CN1443773A - 钩端螺旋体致病相关蛋白及其编码序列 - Google Patents
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Abstract
本发明提供了一类新的钩端螺旋体致病相关蛋白，编码致病相关蛋白的多核苷酸和经重组技术产生这类致病相关蛋白的方法。本发明还公开了这类致病相关蛋白及其编码序列的用途。
Description
技术领域
本发明属于生物技术和医学领域，具体地说，本发明涉及新的编码钩端螺旋体致病相关蛋白的多核苷酸，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。
背景技术
钩端螺旋体病是一种分布广泛的人兽共患病，主要分布在中国南方以及东南亚、南亚等发展中国家和地区。在中国，本病1995年列入法定传染病。1955-1993年，年平均发病率为十万分之七，年平均死亡率占发病人数的1.02％。
中国钩端螺旋体病的的流行菌群以黄疸出血群和波摩那群为主。黄疸出血群有15个血清型，其中以赖型最为常见。
中国虽在该病的防治方面取得了巨大的成功，但对该病的致病免疫的机理、疫苗的高效保护等问题仍未解决。
钩端螺旋体(Leptospira sp.)属格兰氏阴性菌。人类感染致病性钩端螺旋体后有发热、头痛、肌肉酸痛、黄疸、肝肾功能损伤及体内各脏器的弥漫性出血，严重者导致死亡。急性症状患者常出现肺出血而暴死。
对于组织出血机制进行研究发现，人体内出血、凝血处于一个相互平衡的体系中。在正常情况下，参与凝血的因子有PAF(促进血小板凝集)，vWF(借导血小板凝集在血管内皮细胞破损后暴露出来的胶原蛋白上的一个蛋白)，凝血酶(促进血小板凝集)等因子，它们能促进血小板的凝集以及促进血小板堵塞在血管内皮组织破损处，而阻止出血。
然而目前为止，仍没有公开过钩端螺旋体致病相关蛋白。因此，本领域迫切需要开发新的钩端螺旋体致病相关蛋白基因，以便为研究钩端螺旋体病的致病机制及诊断提供了基础。
发明内容
本发明目的是提供新的钩端螺旋体致病相关蛋白以及其片段、类似物和衍生物。
本发明另一目的是提供编码这些多肽的多核苷酸。
本发明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致病相关蛋白多肽，它包含：具有SEQ ID NO：2、4、6、8、10、12、14、16、18、20、22、24、26、28、30、32、34、36、38、40、42、44、46、48、50、52、54氨基酸序列的多肽、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SEQ IDNO：2、4、6、8、10、12、14、16、18、20、22、24、26、28、30、32、34、36、38、40、42、44、46、48、50、52、54氨基酸序列的多肽，更佳地该多肽不含信号肽序列。
在本发明的第二方面，提供分离的编码上述多肽的多核苷酸，该多核苷酸包含一核苷酸序列，该核苷酸序列与选自下组的一种核苷酸序列有至少70％相同性：(a)编码上述钩端螺旋体致病相关蛋白多肽的多核苷酸；和(b)与多核苷酸(a)互补的多核苷酸。较佳地，该多核苷酸编码具有SEQ ID NO：2、4、6、8、10、12、14、16、18、20、22、24、26、28、30、32、34、36、38、40、42、44、46、48、50、52、54所示氨基酸序列的多肽。更佳地，该多核苷酸的序列是选自下组的一种：(a)具有SEQ ID NO：1、3、5、7、9、11、13、15、17、19、21、23、25、27、29、31、33、35、37、39、41、43、45、47、49、51、53。
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，提供了制备钩端螺旋体致病相关蛋白多肽的方法，该方法包含：(a)在适合表达钩端螺旋体致病相关蛋白的条件下，培养上述被转化或转导的宿主细胞；(b)从培养物中分离出钩端螺旋体致病相关蛋白多肽。
在本发明的第五方面，提供了与上述的钩端螺旋体致病相关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。
在本发明的第六方面，提供了检测样品中是否存在致病相关蛋白的方法，它包括：将样品与致病相关蛋白的特异性抗体接触，观察是否形成抗体复合物，形成了抗体复合物就表示样品中存在致病相关蛋白。
在本发明的第七方面，提供了本发明多肽和编码序列的用途。例如本发明的钩端螺旋体致病相关蛋白的编码序列或其片段，可被作为引物用于PCR扩增反应，或者作为探针用于杂交反应，或者用于制造基因芯片或微阵列。
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。
附图说明
下列附图用于说明本发明的具体实施方案，而不用于限定由权利要求书所界定的本发明范围。
图1A和B显示了血小板激活因子乙酰水解酶PAF_ah和已知蛋白1WAB的三维结构。
图2显示了本发明血小板激活因子乙酰水解酶功能结构域。图中的PD009782是乙酰水解酶PAF-AHγ亚基。
具体实施方式
本发明人经过广泛而深入的研究，从问号钩端螺旋体(Leptospirainterrogans)黄疸出血群赖型赖株中分离纯化和鉴定了一类新的致病相关蛋白。在此基础上完成了本发明。
本发明人在钩端螺旋体赖株基因组数据库中鉴别出下列致病相关蛋白，包括(1)ColA(胶原酶，水解胶原蛋白)，(2)PAF-AH(血小板激活因子乙酰水解酶，抑制血小板凝集)，(3)vWF A结构域，以及(4)与凝血酶或者GPIb结合的蛋白。这些基因的产物均抑制血小板的凝集而破坏凝血系统，从而出现大出血现象。通过这四种因子的生物活性进行研究，有助于研究钩端螺旋体病的致病机制。此外，本发明的蛋白还可作为筛选抗凝血的药物靶点，开发抗凝血、抗血栓药物，以及用于诊断出血性疾病。
在本发明中，术语“钩端螺旋体致病相关蛋白多肽”或“钩端螺旋体致病相关蛋白”可互换使用，都指具有钩端螺旋体致病相关蛋白氨基酸序列(SEQ ID NO：2、4、6、8、10、12、14、16、18、20、22、24、26、28、30、32、34、36、38、40、42、44、46、48、50、52、54)的蛋白或多肽。它们包括含有或不含起始甲硫氨酸的钩端螺旋体致病相关蛋白，以及含有或不含有信号肽的成熟蛋白。
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，“分离的致病相关蛋白或多肽”是指致病相关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致病相关蛋白。
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，本发明多肽可以是糖基化的，但优选是非糖基化的。
本发明还包括钩端螺旋体致病相关蛋白的片段、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钩端螺旋体致病相关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，或与抗原IgG片段的形成的融合蛋白)。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。
在本发明中，术语“钩端螺旋体致病相关蛋白多肽”指具有钩端螺旋体致病相关蛋白活性或免疫原性的SEQ ID NO：2、4、6、8、10、12、14、16、18、20、22、24、26、28、30、32、34、36、38、40、42、44、46、48、50、52、54序列的多肽。该术语还包括具有与钩端螺旋体致病相关蛋白相同功能的、上述序列的变异形式。这些变异形式包括(但并不限于)：若干个(通常为1-50个，较佳地1-30个，更佳地1-20个，最佳地1-10个)氨基酸的缺失、插入和/或取代，以及在C末端和/或N末端添加一个或数个(通常为10个以内，更佳地为5个以内)氨基酸。该术语还包括钩端螺旋体致病相关蛋白的活性片段和活性衍生物。
该多肽的变异形式包括：同源序列、保守性变异体、等位变异体、天然突变体、诱导突变体、在高或低的严紧度条件下能与钩端螺旋体致病相关蛋白DNA杂交的DNA所编码的蛋白、以及利用抗钩端螺旋体致病相关蛋白多肽的抗血清获得的多肽或蛋白。本发明还提供了其他多肽，如包含钩端螺旋体致病相关蛋白多肽或其片段的融合蛋白。除了几乎全长的多肽外，本发明还包括了钩端螺旋体致病相关蛋白多肽的可溶性片段。通常，该片段具有钩端螺旋体致病相关蛋白多肽序列的至少约10个连续氨基酸，通常至少约30个连续氨基酸，较佳地至少约50个连续氨基酸，更佳地至少约80个连续氨基酸，最佳地至少约100个连续氨基酸。这些片段也可用作免疫原以诱导针对钩端螺旋体的免疫应答反应。
发明还提供钩端螺旋体致病相关蛋白或多肽的类似物。这些类似物与天然钩端螺旋体致病相关蛋白多肽的差别可以是氨基酸序列上的差异，也可以是不影响序列的修饰形式上的差异，或者兼而有之。这些多肽包括天然或诱导的遗传变异体。诱导变异体可以通过各种技术得到，如通过辐射或暴露于诱变剂而产生随机诱变，还可通过定点诱变法或其他已知分子生物学的技术。类似物还包括具有不同于天然L-氨基酸的残基(如D-氨基酸)的类似物，以及具有非天然存在的或合成的氨基酸(如β、γ-氨基酸)的类似物。应理解，本发明的多肽并不限于上述例举的代表性的多肽。
修饰(通常不改变一级结构)形式包括：体内或体外的多肽的化学衍生形式如乙酰化或羧基化。修饰还包括糖基化，如那些在多肽的合成和加工中或进一步加工步骤中进行糖基化修饰而产生的多肽。修饰形式还包括具有磷酸化氨基酸残基(如磷酸酪氨酸，磷酸丝氨酸，磷酸苏氨酸)的序列。还包括被修饰从而提高了其抗蛋白水解性能或优化了溶解性能的多肽。
在本发明中，“钩端螺旋体致病相关蛋白保守性变异多肽”指与SEQ ID NO：2、4、6、8、10、12、14、16、18、20、22、24、26、28、30、32、34、36、38、40、42、44、46、48、50、52、54的氨基酸序列相比，有至多10个，较佳地至多8个，更佳地至多5个，最佳地至多3个氨基酸被性质相似或相近的氨基酸所替换而形成多肽。这些保守性变异多肽最好根据表1进行氨基酸替换而产生。
表1
| 最初的残基 | 代表性的取代 | 优选的取代 |
| Ala(A) | Val；Leu；Ile | Val |
| Arg(R) | Lys；Gln；Asn | Lys |
| Asn(N) | Gln；His；Lys；Arg | Gln |
| Asp(D) | Glu | Glu |
| Cys(C) | Ser | Ser |
| Gln(Q) | Asn | Asn |
| Glu(E) | Asp | Asp |
| Gly(G) | Pro；Ala | Ala |
| His(H) | Asn；Gln；Lys；Arg | Arg |
| Ile(I) | Leu；Val；Met；Ala；Phe | Leu |
| Leu(L) | Ile；Val；Met；Ala；Phe | Ile |
| Lys(K) | Arg；Gln；Asn | Arg |
| Met(M) | Leu；Phe；Ile | Leu |
| Phe(F) | Leu；Val；Ile；Ala；Tyr | Leu |
| Pro(P) | Ala | Ala |
| Ser(S) | Thr | Thr |
| Thr(T) | Ser | Ser |
| Trp(W) | Tyr；Phe | Tyr |
| Tyr(Y) | Trp；Phe；Thr；Ser | Phe |
| Val(V) | Ile；Leu；Met；Phe；Ala | Leu |
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。以致病相关蛋白-胶原蛋白酶lep930-167为例，编码成熟多肽的编码区序列可以与SEQ ID NO：1所示的编码区序列相同或者是简并的变异体。如本文所用，以lep930-167为例，“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：1所示的编码区序列有差别的核酸序列。本发明的其他蛋白依此类推。本发明的核苷酸序列可以根据密码子的对应关系，根据SEQ ID NO：2、4、6、8、10、12、14、16、18、20、22、24、26、28、30、32、34、36、38、40、42、44、46、48、50、52、54的氨基酸推导出。
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90％以上，更好是95％以上时才发生杂交。并且，可杂交的多核苷酸编码的多肽与成熟的钩端螺旋体致病相关蛋白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/或分离编码致病相关蛋白的多聚核苷酸。
本发明的钩端螺旋体致病相关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DNA文库或按本领域技术人员已知的常规方法所制备的DNA文库作为模板，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得到编码本发明蛋白(或其片段，或其衍生物)的DNA序列。然后可将该DNA序列引入本领域中已知的各种现有的DNA分子(或如载体)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。
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致病相关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。
通过常规重组DNA技术(Science，1984；224：1431)，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致病相关蛋白多肽。一般来说有以下步骤：
(1).用本发明的编码钩端螺旋体致病相关蛋白多肽的多核苷酸(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，钩端螺旋体致病相关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；在哺乳动物细胞中表达的pMSXND表达载体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钩端螺旋体致病相关蛋白编码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。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。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的动物细胞等。
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。另一种方法是使用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在细胞内、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。
另一方面，本发明还包括对钩端螺旋体致病相关蛋白DNA或是其片段编码的多肽具有特异性的多克隆抗体和单克隆抗体，尤其是单克隆抗体。这里，“特异性”是指抗体能结合于钩端螺旋体致病相关蛋白基因产物或片段。较佳地，指那些能与钩端螺旋体致病相关蛋白基因产物或片段结合但不识别和结合于其它非相关抗原分子的抗体。本发明还包括那些能与修饰或未经修饰形式的钩端螺旋体致病相关蛋白基因产物结合的抗体。
本发明不仅包括完整的单克隆或多克隆抗体，而且还包括具有免疫活性的抗体片段，如Fab′或(Fab)2片段；抗体重链；抗体轻链；或嵌合抗体。
本发明的抗体可以通过本领域内技术人员已知的各种技术进行制备。例如，纯化的钩端螺旋体致病相关蛋白基因产物或者其具有抗原性的片段，可被施用于动物以诱导多克隆抗体的产生。与之相似的，表达钩端螺旋体致病相关蛋白或其具有抗原性的片段的细胞可用来免疫动物来生产抗体。本发明的抗体也可以是单克隆抗体。此类单克隆抗体可以利用杂交瘤技术来制备。
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：ColdSpring Harbor Laboratory Press，1989)中所述的条件，或按照制造厂商所建议的条件。
实施例1
菌体培养
1)培养基：使用EMJH液体培养基，配制5×的EMJH培养基1000毫升，称取BSA50g。无水醋酸钠0.5g，丙酮酸钠1g，硫酸亚铁0.25g，1.5％的氯化镁3.5ml，0.4％的硫酸锌5ml，0.15的硫酸铜1ml，磷酸氢二钠12.6g，磷酸二氢钾1.5g，氯化钠5g，氯化铵1.25g，0.05％的VB122ml，VB10.025g，调pH为7.2-7.6，定容至1000ml分别以0.7、0.45、0.2nm孔径的硝酸纤维素膜过滤除菌。使用时，用无菌水其5倍稀释。
2)培养条件：30℃，100转/分钟摇动培养40小时。
3)菌株：问号钩端螺旋体黄疸出血群赖型赖株。
实施例2
致病相关蛋白的核酸序列和氨基酸序列
用常规方法抽提钩端螺旋体DNA，构建文库。测定基因组全序列。根据计算机分析结果，合成引物，以抽提的钩端螺旋体DNA为模板，进行PCR扩增，获得致病相关蛋白的ORF序列。再次测序验证，与基因组序列相符。这些ORF编码具有SEQID NO：2、4、6、8、10、12、14、16、18、20、22、24、26、28、30、32、34、36、38、40、42、44、46、48、50、52、54所示氨基酸序列的多肽。本发明的8种致病相关蛋白的基本情况如下表所示。
表A致病相关蛋白的核酸序列和氨基酸序列
| 名称 | ORF序列SEQID NO： | 长度(bp) | 蛋白序列SEQ ID NO： | 长度(aa) | 信号肽* | |
| 胶原蛋白酶 | lep930-167 | 1 | 2667 | 888 | 2 | 1-27 |
| 血小板激活因子乙酰水解酶 | Paf_ah | 3 | 690 | 229 | 4 | 无 |
| vWF A Domain No.1 | LB54 | 5 | 963 | 320 | 6 | 1-27 |
| vWF A Domain No.2 | LB55 | 7 | 1044 | 347 | 8 | 无 |
| 与凝血酶或者GPIb结合的蛋白No.1 | LA699 | 9 | 2826 | 941 | 10 | 无 |
| 与凝血酶或者GPIb结合 | LA703 | 11 | 504 | 167 | 12 | 1-33 |
| 的蛋白No.2 | ||||||
| 与凝血酶或者GPIb结合的蛋白No.3 | LA871 | 13 | 1272 | 423 | 14 | 1-23 |
| 与凝血酶或者GPIb结合的蛋白No.4 | LA1174 | 15 | 369 | 122 | 16 | 1-23 |
| 与凝血酶或者GPIb结合的蛋白No.5 | LA1321 | 17 | 654 | 217 | 18 | 1-29 |
| 与凝血酶或者GPIb结合的蛋白No.6 | LA1350 | 19 | 531 | 176 | 20 | 1-9 |
| 与凝血酶或者GPIb结合的蛋白No.7 | LA1351 | 21 | 300 | 99 | 22 | 无 |
| 与凝血酶或者GPIb结合的蛋白No.8 | LA1572 | 23 | 282 | 93 | 24 | 1-10 |
| 与凝血酶或者GPIb结合的蛋白No.9 | LA2435 | 25 | 1581 | 526 | 26 | 52-53 |
| 与凝血酶或者GPIb结合的蛋白No.10 | LA2438 | 27 | 234 | 77 | 28 | 无 |
| 与凝血酶或者GPIb结合的蛋白No.11 | LA2439 | 29 | 1497 | 498 | 30 | 1-29 |
| 与凝血酶或者GPIb结合的蛋白No.12 | LA2440 | 31 | 1287 | 428 | 32 | 1-29 |
| 与凝血酶或者GPIb结合的蛋白No.13 | LA2442 | 33 | 1902 | 633 | 34 | 1-29 |
| 与凝血酶或者GPIb结合的蛋白No.14 | LA2444 | 35 | 870 | 289 | 36 | 1-53 |
| 与凝血酶或者GPIb结合的蛋白No.15 | LA2851 | 37 | 867 | 288 | 38 | 无 |
| 与凝血酶或者GPIb结合的蛋白No.16 | LA2953 | 39 | 1281 | 426 | 40 | 1-29 |
| 与凝血酶或者GPIb结合的蛋白No.17 | LA2955 | 41 | 1227 | 408 | 42 | 1-30 |
| 与凝血酶或者GPIb结合的蛋白No.18 | LA3017 | 43 | 2058 | 685 | 44 | 无 |
| 与凝血酶或者GPIb结合的蛋白No.19 | LA3307 | 45 | 1134 | 377 | 46 | 1-29 |
| 与凝血酶或者GPIb结合的蛋白No.20 | LA3308 | 47 | 1359 | 452 | 48 | 1-29 |
| 与凝血酶或者GPIb结合的蛋白No.21 | LA3309 | 49 | 855 | 284 | 50 | 1-28 |
| 与凝血酶或者GPIb结合的蛋白No.22 | LA3310 | 51 | 936 | 311 | 52 | 1-29 |
| 与凝血酶或者GPIb结合的蛋白No.23 | LA3311 | 53 | 990 | 329 | 54 | 无 |
*信号肽一栏中给出的数字是信号肽的起始和终止氨基酸位置。
本发明的钩端螺旋体致病相关蛋白中，Paf_ah与IWAB的氨基酸序列在一级结构上有41/128(32％)相同，70/128(54％)相似。在三维结构上也极其相似(图1A和1B)。模拟PDB数据库中的1WAB(已知的牛脑的PAF-AH)，发现钩端螺旋体的PAF-AH的N-端Ser27，Asp171，His174与1WAB的Ser47，Asp192，His195的活性位点完全相同。
vWF A Domain LB54与LB55有较高的同源性。
胶原蛋白酶在一级结构上与PD043590(最相近的功能域：COLA_CLOPE 90-760)有193/675(28％)相同性，324/675(47％)相似性。
实施例3
蛋白表达
用常规基因工程方法将钩端螺旋体致病相关蛋白基因克隆入融合表达载体pET28b(Novagene公司)，融合表达的受体菌选用大肠杆菌BL21(DE3)。常规条件发酵后，提取重组菌体蛋白，然后进行SDS-PAGE电泳，分子量与预测值基本相同。大肠杆菌中表达的致病相关蛋白(带有His-tag的融合蛋白)，通过Ni-NTA柱分离纯化，得到纯的钩端螺旋体致病相关蛋白(融合蛋白)。
实施例4
制备抗体
将实施例3中获得的纯化的、重组His-融合蛋白用来免疫动物以产生抗体，具体方法如下：将融合蛋白用等量的完全Freund′s佐剂乳化。用200μg乳化过的蛋白，对家兔进行淋巴结注射。一个月后，用等量的非完全Freund′s佐剂乳化的同样抗原，对家兔以200μg的剂量进行淋巴结注射以加强免疫。每隔14天进行一次加强免疫，至少进行三次。获得的抗血清的特异反应活性用ELISA检验抗体滴度。结果发现，抗体可特异性地与本发明蛋白发生结合。
在本发明提及的所有文献都在本申请中引用作为参考，就如同每一篇文献被单独引用作为参考那样。此外应理解，在阅读了本发明的上述讲授内容之后，本领域技术人员可以对本发明作各种改动或修改，这些等价形式同样落于本申请所附权利要求书所限定的范围。
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180 185 190Tyr Val Lys His Ser His Pro Ser Leu Pro Phe Asp Thr Asn Gln Ile
195 200 205Ser Gln Ala Leu Val Thr Pro Met Gln Ile Phe Ala Ser Ser Ala His
210 215 220Phe Leu Asp Gly Thr Asn Asp Ala Gly Lys Val Leu Glu Phe Phe Phe225 230 235 240Thr Val Ala Asp Ser Thr Lys Ile Gly His Thr Ile Tyr Pro Arg Ile
