重组人乳头瘤病毒l1衣壳蛋白的氨基酸序列及其应用的制作方法

专利名称：：重组人乳头瘤病毒l1衣壳蛋白的氨基酸序列及其应用的制作方法
技术领域：
：本发明涉及人乳头瘤病毒感染的预防和治疗领域。具体地，本发明涉及重组人乳头瘤病毒Ll衣壳蛋白的氨基酸序列，编码该氨基酸序列的核苦酸序列，以及包含这些核苷酸序列的载体和宿主细胞，本发明还涉及由该氨基酸序列所组成的HPVL1蛋白五聚体在制备疫苗、药物组合物、诊断抗原或抗体中的应用。背条技术乳头瘤病毒(P^z7/o/^Wn^)是一大类侵染人类和其他动物的DNA病毒。其中人乳头瘤病毒(/f"加cmp印/Z/o加av/nw;HPV)与人类的多种疾病相关，这些疾病包括良性疣和癌症。乳头瘤病毒含有两个用于保护病毒染色体的结构蛋白(或衣壳蛋白)，分别为Ll和L2。360个L1蛋白分子或72个L1五聚体形成病毒颗粒的外壳；L2位于L1外壳的内部，又称内衣壳蛋白，通常每一个L2分子与一个L1五聚体相联。因此，L1和L2是重要的候选免疫原。由于乳头瘤病毒在长期进化过程中所形成的寄主专化性，该病毒很难通过常规的细胞培养来繁殖扩增，必须用基因重组的方法来生产该病毒衣壳蛋白。到目前为止，通过基因重组规模表达目标基因有两个基本选择原核表达与真核表达。其中，原核表达相对于真核表达而言，具有产率高、成本低、系统相对简单和技术相对成熟的优点。但是，原核表达技术也存在着缺陷分子量大的生物大分子的活性表达相对困难，尤其是高等动物及人类的很多基因的生物活性表达更为困难。困难的根本原因在于原核表达的宿主是低等的大肠杆菌等原核生物，基因表达时对表达产物多肽的加工、修饰元件及修饰过程与髙等生物有着重要区别，表达人类等髙等生物基因时，容易导致目标蛋白分子的二级或髙级结构发生错误折叠，造成不正确的分子构象，从而影响或丧失表达产物的生物活性。人乳头瘤病毒是人类及其它高等哺乳动物的专性寄生病毒，自身没有基因表达系统，其全部基因的表达完全依赖其寄主一_髙等真核生物的蛋白表达、翻译、加工和修饰体系。因此，在原核生物中表达象L1衣壳蛋白这样的基因时，往往因为分子折叠及构象发生错误而丧失生物活性。例如，未加修饰的L1蛋白在大肠杆菌中表达时，表达产物常常形成不溶于水的包含体而沉淀。沉淀的L1蛋白目前无法使其回复生物活性，即无法恢复成分子构象正确的可溶性蛋白。原核系统表达髙等生物基因的这些缺陷在一定程度上阻止了其巨大优势的发挥，是目前发酵工业还没有全部使用原核发酵的主要原因之一。为了生产一些具有重要生物活性的多态类药物或生物制剂，人们不得不釆用成本较高的、也较复杂的真核发酵方法。解决上述问题的办法之一是在深入研究的基础上，对目标基因表达产物的序列进行仔细分析，在不影响和改变蛋白分子三维结构的基础上，对目标基因进行一定的修饰，使其能够在大肠杆菌等原核细胞中实现可溶性表达。在实现乳头瘤病毒Ll蛋白原核活性表达的基础上，表达的Ll蛋白分子能够形成正确的分子构象而可溶于水。可溶的L1分子由于三维构象的缘故能够自动进行分子聚合，形成Ll蛋白分子的多聚体。其中一种聚合方式是形成Ll蛋白分子的五聚体。在自然条件下，寄生于人体或其它哺乳动物上皮细胞的乳头瘤病毒在寄主细胞衰老及行将死亡前大量繁殖。繁殖时，病毒在启动自身基因组DNA复制的同时启动L1衣壳蛋白基因在寄主细胞中的表达，表达后的L1蛋白自动形成五聚体结构，然后这些五聚体自身进一步自动聚合，最后由72个五聚体形成一个野生病毒颗粒，每个病毒颗粒内部储藏一套病毒的DNA分子。人体对任何入侵病毒的识别主要是识别病毒表面的衣壳蛋白分子，没有例外。人乳头瘤病毒L1衣壳蛋白的五聚体分子表面构成了人体的抗原识别表位(即抗原表位)。在病毒被识别后，人体产生特异抗体来中和病毒，对抗病毒的感染和危害。为了诱导人体产生对抗乳头瘤病毒的特异抗体，我们通过生物工程的方法表达和纯化病毒的衣壳蛋白，并以纯化的Ll蛋白作为抗原来生产疫苗。重组L1蛋白五聚体本身就具备完整的抗原表位，因此就可以作为抗原用来制备疫苗。但在生产的过程中，部分L1五聚体可以自动聚合形成与野生病毒颗粒大小完全一样的类病毒颗粒(VLP，viruslikeparticle)。VLP的免疫原性及抗原性与Ll五聚体没有实质区别。为了降低生产成本，实现乳头瘤病毒抗原的原核表达，有必要通过基因改造控制L1蛋白VLP的形成，这样有利于使抗原分子的大小均勾一致，便于质量控制，也有利于提高产率。
发明内容(一)要解决的技术问题本发明的目的是提供一种重组人乳头瘤病毒L1衣壳蛋白的氨基酸序列，使得在原核系统中表达的人乳头瘤病毒Ll衣壳蛋白能溶于水，同时控制Ll五聚体的进一步聚合；本发明的又一目的是提供上述重组人乳头瘤病毒L1衣壳蛋白氨基酸序列的应用。(二)技术方案本发明所述的人乳头瘤病毒衣壳蛋白L1的重组氨基酸序列，是将野生型HPVLl蛋白的氨基酸序列N-末端的保守序列VYLPP或VYVPP上游的氨基酸序列替换成GSGGG，同时在其C-末端的保守序列LGRKFL下游第5个氨基酸处通过引入蛋白质翻译的终止密码子形成相对于野生型的截短型序列，其中所述的HPV型选自HPV6、HPVll、HPV16、HPV18、HPV26、HPV31、HPV33、HPV35、HPV39、HPV42、HPV45、HPV51、HPV52、HPV53、HPV56、HPV58、HPV59、HPV66、HPV73、HPV82中的一种或多种。上述不同型HPV的野生型Ll蛋白的氨基酸序列对应的Genebank登陆号分别为:HPV6-NC—000904,HPVll-NC一OO1525HPV26-NC—001583，HPV31-NC—001527，HPV39-NC一001535,HPV42-NC—00153,HPV52-NC001592,HPV53-NC001593,HPV16-AF402678,HPV18-AY262282HPV33-NC—001528,HPV35-NC—001529HPV45-NC—001590,HPV51-NC_001533HPV56-NC001594,HPV58-NC001443HPV59-NC—001635,HPV66-NC—001695，HPV73-X94165,HPV82-AF293961。采用上述策略进行改造后，不同型HPV对应的重组氨基酸序列分别为HPV6对应的氨基酸序列如SEQIDN0:1所示，HPV11对应的氨基酸序列如SEQIDN0:2所示，HPV16对应的氨基酸序列如SEQIDNO:3所示，HPVl8对应的氨基酸序列如SEQIDNO:4所示，HPV26对应的氨基酸序列如SEQIDNO:5所示，HPV31对应的氨基酸序列如SEQIDNO:6所示，HPV33对应的氨基酸序列如SEQIDNO:7所示，HPV35对应的氨基酸序列如SEQIDNO:8所示,HPV39对应的氨基酸序列如SEQIDNO:9所示，HPV42对应的氨基酸序列如SEQIDNO:10所示，HPV45对应的氨基酸序列如SEQIDNO:11所示，HPV51对应的氨基酸序列如SEQIDNO:12所示，HPV52对应的氨基酸序列如SEQIDNO:13所示，HPV53对应的氨基酸序列如SEQIDNO:14所示，HPV56对应的氨基酸序列如SEQIDNO:15所示，HPV58对应的氨基酸序列如SEQIDNO:16所示，HPV59对应的氨基酸序列如SEQIDNO:17所示，HPV66对应的氨基酸序列如SEQIDNO:18所示，HPV73对应的氨基酸序列如SEQIDNO:19所示，HPV82对应的氨基酸序列如SEQIDNO:20所示。采用上述策略进行改造后，在原核系统中表达时得到的是HPVL1蛋白的五聚体，而且该五聚体能溶于水，即HPVL1蛋白实现了正确的空间折叠，具备了生物活性。由于编码不同型HPVL1蛋白的DNA序列均具有较高的保守性，所以上述改造策略适用于所有的HPV型，得到的表达产物均为具备生物活性的HPVLl蛋白五聚体。本发明还涉及包含编码上述氨基酸序列的核苷酸序列的核酸分子，编码上述氨基酸序列的核苷酸序列的共同特征是表达得到的产物具有与野生型HPVLl蛋白相同的免疫特性。所述核酸分子包括但不限于以下形式编码上述氨基酸序列的核苷酸序列与编码HPV前期蛋白(如E6/E7)的核酸序列组成的融合序列。本发明也涉及包含上述核苷酸序列的表达载体，这些载体是通过将上述核酸序列克隆到包含合适启动子和其它合适转录表达调控原件的连接载体中得到的，其中用于连接的载体包括商业上可获得的质粒、细菌噬菌体和粘粒。优选地，用于连接的载体选自pGEX-4T-l、pGEX-4T-2、pGEX-4T-3、pET-28a、pcDNA3.1或任何其它可用于原核表达的表达质粒。更优选地，用于连接的载体选自pGEX-4T-2。这里用到的分子克隆方法具体操作参见《分子克隆》(第三版，科学出版社，2002年8月出版)。本发明还涉及包含上述表达载体的宿主细胞，这些宿主细胞可选自大肠杆菌、昆虫细胞、酵母细胞中的一种或几种。优选地，宿主细胞选自大肠杆菌。本发明进一步涉及HPVL1蛋白五聚体，该五聚体蛋白的单体的一级结构由上述重组氨基酸序列所组成，其二级结构和高级结构由其一级结构所决定的。本发明还提供了生产上述HPVL1蛋白五聚体的方法，它包括如下步骤(1)将编码上述重组氨基酸序列的核苷酸序列克隆到连接载体上，转化大肠杆菌；(2)发酵培养上述大肠杆菌，并表达HPVL1蛋白；(3)分离、纯化表达产物，得到HPVL1蛋白五聚体。本发明进一步涉及包含一种或多种上述重组HPVL1蛋白五聚体的疫苗，该疫苗可应用于制备预防HPV感染的疫苗。优选地，本发明所述的疫苗包含HPV16的重组HPVLl蛋白五聚体，该五聚体蛋白的单体的一级结构由SEQIDN0:3所示的氨基酸序列所组成。更优选地，本发明所述的疫苗包含五种重组HPVLl蛋白五聚体，对应的HPV型分别为HPV6、HPVll、HPV16、HPV18和HPV33,这五种HPVLl蛋白五聚体的单体的一级结构分别由SEQIDNO:l、SEQIDNO:2、SEQIDNO:3、SEQIDNO:4、SEQIDNO:7所示的氨基酸序列所组成。本发明所述的疫苗的形式也可以是包含HPVLl蛋白五聚体+L2蛋白的组合蛋白。上述疫苗的免疫组分也包括一种生理上可接受的载体，包括但不限于能保持HPVL1蛋白五聚体完整性的简单低浓度盐溶液，例如10mMNaCl，O.lmMEDTA;或者更大范围上讲，载体包括代谢缓慢的大分子如蛋白质、多糖、聚乳糖酸、聚甘油酸、复合氨基酸、以及失活病毒粒子等。药理上适用的盐也可在HPVL1蛋白五聚体的复合物中使用。例如，矿物盐象盐酸盐，溴化盐、磷酸盐、硫酸盐等；有机盐类如乙酸盐、甘露糖、苯甲酸等。免疫组分还包括液体，如水、生理盐水、甘油、酒精，以及其他一些物质如可湿剂、乳化剂、和pH缓冲剂。将上述重组HPVL1蛋白五聚体或HPVL1蛋白五聚体+L2蛋白的组合蛋白以免疫有效剂量接种给人体后，可引诱人体产生对这些重组蛋白的免疫反应。人体的这种特异免疫反应可帮助人体预防人乳头瘤病毒的入侵或中和清除已经入侵的人乳头瘤病毒。由重组HPVLl蛋白五聚体或HPVLl蛋白五聚体+L2蛋白的组合蛋白制备的疫苗可采用多种方法作用于人体，包括静脉、肌肉和皮下注射。本发明所述的疫苗在能引发对L1蛋白免疫反应的有效剂量的范围使用。用于增强免疫反应的复合物的特定剂量依照Ll复合物不同而变化。一般而言，Ll五聚体的用量大约在l-500pg/Kg体重之间。上述剂量范围并不排除更高或更低剂量的可能。例如，具体的剂量会根据是否环伴有其他药物剂量而定，或取决于个人的药代动力学、药物积累和代谢速率。本发明还涉及包含上述重组HPVL1蛋白五聚体的药物组合物。该药物组合物包括但不限于下列形式例如包含重组HPVL1蛋白五聚体、HPVE6/E7蛋白及其它药物上可接受的辅料的药物组合物。本发明还进一步公开了上述重组HPVLl蛋白五聚体在制备HPV病毒免疫诊断抗原或抗体中的应用。上述重组HPVLl蛋白五聚体可以用来产生对HPVLl蛋白有特异亲和力的抗体。这些抗体可以被用来直接检测生物样品中是否存在或与特定病理阶段相联系的特定HPV型。HPVLl蛋白五聚体还能在免疫测定中用来检测生物样品中是否存在HPV病毒的抗体或抗原。包括特异结合到Ll蛋白的单链抗体在内的多克隆或单克隆抗体能够被本领域技术人员用成熟的技术生产出来。这些由特异抗原例如HPV不同亚型引诱产生的抗体具有识别特定的HPV亚型的特异性，例如HPV16和HPV18型。利用抗体反应的免疫测定来检测Ll蛋白包括ELISAs、Westernblot、放射免疫测定、免疫组化测定、免疫沉淀或其他方法。抗体和Ll五聚体都可作为检测标记，例如酶联免疫、放射标记、荧光或化学荧光。