一种遗传工程化的宿主细胞,其特征在于,它是选自下组的一种宿主细胞:
(a)用权利要求4所述的载体转化或转导的宿主细胞;
(b)用权利要求1所述的多核苷酸转化或转导的宿主细胞。
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