245 250 255Leu Ser Phe Leu Glu Ala Thr Ile Asn Asp Pro Gln Arg Leu Arg Asn
260 265 270Asn Leu Ser Gln Ala Ile Ala Leu Asn Ala Val Phe Arg Leu Phe Gln
275 280 285Arg His Ile His Ser Asn Ser Asn Glu Phe Leu Thr Met Ile Asp Tyr
290 295 300Arg Leu Ile Ser Lys Leu Arg Arg Leu Ala Leu Asp Thr Ser Leu Asn305 310 315 320Thr Asp Ser Gln Val Trp Ile Ile Asn Asn Ala Ile Phe Gly Leu Asp
325 330 335Arg Ile Tyr Glu Tyr Leu Pro Ser Phe Gln Pro Val Ile Ala Ser Val
340 345 350Met Thr Asp Val Leu Glu Thr Tyr Pro Tyr Ile Ser Glu Pro Tyr Leu
355 360 365Leu Gly Ile Lys Ala Leu Thr Arg His Ser Asp Cys Ala Asn Leu Arg
370 375 380Ile Gly Arg Ile Cys Leu Ser Asp Ile Lys Glu Thr Val Lys Lys Ala385 390 395 400Val Leu Ser Asn Thr Tyr Tyr Phe Asp Asp Lys Thr Gln Ile Val His
405 410 415Thr Ala Leu Ser Ile Asp Glu Ile Gln Pro Leu Tyr His Ala Leu Lys
420 425 430Gln Val Glu Ser Gln Phe Phe Arg Leu Ile Gly Ile His Ala Pro Val
435 440 445Ser Gly Asp Thr Thr Asp Ser Ile Thr Met Tyr Val Tyr Lys Ser Arg
450 455 460Lys Asp Tyr Glu Thr Phe His Pro Phe Leu Phe Asp Leu Ser Thr Asp465 470 475 480Asn Gly Gly Ile Tyr Ile Glu Arg Asp Lys Thr Leu Tyr Thr Tyr Gln
485 490 495Arg Thr Pro Ala Glu Ser Ile Tyr Thr Leu Glu Glu Leu Leu Arg His
500 505 510Glu Tyr Ser His Tyr Leu Val Gly Arg Phe Ile Ile Pro Gly Leu Trp
515 520 525Ser Gln Thr Ser Ala His Asp Asn Glu Arg Leu Thr Trp Phe Glu Glu
530 535 540Gly Ile Ala Glu Phe Leu Thr Gly Ser Ser Ser Arg Glu Ile His Pro545 550 555 560Arg Lys Ser Leu Ile Ser Glu Ile Lys Arg Asp Gly Ser Ser Arg Met
565 570 575Asp Val Ser Lys Ile Leu Thr Ala Arg Tyr Gly Asp Phe Lys Phe Tyr
580 585 590Arg Tyr Ser Gly Asn Phe Phe Asn Tyr Leu Tyr Thr Tyr Lys Lys Asp
595 600 605Thr Leu Lys Asp Leu Ile Ser Ala Leu Arg Asp Ser Asn Ile Gln Thr
610 615 620Phe Asp Phe Leu Ile Asp Gln Met Ser His Asp Thr Gly Leu Asn Thr625 630 635 640Ser Phe Gln Ala Tyr Leu Asp Tyr Leu Val Ser Asn Val Asp Asn Leu
645 650 655Thr Asp Pro Ser Thr Pro Ala Pro Asp Leu Ala Asn Leu Ser Thr Asn
660 665 670Asp Pro Lys Val Ile Arg Ser Phe Phe Arg Lys Thr Lys Thr Gly Leu
675 680 685Lys Ala Lys Cys Thr Thr Ala Ala Phe Gly Leu Asn Ser Arg Phe Ser
690 695 700Cys Arg Gly Leu Ile Phe Gly Asn Ala Thr Tyr Ser Pro Asp Trp Asn705 7l0 715 720Ser Ala Trp Ser Asp Phe Asn Ser Arg Ile Asn Asp Leu Met Ser Thr
725 730 735Leu Glu Ser Ser Gly Val Asn Asn Phe Gln Ser Met Asn Cys Arg Ile
740 745 750Gly Glu Ile Arg Phe Asp Lys His Leu Asn Leu Phe Arg Pro Phe Thr
755 760 765Ile Tyr Ser Cys Glu Gly Pro Leu Ala Tyr Arg Ser Arg Ile Ser Tyr
770 775 780Arg His Pro Gln Arg Gly Gln Thr Asp Phe Arg Asn Thr Arg Ile Gly785 790 795 800Ile His Ser Thr Cys Phe Ala Thr Pro Ser Asn Gly Ser Ser Thr Thr
805 8l0 815Cys Asn Thr Pro Ile Val Thr Asn Ala Phe Ser Asn Thr Ala Thr Tyr
820 825 830Glu Glu Met Tyr Gln Lys Leu Asn Gly Glu Leu Asn Glu Leu Lys Ser
835 840 845Glu Val Tyr Thr Met Arg Pro Ser Phe Tyr Lys Lys Leu Asp Cys Asn
850 855 860Phe Asp Ser Ile Asp Thr Ile His Leu Pro Asp Gly Arg Lys Tyr Leu865 870 875 880Ala Ala Asn Ser Ser Cys Asn Phe
885<210>3<211>690<212>DNA<213>钩端螺旋体(Leptospira interrogans)<400>3ttgattaaga aatcatatac ggacgattat tctagtccta attttgaatg ttggtctgga 60tccggttttc gcgattccga aacgttcgga ttatacaaat ctgcatgggc cgagcttcga 120aatttttata aaattgaaaa tcagaaaatt aaatctgcaa atatagtctt tgtaggagat 180tctttgattc aattgtttcc caaagagctg atggtccaag aatttccagg tgcggttaat 240cgcggtattg gtggagattt gacggaaact cttctggaaa gaatagaaga agacgtttta 300agtttaaatc caaaagtaat cgttttagag attggtggaa atgatctcat ccaaggaaaa 360tgtctacata ttacagaaac gaatttaaat cgaattttag aaaaaattct caagttcaac 420ccgaatataa aaatagttat tttaggaatt cctcctgttc ggaatcaaac tgtaaataga 480gtttctcccg tactcaatct tacctggatt tctatcattc gaccttataa gaatgtggaa 540tttttagacg gttggcaatt cttgagagaa aaggatcgac ccgttttgga ttcggaattt 600tggccaggcc aggataaaat tcatgtgaac gaaaaagcat atcgagcctg gattcataaa 660ttaaaaccga tacttttacc ctatctttaa 690<210>4<211>229<212>PRT<213>钩端螺旋体(Leptospira interrogans)<400>4Leu Ile Lys Lys Ser Tyr Thr Asp Asp Tyr Ser Ser Pro Asn Phe Glu1 5 10 15Cys Trp Ser Gly Ser Gly Phe Arg Asp Ser Glu Thr Phe Gly Leu Tyr
20 25 30Lys Ser Ala Trp Ala Glu Leu Arg Asn Phe Tyr Lys Ile Glu Asn Gln
35 40 45Lys Ile Lys Ser Ala Asn Ile Val Phe Val Gly Asp Ser Leu Ile Gln
50 55 60Leu Phe Pro Lys Glu Leu Met Val Gln Glu Phe Pro Gly Ala Val Asn65 70 75 80Arg Gly Ile Gly Gly Asp Leu Thr Glu Thr Leu Leu Glu Arg Ile Glu
85 90 95Glu Asp Val Leu Ser Leu Asn Pro Lys Val Ile Val Leu Glu Ile Gly
100 105 110Gly Asn Asp Leu Ile Gln Gly Lys Cys Leu His Ile Thr Glu Thr Asn
115 120 125Leu Asn Arg Ile Leu Glu Lys Ile Leu Lys Phe Asn Pro Asn Ile Lys130 135 140Ile Val Ile Leu Gly Ile Pro Pro Val Arg Asn Gln Thr Val Asn Arg145 150 155 160Val Ser Pro Val Leu Asn Leu Thr Trp Ile Ser Ile Ile Arg Pro Tyr
165 170 175Lys Asn Val Glu Phe Leu Asp Gly Trp Gln Phe Leu Arg Glu Lys Asp
180 185 190Arg Pro Val Leu Asp Ser Glu Phe Trp Pro Gly Gln Asp Lys Ile His
195 200 205Val Asn Glu Lys Ala Tyr Arg Ala Trp Ile His Lys Leu Lys Pro Ile
210 215 220Leu Leu Pro Tyr Leu225<210>5<211>963<212>DNA<213>钩端螺旋体(Leptospira interrogans)<400>5ttggaaagga atctgggatc aaaaatgaat tttcaatatc cttacgcgct tttacttttg 60attccagttt ggatctgggc ttttatatat tataaaaaca aaatatactt aaaaaaaatg 120gaaattcgtt ttccaggaag aagagaaagt actttttcca aatttgaaaa cttagggaaa 180tttttaccga tacttcgccc gattgcgatt agtttgatga tactttctct ttccggacca 240ggtaaaaaaa ttacctttct acccgacgaa aaagaaggag tagatatatt aatcgccttg 300gacgtctcag gttccatgtc tagaagtaga gactttcttc cggaaactag attaggagtt 360tctaaaaaac tattaaaaag atttatcgaa aaaagaaata atgatcgtct tggtttagtg 420gtttttgcag gtgcggctta tttacaagct cctcttacag gtgatcgaga atctttgagc 480gaaattttag aaaccataga agaagaaaca gtgacggaac aagggaccgc aatcggagac 540gcgatcattc tttctacgta tcgattgagg aattctaagg ctcgttctaa agtgattgta 600ctcatcaccg atggagtttc caatacgggt aaaatcgatc cagtaacagc gactgatctt 660gcggaacaaa ttggagcgaa aatttattcc gtcggaattg gaaaagaaga cggttcttat 720gaaatcaatt tcgaaatttt gcaagaactt tctgccaata ctggaggcag attttttagg 780gcagaagatc cggaggaaat gaaagcggtt ctctcttcga tcgattctct tgaaaaagat 840cctctgcaaa gtcctcccaa agaaataaga gaaactgaat atgaaatctg gctctataga 900gccttagcgg ttttactttt agatttaata ttacgtgcgt tttttctgag atattatgta 960tga 963<210>6<211>320<212>PRT<213>钩端螺旋体(Leptospira interrogans)<400>6Met Glu Arg Asn Leu Gly Ser Lys Met Asn Phe Gln Tyr Pro Tyr Ala1 5 10 15Leu Leu Leu Leu Ile Pro Val Trp Ile Trp Ala Phe Ile Tyr Tyr Lys
20 25 30Asn Lys Ile Tyr Leu Lys Lys Met Glu Ile Arg Phe Pro Gly Arg Arg
35 40 45Glu Ser Thr Phe Ser Lys Phe Glu Asn Leu Gly Lys Phe Leu Pro Ile
50 55 60Leu Arg Pro Ile Ala Ile Ser Leu Met Ile Leu Ser Leu Ser Gly Pro65 70 75 80Gly Lys Lys Ile Thr Phe Leu Pro Asp Glu Lys Glu Gly Val Asp Ile
85 90 95Leu Ile Ala Leu Asp Val Ser Gly Ser Met Ser Arg Ser Arg Asp Phe
100 105 110Leu Pro Glu Thr Arg Leu Gly Val Ser Lys Lys Leu Leu Lys Arg Phe
115 120 125Ile Glu Lys Arg Asn Asn Asp Arg Leu Gly Leu Val Val Phe Ala Gly
130 135 140Ala Ala Tyr Leu Gln Ala Pro Leu Thr Gly Asp Arg Glu Ser Leu Ser145 150 155 160Glu Ile Leu Glu Thr Ile Glu Glu Glu Thr Val Thr Glu Gln Gly Thr
165 170 175Ala Ile Gly Asp Ala Ile Ile Leu Ser Thr Tyr Arg Leu Arg Asn Ser
180 185 190Lys Ala Arg Ser Lys Val Ile Val Leu Ile Thr Asp Gly Val Ser Asn
195 200 205Thr Gly Lys Ile Asp Pro Val Thr Ala Thr Asp Leu Ala Glu Gln Ile
210 215 220Gly Ala Lys Ile Tyr Ser Val Gly Ile Gly Lys Glu Asp Gly Ser Tyr225 230 235 240Glu Ile Asn Phe Glu Ile Leu Gln Glu Leu Ser Ala Asn Thr Gly Gly
245 250 255Arg Phe Phe Arg Ala Glu Asp Pro Glu Glu Met Lys Ala Val Leu Ser
260 265 270Ser Ile Asp Ser Leu Glu Lys Asp Pro Leu Gln Ser Pro Pro Lys Glu
275 280 285Ile Arg Glu Thr Glu Tyr Glu Ile Trp Leu Tyr Arg Ala Leu Ala Val
290 295 300Leu Leu Leu Asp Leu Ile Leu Arg Ala Phe Phe Leu Arg Tyr Tyr Val305 310 315 320<210>7<211>1044<212>DNA<213>钩端螺旋体(Leptospira interrogans)<400>7atgaatcaag agttttccgc taatttaaaa aacatttttt tcgcccttgt aattgtatgg 60attatatatt caactattag aatatttttg atttctaaaa taagggaatg gaaaattttt 120tatcctggtt tagaacgttc ttcttcgttt ccggatgttc gttgggtatt ggtaagaatt 180ttattaattt ttgtggtttt agtctgtgtt ttttttgcag gtctaaaaac ggaaatgaaa 240gacgaaaaaa aggaagaatc tttcaaagga gtggatatac tctttttagt ggatgtaagt 300ctttctatgc aggcgataga tagttctcca actcggttgg ctaaatttaa agaagtgtta 360cttaggatgt tgccctcttt atctggaaac agatttggaa tgattgtatt tgccggaagt 420ccgtttttat attgtccgat gactacggat gtgtccgctt tttcggatta tgttcgtgga 480ttggatgtgg atatggtagg ggatcgcgga accgatttga gtcaggcttt tacaaaggcg 540gaagcgttgc ttcgttccga aaaagtattt cgaaatcgaa ttttgatttt agtaaccgat 600ggagaagatc aaaatgatcc acaagcgatt tcctttccgg cgagttttca agtttgggct 660gccggaacgg aaagcggggg gccgatagtt tataacgatg aaaattctgg tttgaacggt 720tttcttttga aggatggaac tctgacttca aactctaatt ctcctggaat cattcattct 780aaaatgaatc gaactttttt aaaaaatcta gcagataaaa acgacgggaa tttttattca 840ctggattcaa atccacccga ttctacaatt ctacaaaagg aaattttttc tttagaggaa 900aatttatatt ctagaaaaaa agatctaaaa cgcgcggaag gttctggcaa atttttattt 960gcggcagttt ttctactatt attggattgg attttagtgg aatctttgtt atttcccaaa 1020cggaaatcgg tttctgtaat atga 1044<210>8<211>347<212>PRT<213>钩端螺旋体(Leptospira interrogans)<400>8Met Asn Gln Glu Phe Ser Ala Asn Leu Lys Asn Ile Phe Phe Ala Leu1 5 10 15Val Ile Val Trp Ile Ile Tyr Ser Thr Ile Arg Ile Phe Leu Ile Ser
20 25 30Lys Ile Arg Glu Trp Lys Ile Phe Tyr Pro Gly Leu Glu Arg Ser Ser
35 40 45Ser Phe Pro Asp Val Arg Trp Val Leu Val Arg Ile Leu Leu Ile Phe
50 55 60Val Val Leu Val Cys Val Phe Phe Ala Gly Leu Lys Thr Glu Met Lys65 70 75 80Asp Glu Lys Lys Glu Glu Ser Phe Lys Gly Val Asp Ile Leu Phe Leu
85 90 95Val Asp Val Ser Leu Ser Met Gln Ala Ile Asp Ser Ser Pro Thr Arg
100 105 110Leu Ala Lys Phe Lys Glu Val Leu Leu Arg Met Leu Pro Ser Leu Ser
115 120 125Gly Asn Arg Phe Gly Met Ile Val Phe Ala Gly Ser Pro Phe Leu Tyr
130 135 140Cys Pro Met Thr Thr Asp Val Ser Ala Phe Ser Asp Tyr Val Arg Gly145 150 155 160Leu Asp Val Asp Met Val Gly Asp Arg Gly Thr Asp Leu Ser Gln Ala
165 170 175Phe Thr Lys Ala Glu Ala Leu Leu Arg Ser Glu Lys Val Phe Arg Asn
180 185 190Arg Ile Leu Ile Leu Val Thr Asp Gly Glu Asp Gln Asn Asp Pro Gln
195 200 205Ala Ile Ser Phe Pro Ala Ser Phe Gln Val Trp Ala Ala Gly Thr Glu
210 215 220Ser Gly Gly Pro Ile Val Tyr Asn Asp Glu Asn Ser Gly Leu Asn Gly225 230 235 240Phe Leu Leu Lys Asp Gly Thr Leu Thr Ser Asn Ser Asn Ser Pro Gly
245 250 255Ile Ile His Ser Lys Met Asn Arg Thr Phe Leu Lys Asn Leu Ala Asp
260 265 270Lys Asn Asp Gly Asn Phe Tyr Ser Leu Asp Ser Asn Pro Pro Asp Ser
275 280 285Thr Ile Leu Gln Lys Glu Ile Phe Ser Leu Glu Glu Asn Leu Tyr Ser
290 295 300Arg Lys Lys Asp Leu Lys Arg Ala Glu Gly Ser Gly Lys Phe Leu Phe305 310 315 320Ala Ala Val Phe Leu Leu Leu Leu Asp Trp Ile Leu Val Glu Ser Leu