抗体或L1五聚体可以固定在某个固体支持物上，例如玻璃或塑料波片，组织培养板、多孔板、试管、交换拄、交换柱填充物、蛋白；或者颗粒上如微球体，包括但不限于乳胶、聚苯乙希或玻璃球；或者薄膜上如醋酸纤维或尼龙膜。偶联蛋白组分的方法为本领域技术人员所熟知。只要不防碍抗体原抗体特异结合的能力，任何偶联方式都可使用.生物样品可以是任何被怀疑含有HPV病毒的样品，例如组织活检、涂片、组织切样如皮肤、子宫、生殖上皮细胞、喉、上呼吸道、结膜或口腔内组织。(三)有益效果本发明通过对HPVL1野生型序列的改造，使得在原核系统中表达的重组HPVL1衣壳蛋白实现了正确的空间折叠，能溶于水，解决了目前利用原核系统中表达的HPVL1衣壳蛋白都是不溶于水的包含体(没有生物活性)的难题，打破了用原核表达系统生产HPVLl衣壳蛋白的瓶颈。不仅如此，本发明还有效控制了HPVL1蛋白的聚合程度，表达得到的是与HPVL1的VLP具有相同免疫原性和抗原性的Ll五聚体，Ll五聚体相对于L1的VLP而言具有结构更稳定、大肠杆菌表达更容易实现的优点。本发明使得利用原核表达系统工业生产HPVL1衣壳蛋白成为现实，相对于目前釆用的真核表达系统具有产品质量更稳定、产率更髙、成本低、质量控制方便的优点，具有重大的经济效益和社会效应。图1是重组改造后得到的HPV16Ll衣壳蛋白的SDS-聚丙烯酰胺凝胶电泳，图中泳道l表示marker,泳道2和泳道3表示蛋白清液，泳道4和泳道5表示树脂吸附的目的蛋白，泳道6、泳道7和泳道8表示酶切后的树脂，泳道9表示目的蛋白洗脱液；图2是重组改造后得到的HPV16L1衣充蛋白的液相色谱图；图3是HPV16L1蛋白的电镜照片比较，其中图3(a)表示HPV16Ll的VLP电镜照片，图3(b)表示重组改造后得到的HPV16L1五聚体电镜照片。具体实施方式以下实施例用于说明本发明，但不用来限制本发明的范围。实施例1以HPV16型为例说明对HPVLl衣壳蛋白氨基酸序列的重组改造1、人乳头瘤病毒16型(HPV16)DNA的检测和分离可能含有野生型HPV16病毒的临床细胞样本废弃物(用于常规细胞形态学检查的宫颈脱落细胞的剩余部分)购自北京市安贞医院妇科门诊。悬浮在1.0毫升水中的(约lxl()S个)脱落细胞经1000单位的蛋白酶K(购自华美生物制品有限公司)在摄氏55度处理300分钟，经10000g高速离心30分钟，所得上清液可用于检测是否存在HPV病毒。受感染的HPV16阳性细胞上清液可直接用于PCR扩增HPV16病毒DNA序列的目标基因片段。2、HPV16型L1蛋白DNA的分子克隆HPV16型L1衣壳蛋白基因由目标基因特异寡核苷酸引物对经DNA聚合酶链式反应(PCR)获得。经典的PCR反应体系包含20ugDNA模板、lxPCR缓冲液、正向和反向特异引物(浓度均为0,2^iM)、1.5mM镁离子、1.0单位的TAQDNA聚合酶。反应条件为摄氏95度变性5分钟，经36个PCR循环放大(每一循环为94度30秒，55度30秒，72度2分钟)，反应产物在72度下温育10分钟，然后停止反应。PCR反应的正向引物序列和反向引物序列见表l。PCR反应扩增放大的HPV16LlcDNA片段经TA克隆(具体操作参见《分子克隆》第三版，科学出版社，2002年8月出版)克隆到一个T-Easyvector质粒(购自美国Promega生物试剂公司)上。3、HPV16L1蛋白基因的修饰HPV16Ll蛋白的免疫源性及抗原性依赖于其正确的分子折叠所产生的正确三维构象。对HPV16L1蛋白基因序列进行修饰的目的是增加重组目标基因原核表达时的可溶性，从而利用￡.c^/生产出具有生物活性的Ll蛋白。基因改造的基本步奏是设计特异引物，然后利用以特异引物为模板PCR扩增放大L1目的基因片段。改造后的重组HPV16Ll蛋白基因DNA序列在其5，-端包含有5cwnHI限制性酶切位点，同时含有编码GSGGG的DNA密码子以取代编码氨基酸序列N-末端的保守序列VYLPP上游氨基酸的密码子。用于改造的正向引物序列和反向引物序列见表l。下游引物引入终止密码子UAA，导致L1cDNA在3，-端减少约25个氨基酸。PCR反应体系包含20ugDNA模板、lxPCR缓冲液、正向和反向特异引物(浓度均为0.2pM)、1.5mM镁离子、1.0单位的TAQDNA聚合酶。反应条件为摄氏95度变性5分钟，经36个PCR循环放大(每一循环为94度30秒，55度30秒，72度2分钟)，反应产物在72度下温育IO分钟，然后停止反应。PCR反应扩增放大的HPV16LlcDNA产物经SawHI和屈oI限制性酶酶切形成黏性末端，同时表达质粒pGEX-4T-2(购自华美生物制品有限公司)DNA用Bamffl和XhoI酶切消化，再用T4连接酶(购自美国Promega生物试剂公司)连接PCR产物和表达质粒。连接产物经电转化转移到大肠杆菌BL21(购自华美生物制品有限公司)宿主细胞内(电转化的具体操作参见《分子克隆》第三版，科学出版社，2002年8月出版)。4、Ll蛋白的原核表达和检测载有HPV16L1基因插入片段的表达质粒导入大肠杆菌BL21细胞之后，使用LB培养基(配方见《分子克隆》第三版，科学出版社，2002年8月出版)进行37度培养，菌种接入后生长12小时开始用表达诱导剂IPTG(购自美国Promega公司)诱导，9小时后收获细胞，细胞用NiroSoaviNS2006型髙压勻质机在800bar下重复破碎细胞2次，显微镜检查细胞破碎率达到90%以上。上清溶液中的L1蛋白用经亲和色谱纯化蛋白预装谷胱甘肽-琼脂糖树脂(Amersham公司生产的GlutathioneSepharose4B)色谱柱，取浓度为50%的GlutathioneSepharose4B勾浆放入色谱柱中(每200ml蛋白清液需要5-10ml匀浆)。用5-10倍的柱床体积的缓冲液A(组分为50mmol/LTric-HCl，200mmol/LNaCl，lmmol/LEDTA,pH8.0)洗涤树脂，将蛋白清液加入色谱柱中，与树脂混合均句并在室温下作用20分钟后放出滤过液，用10倍柱床体积的缓冲液A洗涤树脂柱。检测用SDS-聚丙烯酰胺凝胶电泳经密度扫描(具体操作见《分子克隆》第三版，科学出版社，2002年8月出版)检测目的蛋白的纯度(凝胶电泳结果见附图l),结果表达得到的HPV16Ll蛋白纯度高达95%。再用液相分子排阻色谱法[用AmershamBiosciences生产的AKTAFPLC过滤系统进行凝胶过滤色谱，凝胶过滤介质为Amersham生产的HiloadSuperdex200,流动相为pH7.4的磷酸盐缓冲盐溶液(成分见《分子克隆》第三版，科学出版社，2002年8月出版)]检测，结果见附图2，该图表明表达得到的HPV16L1蛋白是可溶于水的，即具备了生物活性。然后用电镜检査纯化的HPV16L1蛋白，结果见附图3(b),对比附图3(a)[HPV16L1的VLP电镜照片]，可见得到的是HPV16L1五聚体。实施例2其余HPV型的L1衣壳蛋白氨基酸序列的重组改造其余HPV型的L1衣壳蛋白氨基酸序列的重组改造(包括HPV6、HPVll、HPV18、HPV26、HPV31、HPV33、HPV35、HPV39、HPV42、HPV45、HPV51、HPV52、HPV53、HPV56、HPV58、HPV59、HPV66、HPV73、HPV82)的操作步骤与检测方法均与实施例1相同，不同之处见表l。表l<table>tableseeoriginaldocumentpage10</column></row><table><table>tableseeoriginaldocumentpage11</column></row><table>上述HPV型的L1衣壳蛋白氨基酸序列经重组改造后，得到的结果均与实施例l中得到的HPV16Ll衣壳蛋白相同，即均得到了可溶于水的L1五聚体。序列表<110>马润林陈小江<120>重组人乳头瘤病毒L1衣壳蛋白的氨基酸序列及其应用<130>MA0702<160>20<170>Patentlnversion3.3<210>1<211>465<212>PRT<213>人工序列<220><223>HPV6Ll<400>1GlySerGlyGlyGlyValTyrValProProProAsnProValSerLys151015ValValAlaThrAspAlaTyrValThrAigThrAsnliePheTyrHis202530AlaSerSerSerArgLeuLeuAlaValGlyHisProTjtPheSerlie354045LysArgAlaAsnLysThrValValProLysValSerGlyTyrGinTyr505560ArgValPheLysValValLeuProAspProA幼LysPheAlaLeuPro65707580AspSerSerLeuPheAspProThrThrGinArgLeuValTrpAlaCys859095ThrGlyLeuGluValGlyA屯GlyGinProLeuGlyValGlyValSer100105110GlyHisProPheLeuAsnLysTyrAspAspValGluAsnSerGlySer115120125GlyGlyAsnProGlyGinAspAsnArgValAsnValGlyMetAspTyr130135140LysGinThrGinLeuCysMetValGlyCysAlaProProLeuGlyGlu145150155160HisTrpGlyLysGlyLysGinCysThrAsnThrProValGinAlaGly165170175AspCysProProLeuGluLeuHeThrSerVallieGinAspGlyAsp180185l卯MetValAspThrGlyPheGlyAlaMetAsnPheAlaAspLeuGinThr195200205<image>imageseeoriginaldocumentpage13</image>ValValAlaThrAspAlaTyrValLysArgThrAsnHePheTyrHis202530AlaSerSerSerAtgLeuLeuAlaValGlyHisProTyrTyrSerlie354045LysLysValAsnLysThrValValProLysValSerGlyTyrGinTyr505560ArgValPheLysValValLeuProAspProAsnLysPheAlaLeuPro65707580AspSerSerLeuPheAspProThrThrGinArgLeuValTrpAlaCys859095ThrGlyLeuGluValGlyA堪GlyGinProLeuGlyValGlyValSer100105110GlyHisProLeuLeuAsnLysTyrAspAspValGluAsnSerGlyGly115120125TyrGlyGlyAsnProGlyGinAspAsnArgValAsnValGlyMetAsp130135140TyrLysGinThrGinLeuCysMetValGlyCysAlaProProLeuGly145150155160GluHisTrpGlyLysGlyThrGinCysSerAsnThrSerValGinAsn165170175GlyAspCysProProLeuGluLeuHeThrSerValHeGinAspGly180185190AspMetValAspThrGlyPheGlyAlaMetAsnPheAlaAspLeuGin195200205ThrAsnLysSerAspValProLeuAsplieCysGlyThrValCysLys210215220TyrProAspTyrLeuGinMetAlaAlaAspProTyrGlyAspArgLeu225230235240PhePheTyrLeuArgLysGluGinMetPheAlaArgHisPhePheAsn245250255AigAlaGlyThrValGlyGluProValProAspAspLeuLeuValLys260265270GlyGlyAsnAsnArgSerSerValAlaSerSerlieTyrValHisThr275280285ProSerGlySerLeuValSerSerGluAlaGinLeuPheAsnLysPro290295300TyrTrpLeuGinLysAlaGinGlyHisAsnAsnGlyHeCysTrpGly305310315320AsnHisLeuPheValThrValValAspThrThrArgSerThrAsnMet325330335ThrLeuCysAlaSerValSerLysSerAlaThrTyrThrAsnSerAsp340345350TyrLysGluT\rMetArgHisValGluGluPheAspLeuGinPhelie355360365PheGinLeuCysSerHeThrLeuSerAlaGluValMetAlaTyrlie370375380HisThrMetAsnProSerValLeuGluAspTrpAsnPheGlyLeuSer385390395400ProProProAsnGlyThrLeuGluAspThrArgTyrValGinSerGin405410415AlalieThrCysGinLysProThrProGluLysGluLysGinAspPro420425430TyrLysAspMetSerPheTrpGluValAsnLeuLysGluLysPheSer435440445SerGluLeuAspGinPheProLeuGlyArgLysPheLeuLeuGinSer450455460Gly465<210>3<211>467<212>PRT<213>人工序列<220><223>HPV16LI<400>3GlySerGlyGlyGlyValTyrLeuProProValProValSerLysVal151015ValSerThrAspGluTyrValAlaArgThrAsnHeTyrTyrHisAla202530GlyThrSerArgLeuLeuAlaValGlyHisProTyrPheProHeLys354045LysProAsnAsnAsnLyslieLeuValProLysValSerGlyLeuGin505560TyrArgValPheArglieHisLeuProAspProAsnLysPheGlyPhe65707580ProAspThrSerPheTyrAsnProAspThrGinAi"gLeuValTrpAla859095CysValGlyValGluValGlyArgGlyGinProLeuGlyValGlylie100105110SerGlyHisProLeuLeuAsnLysLeuAspAspThrGluAsnAlaSer115120125AlaTyrAlaAlaAsnAlaGlyValAspAsnArgGluCyslieSerMet130135140AspTyrLysGinThrGinLeuCysLeulieGlyCysLysProProlie145150155160GlyGluHisTrpGlyLysGlySerProCysThrAsnValAlaValAsn165170175ProGlyAspCysProProLeuGluLeulieAsnThrVallieGinAsp180185190GlyAspMetValAspThrGlyPheGlyAlaMetAspPheThrThrLeu195200205GinAlaAsnLysSerGluValProLeuAsplieCysThrSerlieCys210215220LysTyrProAspTyrlieLysMetValSerGluProTyrGlyAspSer225230235240LeuPhePheTyrLeuArgA堪GluGinMetPheValArgHisLeuPhe245250255AsnArgAlaGlyAlaValGlyGluAsnValProAspAspLeuTyrlie260265270LysGlySerGlySerThrAlaAsnLeuAlaSerSerAsnTyrPhePro275280285ThrProSerGlySerMetValThrSerAspAlaGinliePheAsnLys2卯295300ProT>rTrpLeuGinArgAlaGinGlyHisAsnAsnGlylieCysTrp305310315320GlyAsnGinLeuPheValThrValValAspThrThrArgSerThrAsn325330335MetSerLeuCysAlaAlalieSerThrSerGluThrThrTyrLysAsn340345350ThrAsnPheLysGluTyrLeuArgHisGlyGluGluTyrAspLeuGin355360365PheHePheGinLeuCysLyslieThrLeuThrAlaAspValMetThr370375380TyrlieHisSerMetAsnSerThrlieLeuGluAspTrpAsnPheGly385390395400LeuGinProProProGlyGlyThrLeuGluAspThrTyrArgPheVal405410415ThrGinAlaHeAlaCysGinLysHisThrProProAlaProLysGlu420425430AspSerLeuLysLysTyrThrPheTrpGluValAsnLeuLysGluLys435440445PheSerAlaAspLeuAspGinPheProLeuGlyArgLysPheLeuLeu450455460GinAlaGly465<210>4<211>469<212>PRT<213>人工序列<220><223>HPV18L1<400>4GlySerGlyGlyGlyValTyrLeuProProProSerValAlaArgVal151015ValAsnThrAspAspTyrValThrArgThrSerliePheTyrHisAla202530GlySerSerArgLeuLeuThrValGlyAsnProTyrPheArgValPro354045AlaGlyGlyGlyAsnLysGinAsplieProLysValSerAlaTyrGin505560TyrArgValPheArgValGinLeuProAspProAsnLysPheGlyLeu65707580ProAspThrSerHeTyrAsnProGluThrGinArgLeuValTrpAla85邻95CysAlaGlyValGlulieGlyArgGlyGinProLeuGlyValGlyLeu100105noSerGlyHisProPheTyrAsnLysLeuAspAspThrGluSerSerHis115120125AlaAlaThrSerAsnValSerGluAspValArgAspAsnValSerVal130135140AspTyrLysGinThrGinLeuCysHeLeuGlyCysAlaProAlalie145150155160GlyGluHisTrpAlaLysGlyThrAlaCysLysSerArgProLeuSer165170175GinGlyAspCysProProLeuGluLeuLysAsnThrValLeuGluAsp180185190GlyAspMetValAspThrGlyTyrGlyAlaMetAspPheSerThrLeu195200205GinAspThrLysCysGluValProLeuAsplieCysGinSerlieCys210215220LysTyrProAspTyrLeuGinMetSerAlaAspProTyrGlyAspSer225230235240MetPhePheCysLeuArgArgGluGinLeuPheAlaArgHisPheTrp245250255AsnA屯AlaGlyThrMetGlyAspThrValProGinSerLeuTyrlie260265270LysGlyThrGlyMetArgAlaSerProGlySerCysValTyrSerPro275280285SerProSerGlySerlieValThrSerAspSerGinLeuPheAsnLys2卯295300ProTyrTrpLeuHisLysAlaGinGlyHisAsnAsnGlyValCysTrp305310315320HisA幼GinLeuPheThrValValAspThrThrArgSerThrAsn325330335LeuThrlieCysAlaSerThrGinSerProValProGlyGinTyrAsp340345350AlaThrLysPheLysGinTyrSerArgHisValGluGluTyrAspLeu355360365GinPheliePheGinLeuCysThrlieThrLeuThrAlaAspValMet370375380SerTyrlieGinSerMetAsnSerSerlieLeuGluAspTrpAsnPhe385390395400GlyValProProProProThrThrSerLeuValAspThrTyrArgPhe405410415ValGinSerValAlalieThrCysGinLysAspAlaAlaProAlaGlu420425430AsnLysAspProTyrAspLysLeuLysPheTrpAsnValAspLeuLys435440445GluLysPheSerLeuAspLeuAspGinTyrProLeuGlyArgLysPhe450455460LeuValGinAlaGly465<210>5<211>469<212>PRT<213>人工序列<220><223>HPV26L1<400>5GlySerGlyGlyGlyValTyrLeuProProThrProValSerArgVal151015ValAsnThrAspGluTyrValThrArgThrGlylieTyrTyrTyrAla202530GlySerSerArgLeuLeuThrLeuGlyHisProTyrPheSerliePro354045LysThrGlyGinLysAlaGlulieProLysValSerAlaTyrGinTyr505560ArgValPheArgValHisLeuProAspProAsnLysPheGlyLeuPro65707580AspProGinLeuTyrAsnProAspThrGluArgLeuValTrpAlaCys859095ValGlyValGluValGlyArgGlyGinProLeuGlylieGlyLeuSerGlyHisProLeuPheAsnLysLeuAspAspThrGluAsnSerHisLeu115120125AlaThrValAsnAlaAspThrAspAsnArgAspAsnValSerValAsp130135140AsnLysGinThrGinLeuCyslielieGlyCysThrProProLeuGly145150155160GluHisTrpGlyHeGlyThrlieCysLysAsnThrGinThrGinA唯165170175GlyAspCysProProLeuGluLeulieSerSerHelieGluAspGly180185190AspMetHeAspThrGlyPheGlyAlaMetAspPheThrAlaLeuGin195200205AlaThrLysSerAspValProlieAsplieSerGinSerThrCysLys210215220TyrProAspTyrLeuLysMetSerAlaAspThrTyrGlyAsnSerMet225230235240PhePhePheLeuArgArgGluGinLeuPheAlaArgHisPheTyrAsn245250255LysAlaGlyAlaValGlyAspAlaHeProThrThrLeuTyrlieLys260265270GlyAlaGluSerGlyArgGluProProThrSerSerlieTyrSerAla275280285ThrProSerGlySerMetValThrSerAspAlaGinLeuPheAsnLys2卯295300ProTyrTrpLeuGinArgAlaGinGlyHisAsnAsnGlyHeCysTrp305310315320GlyAsnGinLeuPheValThrCysValAspThrThrArgSerThrAsn325330335LeuThrlieSerThrLeuSerAlaAlaSerAlaSerThrProPheLys340345350ProSerAspT>rLysGinPhelieArgHisGlyGluGluTyrGluLeu355360365GinPheliePheGinLeuCysLyslieThrLeuThrThrAspValMet370375380AlaTyrlieHisLeuMetAsnAlaSerlieLeuGluAspTrpAsnPhe385390395400GlyLeuThrLeuProProThrAlaSerLeuGluAspAlaTyrArgPhe405410415lieLysAsnSerAlaThrThrCysGinAtgAsnAlaProProValPro420425430LysGluAspProPheGinLysPheLysPheTrpAspValAspLeuLys435440445GluLysPheSerHeAspLeuAspGinPheProLeuGlyArgLysPhe450455460MetLeuGinAlaGly465<210>6<211>469<212>PRT<213>人工序列<220><223>HPV31LI<400>6GlySerGlyGlyGlyValTyrLeuProProValProValSerLysVal15'1015ValSerThrAspGluTyrValThrA@ThrAsnlieTyrTyrHisAla202530GlySerAlaArgLeuLeuThrValGlyHisProTyrTyrSerliePro354045LysSerAspAsnProLysLysHeValValProLysValSerGlyLeu505560GinTyrArgValPheArgValLeuProAspProAsnLysPheGly65707580PheProAspThrSerPheTyrAsnProGluThrGinArgLeuValTrp859095AlaCysValGlyLeuGluValGlyArgGlyGinProLeuGlyValGly100105110lieSerGlyHisProLeuLeuAsnLysPheAspAspThrGluAsnSer115120125AsnArgTyrAlaGlyGlyProGlyThrAspAsnArgGluCyslieSer130135140MetAspTyrLysGinThrGinLeuCysLeuLeuGlyCysLysProPro145150155160lieGlyGluHisTrpGlyLysGlySerProCysSerAsnAsnAlalie165170175ThrProGlyAspCysProProLeuGluLeuLysAsnSerVallieGin180185190AspGlyAspMetValAspThrGlyPheGlyAlaMetAspPheThrAla195200205LeuGinAspThrLysSerAsnValProLeuAsplieCysAsnSerlie210215220CysLysTyrProAspTyrLeuLysMetValAlaGluProTyrGlyAsp225230235240ThrLeuPhePheTyrLeuArgArgGluGinMetPheValArgHisPhe245250255PheAsnArgSerGlyThrValGlyGluSerValProThrAspLeuTyr260265270lieLysGlySerGlySerThrAlaThrLeuAlaAsnSerThrTyrPhe275280285ProThrProSerGlySefMetValThrSerAspAlaGinliePheAsn290295300LysProTyrTrpMetGinArgAlaGinGlyHisAsnAsnGlylieCys305310315320TrpGlyAsnGinLeuPheValThrValValAspThrThrArgSerThr325330335AsnMetSerValCysAlaAlalieAlaAsnSerAspThrThrPheLys340345350SerSerAsnPheLysGluTyrLeuArgHisGlyGluGluPheAspLeu355360365GinPheliePheGinLeuCysLyslieThrLeuSerAlaAsplieMet370375380ThrTyrHeHisSerMetAsnProAlalieLeuGluAspTrpAsnPhe385390395400GlyLeuThrThrProProSerGlySerLeuGluAspThrTyrArgPhe405410415ValThrSerGinAlalieThrCysGinLysThrAlaProGinLysPro420425430LysGluAspProPheLysAspTyrValPheTrpGluValAsnLeuLys435440445GluLysPheSerAlaAspLeuAspGinPheProLeuGlyArgLysPhe450455460LeuLeuGinAlaGly465<220><223>HPV33LI<400>7GlySerGlyGlyGlyValTyrLeuProProValProValSerLysVal151015ValSerThrAspGluTyrValSerArgThrSerlieTyrTyrTyrAla202530GlySerSerArgLeuLeuAlaValGlyHisProTyrPheSerHeLys354045<210><211><212><213>200710005099.4AsnProThrAsnAlaLysLysLeuLeuValProLysValSerGlyLeu505560GinTyrArgValPheA屯ValArgLeuProAspProAsnLysPheGly65707580PheProAspThrSerPheTyrAsnProAspThrGinArgLeuValTrp859095AlaCysValGlyLeuGlulieGlyArgGlyGinProLeuGlyValGly100105110HeSerGlyHisProLeuLeuAsnLysPheAspAspThrGluThrGly115120125AsnLysTyrProGlyGinProGlyAlaAspAsnArgGluCysLeuSer130135140MetAspTyrLysGinThrGinLeuCysLeuLeuGlyCysLysProPro145150155160ThrGlyGluHisTrpGlyLysGlyValAlaCysThrAsnAlaAlaPro165170175AlaAsnAspCysProProLeuGluLeuHeAsnThrlielieGluAsp180185190GlyAspMetValAspThrGlyPheGlyCysMetAspPheLysThrLeu195200205GinAlaAsnLysSerAspValProHeAsplieCysGlySerThrCys210215220LysTyrProAspTyrLeuLysMetThrSerGluProTyrGlyAspSer225230235240LeuPhePhePheLeuArgArgGluGinMetPheValArgHisPhePhe245250255AsnArgAlaGlyThrLeuGlyGluAlaValProAspAspLeuTyrlie260265270LysGlySerGlyThrThfAlaSerlieGinSerSerAlaPhePhePro275280285ThrProSerGlySerMetValThrSerGluSerGinUuPheAsnLys290295300ProTyrTrpLeuGinArgAlaGinGlyHisAsnAsnGlylieCysTrp305310315320GlyAsnGinValPheValThrValValAspThrThrArgSerThrAsn325330335M改ThrLeuCysThrGinValThrSerAspSerThrTyrLysAsnGlu340345350AsnPheLysGluTyrlieAfgHisValGluGluTyrAspLeuGinPhe355360365ValPheGinLeuCysLysValThrLeuThrAlaGluValMetThrTyr370375380lieHisAlaMetAsnProAspHeLeuGluAspTrpGinPheGlyLeu385390395400ThrProProProSerAlaSerLeuGinAspThrTyrArgPheValThr405410415SerGinAlalieThrCysGinLysThrValProProLysGluLysGlu420425430AspProLeuGlyLysTyrThrPheTipGluValAspLeuLysGluLys435440445PheSerAlaAspLeuAspGinPheProLeuGlyLysPheLeuLeu450455460GinAlaGly465<210>8<211>469<212>PRT<213>人工序列<220><223>HPV35LI<400>8GlySerGlyGlyGlyValTyrLeuProProValSerValSerLysVal151015ValSerThrAspGluTyrValThrArgThrAsnlieTyrTyrHisAla202530GlySerSerArgLeuLeuAlaValGlyHisProTyrTyrAlalieLys354045LysGinAspSerAsnLyslieAlaValProLysValSerGlyLeuGin505560TyrAigValPheArgValLysLeuProAspProAsnLysPheGlyPhe65707580ProAspThrSerPheTyrAspProCysLeuGinArgLeuValTipAla859095CysThrGlyValGluVa!GlyA屯GlyGinProLeuGlyValGlylie100105110SerGlyHisProLeuLeuAsnLysLeuAspAspThrGluAsnLeuAsn115120125LysTyrValGlyAsnSerGlyAsnSerGlyThrAspAsnArgGluCys130135140HeSerMetAspTyrLysGinThrGinLeuCysLeuHeGlyCysArg145150155160ProProlieGlyGluHisTipGlyLysGlyThrProCysAsnAlaAsn165170175GinValLysAlaGlyGluCysProProLeuGluLeuLeuAsnThrVal180185190LeuGinAspGlyAspMetValAspThrGlyPheGlyAlaMetAspPhe195200205ThrThrLeuGinAlaAsnLysSerAspValProLeuAsplieCysSer210215220SerlieCysLysT^rProAspTyrLeuLysMetValSerGluProTyr225230235240GlyAspMetLeuPhePheT^rLeuArgA堪GluGinMetPheValArg245250255HisLeuPheAsnArgAlaGlyThrValGlyGluThrValProAlaAsp260265270LeuTyrHeLysGlyThrThrGlyThrLeuProSerThrSerTyrPhe275280285ProThrProSerGlySerMetValThrSerAspAlaGinliePheAsn2卯295300LysProTyrTipLeuGinArgAlaGinGlyHisAsnAsnGlylieCys305310315320TipSerAsnGinLeuPheValThrValValAspThrThrArgSerThr325330335AsnMetSerValCysSerAlaValSerSerSerAspSerThrTyrLys340345350AsnAspAsnPheLysGluTyrLeuArgHisGlyGluGluTyrAspLeu355360365GinPheliePheGinLeuCysLyslieThrLeuThrAlaAspValMet370375380ThriyrlieHisSerMetAsnProSerlieLeuGluAspTipAsnPhe385390395400GlyLeuThrProProProSerGlyThrLeuGluAspThrTyrArgTyr405410415ValThrSerGinAlaValThrCysGinLysProSerAlaProLysPro420425430LysAspAspProLeuLysAsnTyrThrPheTipGluValAspLeuLys435440445GluLysPheSerAlaAspLeuAspGinPheProLeuGlyArgLysPhe450455460LeuLeuGinAlaGly465<210>9<211>468<212>PRT<213>人工序列<220><223>HPV39LI<400>9GlySerGlyGlyGlyValTyrLeuProProProSerValAlaLysVal151015ValAsnThrAspAspTyrValThrArgThrGlyHeTyrTyrTyrAla202530GlySerSerArgLeuLeuThrValGlyHisProTyrPheLysValGly354045MetAsnGlyGlyArgLysGinAsplieProLysValSerAlaTyrGin505560TyrArgValPheArg\yThrLeuProAspProAsnLysPheSerlie65707580ProAspAlaSerLeuTyrAsnProGluThrGinArgLeuValTrpAla859095CysValGlyValGluValGlyArgGlyGinProLeuGlyValGlyHe100105noSerGlyHisProLeuTyrAsnArgGinAspAspThrGluAsnSerPro115120125PheSerSerThrThrAsnLysAspSerArgAspAsnValSerValAsp130135140TyrLysGinThrGinLeUCysHelieGlyCysValProAlalieGly145150155160GluHisTrpGlyLysGlyLysAlaCysLysProAsnAsnValSerThr165170175GlyAspCysProProLeuGluLeuValAsnThrProlieGluAspGly180185190AspMetlieAspThrGlyTyrGlyAlaMetAspPheGlyAlaLeuGin195200205GluThrLysSerGluValProLeuAsplieCysGinSerlieCysLys210215220TyrProAspTyrLeuGinMetSerAlaAspValTyrGlyAspSerMet225230235240PhePheCysLeuArgArgGluGinLeuPheAlaArgHisPheTrpAsn245250255A屯GlyGlyMetValGlyAspAlalieProAlaGinLeuTyrHeLys260265270GlyThrAspHeArgAlaAsnProGlySerSerValTyrCysProSer275280285ProSerGlySerMetValThrSerAspSerGinLeuPheAsnLysPro290295300TyrTrpLeuHisLysAlaGinGlyHisAsnAsnGlylieCysTrpHis305310315320AsnGinLeuPheLeuThrValValAspThrThrArgSerThrAsnPhe325330335ThrLeuSerThrSerlie'GluSerSerlieProSerThrTyrAspPro340345350SerLysPheLysGluTyrThrArgHisValGluGluTyrAspLeuGin355360365PheliePheGinLeuCysThrValThrLeuThrThrAspValMetSer370375380TyrHeHisThrMetAsnSerSerlieLeuAspAsnTrpAsnPheAla385390395400ValAlaProProProSerAlaSerLeuValAspThrTyrArgTyrLeu405410415GinSerAlaAlalieThrCysGinLysAspAlaProAlaProGluLys420425430LysAspProTyrAspGlyLeuLysPheTrpAsnValAspLeuArgGlu435440445LysPheSerLeuGluLeuAspGinPheProLeuGlyArgLysPheLeu450455460LeuGinAlaArg465<210>10<211>467<212>PRT<213>人工序列<220><223>HPV42L1<400>10GlySerGlyGlyGlyValTyrLeuProProProProValSerLysVal151015ValSerThrAspGluTyrValGinArgThrAsnTyrPheTyrHisAla202530SerSerSerArgLeuLeuValValGlyHisProTyrTyrSerlieThr354045LysArgProAsnLysThrSerHeProLysValSerGlyLeuGinTyr505560A堪ValPheAigValArgLeuProAspProAsnLysPheThrLeuPro65707580GluThrAsnLeuTyrAsnProGluThrGinArgMetValTrpAlaCys85卯95ValGlyLeuGluValGlyArgGlyGinProLeuGlyValGlylieSer100105110GlyHisProLeuLeuAsnLysLeuAspAspThrGluAsnAlaProThr115120125TyrGlyGlyGlyProGlyThrAspAsnArgGluAsnValSerMetAsp130135140TyrLysGinThrGinLeuCysLeuValGlyCysLysProAlalieGly145150155160GluHisTrpGlyLysGlyThrAlaCysThrProGinSerAsnGlyAsp165170175CysProProLeuGluLeuLysAsnSerPheHeGinAspGlyAspMet180185190ValAspValGlyPheGlyAlaLeuAspPheGlyAlaLeuGinSerSer195200205LysAlaGluValProLeuAspHeValAsnSerHeThrLysTyrPro210215220AspTyrLeuLysMetSerAlaGluAlaTyrGlyAspSerMetPhePhe225230235240PheLeuArgArgGluGinMetPheValA屯HisLeuPheAsnArgAla245250255GlyAlalieGlyGluProValProAspGluLeuTyrThrLysAlaAla260265270AsnAsnAlaSerGlyAigHisAsnLeuGlySerSerlieTyrTyrPro275280285ThrProSerGlySerMetValThrSerAspAlaGinLeuPheAsnLys290295300ProiyrTrpLeuGinGinAlaGinGlyHisAsnAsnGlyHeCysTrp305310315320GlyAsnGinLeuPheUuThrValValAspThrThrArgSerThrAsn325330335MetThrLeuCysAlaThrAlaThrSerGlyAspThrTyrThrAlaAla340345350AsnPheLysGluTyrLeuArgHisAlaGluGluTyrAspValGinPhe355360365liePheGinLeuCysLyslieThrLeuThrValGluValMetSerTyr370375380lieHisAsnMetAsnProAsnlieLeuGluGluTrpAsnValGlyVal385390395400AlaProProProSerGlyThrLeuGluAspSerTyrArgTyrValGin405410415SerGluAlalieArgCysGinAlaLysValThrThrProGluLysLys420425430AspProTyrSerAspPheTipPheTrpGluValAsnLeuSerGluLys435440445PheSerThrAspLeuAspGinPheProLeuGlyArgLysPheLeuLeu450455460GinAlaGly465<210>11<211>472<212>PRT<213>人工序列<220><223>HPV45Ll<400>11GlySerGlyGlyGlyValTyrLeuProProProSerValAlaArgVal151015ValSerThrAspAspTjtValSerArgThrSerliePheTyrHisAla202530GlySerSerArgLeuLeuThrValGlyAsnProTyrPheArgValVal354045ProAsnGlyAlaGlyAsnLysGinAlaValProLysValSerAlaTyr505560GinTyrArgValPheArgValAlaLeuProAspProAsnLysPheGly65707580LeuProAspSerThrHeTyrAsnProGluThrGinArgLeuValTrp85卯95AlaCysValGlyMetGlulieGlyAigGlyGinProLeuGlylieGly100105110LeuSerGlyHisProPheTyrAsnLysLeuAspAspThrGluSerAla115120125HisAlaAlaThrAlaVallieThrGinAspValA*AspAsnValSer130135140ValAspTyrLysGinThrGinLeuCyslieLeuGlyCysValProAla145150155160HeGlyGluHisTrpAlaLysGlyThrLeuCysLysProAlaGinLeu165170175GinProGlyAspCysProProLeuGluLeuLysAsnThrlielieGlu180185l卯AspGlyAspMetValAspThrGlyTyrGlyAlaMetAspPheSerThr195200205LeuGinAspThrLysCysGluValProLeuAsplieCysGinSerlie210215220CysLysTyrProAspTyrLeuGinMetSerAlaAspProTyrGlyAsp225230235240SerMetPhePheCysLeuArgArgGluGinLeuPheAlaArgHisPhe245250255TrpAsnArgAlaGlyValMetGlyAspThrValProThrAspLeuTyr260265270lieLysGlyThrSerAlaAsnMetArgGluThrProGlySerCysVal275280285TyrSerProSerProSerGlySerHeHeThrSerAspSerGinLeu290295300PheAsnLysProTyrTrpLeuHisLysAlaGinGlyHisAsnAsnGly305310315320lieCysTrpHisAsnGinLeuPheValThrValValAspThrThrArg325330335SerThrAsnLeuThrLeuCysAlaSerThrGinAsnProValProSer340345350ThrTyrAspProThrLysPheLysGinTyrSerArgHisValGluGlu355360365TyrAspLeuGinPheliePheGinLeuCysThrlieThrLeuThrAla370375380GluValMetSerTyrHeHisSerMetAsnSerSerHeLeuGluAsn385390395400TrpAsnPheGlyValProProProProThrThrSerLeuValAspThr405410415iyArgPheValGinSerValAlaValThrCysGinLysAspThrThr420425430ProProGluLysGinAspProTyrAspLysLeuLysPheTrpThrVal435440445AspLeuLysGluLysPheSerSerAspLeuAspGinTyrProLeuGly450455460ArgLysPheLeuValGinAlaGly465470<210>12<211>468<212>PRT<213>人工序列<220><223>HPV51L1<400>12GlySerGlyGlyGlyValTyrLeuProProAlaProValSerArglie151015ValAsnThrGluGluTyrlieThrArgThrGlylieTyrTyrTyrAla202530GlySerSerArgLeuHeThrLeuGlyHisProTyrPheProliePro354045LysThrSerThrArgAlaAlalieProLysValSerAlaPheGinTyr505560A屯ValPheArgValGinLeuProAspProAsnLysPheGlyLeuPro65707580AspProAsnLeuTyrAsnProAspThrAspArgLeuValTrpGlyCysValGlyValGluValGlyArgGlyGinProLeuGlyValGlyLeuSer100105110GlyHisProLeuPheAsnLysTyrAspAspThrGluAsnSerArgHe115120125AlaAsnGlyAsnAlaGinGinAspValA屯AspAsnThrSerValAsp130135140AsnLysGinThrGinLeuCyslielieGlyCysAlaProProlieGly145150155160GluHisTrpGlylieGlyThrThrCysLysAsnThrProValProPro165170175GlyAspCysProProLeuGluLeuValSerSerVallieGinAspGly180185190AspMetHeAspThrGlyPheGlyAlaMetAspPheAlaAlaLeuGin195200205AlaThrLysSerAspValProLeuAsplieSerGinSerValCysLys210215220TyrProAspT^rLeuLysMetSerAlaAspThrTyrGlyAsnSerMet225230235240PhePheHisLeuArgArgGluGinliePheAlaHisTyrTyrAsn245250255LysLeuValGlyValGlyGluAsplieProAsnAspTyrTyrlieLys260265270GlySerGlyAsnGlyArgAspProIkGluSerTyrlieTyrSerAla275280285ThrProSerGlySerMetlieThrSerAspSerGinliePheAsnLys2卯295300ProTyrTrpLeuHisA堪AlaGinGlyHisAsnAsnGlylieCysTrp305310315320AsnAsnGinLeuPhelieThrCysValAspThrThrArgSerThrAsn325330335LeuThrHeSerThrAlaThrAlaAlaValSerProThrPheThrPro340345350SerAsnPheLysGinTyrHeArgHisGlyGluGluTyrGluLeuGin355360365PheHePheGinLeuCysLyslieThrLeuThrThrGluValMetAla370375380TyrLeuHisThrMetAspProThrlieLeuGluGinTrpAsnPheGly385390395400LeuThrLeuProProSerAlaSerLeuGluAspAlaTyrArgPheVal405410415A堪AsnAlaAlaThrSerCysGinLysAspThrProProGinAlaLys420425430ProAspProLeuAlaLysTyrLysPheTrpAspValAspLeuLysGlu435440445AigPheSerLeuAspLeuAspGinPheAlaLeuGlyArgLysPheLeu450455460LeuGinValGly465<210>13<211>472<212>PRT<213>人工序列<220><223>HPV52L1<400>13GlySerGlyGlyGlyValTyrLeuProProValProValSerLysVal151015ValSerThrAspGluTyrValSerArgThrSerlieTyrTyrTyrAla202530GlySerSerAigLeuLeuThrValGlyHisProTyrPheSerlieLys354045AsnThrSerSerGlyAsnGlyLysLysValLeuValProLysValSer505560GlyLeuGinTyrArgValPheArglieLysLeuProAspProAsnLys65707580PheGlyPheProAspThrSerPheTyrAsnProGluThrGinArgLeu859095ValTrpAlaCysThrGlyLeuGlulieGlyArgGlyGinProLeuGly100105110ValGlylieSerGlyHisProLeuLeuAsnLysPheAspAspThrGlu115120125ThrSerAsnLysT^rAlaGlyLysProGlylieAspAsnArgGluCys130135140LeuSerMetAspTyrLysGinThrGinLeuCyslieLeuGlyCysLys145150155160ProProlieGlyGluHisTrpGlyLysGlyThrProCysAsnAsnAsn165170175SerGlyAsnProGlyAspCysProProLeuGinLeulieAsnSerVal180185190HeGinAspGlyAspM改ValAspThrGlyPheGlyCysMetAspPhe195200205AsnThrLeuGinAlaSerLysSerAspValProHeAsplieCysSer210215220GlyAspSerLeuPhePhePlteLeuArgArgGluGinMetPheValArg245250255HisPhePheAsnArgAlaGlyThrLeuGlyAspProValProGlyAsp260265270LeuTyrlieGinGlySerAsnSerGlyAsnThrAlaThrValGinSer275280285SerAlaPhePheProThrProSerGlySerMetValThrSerGluSer2卯295300GinLeuPheAsnLysProTyrTrpLeuGinAlaGinGlyHisAsn305310315320AsnGlylieCysTrpGlyAsnGinLeuPheValThrValValAspThr325330335ThrArgSerThrAsnMetThrLeuCysAlaGluValLysLysGluSer340345350ThrTyrLysAsnGluAsnPheLysGluTyrLeuArgHisGlyGluGlu355360365PheAspLeuGinPheliePheGinLeuCysLyslieThrLeuThrAla370375380AspValMetThrlyleHisLysMetAspAlaThrlieLeuGluAsp385390395400TrpGinPheGlyLeuThrProProProSerAlaSerLeuGluAspThr405410415TyrAigPheValThrSerThrAlalieThrCysGinLysAsnThrPro420425430ProLysGlyLysGluAspProLeuLysAspTyrMetPheTrpGluVal435440445AspLeuLysGluLysPheSerAlaAspLeuAspGinPheProLeuGly450455460A堪LysPheLeuLeuGinAlaGly465470<210>14<211>465<212>PRT<213>人工序列<220><223>HPV53L1<400>14GlySerGlyGlyGlyValTyrLeuProProThrProValSerLysVal151015HeThrThrAspAlaTyrValLysArgThrThrliePheTyrHisAla202530GlySerSerArgLeuLeuThrValGlyHisProTyrTyrProHeSer354045LysSerGlyLysAlaAsplieProLysValSerAlaPheGinTyrArg505560ValPheArgValLeuProAspProAsnLysPheGlyLeuProAsp65707580ThrAsnHePheAsnProAspGinGluArgLeuValTrpAlaCysVal859095GlyLeuGlulieGlyArgGlyGinProLeuGlyValGlyValSerGly100105110HisProLeuPheAsnArgLeuAspAspThrGluSerSerSerlieAla115120125HeGinAspThrAlaProAspSerArgAspAsnValSerValAspPro130135140LysGinThrGinLeuCysHelieGlyCysAlaProAlalieGlyGlu145150155160HisTrpThrLysGlyThrAlaCysArgSerThrProThrThrAlaGly165170175AspCysProProLeuGluLeuHeAsnSerProlieGluAspGlyAsp180185l卯MetValAspThrGlyPheGlyAlaLeuAsnPheLysAlaLeuGinGlu195200205SerLysSerAspValPro'LeuAsplieValGinSerThrCysLysTyr210215220ProAspTyrLeuLysMetSerAlaAspAlaTyrGlyAspSerMetTrp225230235240PheTyrLeuArgArgGluGinLeuPheThrArgHisPhePheAsnArg245250255AlaGlyVallieGlyGluGluHeProAsnAspLeuTyrHeLysGly260265270SerAsnGlyArgAspProProProSerSerValTyrValAlaThrPro275280285SerGlySerMetlieThrSerGluAlaGinLeuPheAsnLysProTyr290295300TrpLeuGinArgAlaGinGlyHisAsnAsnGlylieCysTrpAsnAsn305310315320GinLeuPheValThrValValAspThrThrArgAsnThrAsnMetThr325330335UuSerAlaThrThrGinSerMetSerThrTyrAsnSerLysGinlie340345350LysGinTyrValArgHisAlaGluGluTyrGluLeuGinPheValPhe355360365GinLeuCysLyslieSerLeuSerAlaGluValMetAlaTyrLeuHis370375380ThrMetAsnSerThrLeuLeuGluAspTrpAsnlieGlyLeuSerPro385390395400ProValAlaThrSerLeuGluAspLysTyrArgTyrValLysSerAla405410415AlalieThrCysGinLysAspGinProProProGluLysGinAspPro420425430LeuSerLysTyrLysPheTrpGluValAsnLeuGinAsnSerPheSer435440445AlaAspLeuAspGinPheProLeuGlyArgLysPheLeuMetGinVal450455460Gly465<210>15<211>466<212>PRT<213>人工序列<220><223>HPV56U<400>15GlySerGlyGlyGlyVdTyrLeuProProThrProValSerLysVal151015ValAlaThrAspSerTyrValLysArgThrSerliePheTyrHisAla202530GlySerSerArgLeuLeuAlaValGlyHisProTyrTyrSerValThr354045LysAspAsnThrLysThrAsnHeProLysValSerAlaTyrGinTyr505560ArgValPheArgValArgLeuProAspProAsnLysPheGlyLeuPro65707