325 330 335Leu Phe Pro Lys Arg Lys Ser Val Ser Val Ile
340 345<210>9<211>2826<212>DNA<213>钩端螺旋体(Leptospira interrogans)<400>9atgatcaaag tatggccggg agaatcccaa caccaacgtg cggtcttagg tttagaaatt 60ttaaaaacga tcggttcgga tacggccctt atgcagttaa acggaatcgc tcaaaaagtg 120aaattcaaag gcctgaaaga tatggccaac acgtttatgg aatctatcgc caagaaaaag 180gggcttagaa aatccgaatt agaagatcgt gtggttccag attgtggttt ggacgaagac 240ggtaaaaggg aatttaattt cggtccacga aaattccatt ttgtatttgg tccggatatg 300aagcctatga tcaaagacga agacggaaaa atcaaagacg atcttcccaa accaaattca 360aaagatgact cagacctggc aaacacttcc gtagaagaat ggaagttaat gaaaaaacaa 420atccgagaaa tcggcaaaat ccatatacat agaatggaac aggcaatggt caccggaaga 480agatggaaag tagaagaatg ggaaatgctc atcgccaaac acccattaat gacacatatt 540gccaaaaccc tactttggtg ggtttatttt ccagatcaaa atcaatcgat caatgtattt 600cgactaacgg aagaaaaaga ctacgcggat atacatgata attctttaaa ccttcaagga 660ggagcctata tcggaatcgt tcatccattg ctgctttccc aagaggaaaa aaaatcctgg 720ggacaattgt tcagcgatta tgaaatcatt tctccgtttt ctcaaatttc aagaccggta 780tacgtccttt ctggggaaga taaaaacaaa caagaaatac ccggatttac aaaccaaaaa 840gtaaaagcgg aacagctagt atttggactc gaaaaaatgg gctggtccag aggatcggca 900ggagacggag gaggaatcga cgaacactcc atacaatttc ttggtgatga tgtaacagcg 960gttatacgat acgacggaga cgatctttcc tacggaaaca taggtggtca ggatctagat 1020ttagaagaag cttattttgt aaagggcctc cgagaacctt ctttttccgt aggaaaggaa 1080acaaaactaa ctttggaaga aatcaatccg gtagctttta gcgaaactct acatagtcta 1140ttacaagttt ctggattttc ttacccagca tctaacgaaa atagtttgaa agaagccaga 1200gaaaccttac tcaaaaatct acaaacacag gaaaagaaag aagaagtatt cgatgaagtc 1260gactacaccg aaatttacaa cggattttca acggcttgga aaaaatttct ttccaatgat 1320caccaaatca caagatccaa aaatcacaaa agcatctcta aaattacaaa aatgtatatt 1380agaagttccg ataaaattac ttctatccaa gaacttaagt tttttactaa attagaagaa 1440ctaacactta acgggccggt aacagattct tctttattat cggaattaaa aaatctgaaa 1500aaaatcgagc tccaaaattg gaacctaaaa gatctaaacg ttctaaactc ctgcacaaac 1560ttagaaaaag tagaacttgt ggatattcaa ggttttgaaa ctgatttcga ctgtagtgga 1620ttgttaaacg aatccaaagc gacgattcat ttaaatctat ccggaaccaa attcgaacgt 1680tttccaattt cagtgaccag gtttcaaaac ctaacctccc tttctttaag agactgtaaa 1740ctttcggaag ttccagaaag tataggcaat ctaaaacgac tcatcaattt gtacttggat 1800aaaaatcaac tgaccacatt acctacaagt ttaggaacct tagaacaact aacccaactc 1860catatagact caaatccatt tactacgatc ccggatgcgg tgttgtctct taaaaatctc 1920aaaactcttt tagcacgatg gaatcagatt tctactttac caaatgaaat cggaaatctt 1980acctctctgg aagatctaaa tttacatgac aaccaacttt ccagtcttcc aacaacgatt 2040caaaatctaa gttctttaac taagatcggt ttatcaaaaa ataagttttc agaatttccg 2100gagccaattc tatatttaaa aaatttaaaa catctagacg taggcgaaaa taaaattcgt 2160cagttaccgg aaacgatcgg aaatttaagt aatttaaaat ctctagatat aaaagagact 2220tggatagaat ctttgccgca gagtatccaa aatctgactc aactagaaac tatctacttg 2280ccaaaagcca aattcagaaa ccttccggat tttttggcaa atatggaatc tcttaaaaag 2340attaaatttg aaagcgagga atacaatcaa ctcacaaaat ggtgtgaatt tgaatattca 2400aaatatataa agcttctaca cggaggaaaa tatccggagg caagagataa aatcacaaat 2460ttgtttacca caaagggaga ggatttttta aaactcaacc aatgggaagt aaaactaaaa 2520atctccgaaa gttactatcg taaggccgaa gcctttgcaa ccgtcgtata cgcattagcc 2580acaattctaa aggacgagga aatctattcc attgcgtccc aaatgacggg ggaccaatac 2640atccacccag ctcttccgtt caataaagcc tgttactttg cggtgaccgg tcaaaaggaa 2700cctatgctcc aatcgattcg aaaatccgtc aaattaggaa caaaagcgga tgaatttacg 2760aaggaaaagg attttgcttc ttatttaaaa gatcctgact ttttggaggc aatccgtaaa 2820aattaa 2826<210>10<211>941<212>PRT<213>钩端螺旋体(Leptospira interrogans)<400>10Met Ile Lys Val Trp Pro Gly Glu Ser Gln His Gln Arg Ala Val Leu1 5 10 15Gly Leu Glu Ile Leu Lys Thr Ile Gly Ser Asp Thr Ala Leu Met Gln
20 25 30Leu Asn Gly Ile Ala Gln Lys Val Lys Phe Lys Gly Leu Lys Asp Met
35 40 45Ala Asn Thr Phe Met Glu Ser Ile Ala Lys Lys Lys Gly Leu Arg Lys
50 55 60Ser Glu Leu Glu Asp Arg Val Val Pro Asp Cys Gly Leu Asp Glu Asp65 70 75 80Gly Lys Arg Glu Phe Asn Phe Gly Pro Arg Lys Phe His Phe Val Phe
85 90 95Gly Pro Asp Met Lys Pro Met Ile Lys Asp Glu Asp Gly Lys Ile Lys
100 105 110Asp Asp Leu Pro Lys Pro Asn Ser Lys Asp Asp Ser Asp Leu Ala Asn
115 120 125Thr Ser Val Glu Glu Trp Lys Leu Met Lys Lys Gln Ile Arg Glu Ile
130 135 140Gly Lys Ile His Ile His Arg Met Glu Gln Ala Met Val Thr Gly Arg145 150 155 160Arg Trp Lys Val Glu Glu Trp Glu Met Leu Ile Ala Lys His Pro Leu
165 170 175Met Thr His Ile Ala Lys Thr Leu Leu Trp Trp Val Tyr Phe Pro Asp
180 185 190Gln Asn Gln Ser Ile Asn Val Phe Arg Leu Thr Glu Glu Lys Asp Tyr
195 200 205Ala Asp Ile His Asp Asn Ser Leu Asn Leu Gln Gly Gly Ala Tyr Ile
210 215 220Gly Ile Val His Pro Leu Leu Leu Ser Gln Glu Glu Lys Lys Ser Trp225 230 235 240Gly Gln Leu Phe Ser Asp Tyr Glu Ile Ile Ser Pro Phe Ser Gln Ile
245 250 255Ser Arg Pro Val Tyr Val Leu Ser Gly Glu Asp Lys Asn Lys Gln Glu
260 265 270Ile Pro Gly Phe Thr Asn Gln Lys Val Lys Ala Glu Gln Leu Val Phe
275 280 285Gly Leu Glu Lys Met Gly Trp Ser Arg Gly Ser Ala Gly Asp Gly Gly
290 295 300Gly Ile Asp Glu His Ser Ile Gln Phe Leu Gly Asp Asp Val Thr Ala305 310 315 320Val Ile Arg Tyr Asp Gly Asp Asp Leu Ser Tyr Gly Asn Ile Gly Gly
325 330 335Gln Asp Leu Asp Leu Glu Glu Ala Tyr Phe Val Lys Gly Leu Arg Glu
340 345 350Pro Ser Phe Ser Val Gly Lys Glu Thr Lys Leu Thr Leu Glu Glu Ile
355 360 365Asn Pro Val Ala Phe Ser Glu Thr Leu His Ser Leu Leu Gln Val Ser
370 375 380Gly Phe Ser Tyr Pro Ala Ser Asn Glu Asn Ser Leu Lys Glu Ala Arg385 390 395 400Glu Thr Leu Leu Lys Asn Leu Gln Thr Gln Glu Lys Lys Glu Glu Val
405 4l0 415Phe Asp Glu Val Asp Tyr Thr Glu Ile Tyr Asn Gly Phe Ser Thr Ala
420 425 430Trp Lys Lys Phe Leu Ser Asn Asp His Gln Ile Thr Arg Ser Lys Asn
435 440 445His Lys Ser Ile Ser Lys Ile Thr Lys Met Tyr Ile Arg Ser Ser Asp
450 455 460Lys Ile Thr Ser Ile Gln Glu Leu Lys Phe Phe Thr Lys Leu Glu Glu465 470 475 480Leu Thr Leu Asn Gly Pro Val Thr Asp Ser Ser Leu Leu Ser Glu Leu
485 490 495Lys Asn Leu Lys Lys Ile Glu Leu Gln Asn Trp Asn Leu Lys Asp Leu
500 505 510Asn Val Leu Asn Ser Cys Thr Asn Leu Glu Lys Val Glu Leu Val Asp
515 520 525Ile Gln Gly Phe Glu Thr Asp Phe Asp Cys Ser Gly Leu Leu Asn Glu
530 535 540Ser Lys Ala Thr Ile His Leu Asn Leu Ser Gly Thr Lys Phe Glu Arg545 550 555 560Phe Pro Ile Ser Val Thr Arg Phe Gln Asn Leu Thr Ser Leu Ser Leu
565 570 575Arg Asp Cys Lys Leu Ser Glu Val Pro Glu Ser Ile Gly Asn Leu Lys
580 585 590Arg Leu Ile Asn Leu Tyr Leu Asp Lys Asn Gln Leu Thr Thr Leu Pro
595 600 605Thr Ser Leu Gly Thr Leu Glu Gln Leu Thr Gln Leu His Ile Asp Ser
610 615 620Asn Pro Phe Thr Thr Ile Pro Asp Ala Val Leu Ser Leu Lys Asn Leu625 630 635 640Lys Thr Leu Leu Ala Arg Trp Asn Gln Ile Ser Thr Leu Pro Asn Glu
645 650 655Ile Gly Asn Leu Thr Ser Leu Glu Asp Leu Asn Leu His Asp Asn Gln
660 665 670Leu Ser Ser Leu Pro Thr Thr Ile Gln Asn Leu Ser Ser Leu Thr Lys
675 680 685Ile Gly Leu Ser Lys Asn Lys Phe Ser Glu Phe Pro Glu Pro Ile Leu
690 695 700Tyr Leu Lys Asn Leu Lys His Leu Asp Val Gly Glu Asn Lys Ile Arg705 710 715 720Gln Leu Pro Glu Thr Ile Gly Asn Leu Ser Asn Leu Lys Ser Leu Asp
725 730 735Ile Lys Glu Thr Trp Ile Glu Ser Leu Pro Gln Ser Ile Gln Asn Leu
740 745 750Thr Gln Leu Glu Thr Ile Tyr Leu Pro Lys Ala Lys Phe Arg Asn Leu
755 760 765Pr0 Asp Phe Leu Ala Asn Met Glu Ser Leu Lys Lys Ile Lys Phe Glu
770 775 780Ser Glu Glu Tyr Asn Gln Leu Thr Lys Trp Cys Glu Phe Glu Tyr Ser785 790 795 800Lys Tyr Ile Lys Leu Leu His Gly Gly Lys Tyr Pro Glu Ala Arg Asp
805 810 815Lys Ile Thr Asn Leu Phe Thr Thr Lys Gly Glu Asp Phe Leu Lys Leu
820 825 830Asn Gln Trp Glu Val Lys Leu Lys Ile Ser Glu Ser Tyr Tyr Arg Lys
835 840 845Ala Glu Ala Phe Ala Thr Val Val Tyr Ala Leu Ala Thr Ile Leu Lys
850 855 860Asp Glu Glu Ile Tyr Ser Ile Ala Ser Gln Met Thr Gly Asp Gln Tyr865 870 875 880Ile His Pro Ala Leu Pro Phe Asn Lys Ala Cys Tyr Phe Ala Val Thr
885 890 895Gly Gln Lys Glu Pro Met Leu Gln Ser Ile Arg Lys Ser Val Lys Leu
900 905 910Gly Thr Lys Ala Asp Glu Phe Thr Lys Glu Lys Asp Phe Ala Ser Tyr
915 920 925Leu Lys Asp Pro Asp Phe Leu Glu Ala Ile Arg Lys Asn
930 935 940<210>1l<211>504<212>DNA<213>钩端螺旋体(Leptospira interrogans)<400>11gtggttggaa agaaaagcgg agaaattttt atgaagtgcc actttatatt tgttagcatt 60ggattgatct tgtctttgta ctgtagtctg aaaatgacaa aaacgtatca taatttgcag 120gatgctcttg ataacccttt ggacgtttta attcttgaga tgagtatgaa tacgcgtata 180agtatgggtt tgcatgaact cgaatctctt ccaagggtaa ttgggttatt tcaaaattta 240gaaaaattga atttagatgg taatcaactt acgtctcttc cgaaagaaat tggacaactt 300caaaatttat tcgaattaaa tctacaagac aacaaactca agactcttcc gaaagaaatc 360gagcaacttc aaaatttaca agtgctgcgg ttgtattcga actctttttc attaaaagaa 420aaacagaaaa ttcaagagct acttcctaac tgtgaaatcg attttgaaag tgaaggtaaa 480agcgaatctt ctttgactga atga 504<210>12<211>167<212>PRT<213>钩端螺旋体(Leptospira interrogans)<400>12Met Val Gly Lys Lys Ser Gly Glu Ile Phe Met Lys Cys His Phe Ile1 5 10 15Phe Val Ser Ile Gly Leu Ile Leu Ser Leu Tyr Cys Ser Leu Lys Met
20 25 30Thr Lys Thr Tyr His Asn Leu Gln Asp Ala Leu Asp Asn Pro Leu Asp
35 40 45Val Leu Ile Leu Glu Met Ser Met Asn Thr Arg Ile Ser Met Gly Leu
50 55 60His Glu Leu Glu Ser Leu Pro Arg Val Ile Gly Leu Phe Gln Asn Leu65 70 75 80Glu Lys Leu Asn Leu Asp Gly Asn Gln Leu Thr Ser Leu Pro Lys Glu
85 90 95Ile Gly Gln Leu Gln Asn Leu Phe Glu Leu Asn Leu Gln Asp Asn Lys
100 105 110Leu Lys Thr Leu Pro Lys Glu Ile Glu Gln Leu Gln Asn Leu Gln Val
115 120 125Leu Arg Leu Tyr Ser Asn Ser Phe Ser Leu Lys Glu Lys Gln Lys Ile
130 135 140Gln Glu Leu Leu Pro Asn Cys Glu Ile Asp Phe Glu Ser Glu Gly Lys145 150 155 160Ser Glu Ser Ser Leu Thr Glu
165<210>13<211>1272<212>DNA<213>钩端螺旋体(Leptospira interrogans)<400>13atgaattata gatttacaaa catcctaaag gttataattt tcttttgttt tattttcgaa 60atacaatcag aagaaataca aaaaccttat aaaaatttag caaaggctct tcaaaatcca 120gcagacgttc ggaacttaga tttgagtttc caagggttga agacgttgcc aaacaaaata 180ggacaactta agaatttaca aaaattggat ttaggcggaa acgaacctac gattttgtct 240aaagaaatct ggcagttaaa agatctgcaa aaattgaatt tgaacaataa caaacttacg 300gttctcccta aggaaatcgg acaattacaa aacttgcaag aattaagttt gcatagcaat 360gagctcgtaa atcttccgaa agagattgga cagtttaaga atttgcaaaa actaaatttg 420gacaataata aacttacggt tctccctaag gaaatcggac aattacaaaa cttgcaagaa 480ttgagtttac ttagtaacaa actcataagt cttcccacag aaattgaaca acttaaaagt 540ctgaagaatc tagatttgaa tcacaatgaa ttaactactg tttcgaaaga agttatgcta 600cttgaaactt tagaaaattt agatctaaga tccaacaaac tgaaaacaat tcctaaagag 660ataagacaac ttaaaagttt gaaagtgttg atgttaactg gcaatcaact tacttctttg 720ccgaaagaaa tcgaacaact ccaaaatcta aaaacattaa atttgggaga aaatcgtttt 780caaatacttc ccgtagaaat tttagaactt aagaatttac tggaattgaa tttgtattat 840aaccaacttg tagagtttcc gaaggaagtt ggacaactta aaagtttaaa gtatttgagt 900ttgtatcaca atcaaattac aactcttccg gtagaagtta cacaattgcc ggatttgcaa 960gagctgcatt tgtctgggaa taaaattacg attcttccaa aagagattct acaacttaaa 1020aatttagaat ggctgtcatt gagtaacaat aaactcaatg cgcttccgaa agaaatcgga 1080cagttgaaaa aattacaaag gttggaatta ggaaacaatc agcttacgac ccttccgaaa 1140gaaatagaac aacttaaaaa tttacaaaga ttagaattag attccaatcc catttctcca 1200aaagaaaaag aaagaatccg aaaacttctt cctaaatgtg aaatcgattt tgaaggaggg 1260ggaggagaat ag 1272<210>14<211>423<212>PRT<213>钩端螺旋体(Leptospira interrogans)<400>14Met Asn Tyr Arg Phe Thr Asn Ile Leu Lys Val Ile Ile Phe Phe Cys1 5 10 15Phe Ile Phe Glu Ile Gln Ser Glu Glu Ile Gln Lys Pro Tyr Lys Asn
20 25 30Leu Ala Lys Ala Leu Gln Asn Pro Ala Asp Val Arg Asn Leu Asp Leu
35 40 45Ser Phe Gln Gly Leu Lys Thr Leu Pro Asn Lys Ile Gly Gln Leu Lys
50 55 60Asn Leu Gln Lys Leu Asp Leu Gly Gly Asn Glu Pro Thr Ile Leu Ser65 70 75 80Lys Glu Ile Trp Gln Leu Lys Asp Leu Gln Lys Leu Asn Leu Asn Asn
85 90 95Asn Lys Leu Thr Val Leu Pro Lys Glu Ile Gly Gln Leu Gln Asn Leu
100 105 110Gln Glu Leu Ser Leu His Ser Asn Glu Leu Val Asn Leu Pro Lys Glu
115 120 125Ile Gly Gln Phe Lys Asn Leu Gln Lys Leu Asn Leu Asp Asn Asn Lys
130 135 140Leu Thr Val Leu Pro Lys Glu Ile Gly Gln Leu Gln Asn Leu Gln Glu145 150 155 160Leu Ser Leu Leu Ser Asn Lys Leu Ile Ser Leu Pro Thr Glu Ile Glu
165 170 175Gln Leu Lys Ser Leu Lys Asn Leu Asp Leu Asn His Asn Glu Leu Thr
180 185 190Thr Val Ser Lys Glu Val Met Leu Leu Glu Thr Leu Glu Asn Leu Asp
195 200 205Leu Arg Ser Asn Lys Leu Lys Thr Ile Pro Lys Glu Ile Arg Gln Leu
210 215 220Lys Ser Leu Lys Val Leu Met Leu Thr Gly Asn Gln Leu Thr Ser Leu225 230 235 240Pro Lys Glu Ile Glu Gln Leu Gln Asn Leu Lys Thr Leu Asn Leu Gly
245 250 255Glu Asn Arg Phe Gln Ile Leu Pro Val Glu Ile Leu Glu Leu Lys Asn
260 265 270Leu Leu Glu Leu Asn Leu Tyr Tyr Asn Gln Leu Val Glu Phe Pro Lys
275 280 285Glu Val Gly Gln Leu Lys Ser Leu Lys Tyr Leu Ser Leu Tyr His Asn
290 295 300Gln Ile Thr Thr Leu Pro Val Glu Val Thr Gln Leu Pro Asp Leu Gln305 310 315 320Glu Leu His Leu Ser Gly Asn Lys Ile Thr Ile Leu Pro Lys Glu Ile
325 330 335Leu Gln Leu Lys Asn Leu Glu Trp Leu Ser Leu Ser Asn Asn Lys Leu
340 345 350Asn Ala Leu Pro Lys Glu Ile Gly Gln Leu Lys Lys Leu Gln Arg Leu
355 360 365Glu Leu Gly Asn Asn Gln Leu Thr Thr Leu Pro Lys Glu Ile Glu Gln
370 375 380Leu Lys Asn Leu Gln Arg Leu Glu Leu Asp Ser Asn Pro Ile Ser Pro385 390 395 400Lys Glu Lys Glu Arg Ile Arg Lys Leu Leu Pro Lys Cys Glu Ile Asp
405 410 415Phe Glu Gly Gly Gly Gly Glu
420<210>15<211>369<212>DNA<213>钩端螺旋体(Leptospira interrogans)<400>15atgaaaatca tgaatttaca aaaaatagga accttaatct ttctttgttt tttgagccaa 60ctcaaagccg aagagaaagg tcactatcaa aatctaacca aggctcttaa aaatccaacg 120gatgtccaaa ctttagattt aagtaataat caactaataa ctcttccaaa agaaattgga 180caattgaaag agttagaatg gctaagttta agcaagaatc aactcaaaac tcttcctaaa 240gaaatcgaaa ctttgaaaaa gttagaagaa ttatttttag atgatatacc cgttttaaag 300tctcaagaaa aaaagattca gaaattactt cccaaggcgc aaattgactt tatcgaaata 360aaaaagtaa 369<210>16<211>122<212>PRT<213>钩端螺旋体(Leptospira interrogans)<400>16Met Lys Ile Met Asn Leu Gln Lys Ile Gly Thr Leu Ile Phe Leu Cys1 5 10 15Phe Leu Ser Gln Leu Lys Ala Glu Glu Lys Gly His Tyr Gln Asn Leu
20 25 30Thr Lys Ala Leu Lys Asn Pro Thr Asp Val Gln Thr Leu Asp Leu Ser
35 40 45Asn Asn Gln Leu Ile Thr Leu Pro Lys Glu Ile Gly Gln Leu Lys Glu
50 55 60Leu Glu Trp Leu Ser Leu Ser Lys Asn Gln Leu Lys Thr Leu Pro Lys65 70 75 80Glu Ile Glu Thr Leu Lys Lys Leu Glu Glu Leu Phe Leu Asp Asp Ile
85 90 95Pro Val Leu Lys Ser Gln Glu Lys Lys Ile Gln Lys Leu Leu Pro Lys
100 105 110Ala Gln Ile Asp Phe Ile Glu Ile Lys Lys
115 120<210>17<211>654<212>DNA<213>钩端螺旋体(Leptospira interrogans)<400>17atgaatttcc tatctttaaa aaaaatcttt aagctctatt tctttctttg ggtttcgttt 60tttgggatcg attgtaaaaa aaacgcggta gagattttag aagaagcaaa aagaaaatcg 120gaatcgattc aaatcttaga tttaggaatg caaaagttaa cctcgattcc agaaggagtt 180tgctcttttc cgaatttgac tcagctggat ttacgtttga acagtttgaa ttctcttcct 240ggttggattg gagcttgtaa aaatctggaa caaattaacc ttttcggaaa tgatctaaat 300acggttccat cttctttttc taaattaaag aatttgaaag tattactctt gggtaataac 360gattttactt ttctaccttc ggaacttttg tttctgcctt tgcttaaaac cctgtatttg 420gatcaaaaca agcttacgtt gactgaaacg gatgttgaaa ttctagcttc tctttccagt 480ttggaagaat tggatcttaa tttgaactct ggaatcaaag tgcttccttc taactataat 540aaattgaaaa atctgactca tttaaaaaga ttgaatatta aaaaaacttc attaaagggg 600gaggatgctg ataaattaca agcagttctc ccaaatacaa gaatagacta ttga 654<210>18<211>217<212>PRT<213>钩端螺旋体(Leptospira interrogans)<400>18Met Asn Phe Leu Ser Leu Lys Lys Ile Phe Lys Leu Tyr Phe Phe Leu1 5 10 15Trp Val Ser Phe Phe Gly Ile Asp Cys Lys Lys Asn Ala Val Glu Ile
20 25 30Leu Glu Glu Ala Lys Arg Lys Ser Glu Ser Ile Gln Ile Leu Asp Leu
35 40 45Gly Met Gln Lys Leu Thr Ser Ile Pro Glu Gly Val Cys Ser Phe Pro
50 55 60Asn Leu Thr Gln Leu Asp Leu Arg Leu Asn Ser Leu Asn Ser Leu Pro65 70 75 80Gly Trp Ile Gly Ala Cys Lys Asn Leu Glu Gln Ile Asn Leu Phe Gly
85 90 95Asn Asp Leu Asn Thr Val Pro Ser Ser Phe Ser Lys Leu Lys Asn Leu
100 105 110Lys Val Leu Leu Leu Gly Asn Asn Asp Phe Thr Phe Leu Pro Ser Glu
115 120 125Leu Leu Phe Leu Pro Leu Leu Lys Thr Leu Tyr Leu Asp Gln Asn Lys
130 135 140Leu Thr Leu Thr Glu Thr Asp Val Glu Ile Leu Ala Ser Leu Ser Ser145 150 155 160Leu Glu Glu Leu Asp Leu Asn Leu Asn Ser Gly Ile Lys Val Leu Pro
165 170 175Ser Asn Tyr Asn Lys Leu Lys Asn Leu Thr His Leu Lys Arg Leu Asn
180 185 190Ile Lys Lys Thr Ser Leu Lys Gly Glu Asp Ala Asp Lys Leu Gln Ala
195 200 205Val Leu Pro Asn Thr Arg Ile Asp Tyr
210 215<210>19<211>531<212>DNA<213>钩端螺旋体(Leptospira interrogans)<400>19gtgtggatgt cttcttatgt ttcggcattc agaacttgga acaaaaagcg cgatcgctta 60aagtattgta aaattacttc tttcattgaa aaaaatattt cgctgaaaca acgcgtatcc 120ttttttgttc attgcaataa aattttgatt tggaaaatcc gtcgcatgaa cttttttaca 180acacagattc gttttaaacg aatcggaatg gttcttttga tttctatctt ttttttcttc 240aacgaggtag aagcacaaga gcaacttgtg gaagtaaaag cttacacaaa tttaatagag 300gctttccaaa attcttcgga tgttcgtatt ctgcatttga acggtcaagg atttaaaaat 360cttccgagac aaatcggaaa tctacaaaat ttaacagagt tgaatttagg gagcaattca 420cttacgacag ttcctaaaga aatcggtcgg ctacgcaact tacaaagttt gtggttgttt 480ggaaactggc tggcaacctt cctgaagaag tcggggaact tcacaattta g 531<210>20<211>176<212>PRT<213>钩端螺旋体(Leptospira interrogans)<400>20Met Trp Met Ser Ser Tyr Val Ser Ala Phe Arg Thr Trp Asn Lys Lys1 5 10 15Arg Asp Arg Leu Lys Tyr Cys Lys Ile Thr Ser Phe Ile Glu Lys Asn
20 25 30Ile Ser Leu Lys Gln Arg Val Ser Phe Phe Val His Cys Asn Lys Ile
35 40 45Leu Ile Trp Lys Ile Arg Arg Met Asn Phe Phe Thr Thr Gln Ile Arg
50 55 60Phe Lys Arg Ile Gly Met Val Leu Leu Ile Ser Ile Phe Phe Phe Phe65 70 75 80Asn Glu Val Glu Ala Gln Glu Gln Leu Val Glu Val Lys Ala Tyr Thr
85 90 95Asn Leu Ile Glu Ala Phe Gln Asn Ser Ser Asp Val Arg Ile Leu His
100 105 110Leu Asn Gly Gln Gly Phe Lys Asn Leu Pro Arg Gln Ile Gly Asn Leu
115 120 125Gln Asn Leu Thr Glu Leu Asn Leu Gly Ser Asn Ser Leu Thr Thr Val
130 135 140Pro Lys Glu Ile Gly Arg Leu Arg Asn Leu Gln Ser Leu Trp Leu Phe145 150 155 160Gly Asn Trp Leu Ala Thr Phe Leu Lys Lys Ser Gly Asn Phe Thr Ile
165 170 175<210>21<211>300<212>DNA<213>钩端螺旋体(Leptospira interrogans)<400>21ttgaatctat ctgacaatcg tcccactttc accgttcttc caaaagaaat cggagaacta 60cggaatttaa aagagttgaa tttagagaac aattcactta cgacaattcc taaagaaatc 120ggaaaactac aaaatttaaa atggttgcgt ttagggagca attcacttac gacgcttcca 180aaagaaatcg gaaaactacg gaatttaaaa gagttggatt tgagttcgaa ttctctttcg 240gtcaaagaga aaaagagaat tcgaaagtta cttccaaaat gttctatcat ctatttttga 300<210>22<211>99<212>PRT<213>钩端螺旋体(Leptospira interrogans)<400>22Met Asn Leu Ser Asp Asn Arg Pro Thr Phe Thr Val Leu Pro Lys Glu1 5 10 15Ile Gly Glu Leu Arg Asn Leu Lys Glu Leu Asn Leu Glu Asn Asn Ser
20 25 30Leu Thr Thr Ile Pro Lys Glu Ile Gly Lys Leu Gln Asn Leu Lys Trp
35 40 45Leu Arg Leu Gly Ser Asn Ser Leu Thr Thr Leu Pro Lys Glu Ile Gly
50 55 60Lys Leu Arg Asn Leu Lys Glu Leu Asp Leu Ser Ser Asn Ser Leu Ser65 70 75 80Val Lys Glu Lys Lys Arg Ile Arg Lys Leu Leu Pro Lys Cys Ser Ile
85 90 95Ile Tyr Phe<210>23<211>282<212>DNA<213>钩端螺旋体(Leptospira interrogans)<400>23atggatcgcc gttctgggct caaaaaggca catttttcaa gtgttccgat aaatgagtct 60ttttacttgc aaaaagtatg tttttctgat agagaaaaat tttccgaacc ttaccgcctc 120ccaaaaatag agaaaactca acacaacgct ctttatgaat cgtgttgtgg agtgggaact 180aagttttaca gaagattttg tcgtaattcc gacagattta tattgagatc aaatacttgt 240ggggttggtg atgaatttgt agaaactact gcattttatt aa 282<210>24<211>93<212>PRT<213>钩端螺旋体(Leptospira interrogans)<400>24Met Asp Arg Arg Ser Gly Leu Lys Lys Ala His Phe Ser Ser Val Pro1 5 10 15Ile Asn Glu Ser Phe Tyr Leu Gln Lys Val Cys Phe Ser Asp Arg Glu
20 25 30Lys Phe Ser Glu Pro Tyr Arg Leu Pro Lys Ile Glu Lys Thr Gln His
35 40 45Asn Ala Leu Tyr Glu Ser Cys Cys Gly Val Gly Thr Lys Phe Tyr Arg
50 55 60Arg Phe Cys Arg Asn Ser Asp Arg Phe Ile Leu Arg Ser Asn Thr Cys65 70 75 80Gly Val Gly Asp Glu Phe Val Glu Thr Thr Ala Phe Tyr
85 90<210>25<211>1581<212>DNA<213>钩端螺旋体(Leptospira interrogans)<400>25atgaaggatt attctgtaat tctttctagt ttatcgatta tgaaaataca atacacggct 60ttaaatagga atcgatctat gttcttcggt ttccgacgac atgttcggat ttttttgacg 120attgtctttg gtctgttttc cgttcaactt caggcaaacg atggttatga gacgtttacc 180aatctaaagg aagcgcttga acacgcgtcc tcgttttctt cgtcgaacgt tcgcatcttg 240gatttgagtc ggcaaaaatt tgcggttttt cccaaagaaa tctgggaact cgaatattta 300gaaattttga aactggagga aaatcggatc acaactcttc cgcgcgaaat aaacaaactg 360aaaaacttaa aagaattata tttgaatggt aataaactaa caatcgttcc taaagaaatc 420tgggagcttg aaaatttaac aatcttacga ctggagaata atagaatttc aacccttccg 480aaagaaatag aaaaatcaaa gaatctccaa gaattgaact tacgtggaaa tcgactcgta 540actcttccgg gagaaatagg cgaacttaaa cttttggaag aactaaacct ggaaaacaac 600aggatcaaaa ttcttccgaa tgaaatcgga gccttagaaa acttatggat attcaatttg 660agtggaaaca agctcgcttc aattccgaag gagataggaa acttgcaaaa tttgcgaatg 720ttatatttgg aaaacaatca actcaaaact ctccctcgac aaatggagaa gcttcaggat 780ttagaggttt taaatttact tatcaacccg cttctatcca aagagagaaa aaaaattcaa 840gcgctactcc ccgattgcga catagatttg agagatgtag aagagggtga aacctatcga 900aacttaaatc tggctttgga acaacctcta aagattctga gtttaagctt ggaatatcaa 960caattccccc ttttttcaaa agaaatatta aggctcaaaa atctacggag tttgagttta 1020tacgatacca gcctcgtagc tctcccgaaa gaaatcgttc gtctcaaaca tttagaacat 1080ctgtcattag gtctcaatca gcttaaaagt cttcctaaag aaatcggatt actcaggaat 1140ttaagaagtt tggatatagg agccaataac gagtttgaag ttcttccgaa ggaaatcgca 1200cggcttcaaa acttgagatc gttactatta aatcaaaatc gattcaaaat ctttcccaaa 1260gaaatctggg aacttaagaa gttggtcatc ctaaacgtaa atacgaatca actcgatgcg 1320cttccggaaa aaatcggacg actaaagggc ttacagatgt tggatttgtc tcataacaga 1380ctcacgactc ttcctagtga gatcggacaa ctacacaatc taactgaatt gtatttacaa 1440tacaatcgaa tcaagacgtt gcccgaagaa atcgcgcggc ttcaaaactt gcggaagctt 1500acgttgtatg aaaatccgat tccgccgcaa gaactggaca aaatacgaaa gcttcttcca 1560aattgtgaga ttcgatttta a 1581<210>26<211>526<212>PRT<213>钩端螺旋体(Leptospira interrogans)<400>26Met Lys Asp Tyr Ser Val Ile Leu Ser Ser Leu Ser Ile Met Lys Ile1 5 10 15Gln Tyr Thr Ala Leu Asn Arg Asn Arg Ser Met Phe Phe Gly Phe Arg