580AspThrAsnlieTyrAsnProAspGinGluArgLeuValTrpAlaCys859095ValGlyLeuGluValGlyArgGlyGinProLeuGlyAlaGlyLeuSer100105110GlyHisProLeuPheAsnArgLeuAspAspThrGluSerSerAsnLeu115120125AlaAsnAsnAsnVallieGluAspSerAtgAspAsnlieSerValAsp130135140GlyLysGinThrGinLeuCyslieValGlyCysThrProAlaMetGly145150155160GluHisTrpThrLysGlyAlaValCysLysSerThrGinValThrThr165170175GlyAspCysProProLeuAlaLeulieAsnThrProlieGluAspGly180185190AspMetlieAspThrGlyPheGlyAlaMetAspPheLysValLeuGin195200205GluSerLysAlaGluValProLeuAsplieValGinSerThrCysLys210215220TyrProAspT>rLeuLysMetSerAlaAspAlaTyrGlyAspSerMet225230235240TrpPheTyrLeuArgAigGluGinLeuPheAlaArgHisTyrPheAsn245250255AigAlaGlyLysValGlyGluThrlieProAlaGluLeuTyrLeuLys260265270GlySerAsnGlyArgGluProProProSerSerValTyrValAlaThr275280285ProSerGlySerMetHeThrSerGluAlaGinLeuPheAsnLysPro2卯295300TyrTrpLeuGinArgAlaGinGlyHisAsnAsnGlylieCysTrpGly305310315320AsnGinLeuPheValThrValValAspTTirThrArgSerThrAsnMet325330335ThrlieSerThrAlaThrGluGinLeuSerLysTyrAspAlaArgLys340345350lieAsnGinTyrLeuArgHisValGluGluTyrGluLeuGinPheVal355360365PheGinLeuCysLysHeThrLeuSerAlaGluValMetAlaTyrLeu370375380HisAsnMetAsnAlaAsnLeuLeuGluAspTrpAsnlieGlyLeuSer385390395400ProProValAlaThrSerLeuGluAspLysTyrArgTyrValArgSer405410415ThrAlalieThrCysGinArgGluGinProProThrGluLysGinAsp420425430ProLeuAlaLysTyrLysPheTrpAspValAsnLeuGinAspSerPhe435440445SerThrAspLeuAspGinPheProLeuGlyArgLysPheLeuMetGin450455460LeuGly465<210>16<211>467<212>PRT<213>人工序列<220><223>HPV58Ll<400>16GlySerGlyGlyGlyValTyrLeuProProValProValSerLysVal151015ValSerThrAspGluTyrValSerArgThrSerHeTyrTyrTyrAla202530GlySerSerArgLeuLeuAlaValGlyAsnProTyrPheSerlieLys354045SerProAsnAsnAsnLysLysValLeuValProLysValSerGlyLeu505560GinTyrArgValPheArgValArgLeuProAspProAsnLysPheGly65707580PheProAspThrSerPheTyrAsnProAspThrGinArgLeuValTrp859095AlaCysValGlyLeuGlulieGlyArgGlyGinProLeuGlyValGly100105110ValSerGlyHisProTyrLeuAsnLysPheAspAspThrGluThrSer115120125AsnArgTyrProAlaGinProGlySerAspAsnArgGluCysLeuSer130135140MetAspTyrLysGinThrGinLeuCysLeulieGlyCysLysProPro145150155160ThrGlyGluHisTrpGlyLysGlyValA墓aCysAsnAsnAsnAlaAla165170175AlaThrAspCysProProLeuGluLeuPheAsnSerlielieGluAsp180185190GlyAspMetValAspThrGtyPheGlyCysMetAspPheGlyThrLeu195200205GinAlaAsnLysSerAspValProlieAsplieCysAsnSerThrCys210215220LysTyrProAspTyrLeuLysMetAlaSerGluProTyrGlyAspSer225230235240LeuPhePhePheLeuArgAigGluGinMetPheValArgHisPhePhe245250255AsnArgAlaGlyLysLeuGlyGluAlaValProAspAspLeuTyrlie260265270LysGlySerGlyAsnThrAlaValHeGinSerSerAlaPhePhePro275280285ThrProSerGlySerlieValThrSerGluSerGinLeuPheAsnLys290295300ProTyrTrpLeuGinArgAlaGinGlyHisAsnAsnGlylieCysTrp305310315320GlyAsnGinLeuPheValThrValValAspThrThrArgSerThrAsn325330335MetThrLeuCysThrGluValThrLysGluGlyThrTyrLysAsnAsp340345350AsnPheLysGluTyrValArgHisValGluGluTyrAspLeuGinPhe355360365ValPheGinLeuCysLyslieThrLeuThrAlaGluHeMetThrTyr370375380lieHisThrMetAspSerAsnlieLeuGluAspTrpGinPheGlyLeu385390395400ThrProProProSerAlaSerLeuGinAspThrTyrArgPheValThr405410415SerGinAlalieThrCysGinLysThrAlaProProLysGluLysGlu420425430AspProLeuAsnLysTyrThrPheTrpGluValAsnLeuLysGluLys435440445PheSerAlaAspLeuAspGinPheProLeuGlyArgLysPheLeuLeu450455460GinSerGly465<210>17<211>469<212>PRT<213>人工序列<220><223>HPV59L1<400>17GlySerGlyGlyGlyValTyrLeuProProProSerValAlaLysVal151015ValSerThrAspGluTyrValThrArgThrSerliePheTyrHisAla202530GlySerSerArgLeuLeuThrValGlyHisProTyrPheLysValPro354045LysGlyGlyAsnGlyArgGinAspValProLysValSerAlaTyrGin505560TyrArgValPheArgViedLysLeuProAspProAsnLysPheGlyLeu65707580ProAspAsnThrValT^rAspProAsnSerGinArgLeuValTrpAla859095CysValGlyValGlulieGlyAigGlyGinProLeuGlyValGlyLeul(W105110SerGlyHisProLeuTyrAsnLysLeuAspAspThrGluAsnSerHis115120125ValAlaSerAlaValAspThrLysAspThrA$AspAsnValSerVal130135140AspTyrLysGinThrGinLeuCyslieleGlyCysValProAlalie145150155160GlyGluHisTrpThrLysGlyThrAlaCysLysProThrThrValVal165170175GinGlyAspCysProProLeuGluLeulieAsnThrProlieGluAsp180185190GlyAspMetValAspThrGlyTyrGlyAlaMetAspPheLysLeuLeu195200205GinAspAsnLysSerGluValProLeuAsplieCysGinSerlieCys210215220LysTyrProAspTyrLeuGinMetSerAlaAspAlaTyrGlyAspSer225230235240MetPhePheCysLeuArgGluGinValPheAlaArgHisPheTrp245250255AsnArgSerGlyThrMetGlyAspGinLeuProGluSerLeuTyrlie260265270LysGlyThrAsplieAi"gAlaAsnProGlySerTyrLeuTyrSerPro275280285SerProSerGlySerValValThrSerAspSerGinLeuPheAsnLys2卯295300ProTyrTrpLeuHisLysAlaGinGlyLeuAsnAsnGlylieCysTrp305310315320HisAsnGinLeuPheLeuThrValValAspThrThrArgSerThrAsn325330335LeuSerValCysAlaSerThrThrSerSerlieProAsnValTyrThr340345350ProThrSerPheLysGluTyrAlaArgHisValGluGluPheAspLeu355360365GinPheliePheGinLeuCysLyslieThrLeuThrThrGluValMet370375380SerTyrlieHisAsnMetAsnThrThrlieLeuGluAspTrpAsnPhe385390395400GlyValThrProProProThrAlaSerLeuValAspThrTyrArgPhe405410415ValGinSerAlaAlaValThrCysGinLysAspThrAlaProProVal420425430LysGinAspProTyrAspLysLeuLysPheTipProValAspLeuLys435440445GluArgPheSerAlaAspLeuAspGinPheProLeuGlyArgLysPhe450455460<image>imageseeoriginaldocumentpage39</image>GlyGlyAsnGlyArgAspProProProSerSerValTyrValAlaThr275280285ProSerGlySerMetlieThrSerGluAlaGinLeuPheAsnLysPro290295300T>rTrpLeuGinArgAlaGinGlyHisAsnAsnGlyHeCysTrpGly305310315320AsnGinValPheValThrValValAspThrThrArgSerThrAsnMet325330335ThrlieAsnAlaAlaLysSerThrLeuThrLysTyrAspAlaArgGlu340345350lieAsnGinTyrLeuArgHisValGluGluTyrGluLeuGinPheVal355360365PheGinLeuCysLysHeThrLeuThrAlaGluValMetAlaTyrLeu370375380HisAsnMetAsnAsnThrLeuLeuAspAspTrpAsnlieGlyLeuSer385390395400ProProValAlaThrSerLeuGluAspLysTyrArgTyrlieLysSer405410415ThrAlalieThrCysGinArgGluGinProProAlaGluLysGinAsp420425430ProLeuAlaLysTyrLysPheTrpGluValAsnLeuGinAspSerPhe435440445SerAlaAspLeuAspGinPheProLeuGlyArgLysPheLeuMetGin450455460LeuGly465<210>19<211>470<212>PRT<213>人工序列<220><223>HPV73L1<400>19GlySerGlyGlyGlyValTyrLeuProProValSerValSerLysVal151015ValSerThrAspGluTyrValThrArgThrAsnlieTyrTyrTyrAla202530GlySerThrArgLeuLeuAlaValGlyHisProTyrPheProHeLys354045AspSerGinLysArgLysThrleValProLysValSerGlyLeuGin505560TyrArgValPheArgLeuArgLeuProAspProAsnLysPheGlyPhe65707580ProAspAlaSerPheTyrAsnProAspLysGluArgLeuValTrpAla859095CysSerGlyValGluValGlyArgGlyGinProLeuGlylieGlyThr100105110SerGlyAsnProPheMetAsnLysLeuAspAspThrGluAsnAlaPro115120125LysiyrlieAlaGlyGinAsnThrAspGlyArgGluCysMetSerVal130135140AspTyrLysGinThrGinLeuCyslieLeuGlyCysArgProProLeu145150155160GlyGluHisTrpGlyProGlyThrProCysThrSerGinThrValAsn165170175ThrGlyAspCysProProLeuGluLeuLysAsnThrProlieGinAsp180185190GlyAspMetlieAspValGlyPheGlyAlaMetAspPheLysAlaLeu195200205GinAlaAsnLysSerAspValProlieAsplieSerAsnThrThrCys210215220LysTyrProAspTyrLeuGlyMetAlaAlaAspProTyrGlyAspSer225230235240MetTrpPheTyrLeuArgArgGluGinMetPheValArgHisLeuPhe245250255AsnArgAlaGlyAspThrGlyAspLyslieProAspAspLeuMetlie260265270LysGlyThrGlyAsnThrAlaThrProSerSerCysValPheTyrPro275280285ThrProSerGlySerMetValSerSerAspAlaGinLeuPheAsnLys2卯295300ProTyrTrpLeuGinLysAlaGinGlyGinAsnAsnGlyHeCysTrp305310315320HisAsnGinLeuPheLeuThrValValAspThrThrArgSerThrAsn325330335PheSerValCysValGlyThrGinAlaSerSerSerThrThrThrTyr340345350AlaAsnSerAsnPheLysGluTyrLeuArgHisAlaGluGluPheAsp355360365LeuGinPheValPheGinLeuCysLysHeSerLeuThrThrGluVal370375380MetThrTyrlieHisSerMetAsnSerThrHeLeuGluGluTrpAsn385390395400PheGlyLeuThrProProProSerGlyThrLeuGluGluThrTyrArg405410415TyrValThrSerGinAlaHeSerCysGinArgProGinProProLys420425430GluThrGluAspProTyrAlaLysLeuSerPheTrpAspValAspLeu435440445LysGluLysPheSerAlaGluLeuAspGinPheProLeuGlyArgLys450455460PheLeuLeuGinLeuGly465470<210>20<211>469<212>PRT<213>人工序列<220><223>HPV83L1<400>20GlySerGlyGlyGlyValTyrLeuProProAlaProValSerArglie151015ValAsnThrGluGluTyrlieThrArgTiffGlyHeTyrTyrTyrAla202530GlySerSerArgLeuHeThrLeuGlyHisProTyrPheSerliePro354045LysThrAsnThrArgAlaGluHeProLysValSerAlaPheGinTyr505560AigValPheArgValGinLeuProAspProAsnLysPheGlyLeuPro65707580AspProAsnLeuPheAsnProAspThrAspArgLeuValTrpGlyCys859095ValGlyValGluValGlyArgGlyGinProLeuGlyValGlyLeuSer100105110GlyHisProLeuPheAsnLysTyrAspAspThrGluAsnSerArgPhe115120125AlaAsnGlyAsnAspGinGinAspValArgAspAsnlieSerValAsp130135140AsnLysGinThrGinLeuCyslielieGlyCysAlaProProlieGly145150155160GluHisTrpAlaThrGlyThrThrCysLysAsnValProValProGin165170175GlyAspCysProProLeuGluLeuValSerThrVallieGluAspGly180185190AspMetValAspThrGlyPheGlyAlaMetAspPheAlaAsnLeuGin195200205AlaThrLysSerAspValProLeuAsplieAlaGinSerValCysLys210215220TyrProAspTyrLeuLysMetSerAlaAspThrTyrGlyAsnSerMet225230235240PhePheHisLeuArgArgGluGinliePheAlaArgHisTyrTyrAsn245250255LysAlaGlyValValGlyAspAlaHeProAspLysAlaTyrlieLys260265270GlyThrGlyAlaGlyA堪AspProlieSerSerTyrlieTyrSerAla275280285ThrProSerGlySerMetlieThrSerAspSerGinliePheAsnLys2卯295300ProTyrTrpLeuHisArgAlaGinGlyHisAsnAsnGlyHeCysTrp305310315320AsnAsnGinLeuPhelieThrCysValAspThrThrLysSerThrAsn325330335LeuThrHeSerThrAlaValThrProSerValAlaGinThrPheThr340345350ProAlaAsnHieLysGinTyrlieArgHisGlyGluGluTyrGluLeu355360365GinPheliePheGinLeuCysLyslieThrLeuThrThrGlulieMet370375380AlaTyrLeuHisThrMetAspSerThrlieLeuGluGinTrpAsnPhe385390395400GlyLeuThrLeuProProSerAlaSerLeuGluAspAlaTyrArgPhe405410415ValLysAsnAlaAlaThrSerCysHisLysAspSerProProGinAla420425430LysGluAspProLeuAlaLysT^rLysPheTrpAsnValAspLeuLys435440445GluArgPheSerLeuAspLeuAspGinPheAlaLeuGlyArgLysPhe450455460LeuLeuGinlieGly46权利要求1.一种重组人乳头瘤病毒L1衣壳蛋白的氨基酸序列，其特征在于将野生型HPVL1蛋白的氨基酸序列N-末端的保守序列VYLPP或VYVPP上游的氨基酸序列替换成GSGGG，同时在其C-末端的保守序列LGRKFL下游第5个氨基酸处通过引入蛋白质翻译的终止密码子形成相对于野生型的截短型序列，其中所述的HPV型选自HPV6、HPV11、HPV16、HPV18、HPV26、HPV31、HPV33、HPV35、HPV39、HPV42、HPV45、HPV51、HPV52、HPV53、HPV56、HPV58、HPV59、HPV66、HPV73、HPV82中的一种或多种。2、根据权利要求1所述的氨基酸序列，其中HPV6对应的氨基酸序列如SEQIDNO:l所示，HPV11对应的氨基酸序列如SEQIDNO:2所示，HPV16对应的氨基酸序列如SEQIDNO:3所示，HPV18对应的氨基酸序列如SEQIDNO:4所示，HPV26对应的氨基酸序列如SEQIDNO:5所示，HPV31对应的氨基酸序列如SEQIDNO:6所示，HPV33对应的氨基酸序列如SEQIDNO:7所示，HPV35对应的氨基酸序列如SEQIDNO:8所示，HPV39对应的氨基酸序列如SEQIDNO:9所示，HPV42对应的氨基酸序列如SEQIDNO:10所示，HPV45对应的氨基酸序列如SEQIDNO:ll所示，HPV51对应的氨基酸序列如SEQIDNO:12所示，HPV52对应的氨基酸序列如SEQIDNO:13所示，HPV53对应的氨基酸序列如SEQIDNO:14所示，HPV56对应的氨基酸序列如SEQIDNO:15所示，HPV58对应的氨基酸序列如SEQIDNO:16所示，HPV59对应的氨基酸序列如SEQIDNO:17所示，HPV66对应的氨基酸序列如SEQIDNO:18所示，HPV73对应的氨基酸序列如SEQIDNO:19所示，HPV82对应的氨基酸序列如SEQIDNO:20所示。3、一种核酸分子，其特征在于它包含编码权利要求1所述的氨基酸序列的核苷酸序列。4、一种表达载体，其特征在于它包含权利要求3所述的核苷酸序列。5、一种宿主细胞，其特征在于它包含权利要求4所述的载体。6、根据权利要求5所述的宿主细胞，其特征在于它为大肠杆菌细胞。7、一种HPVLl蛋白的五聚体，其特征在于它的一级结构由权利要求l所述的氨基酸序列所组成，其二级结构和高级结构由上述一级结构所决定。8、根据权利要求7所述的HPVL1蛋白的五聚体在制备预防HPV感染的疫苗中的应用。9、根据权利要求8所述的应用，其特征在于用于制备所述的预防HPV感染的疫苗的HPV型包括HPV16。10、根据权利要求8所述的应用，其特征在于用于制备所述的预防HPV感染的疫苗的HPV型包括HPV6、HPVll、HPV16、HPV18和HPV33。11、根据权利要求7所述的HPVL1蛋白的五聚体在制备药物组合物中的应用.12、根据权利要求7所述的HPVL1蛋白的五聚体在制备免疫诊断抗原或抗体中的应用。全文摘要本发明涉及重组人乳头瘤病毒L1衣壳蛋白的氨基酸序列，编码该氨基酸序列的核苷酸序列，和包含所述核苷酸序列的重组载体和表达宿主。本发明还涉及由该氨基酸序列所组成的HPVL1蛋白在制备疫苗、药物组合物和诊断抗原或抗体中的应用。本发明通过对HPVL1野生型序列的改造，使得在原核系统中表达的重组HPVL1衣壳蛋白能溶于水，而且表达得到的是与HPVL1的VLP具有相同免疫原性和抗原性的L1五聚体。本发明使得利用原核表达系统工业生产HPVL1衣壳蛋白成为现实，相对于目前采用的真核表达系统具有产品质量更稳定、产率更高、成本低、质量控制方便的优点，具有重大的经济效益和社会效应。文档编号C12N15/63GK101245099SQ200710005099公开日2008年8月20日申请日期2007年2月14日优先权日2007年2月14日发明者陈小江,马润林申请人:马润林;陈小江