20 25 30Arg His Val Arg Ile Phe Leu Thr Ile Val Phe Gly Leu Phe Ser Val
35 40 45Gln Leu Gln Ala Asn Asp Gly Tyr Glu Thr Phe Thr Asn Leu Lys Glu
50 55 60Ala Leu Glu His Ala Ser Ser Phe Ser Ser Ser Asn Val Arg Ile Leu65 70 75 80Asp Leu Ser Arg Gln Lys Phe Ala Val Phe Pro Lys Glu Ile Trp Glu
85 90 95Leu Glu Tyr Leu Glu Ile Leu Lys Leu Glu Glu Asn Arg Ile Thr Thr
100 105 110Leu Pro Arg Glu Ile Asn Lys Leu Lys Asn Leu Lys Glu Leu Tyr Leu
115 120 125Asn Gly Asn Lys Leu Thr Ile Val Pro Lys Glu Ile Trp Glu Leu Glu
130 135 140Asn Leu Thr Ile Leu Arg Leu Glu Asn Asn Arg Ile Ser Thr Leu Pro145 150 155 160Lys Glu Ile Glu Lys Ser Lys Asn Leu Gln Glu Leu Asn Leu Arg Gly
165 170 175Asn Arg Leu Val Thr Leu Pro Gly Glu Ile Gly Glu Leu Lys Leu Leu
180 185 190Glu Glu Leu Asn Leu Glu Asn Asn Arg Ile Lys Ile Leu Pro Asn Glu
195 200 205Ile Gly Ala Leu Glu Asn Leu Trp Ile Phe Asn Leu Ser Gly Asn Lys
210 215 220Leu Ala Ser Ile Pro Lys Glu Ile Gly Asn Leu Gln Asn Leu Arg Met225 230 235 240Leu Tyr Leu Glu Asn Asn Gln Leu Lys Thr Leu Pro Arg Gln Met Glu
245 250 255Lys Leu Gln Asp Leu Glu Val Leu Asn Leu Leu Ile Asn Pro Leu Leu
260 265 270Ser Lys Glu Arg Lys Lys Ile Gln Ala Leu Leu Pro Asp Cys Asp Ile
275 280 285Asp Leu Arg Asp Val Glu Glu Gly Glu Thr Tyr Arg Asn Leu Asn Leu
290 295 300Ala Leu Glu Gln Pro Leu Lys Ile Leu Ser Leu Ser Leu Glu Tyr Gln305 310 315 320Gln Phe Pro Leu Phe Ser Lys Glu Ile Leu Arg Leu Lys Asn Leu Arg
325 330 335Ser Leu Ser Leu Tyr Asp Thr Ser Leu Val Ala Leu Pro Lys Glu Ile
340 345 350Val Arg Leu Lys His Leu Glu His Leu Ser Leu Gly Leu Asn Gln Leu
355 360 365Lys Ser Leu Pro Lys Glu Ile Gly Leu Leu Arg Asn Leu Arg Ser Leu
370 375 380Asp Ile Gly Ala Asn Asn Glu Phe Glu Val Leu Pro Lys Glu Ile Ala385 390 395 400Arg Leu Gln Asn Leu Arg Ser Leu Leu Leu Asn Gln Asn Arg Phe Lys
405 410 415Ile Phe Pro Lys Glu Ile Trp Glu Leu Lys Lys Leu Val Ile Leu Asn
420 425 430Val Asn Thr Asn Gln Leu Asp Ala Leu Pro Glu Lys Ile Gly Arg Leu
435 440 445Lys Gly Leu Gln Met Leu Asp Leu Ser His Asn Arg Leu Thr Thr Leu
450 455 460Pro Ser Glu Ile Gly Gln Leu His Asn Leu Thr Glu Leu Tyr Leu Gln465 470 475 480Tyr Asn Arg Ile Lys Thr Leu Pro Glu Glu Ile Ala Arg Leu Gln Asn
485 490 495Leu Arg Lys Leu Thr Leu Tyr Glu Asn Pro Ile Pro Pro Gln Glu Leu
500 505 510Asp Lys Ile Arg Lys Leu Leu Pro Asn Cys Glu Ile Arg Phe
515 520 525<210>27<211>234<212>DNA<213>钩端螺旋体(Leptospira interrogans)<400>27ttggatttaa atgataatca atttacagtt cttcctaagg aaattggaaa actgaaaaaa 60ttacaaacat tggatttacg gaataaccaa ctcacaaccc ttcctaccga aatcggacaa 120ctacaaaatt tacaatggtt gtatttacag aataatcaac tttcactcaa agaacaagaa 180aggattcgaa agttacttcc actaaaatgt aaaataattt ttgaagttta ttag 234<210>28<211>77<212>PRT<213>钩端螺旋体(Leptospira interrogans)<400>28Met Asp Leu Asn Asp Asn Gln Phe Thr Val Leu Pro Lys Glu Ile Gly1 5 10 15Lys Leu Lys Lys Leu Gln Thr Leu Asp Leu Arg Asn Asn Gln Leu Thr
20 25 30Thr Leu Pro Thr Glu Ile Gly Gln Leu Gln Asn Leu Gln Trp Leu Tyr
35 40 45Leu Gln Asn Asn Gln Leu Ser Leu Lys Glu Gln Glu Arg Ile Arg Lys
50 55 60Leu Leu Pro Leu Lys Cys Lys Ile Ile Phe Glu Val Tyr65 70 75<210>29<211>1497<212>DNA<213>钩端螺旋体(Leptospira interrogans)<400>29atgaacttcc gcataacgtt aatttactta caaaaaataa caattggtct tttgcttctc 60attactcttt cttgcgagat tcaagcagaa gaagtagaac caggaactta tagggattta 120acaaaagcgc tccaaaatcc tttggatgtt cgagttttga atttgagtgg agaaaaactc 180acagctcttc ccaaagaaat cggacagctg aagaatttac aagagttgaa tttaaaatgg 240aacctactca cagttcttcc aaaggaaatt ggacaactgg agaatttaca ggaattagat 300ttacgtgata accaactcgc aacttttcct gccgtaatag tagagttaca gaagttagaa 360agtttggatt tatctgagaa ccgactcata atacttccta acgaaatagg acggctacag 420aatttacaag atttgggttt gtataaaaac aaacttacta cttttccaaa ggaaataggg 480cagttacaga atttacaaaa gttatggtta tctgagaacc gactcacagc acttcctaag 540gaaattggtc aattgaagaa tttacaaaca ttggatttgc aggataacca attcacgata 600cttccaaaag aaataggaca attacagaat ttacaaacat tgaatttgag cgataaccaa 660ctcgcaactc ttcctgtcga aatcggacaa ctacagaatt tacaagagtt gtatttgcgg 720aataaccgac tcacagtttt tccaaaggaa ataggacaac tacagaattt gcaaatgtta 780tgttcacctg aaaaccgtct cacagcactt cctaaagaaa tggggcagct acagaattta 840caaacattga atttggtgaa taaccgactc acagtttttc ctaaggaaat cggacaacta 900cagaatctgc aagatttgga attgctaatg aatcctcttt ccttaaaaga aagaaaaaga 960attcaaaagt tgtttccaga tagtaatctt gatttgagag aagtagcaga gaacggtgtt 1020taccgcaatt taaatctggc tcaggaagaa cctttaaaag tttttgaatt gagtcttgaa 1080tacaaagatt tttctcaatc gtttccaaaa gtaattttga aattccgaaa tttacggggt 1140ttaaatttat acgactgcgg attttcaact cttccgaaag aaatcagccg acttaaaaat 1200ttaaagtatt tagccttagg cttaaatggt cttaaaaata ttccttctga aatcggccaa 1260ctgaagaatt tagaagcttt gaatttagaa gcaaacgaat tggaaagact tcccaaagaa 1320attgggcaac tgcggaattt acagaagtta agtttacatc aaaatacatt gaaaatattt 1380cccgccgaaa tcgaacaact gaaaaaattg caaaagttag atttgagcgt taatcaattc 1440acgacttttc ctaaggaaat cggaaaatta gaaaatttac aaacattgaa tttataa 1497<210>30<211>498<212>PRT<213>钩端螺旋体(Leptospira interrogans)<400>30Met Asn Phe Arg Ile Thr Leu Ile Tyr Leu Gln Lys Ile Thr Ile Gly1 5 10 15Leu Leu Leu Leu Ile Thr Leu Ser Cys Glu Ile Gln Ala Glu Glu Val
20 25 30Glu Pro Gly Thr Tyr Arg Asp Leu Thr Lys Ala Leu Gln Asn Pro Leu
35 40 45Asp Val Arg Val Leu Asn Leu Ser Gly Glu Lys Leu Thr Ala Leu Pro
50 55 60Lys Glu Ile Gly Gln Leu Lys Asn Leu Gln Glu Leu Asn Leu Lys Trp65 70 75 80Asn Leu Leu Thr Val Leu Pro Lys Glu Ile Gly Gln Leu Glu Asn Leu
85 90 95Gln Glu Leu Asp Leu Arg Asp Asn Gln Leu Ala Thr Phe Pro Ala Val
100 105 110Ile Val Glu Leu Gln Lys Leu Glu Ser Leu Asp Leu Ser Glu Asn Arg
115 120 125Leu Ile Ile Leu Pro Asn Glu Ile Gly Arg Leu Gln Asn Leu Gln Asp
130 135 140Leu Gly Leu Tyr Lys Asn Lys Leu Thr Thr Phe Pro Lys Glu Ile Gly145 150 155 160Gln Leu Gln Asn Leu Gln Lys Leu Trp Leu Ser Glu Asn Arg Leu Thr
165 170 175Ala Leu Pro Lys Glu Ile Gly Gln Leu Lys Asn Leu Gln Thr Leu Asp
180 185 190Leu Gln Asp Asn Gln Phe Thr Ile Leu Pro Lys Glu Ile Gly Gln Leu
195 200 205Gln Asn Leu Gln Thr Leu Asn Leu Ser Asp Asn Gln Leu Ala Thr Leu
210 215 220Pro Val Glu Ile Gly Gln Leu Gln Asn Leu Gln Glu Leu Tyr Leu Arg225 230 235 240Asn Asn Arg Leu Thr Val Phe Pro Lys Glu Ile Gly Gln Leu Gln Asn
245 250 255Leu Gln Met Leu Cys Ser Pro Glu Asn Arg Leu Thr Ala Leu Pro Lys
260 265 270Glu Met Gly Gln Leu Gln Asn Leu Gln Thr Leu Asn Leu Val Asn Asn
275 280 285Arg Leu Thr Val Phe Pro Lys Glu Ile Gly Gln Leu Gln Asn Leu Gln
290 295 300Asp Leu Glu Leu Leu Met Asn Pro Leu Ser Leu Lys Glu Arg Lys Arg305 310 315 320Ile Gln Lys Leu Phe Pro Asp Ser Asn Leu Asp Leu Arg Glu Val Ala
325 330 335Glu Asn Gly Val Tyr Arg Asn Leu Asn Leu Ala Gln Glu Glu Pro Leu
340 345 350Lys Val Phe Glu Leu Ser Leu Glu Tyr Lys Asp Phe Ser Gln Ser Phe
355 360 365Pro Lys Val Ile Leu Lys Phe Arg Asn Leu Arg Gly Leu Asn Leu Tyr
370 375 380Asp Cys Gly Phe Ser Thr Leu Pro Lys Glu Ile Ser Arg Leu Lys Asn385 390 395 400Leu Lys Tyr Leu Ala Leu Gly Leu Asn Gly Leu Lys Asn Ile Pro Ser
405 410 415Glu Ile Gly Gln Leu Lys Asn Leu Glu Ala Leu Asn Leu Glu Ala Asn
420 425 430Glu Leu Glu Arg Leu Pro Lys Glu Ile Gly Gln Leu Arg Asn Leu Gln
435 440 445Lys Leu Ser Leu His Gln Asn Thr Leu Lys Ile Phe Pro Ala Glu Ile
450 455 460Glu Gln Leu Lys Lys Leu Gln Lys Leu Asp Leu Ser Val Asn Gln Phe465 470 475 480Thr Thr Phe Pro Lys Glu Ile Gly Lys Leu Glu Asn Leu Gln Thr Leu
485 490 495Asn Leu<210>31<211>1287<212>DNA<213>钩端螺旋体(Leptospira interrogans)<400>31atgaacttcc gcataacgtt gatttactta caaaagatta caattggtct tttgtttctc 60attactcttt cttgcgagat tcaagcagac gaagtaaaac caggaactta tatggattta 120acaaaagcgc tccaaaatcc tttaaatgtt cgagttttgg atttgagtgg acaaaatttc 180acaacacttc ctaaggaaat tgaacaactg aagaacttgc aaaagttgta tttatttgat 240aaccgactca aaactcttcc taaggaaatc ggacaattga aaaatttgca agagttgaat 300ttgagtagta accaactcac gatacttcct aaagaaatcg gaaaactgga aaatttacaa 360agattggatt tatatgataa cagactcacg attcttccta tcgaaatcgg aaaattacaa 420aatttgcaaa cgttatattt aagtagtaac cagctgacaa ctcttcctag ggaaagcgga 480aaactggaga atttgcaaga gttgaattta agtgataatc aactcacaac tcttccacaa 540gaaattggac aactacagaa tttacaaacg ttgaatttga agagtaatca acttacaact 600ctttttaaag aaatagaaca attgaagaat ttacaaacgt tgaatttgag cgataaccaa 660ctcacaactc ttccaattga aatcggaaaa ctacaaaatt tgcatacgtt gaatttgagc 720gataaccaac tcacaactct tccaattgaa atcggaaaac tacaaaattt gcatacgttg 780aatttgagcg ataaccaact cacaactctt ccaattgaaa tcggaaaact acaaaatttg 840catacgttga atttgagtgg taatcaactt acaacgcttt caattgaaat cggaaaacta 900cagaacttac aagacttgaa tttgcatagt aatcaactca caaccctttc taaagaaatt 960gaacagctaa aaaatttaca aacgttgagt ttgagttata accgactcgt aattcttcct 1020aaggaaatcg gacaactaca gaatctgcaa gagttgaatt tatggaataa ccaactcaca 1080gctcttccta tcgaaattgg acaactacaa aatttacaaa cgttgagttt gtataaaaac 1140agactcatga cttttccgaa agaaattgga caattgaaaa acttgcaaac gttgtatttg 1200gggggtcata atcaattttc atctgaagaa aaagaaagaa ttcgaaaact gctaccgaat 1260tgcaaaattt actttggaga taattag 1287<210>32<211>428<212>PRT<213>钩端螺旋体(Leptospira interrogans)<400>32Met Asn Phe Arg Ile Thr Leu Ile Tyr Leu Gln Lys Ile Thr Ile Gly1 5 10 15Leu Leu Phe Leu Ile Thr Leu Ser Cys Glu Ile Gln Ala Asp Glu Val
20 25 30Lys Pro Gly Thr Tyr Met Asp Leu Thr Lys Ala Leu Gln Asn Pro Leu
35 40 45Asn Val Arg Val Leu Asp Leu Ser Gly Gln Asn Phe Thr Thr Leu Pro
50 55 60Lys Glu Ile Glu Gln Leu Lys Asn Leu Gln Lys Leu Tyr Leu Phe Asp65 70 75 80Asn Arg Leu Lys Thr Leu Pro Lys Glu Ile Gly Gln Leu Lys Asn Leu
85 90 95Gln Glu Leu Asn Leu Ser Ser Asn Gln Leu Thr Ile Leu Pro Lys Glu
100 105 110Ile Gly Lys Leu Glu Asn Leu Gln Arg Leu Asp Leu Tyr Asp Asn Arg
115 120 125Leu Thr Ile Leu Pro Ile Glu Ile Gly Lys Leu Gln Asn Leu Gln Thr
130 135 140Leu Tyr Leu Ser Ser Asn Gln Leu Thr Thr Leu Pro Arg Glu Ser Gly145 150 155 160Lys Leu Glu Asn Leu Gln Glu Leu Asn Leu Ser Asp Asn Gln Leu Thr
165 170 175Thr Leu Pro Gln Glu Ile Gly Gln Leu Gln Asn Leu Gln Thr Leu Asn
180 185 190Leu Lys Ser Asn Gln Leu Thr Thr Leu Phe Lys Glu Ile Glu Gln Leu
195 200 205Lys Asn Leu Gln Thr Leu Asn Leu Ser Asp Asn Gln Leu Thr Thr Leu
210 215 220Pro Ile Glu Ile Gly Lys Leu Gln Asn Leu His Thr Leu Asn Leu Ser225 230 235 240Asp Asn Gln Leu Thr Thr Leu Pro Ile Glu Ile Gly Lys Leu Gln Asn
245 250 255Leu His Thr Leu Asn Leu Ser Asp Asn Gln Leu Thr Thr Leu Pro Ile
260 265 270Glu Ile Gly Lys Leu Gln Asn Leu His Thr Leu Asn Leu Ser Gly Asn
275 280 285Gln Leu Thr Thr Leu Ser Ile Glu Ile Gly Lys Leu Gln Asn Leu Gln
290 295 300Asp Leu Asn Leu His Ser Asn Gln Leu Thr Thr Leu Ser Lys Glu Ile305 310 315 320Glu Gln Leu Lys Asn Leu Gln Thr Leu Ser Leu Ser Tyr Asn Arg Leu
325 330 335Val Ile Leu Pro Lys Glu Ile Gly Gln Leu Gln Asn Leu Gln Glu Leu
340 345 350Asn Leu Trp Asn Asn Gln Leu Thr Ala Leu Pro Ile Glu Ile Gly Gln
355 360 365Leu Gln Asn Leu Gln Thr Leu Ser Leu Tyr Lys Asn Arg Leu Met Thr
370 375 380Phe Pro Lys Glu Ile Gly Gln Leu Lys Asn Leu Gln Thr Leu Tyr Leu385 390 395 400Gly Gly His Asn Gln Phe Ser Ser Glu Glu Lys Glu Arg Ile Arg Lys
405 410 415Leu Leu Pro Asn Cys Lys Ile Tyr Phe Gly Asp Asn
420 425<210>33<211>1902<212>DNA<213>钩端螺旋体(Leptospira interrogans)<400>33atgaacttcc gcataacgtt gatttactta caaaagatta caattggtct tttgtttctc 60attactcttt cttgcgagat tcaagcagaa gcaccagaag acgaaccagg aacatatagg 120gatttaacag aagcgctcca aaatccttta aatgttcgag ttttgaattt gagtggacaa 180aatttcacaa cacttcctaa ggaaattgaa caactaaaga atttacagga attagattta 240ggtgataacc aactcgcaac ttttcctgcc gtaatagtag agttacagaa gttagaaagt 300ttggatttat ctgagaaccg gcttgtaatg cttcctaacg aaatagggcg gttacaaaat 360ttacaagagt tgggtttgta taaaaacaaa cttattactt ttccaaagga aatcgggcaa 420ttacagaatt tacaaacatt gaatttgcag gataaccaac tcgcgactct tcctgtcgaa 480ataggacaac tacagaattt agaaaagttg aatttgcgga agaatcgact cacagttctt 540ccaaaggaaa taggacaatt acagaattta caaacattga atttgcagga taaccaactc 600gcgactcttc ctgtcgaaat cggacaacta cagaatttgc aaacgttggg tttgagtgag 660aatcaactca caacatttcc taaggaaatc ggacagctgg aaaatttaca agagttgaat 720ttaaaatgga accgactcac agcacttcca aaggaaatcg gacaattgaa aaatttagaa 780aatttggagt tgagtgagaa tcaactcaca acatttccta aggaaatcgg gcaattgaaa 840aaattacgag atttgggttt gggtaggaac caacttacaa catttccgaa agaaatcgga 900caattgaaaa atttacaaat gttggattta tgttataacc aatttaaaac agtttcgaaa 960gaaattgggc aactgaagaa tttactacag ttgaatttga gttacaatca actcgcaact 1020cttcctgcgg aaatcgggca attgaaaaaa ttacaagatt tgagtttggg taggaaccaa 1080cttacaactc ttccgaaaga aattgggcaa ctgaagaatt tgtataattt ggatttgggt 1140acgaaccagc ttacaactct tccgaaagaa attgggcaat tgaagaattt gtacaatttg 1200ggtttgggta ggaaccaact tgcaacattt ccgaaagaaa tcggacagct ggaaaattta 1260caagagttgg atttatggaa taaccgactc acagcacttc caaaggaaat cggacaattg 1320aaaaatttag aaaatttgga gttgagtgag aatcaactca caacatttcc taaggaaatc 1380gggcaattga aaaaattaca agatttgggt ttgagttata accgactcgt aatacttcct 1440aaggaaatcg gacaactgga aaaattacaa gatttgggtt tgagttataa ccgactcgta 1500atacttccta aggaaatcgg acaactgaaa aatttacaaa tgttggattt atgttataac 1560caatttaaaa cagtttcgaa agaaattggg caactgaaga atttactaca gttgaatttg 1620agttacaatc aactcgcaac tcttcctgcg gaaatcgggc aactgaagaa tttgtacaat 1680ttggatttgg gtacgaacca gcttacaact cttccgaaag aaattgggca attgaagaat 1740ttgtacaatt tgggtttggg tacgaaccaa cttacaactc ttccgaaaga aattgggcaa 1800ttgaaaaaat tacagtggtt acttttgagc gtaaatccta tcttacccca agaaaaagaa 1860agaattcgaa agcttcttcc aaattgcgag attcgatttt ga 1902<210>34<211>633<212>PRT<213>钩端螺旋体(Leptospira interrogans)<400>34Met Asn Phe Arg Ile Thr Leu Ile Tyr Leu Gln Lys Ile Thr Ile Gly1 5 10 15Leu Leu Phe Leu Ile Thr Leu Ser Cys Glu Ile Gln Ala Glu Ala Pro
20 25 30Glu Asp Glu Pro Gly Thr Tyr Arg Asp Leu Thr Glu Ala Leu Gln Asn
35 40 45Pro Leu Asn Val Arg Val Leu Asn Leu Ser Gly Gln Asn Phe Thr Thr
50 55 60Leu Pro Lys Glu Ile Glu Gln Leu Lys Asn Leu Gln Glu Leu Asp Leu65 70 75 80Gly Asp Asn Gln Leu Ala Thr Phe Pro Ala Val Ile Val Glu Leu Gln
85 90 95Lys Leu Glu Ser Leu Asp Leu Ser Glu Asn Arg Leu Val Met Leu Pro
100 105 110Asn Glu Ile Gly Arg Leu Gln Asn Leu Gln Glu Leu Gly Leu Tyr Lys
115 120 125Asn Lys Leu Ile Thr Phe Pro Lys Glu Ile Gly Gln Leu Gln Asn Leu
130 135 140Gln Thr Leu Asn Leu Gln Asp Asn Gln Leu Ala Thr Leu Pro Val Glu145 150 155 160Ile Gly Gln Leu Gln Asn Leu Glu Lys Leu Asn Leu Arg Lys Asn Arg
165 170 175Leu Thr Val Leu Pro Lys Glu Ile Gly Gln Leu Gln Asn Leu Gln Thr
180 185 190Leu Asn Leu Gln Asp Asn Gln Leu Ala Thr Leu Pro Val Glu Ile Gly
195 200 205Gln Leu Gln Asn Leu Gln Thr Leu Gly Leu Ser Glu Asn Gln Leu Thr
210 215 220Thr Phe Pro Lys Glu Ile Gly Gln Leu Glu Asn Leu Gln Glu Leu Asn225 230 235 240Leu Lys Trp Asn Arg Leu Thr Ala Leu Pro Lys Glu Ile Gly Gln Leu
245 250 255Lys Asn Leu Glu Asn Leu Glu Leu Ser Glu Asn Gln Leu Thr Thr Phe
260 265 270Pro Lys Glu Ile Gly Gln Leu Lys Lys Leu Arg Asp Leu Gly Leu Gly
275 280 285Arg Asn Gln Leu Thr Thr Phe Pro Lys Glu Ile Gly Gln Leu Lys Asn
290 295 300Leu Gln Met Leu Asp Leu Cys Tyr Asn Gln Phe Lys Thr Val Ser Lys305 310 315 320Glu Ile Gly Gln Leu Lys Asn Leu Leu Gln Leu Asn Leu Ser Tyr Asn
325 330 335Gln Leu Ala Thr Leu Pro Ala Glu Ile Gly Gln Leu Lys Lys Leu Gln
340 345 350Asp Leu Ser Leu Gly Arg Asn Gln Leu Thr Thr Leu Pro Lys Glu Ile
355 360 365Gly Gln Leu Lys Asn Leu Tyr Asn Leu Asp Leu Gly Thr Asn Gln Leu
370 375 380Thr Thr Leu Pro Lys Glu Ile Gly Gln Leu Lys Asn Leu Tyr Asn Leu385 390 395 400Gly Leu Gly Arg Asn Gln Leu Ala Thr Phe Pro Lys Glu Ile Gly Gln
405 410 415Leu Glu Asn Leu Gln Glu Leu Asp Leu Trp Asn Asn Arg Leu Thr Ala
420 425 430Leu Pro Lys Glu Ile Gly Gln Leu Lys Asn Leu Glu Asn Leu Glu Leu
435 440 445Ser Glu Asn Gln Leu Thr Thr Phe Pro Lys Glu Ile Gly Gln Leu Lys
450 455 460Lys Leu Gln Asp Leu Gly Leu Ser Tyr Asn Arg Leu Val Ile Leu Pro465 470 475 480Lys Glu Ile Gly Gln Leu Glu Lys Leu Gln Asp Leu Gly Leu Ser Tyr
485 490 495Asn Arg Leu Val Ile Leu Pro Lys Glu Ile Gly Gln Leu Lys Asn Leu
500 505 510Gln Met Leu Asp Leu Cys Tyr Asn Gln Phe Lys Thr Val Ser Lys Glu
515 520 525Ile Gly Gln Leu Lys Asn Leu Leu Gln Leu Asn Leu Ser Tyr Asn Gln
530 535 540Leu Ala Thr Leu Pro Ala Glu Ile Gly Gln Leu Lys Asn Leu Tyr Asn545 550 555 560Leu Asp Leu Gly Thr Asn Gln Leu Thr Thr Leu Pro Lys Glu Ile Gly
565 570 575Gln Leu Lys Asn Leu Tyr Asn Leu Gly Leu Gly Thr Asn Gln Leu Thr
580 585 590Thr Leu Pro Lys Glu Ile Gly Gln Leu Lys Lys Leu Gln Trp Leu Leu
595 600 605Leu Ser Val Asn Pro Ile Leu Pro Gln Glu Lys Glu Arg Ile Arg Lys
610 615 620Leu Leu Pro Asn Cys Glu Ile Arg Phe625 630<210>35<211>870<212>DNA<213>钩端螺旋体(Leptospira interrogans)<400>35atgacgtgct ccaatctcta caaaatggat tttacccttc gattcagaat aaaatcctgc 60ggagaatcca cgatgaaatt tcgcataacg ttgatttact tacaaaaaat aacaatcttc 120cttttatttc tcatttattt atcttgcgag attcaagcag aagcaccaga agacgaacca 180ggaacatata gggatttaac agaagcgctc caaaattctt tggatgttcg aattttgatt 240ttgagtgaac aaaaacttac aactcttcct aagaaaattg aacaactaaa gaatttacaa 300atgttggatt tatgttataa ccaatttaaa acagttccta aagaaattga acaactaaag 360aatttacaaa tgttggattt atgttataac caatttaaaa cagttcctaa gaaaattggg 420caactaaaga atttgcaagt attgaatttg agtagtaacc agctcacaac tcttccaaag 480gaaatcggaa aactggaaaa tttgcaagta ttgaatttga gtagtaacca gctcataact 540tttccaaaag aaatcggaaa actggaaaat ttgcaagtat tgaatttggg ttctaaccga 600ctcaaaaccc ttcctaaagg aattgagcaa ttgaaaaact tgcaaacgtt gtatttgaat 660tataaccaac ttacgactct tcctcgtgag atcggtcgat tacaaagtct aactaaatta 720catttgcagc ataatcaaat agcgacactc ccggacgaaa tcatacaact tcaaaattta 780cggaaactta ctttgtatga aaatccgatt ccgccgcaag agctggacaa aatacgaaag 840cttcttccaa attgtgagat tcgattttga 870<210>36<211>289<212>PRT<213>钩端螺旋体(Leptospira interrogans)<400>36Met Thr Cys Ser Asn Leu Tyr Lys Met Asp Phe Thr Leu Arg Phe Arg1 5 10 15Ile Lys Ser Cys Gly Glu Ser Thr Met Lys Phe Arg Ile Thr Leu Ile
20 25 30Tyr Leu Gln Lys Ile Thr Ile Phe Leu Leu Phe Leu Ile Tyr Leu Ser
35 40 45Cys Glu Ile Gln Ala Glu Ala Pro Glu Asp Glu Pro Gly Thr Tyr Arg
50 55 60Asp Leu Thr Glu Ala Leu Gln Asn Ser Leu Asp Val Arg Ile Leu Ile65 70 75 80Leu Ser Glu Gln Lys Leu Thr Thr Leu Pro Lys Lys Ile Glu Gln Leu
85 90 95Lys Asn Leu Gln Met Leu Asp Leu Cys Tyr Asn Gln Phe Lys Thr Val
100 105 110Pro Lys Glu Ile Glu Gln Leu Lys Asn Leu Gln Met Leu Asp Leu Cys
115 120 125Tyr Asn Gln Phe Lys Thr Val Pro Lys Lys Ile Gly Gln Leu Lys Asn
130 135 140Leu Gln Val Leu Asn Leu Ser Ser Asn Gln Leu Thr Thr Leu Pro Lys145 150 155 160Glu Ile Gly Lys Leu Glu Asn Leu Gln Val Leu Asn Leu Ser Ser Asn
165 170 175Gln Leu Ile Thr Phe Pro Lys Glu Ile Gly Lys Leu Glu Asn Leu Gln
180 185 190Val Leu Asn Leu Gly Ser Asn Arg Leu Lys Thr Leu Pro Lys Gly Ile
195 200 205Glu Gln Leu Lys Asn Leu Gln Thr Leu Tyr Leu Asn Tyr Asn Gln Leu
210 215 220Thr Thr Leu Pro Arg Glu Ile Gly Arg Leu Gln Ser Leu Thr Lys Leu225 230 235 240His Leu Gln His Asn Gln Ile Ala Thr Leu Pro Asp Glu Ile Ile Gln
245 250 255Leu Gln Asn Leu Arg Lys Leu Thr Leu Tyr Glu Asn Pro Ile Pro Pro
260 265 270Gln Glu Leu Asp Lys Ile Arg Lys Leu Leu Pro Asn Cys Glu Ile Arg
275 280 285Phe<210>37<211>867<212>DNA<213>钩端螺旋体(Leptospira interrogans)<400>37atgattttta atgattcgaa aaatggtaga aatttgcaaa agattgtata tatcgttttt 60gttttattga gttttttttg taaacgtcca ttgaagccgg gagaatatac cgatctagaa 120gaagcattca aaaatcctaa agatgtgttg gttttagatt acagagataa cgaagaaaat 180ccactgaaaa cacttccaaa ggaaattgga aacttacaaa atttaaaaga attatattta 240agcgcgaacg aaattacaac tctgcctcct gaaataggaa atttaaaaaa cttacaggta 300ttatctttaa atggaaacag acttgaaacg attcctaaag aaattggaaa cttaaaaaac 360ctaaaagaac tgtcaataga atggaataaa cttcaaacac ttcctaaaga aataggaaat 420ttaaaaaact taaaagaatt atatttaagc agaaatcaac tcaaaatatt accgcaagaa 480atcgggaact taagaaaact acagcgtata catttaagta caaatgaacc cacaaaactc 540ccgcaagaaa tcaaaaactt ggaaagttta ttagaaattt atttatatga taatcagttt 600acaacacttc ccaaagagat aggcaatttg aaaaatttac gtaatttagt tttaggcaga 660aatcaactta tatcgctact ccctgaaata ggaaatttaa aaaatctaaa agaattatat 720ttagaagaaa accaacttac gatgttacca aaacaaatcg cagccttaaa acaactggca 780agacttagct taaaagggaa tcaatttcct tcggaagaaa aagaaagaat ccaacgtctt 840ttgcctaagt gcaacatttc tttttaa 867<210>38<211>288<212>PRT<213>钩端螺旋体(Leptospira interrogans)<400>38Met Ile Phe Asn Asp Ser Lys Asn Gly Arg Asn Leu Gln Lys Ile Vall 5 10 15Tyr Ile Val Phe Val Leu Leu Ser Phe Phe Cys Lys Arg Pro Leu Lys
20 25 30Pro Gly Glu Tyr Thr Asp Leu Glu Glu Ala Phe Lys Asn Pro Lys Asp
35 40 45Val Leu Val Leu Asp Tyr Arg Asp Asn Glu Glu Asn Pro Leu Lys Thr
50 55 60Leu Pro Lys Glu Ile Gly Asn Leu Gln Asn Leu Lys Glu Leu Tyr Leu65 70 75 80Ser Ala Asn Glu Ile Thr Thr Leu Pro Pro Glu Ile Gly Asn Leu Lys
85 90 95Asn Leu Gln Val Leu Ser Leu Asn Gly Asn Arg Leu Glu Thr Ile Pro
100 105 110Lys Glu Ile Gly Asn Leu Lys Asn Leu Lys Glu Leu Ser Ile Glu Trp
115 120 125Asn Lys Leu Gln Thr Leu Pro Lys Glu Ile Gly Asn Leu Lys Asn Leu
130 135 140Lys Glu Leu Tyr Leu Ser Arg Asn Gln Leu Lys Ile Leu Pro Gln Glu145 150 155 160Ile Gly Asn Leu Arg Lys Leu Gln Arg Ile His Leu Ser Thr Asn Glu
165 170 175Leu Thr Lys Leu Pro Gln Glu Ile Lys Asn Leu Glu Ser Leu Leu Glu
180 185 190Ile Tyr Leu Tyr Asp Asn Gln Phe Thr Thr Leu Pro Lys Glu Ile Gly
195 200 205Asn Leu Lys Asn Leu Arg Asn Leu Val Leu Gly Arg Asn Gln Leu Ile
210 215 220Ser Leu Leu Pro Glu Ile Gly Asn Leu Lys Asn Leu Lys Glu Leu Tyr225 230 235 240Leu Glu Glu Asn Gln Leu Thr Met Leu Pro Lys Gln Ile Ala Ala Leu
245 250 255Lys Gln Leu Ala Arg Leu Ser Leu Lys Gly Asn Gln Phe Pro Ser Glu
260 265 270Glu Lys Glu Arg Ile Gln Arg Leu Leu Pro Lys Cys Asn Ile Ser Phe
275 280 285<210>39<211>1281<212>DNA<213>钩端螺旋体(Leptospira interrogans)<400>39atgaatttcc aatccataaa cattcgttta aataaaagac tcatgattct tttgattctg 60atctgttttt cctgcaaact acaagcccaa tctaacgaag tgcaaacata ttatcgtaat 120ataacagagg ttcttaaaaa tccacagaat gttcgaattc tgaatttgag cggtagcaaa 180ctcacaaccc ttccgggaga aatcggaaaa ctgcaaaatt tacaactgtt gaatttagat 240gataaccaac ttatagctct tccgaaagaa attggaaaac ttcaaaattt acaacaattg 300catttgagta agaaccaact catggcgctt ccagaagaaa tcggacaact tcagaattta 360caaaagttga aattgtatga aaaccagctt acggccattc ccaaagagat tggacaacta 420caaaatttac aagagctgaa tttggcacat aaccaactcg caactctacc agaagacatc 480gaacaattac aaagattaca aacattgtat ttgggtcata accaatttaa ctctattctc 540aaagagattg gacaattaca aaatttagaa tcattaggtt tggatcataa tcaacttaac 600gtccttccca aagaaattgg acaacttcga aatttagaat cattaggttt ggatcataac 660caacttaacg tccttcccaa agaaattgga caacttcaaa atttacaaat attacatttg 720cgtaataacc aactcacaac tcttcccaaa gaaatcgggc agcttcagaa tttacaaaag 780ttacttttaa ataaaaacaa actcacaact cttcccaaag aaattggtca gcttcagaat 840ttacaaaagt tgaaattgta tgaaaaccaa cttacaaccc ttccgaaaga gattggacaa 900ctacaaaatt tacaagagtt ggatttagat ggtaaccaac tcacaactct accagaaaac 960atcggacaat tacagaggtt acaaacgttg tatttgggta ataatcaact taacttcctt 1020ccaaataaag tagaacaact acaaaattta gaatcattag atctggagca taaccaactt 1080aacgcccttc ccaaagaaat tggaaaattg caaaagttgc aaaccctgaa tttgaaatat 1140aatcaacttg cgactcttcc cgaggagatc aaacaactaa agaacttaaa aaagttgtat 1200ttgcataaca acccacttcc atcggaaaaa atagaaagaa ttcgaaaatt acttccccaa 1260tgtaatattt attttgaatg a 1281<210>40<211>426<212>PRT<213>钩端螺旋体(Leptospira interrogans)<400>40Met Asn Phe Gln Ser Ile Asn Ile Arg Leu Asn Lys Arg Leu Met Ile1 5 10 15Leu Leu Ile Leu Ile Cys Phe Ser Cys Lys Leu Gln Ala Gln Ser Asn
20 25 30Glu Val Gln Thr Tyr Tyr Arg Asn Ile Thr Glu Val Leu Lys Asn Pro
35 40 45Gln Asn Val Arg Ile Leu Asn Leu Ser Gly Ser Lys Leu Thr Thr Leu
50 55 60Pro Gly Glu Ile Gly Lys Leu Gln Asn Leu Gln Leu Leu Asn Leu Asp65 70 75 80Asp Asn Gln Leu Ile Ala Leu Pro Lys Glu Ile Gly Lys Leu Gln Asn
85 90 95Leu Gln Gln Leu His Leu Ser Lys Asn Gln Leu Met Ala Leu Pro Glu
100 105 110Glu Ile Gly Gln Leu Gln Asn Leu Gln Lys Leu Lys Leu Tyr Glu Asn
115 120 125Gln Leu Thr Ala Ile Pro Lys Glu Ile Gly Gln Leu Gln Asn Leu Gln
130 135 140Glu Leu Asn Leu Ala His Asn Gln Leu Ala Thr Leu Pro Glu Asp IIe145 150 155 160Glu Gln Leu Gln Arg Leu Gln Thr Leu Tyr Leu Gly His Asn Gln Phe
165 170 175Asn Ser Ile Leu Lys Glu Ile Gly Gln Leu Gln Asn Leu Glu Ser Leu
180 185 190Gly Leu Asp His Asn Gln Leu Asn Val Leu Pro Lys Glu Ile Gly Gln
195 200 205Leu Arg Asn Leu Glu Ser Leu Gly Leu Asp His Asn Gln Leu Asn Val
210 215 220Leu Pro Lys Glu Ile Gly Gln Leu Gln Asn Leu Gln Ile Leu His Leu225 230 235 240Arg Asn Asn Gln Leu Thr Thr Leu Pro Lys Glu Ile Gly Gln Leu Gln
245 250 255Asn Leu Gln Lys Leu Leu Leu Asn Lys Asn Lys Leu Thr Thr Leu Pro
260 265 270Lys Glu Ile Gly Gln Leu Gln Asn Leu Gln Lys Leu Lys Leu Tyr Glu
275 280 285Asn Gln Leu Thr Thr Leu Pro Lys Glu Ile Gly Gln Leu Gln Asn Leu
290 295 300Gln Glu Leu Asp Leu Asp Gly Asn Gln Leu Thr Thr Leu Pro Glu Asn305 310 315 320Ile Gly Gln Leu Gln Arg Leu Gln Thr Leu Tyr Leu Gly Asn Asn Gln
325 330 335Leu Asn Phe Leu Pro Asn Lys Val Glu Gln Leu Gln Asn Leu Glu Ser
340 345 350Leu Asp Leu Glu His Asn Gln Leu Asn Ala Leu Pro Lys Glu Ile Gly
355 360 365Lys Leu Gln Lys Leu Gln Thr Leu Asn Leu Lys Tyr Asn Gln Leu Ala
370 375 380Thr Leu Pro Glu Glu Ile Lys Gln Leu Lys Asn Leu Lys Lys Leu Tyr385 390 395 400Leu His Asn Asn Pro Leu Pro Ser Glu Lys Ile Glu Arg Ile Arg Lys
405 410 415Leu Leu Pro Gln Cys Asn Ile Tyr Phe Glu
420 425<210>41<211>1227<212>DNA<213>钩端螺旋体(Leptospira interrogans)<400>41atgaaaccga atttaatacg acaacgctgt tttcaaaaag tggaaatcct ttttttaata 60tttgtatgtt cccttacgca attccatgcg gaagaaaatc acacaacgaa agaaggtctt 120tatacaaatt taaccgaggc tcttaaaaat ccgaatgagg ttcggatttt agatttaagt 180agaagcaaac tcaagattct tccgaaagaa atcgggcaac tgcaaaattt acaaatcctg 240aattcagaaa ataatcaact cacgaccctt cccaaagaaa tcggaaaatt gcagaacttg 300caagagttac atctgcagaa taaccaacta acaacccttc cagaagaaat agggcagtta 360cagaatttaa aagtattaca tttgaacaat aatcaactca ctactcttcc agaagaaatc 420ggaaaactac aaaatctaca agagttgaat ttatttgtta atcgacttaa catccttcca 480aaggaaatcg gacgactaca aaatttacaa gagttatatt tgtctcttaa tcggcttaca 540atccttccag aagaaattgg acaattagag agtttacgaa agttgagttt gggtggtaag 600aataagccat tcacgattct tccaaaagaa atcacgcagc ttcagaattt acaagagttg 660catttgaaat ttaaccggct cacggtcctt cccaaagaaa ttggacaact tcaaaatttg 720cggatattag atttgtatca aaaccgactc acgattcttc cgaaagaaat tggacaactc 780aaaaatttac tagtattgga tttgtctggt aaccaactta caattcttcc gaaagaaatc 840acacaacttc agaatttaca agaattgaat ttggaataca atcgattcga agcttttcca 900aaagaaatca cacagtttca gaatttacaa gtgttggatt tgtatcaaaa ccgactcacg 960actcttccaa aagaaattgg acaattgcag aatttacaaa agttgcattt gagtagaaac 1020caacttacga ctcttccgaa agagattgga cgattgcaaa agttagaatc attgggtttg 1080gatcataatc aacttgcgac tcttcccgag gagatcaaac aactaaagaa cttaaaaaag 1140ttgtatttgc ataacaaccc acttctatcg gaaaaaatag aaagaattcg aaaattactt 1200ccccaatgta ttatttattt tgaatga 1227<210>42<211>408<212>PRT<213>钩端螺旋体(Leptospira interrogans)<400>42Met Lys Pro Asn Leu Ile Arg Gln Arg Cys Phe Gln Lys Val Glu Ile1 5 10 15Leu Phe Leu Ile Phe Val Cys Ser Leu Thr Gln Phe His Ala Glu Glu
20 25 30Asn His Thr Thr Lys Glu Gly Leu Tyr Thr Asn Leu Thr Glu Ala Leu
35 40 45Lys Asn Pro Asn Glu Val Arg Ile Leu Asp Leu Ser Arg Ser Lys Leu
50 55 60Lys Ile Leu Pro Lys Glu Ile Gly Gln Leu Gln Asn Leu Gln Ile Leu65 70 75 80Asn Ser Glu Asn Asn Gln Leu Thr Thr Leu Pro Lys Glu Ile Gly Lys
85 90 95Leu Gln Asn Leu Gln Glu Leu His Leu Gln Asn Asn Gln Leu Thr Thr
100 105 110Leu Pro Glu Glu Ile Gly Gln Leu Gln Asn Leu Lys Val Leu His Leu
115 120 125Asn Asn Asn Gln Leu Thr Thr Leu Pro Glu Glu Ile Gly Lys Leu Gln
130 135 140Asn Leu Gln Glu Leu Asn Leu Phe Val Asn Arg Leu Asn Ile Leu Pro145 150 155 160Lys Glu Ile Gly Arg Leu Gln Asn Leu Gln Glu Leu Tyr Leu Ser Leu
165 170 175Asn Arg Leu Thr Ile Leu Pro Glu Glu Ile Gly Gln Leu Glu Ser Leu
180 185 190Arg Lys Leu Ser Leu Gly Gly Lys Asn Lys Pro Phe Thr Ile Leu Pro
195 200 205Lys Glu Ile Thr Gln Leu Gln Asn Leu Gln Glu Leu His Leu Lys Phe
210 215 220Asn Arg Leu Thr Val Leu Pro Lys Glu Ile Gly Gln Leu Gln Asn Leu225 230 235 240Arg Ile Leu Asp Leu Tyr Gln Asn Arg Leu Thr Ile Leu Pro Lys Glu
245 250 255Ile Gly Gln Leu Lys Asn Leu Leu Val Leu Asp Leu Ser Gly Asn Gln
260 265 270Leu Thr Ile Leu Pro Lys Glu Ile Thr Gln Leu Gln Asn Leu Gln Glu
275 280 285Leu Asn Leu Glu Tyr Asn Arg Phe Glu Ala Phe Pro Lys Glu Ile Thr
290 295 300Gln Phe Gln Asn Leu Gln Val Leu Asp Leu Tyr Gln Asn Arg Leu Thr305 310 315 320Thr Leu Pro Lys Glu Ile Gly Gln Leu Gln Asn Leu Gln Lys Leu His
325 330 335Leu Ser Arg Asn Gln Leu Thr Thr Leu Pro Lys Glu Ile Gly Arg Leu
340 345 350Gln Lys Leu Glu Ser Leu Gly Leu Asp His Asn Gln Leu Ala Thr Leu
355 360 365Pro Glu Glu Ile Lys Gln Leu Lys Asn Leu Lys Lys Leu Tyr Leu His
370 375 380Asn Asn Pro Leu Leu Ser Glu Lys Ile Glu Arg Ile Arg Lys Leu Leu385 390 395 400Pro Gln Cys Ile Ile Tyr Phe Glu
405<210>43<211>2058<212>DNA<213>钩端螺旋体(Leptospira interrogans)<400>43atgaaacact acctaacctt ccaagacgat aagtcagata aattctggca aatagaagtt 60tccgaaaatt catttacagt aacctacgga aaaactggca cttctggaca aacacagatt 120aaaacctttg acgataagga aatctgtctt aaagaagcaa aaaaacttct ctctgaaaaa 180ttaaaaaaag gttatgtaga aacagatcca cctatttctt ctacaagttc cgattcttct 240aagaaaaaaa actcaacttc cacagaaact tctgtttctc aaacaaaatc aaaaatagac 300aaaccagcct ctttgacttc agaaacattt catcaatttt taaaagttag aatccaagat 360atttcttctt caaaatattc taaaatactt aaaaatacag attgggaaaa tctagcggga 420gacattttcg attccgttta taaatattgg gaagaactaa atcaaaaaga aaaatctcca 480ataggaattt ttgatataag atgggacgac gcaagtacgg attattgtat agaatttgac 540tatgatttag aaagtaacga tcctaaaaat tctatggaag aaggtgcaat tcgaaacgtc 600cctgtagtcg atttttctaa ttttattaaa aatcaactca aacagaaacc agaaaaaatc 660cgggaagtat tggaagacga agacttttct ctgataaaag acatactgtg taaactaacg 720caggaagtaa ttatacaaac tttaaaagca gaagttttta aacaaatccc aaaacaacaa 780ccgtattttc taatgttctc ctattaccac gatgaagatg cttctgtatt tttcgattcg 840aacgcaaagg aacaaaaaaa tttatactcc atactcaaaa tctcaaaatc ttctctattt 900aaatattttt taagcgggga gcaatcttta atcatttcta atgtttcaga aatagatctt 960accgatataa atctttttga aaatttaaaa aatttcaata tttccaattg cgaaaatata 1020atttctctcc aaagtttaag tgaattaaaa aatatccaaa gtatttccat aaaaggtgca 1080aaattatccg agttccccga ttttttacta aatcttccgt ctcttaaatc tttgtatctc 1140acaaatagca acctatctat agaaaataaa attagtatat tcaattcttc gcaattagaa 1200ctactttgcc taaacgctaa ctccttaacc acaattccag aattcgtatt ccaacttccg 1260caattaaaaa agctactctt tatggacaac caattgacag agttaccaga tcgattagca 1320gatctaaaat ttcttcgaaa tttaaattta tccggcaaca aaatcactca aatttccaat 1380ttaactaagg aattcagcga aatcatagaa cttggtcttt ttgataatcg actcacttct 1440ttggatggaa tttgtcgttt tcctaaattg aatgaacttt taatttgggg caacgaatta 1500gaaacaattt caccggaaat ttttaattta aaaaatctaa ctcggataga tacgactaaa 1560aataaaattt caagttttcc taatatagga attacactcg aatcagtaac caatttgagt 1620ctgagcaata accaattgac tcaaatacca gaaggactaa ctcaatttcc aaatctaaaa 1680tctttgggtt tagatgacaa tttacttaaa gaactacctg acgatctatt taaaaacttt 1740caaaaattgg aaactcttgc attaagcaac aatcggctat ctaatcttcc taaatccatt 1800tcgcagttag aatctttaaa aaacatatat ttaaaaaata atcagtttat tcaaattcct 1860gagattttaa aggagctgaa aaaacttaaa gacgtatctt tgagcggaaa ccagatttca 1920aaattacccg aatttctttc ggaaatgaca gcattgagag aattaaaaat cggcaacaat 1980ccaattgcac aaaacccaga aagtgtagaa accaagatga aagaaatcaa ttcaaaagta 2040gtactttatc tttcttga 2058<210>44<211>685<212>PRT<213>钩端螺旋体(Leptospira interrogans)<400>44Met Lys His Tyr Leu Thr Phe Gln Asp Asp Lys Ser Asp Lys Phe Trp1 5 10 15Gln Ile Glu Val Ser Glu Asn Ser Phe Thr Val Thr Tyr Gly Lys Thr
20 25 30Gly Thr Ser Gly Gln Thr Gln Ile Lys Thr Phe Asp Asp Lys Glu Ile
35 40 45Cys Leu Lys Glu Ala Lys Lys Leu Leu Ser Glu Lys Leu Lys Lys Gly
50 55 60Tyr Val Glu Thr Asp Pro Pro Ile Ser Ser Thr Ser Ser Asp Ser Ser65 70 75 80Lys Lys Lys Asn Ser Thr Ser Thr Glu Thr Ser Val Ser Gln Thr Lys
85 90 95Ser Lys Ile Asp Lys Pro Ala Ser Leu Thr Ser Glu Thr Phe His Gln
100 105 110Phe Leu Lys Val Arg Ile Gln Asp Ile Ser Ser Ser Lys Tyr Ser Lys
115 120 125Ile Leu Lys Asn Thr Asp Trp Glu Asn Leu Ala Gly Asp Ile Phe Asp
130 135 140Ser Val Tyr Lys Tyr Trp Glu Glu Leu Asn Gln Lys Glu Lys Ser Pro145 150 155 160Ile Gly Ile Phe Asp Ile Arg Trp Asp Asp Ala Ser Thr Asp Tyr Cys
165 170 175Ile Glu Phe Asp Tyr Asp Leu Glu Ser Asn Asp Pro Lys Asn Ser Met
180 185 190Glu Glu Gly Ala Ile Arg Asn Val Pro Val Val Asp Phe Ser Asn Phe
195 200 205Ile Lys Asn Gln Leu Lys Gln Lys Pro Glu Lys Ile Arg Glu Val Leu
210 215 220Glu Asp Glu Asp Phe Ser Leu Ile Lys Asp Ile Leu Cys Lys Leu Thr225 230 235 240Gln Glu Val Ile Ile Gln Thr Leu Lys Ala Glu Val Phe Lys Gln Ile
245 250 255Pro Lys Gln Gln Pro Tyr Phe Leu Met Phe Ser Tyr Tyr His Asp Glu
260 265 270Asp Ala Ser Val Phe Phe Asp Ser Asn Ala Lys Glu Gln Lys Asn Leu
275 280 285Tyr Ser Ile Leu Lys Ile Ser Lys Ser Ser Leu Phe Lys Tyr Phe Leu
290 295 300Ser Gly Glu Gln Ser Leu Ile Ile Ser Asn Val Ser Glu Ile Asp Leu305 310 315 320Thr Asp Ile Asn Leu Phe Glu Asn Leu Lys Asn Phe Asn Ile Ser Asn
325 330 335Cys Glu Asn Ile Ile Ser Leu Gln Ser Leu Ser Glu Leu Lys Asn Ile
340 345 350Gln Ser Ile Ser Ile Lys Gly Ala Lys Leu Ser Glu Phe Pro Asp Phe
355 360 365Leu Leu Asn Leu Pro Ser Leu Lys Ser Leu Tyr Leu Thr Asn Ser Asn
370 375 380Leu Ser Ile Glu Asn Lys Ile Ser Ile Phe Asn Ser Ser Gln Leu Glu385 390 395 400Leu Leu Cys Leu Asn Ala Asn Ser Leu Thr Thr Ile Pro Glu Phe Val
405 410 415Phe Gln Leu Pro Gln Leu Lys Lys Leu Leu Phe Met Asp Asn Gln Leu
420 425 430Thr Glu Leu Pro Asp Arg Leu Ala Asp Leu Lys Phe Leu Arg Asn Leu
435 440 445Asn Leu Ser Gly Asn Lys Ile Thr Gln Ile Ser Asn Leu Thr Lys Glu
450 455 460Phe Ser Glu Ile Ile Glu Leu Gly Leu Phe Asp Asn Arg Leu Thr Ser465 470 475 480Leu Asp Gly Ile Cys Arg Phe Pro Lys Leu Asn Glu Leu Leu Ile Trp
485 490 495Gly Asn Glu Leu Glu Thr Ile Ser Pro Glu Ile Phe Asn Leu Lys Asn
500 505 510Leu Thr Arg Ile Asp Thr Thr Lys Asn Lys Ile Ser Ser Phe Pro Asn
515 520 525Ile Gly Ile Thr Leu Glu Ser Val Thr Asn Leu Ser Leu Ser Asn Asn
530 535 540Gln Leu Thr Gln Ile Pro Glu Gly Leu Thr Gln Phe Pro Asn Leu Lys545 550 555 560Ser Leu Gly Leu Asp Asp Asn Leu Leu Lys Glu Leu Pro Asp Asp Leu
565 570 575Phe Lys Asn Phe Gln Lys Leu Glu Thr Leu Ala Leu Ser Asn Asn Arg
580 585 590Leu Ser Asn Leu Pro Lys Ser Ile Ser Gln Leu Glu Ser Leu Lys Asn
595 600 605Ile Tyr Leu Lys Asn Asn Gln Phe Ile Gln Ile Pro Glu Ile Leu Lys
610 615 620Glu Leu Lys Lys Leu Lys Asp Val Ser Leu Ser Gly Asn Gln Ile Ser625 630 635 640Lys Leu Pro Glu Phe Leu Ser Glu Met Thr Ala Leu Arg Glu Leu Lys
645 650 655Ile Gly Asn Asn Pro Ile Ala Gln Asn Pro Glu Ser Val Glu Thr Lys
660 665 670Met Lys Glu Ile Asn Ser Lys Val Val Leu Tyr Leu Ser
675 680 685<210>45<211>1134<212>DNA<213>钩端螺旋体(Leptospira interrogans)<400>45atgaacttcc gcataacgtt aattcactta caaaaaatta caattggtct tttgtttctc 60attcatcttt cttacgagat tcaagcagaa gaaccaggaa cttatagaga tttaacaaaa 120gcgctccaaa atcctttgga ggttcgagtt ttggatttga gtcgtcaaga actcaaaacc 180cttcctatag aaatcggaaa attgaagaat ctgcaaaggt tgtatttaca ttataaccaa 240cttacagttc ttccacaaga aatagaacaa ctgaaaaatt tacaattgtt gtatttgcgt 300tctaaccgac tcactactct tcctaacgaa atcgagcaat tgaagaactt gcaagttttg 360gatttgggtt ctaaccaact tacaattctt ccacaagaaa tagaacagct gaaaaattta 420caattgttgt atttgcattc taaccgactc actactcttt ctaaagacat tgaacaacta 480cagaatttga aatcgttgga tttgagtaat aaccaactta caactcttcc taacgaaatc 540gagcaattga aaaacttgaa atcgttgtat ttgagtgaga accaattcgc aacttttcct 600aaagaaattg gacaactaca gaatttaaaa gtgttgtttt tgaataataa ccaaatcaca 660atacttccca acgaaatcgc aaaactggaa aagctacaat acttatattt aagcgacaat 720caactcataa cccttccgaa agaaatagag caattgaaga acttgcaaac gttggatttg 780agttataacc aacttacaat acttcctaag gaagtcggac aattagaaaa tttgcaaacg 840ttggatttgc gtaataacca actcaaaact cttccgaaag aaatagagca attgaagaac 900ttacaaacgt tgtttttgaa taataaccaa ctcacaatac ttccacaaga aatcggaaaa 960ctgaagaatt tgttatggtt gagtttggtt tataaccaac tcactactct tcctaacgaa 1020atcgagcaat tgaagaactt acaaacattg tatttaaaca acaaccaatt ctcatcccaa 1080gaaaaaaaaa gaattcgaaa acttcttcca aagtgccaaa tttcctttga ataa 1134<210>46<211>377<212>PRT<213>钩端螺旋体(Leptospira interrogans)<400>46Met Asn Phe Arg Ile Thr Leu Ile His Leu Gln Lys Ile Thr Ile Gly1 5 10 15Leu Leu Phe Leu Ile His Leu Ser Tyr Glu Ile Gln Ala Glu Glu Pro
20 25 30Gly Thr Tyr Arg Asp Leu Thr Lys Ala Leu Gln Asn Pro Leu Glu Val
35 40 45Arg Val Leu Asp Leu Ser Arg Gln Glu Leu Lys Thr Leu Pro Ile Glu
50 55 60Ile Gly Lys Leu Lys Asn Leu Gln Arg Leu Tyr Leu His Tyr Asn Gln65 70 75 80Leu Thr Val Leu Pro Gln Glu Ile Glu Gln Leu Lys Asn Leu Gln Leu
85 90 95Leu Tyr Leu Arg Ser Asn Arg Leu Thr Thr Leu Pro Asn Glu Ile Glu
100 105 110Gln Leu Lys Asn Leu Gln Val Leu Asp Leu Gly Ser Asn Gln Leu Thr
115 120 125Ile Leu Pro Gln Glu Ile Glu Gln Leu Lys Asn Leu Gln Leu Leu Tyr
130 135 140Leu His Ser Asn Arg Leu Thr Thr Leu Ser Lys Asp Ile Glu Gln Leu145 150 155 160Gln Asn Leu Lys Ser Leu Asp Leu Ser Asn Asn Gln Leu Thr Thr Leu
165 170 175Pro Asn Glu Ile Glu Gln Leu Lys Asn Leu Lys Ser Leu Tyr Leu Ser
180 185 190Glu Asn Gln Phe Ala Thr Phe Pro Lys Glu Ile Gly Gln Leu Gln Asn
195 200 205Leu Lys Val Leu Phe Leu Asn Asn Asn Gln Ile Thr Ile Leu Pro Asn
210 215 220Glu Ile Ala Lys Leu Lys Lys Leu Gln Tyr Leu Tyr Leu Ser Asp Asn225 230 235 240Gln Leu Ile Thr Leu Pro Lys Glu Ile Glu Gln Leu Lys Asn Leu Gln
245 250 255Thr Leu Asp Leu Ser Tyr Asn Gln Leu Thr Ile Leu Pro Lys Glu Val
260 265 270Gly Gln Leu Glu Asn Leu Gln Thr Leu Asp Leu Arg Asn Asn Gln Leu
275 280 285Lys Thr Leu Pro Lys Glu Ile Glu Gln Leu Lys Asn Leu Gln Thr Leu
290 295 300Phe Leu Asn Asn Asn Gln Leu Thr Ile Leu Pro Gln Glu Ile Gly Lys305 310 315 320Leu Lys Asn Leu Leu Trp Leu Ser Leu Val Tyr Asn Gln Leu Thr Thr
325 330 335Leu Pro Asn Glu Ile Glu Gln Leu Lys Asn Leu Gln Thr Leu Tyr Leu
340 345 350Asn Asn Asn Gln Phe Ser Ser Gln Glu Lys Lys Arg Ile Arg Lys Leu
355 360 365Leu Pro Lys Cys Gln Ile Ser Phe Glu
370 375<210>47<211>1359<212>DNA<213>钩端螺旋体(Leptospira interrogans)<400>47atgaacttcc gcataacgtt aattcactta caaaaaatta caattagcct ttttcttcta 60acctgtttta tttacgaatt gcaagcagaa gaatcagaat caggaactta cacagactta 120gcaaaaacgc ttcaaaatcc tttgaaggtt cgaaccttgg atttgagata ccaaaaactc 180acaactcttc cgaaagaaat cggacaacta cagaatttac aacggttgga tttgagtttt 240aactcactca caactcttcc gaaagaaatc ggacaactac ggaatttaca agagttggat 300ttgagtttta actcacttac aactctcccc aaagaagtcg ggcaattgga gaatttacaa 360cggttggatt tgcatcaaaa ccgactcgca actcttccta tggaaatcgg acagttaaaa 420aatttacaag agttagattt gaatagcaac aaactcacaa cccttccgaa agaaataaga 480caactacgga atttacaaga gttagatttg aatagcaaca aactcacaac tctcccaaaa 540gaaatcggac aactacagaa tttaaagacc ttgaatttga ttgttaccca gctaacaacc 600cttcccaaag aaatagggga gcttcagaat ttaaagacct tgaatttgct tgataaccaa 660cttacaactc ttcccaaaga aataggggag cttcagaatt tagagatctt ggttttgcgt 720gaaaaccgga tcacagctct tccgaaagaa atcggacaac tacagaattt acaatggttg 780gatttgcatc aaaaccaact cacaactctc ccgaaagaaa tcggacaact acagaattta 840caacggttgg atttgcatca aaaccaactc acaactctcc cgaaagaaat cggacagcta 900cagaatttac aagagctctg tttagatgag aatcaactca cgactcttcc gaaagaaatc 960gagcaattac aaaacttacg agtcttggat ttggataata accaactcac gactcttccg 1020aaagaagtcc tacggctaca gagcttgcaa gtgttggctt tgggtagtaa ccgactctca 1080acccttccga aagaaatcgg acaattgcag aacttgcaag tgttgggttt gattagtaac 1140caactcacga ctcttccgaa agaaatcgga caattacaga acttgcaaga gctctgtttg 1200gatgagaatc aactcacaac ttttcctaag gaaatcaggc aactgaaaaa tttacaagag 1260ttgcatttgt atttgaatcc tctctcatcc aaagaaaaaa aaaggattcg aagattactt 1320ccaaaatgtg aaattcattt tgaagaatat catatatag 1359<210>48<211>452<212>PRT<213>钩端螺旋体(Leptospira interrogans)<400>48Met Asn Phe Arg Ile Thr Leu Ile His Leu Gln Lys Ile Thr Ile Ser1 5 10 15Leu Phe Leu Leu Thr Cys Phe Ile Tyr Glu Leu Gln Ala Glu Glu Ser
20 25 30Glu Ser Gly Thr Tyr Thr Asp Leu Ala Lys Thr Leu Gln Asn Pro Leu
35 40 45Lys Val Arg Thr Leu Asp Leu Arg Tyr Gln Lys Leu Thr Thr Leu Pro
50 55 60Lys Glu Ile Gly Gln Leu Gln Asn Leu Gln Arg Leu Asp Leu Ser Phe65 70 75 80Asn Ser Leu Thr Thr Leu Pro Lys Glu Ile Gly Gln Leu Arg Asn Leu
85 90 95Gln Glu Leu Asp Leu Ser Phe Asn Ser Leu Thr Thr Leu Pro Lys Glu
100 105 110Val Gly Gln Leu Glu Asn Leu Gln Arg Leu Asp Leu His Gln Asn Arg
115 120 125Leu Ala Thr Leu Pro Met Glu Ile Gly Gln Leu Lys Asn Leu Gln Glu
130 135 140Leu Asp Leu Asn Ser Asn Lys Leu Thr Thr Leu Pro Lys Glu Ile Arg145 150 155 160Gln Leu Arg Asn Leu Gln Glu Leu Asp Leu Asn Ser Asn Lys Leu Thr
165 170 175Thr Leu Pro Lys Glu Ile Gly Gln Leu Gln Asn Leu Lys Thr Leu Asn
180 185 190Leu Ile Val Thr Gln Leu Thr Thr Leu Pro Lys Glu Ile Gly Glu Leu
195 200 205Gln Asn Leu Lys Thr Leu Asn Leu Leu Asp Asn Gln Leu Thr Thr Leu
210 215 220Pro Lys Glu Ile Gly Glu Leu Gln Asn Leu Glu Ile Leu Val Leu Arg225 230 235 240Glu Asn Arg Ile Thr Ala Leu Pro Lys Glu Ile Gly Gln Leu Gln Asn
245 250 255Leu Gln Trp Leu Asp Leu His Gln Asn Gln Leu Thr Thr Leu Pro Lys
260 265 270Glu Ile Gly Gln Leu Gln Asn Leu Gln Arg Leu Asp Leu His Gln Asn
275 280 285Gln Leu Thr Thr Leu Pro Lys Glu Ile Gly Gln Leu Gln Asn Leu Gln
290 295 300Glu Leu Cys Leu Asp Glu Asn Gln Leu Thr Thr Leu Pro Lys Glu Ile305 310 315 320Glu Gln Leu Gln Asn Leu Arg Val Leu Asp Leu Asp Asn Asn Gln Leu
325 330 335Thr Thr Leu Pro Lys Glu Val Leu Arg Leu Gln Ser Leu Gln Val Leu
340 345 350Ala Leu Gly Ser Asn Arg Leu Ser Thr Leu Pro Lys Glu Ile Gly Gln
355 360 365Leu Gln Asn Leu Gln Val Leu Gly Leu Ile Ser Asn Gln Leu Thr Thr
370 375 380Leu Pro Lys Glu Ile Gly Gln Leu Gln Asn Leu Gln Glu Leu Cys Leu385 390 395 400Asp Glu Asn Gln Leu Thr Thr Phe Pro Lys Glu Ile Arg Gln Leu Lys
405 410 415Asn Leu Gln Glu Leu His Leu Tyr Leu Asn Pro Leu Ser Ser Lys Glu
420 425 430Lys Lys Arg Ile Arg Arg Leu Leu Pro Lys Cys Glu Ile His Phe Glu
435 440 445Glu Tyr His Ile
450<210>49<211>855<212>DNA<213>钩端螺旋体(Leptospira interrogans)<400>49atgaaatttc gcttaacttt gaattactta caaaatataa caattggcct tctgtttctc 60attcatcttt cttgcaagat tcaaacggaa aaagtagaac cagtaactta tagggattta 120acaaaagcgc tccaaaatcc tttgggtgtt cgaattttga atttgagtcg tcaaaaactc 180aaaacttttc ctaaagaaat tgggcaattg aagaacttgc aagagttgca tttgagtagt 240aaccaattca ctactcttcc taaagaaatc gaacaactac aaaatttgaa atcattggat 300ttatgggata atcaacttaa aacccttcct aaggaaatcg gaaaactaca gaacttgaaa 360tcgttggatt tgggtagcaa ccaactcaca atccttccaa aggaaatcgg acaactacag 420aatttgcaaa aattgaattt atggaataat caacttaaaa ctcttcctaa ggaaattgga 480caattacaga acttgcaaaa aatgaattta gataaaaaca ggctcaatac acttcctaat 540gaaattggac aactacagaa tttggaatcg ttgtatttga attataacca actcactata 600cttccaaaag aaattggaca actacagaat ttggaatcgt tgtatttgaa ttataaccaa 660ctcacaatgc ttccacaaga aattggacaa ctacagaatt tggaagggtt gtatctgaaa 720tataatcaac tcacaaccct tcctaaagaa attggacgac tacagaattt gaaaaggttg 780tatctgaaat ataatcaatt ctcatccaaa gaaaaagaaa aaatccaaaa gcttcttcca 840aagtatccag ggtaa 855<210>50<211>284<212>PRT<213>钩端螺旋体(Leptospira interrogans)<400>50Met Lys Phe Arg Leu Thr Leu Asn Tyr Leu Gln Asn Ile Thr Ile Gly1 5 10 15Leu Leu Phe Leu Ile His Leu Ser Cys Lys Ile Gln Thr Glu Lys Val
20 25 30Glu Pro Val Thr Tyr Arg Asp Leu Thr Lys Ala Leu Gln Asn Pro Leu
35 40 45Gly Val Arg Ile Leu Asn Leu Ser Arg Gln Lys Leu Lys Thr Phe Pro
50 55 60Lys Glu Ile Gly Gln Leu Lys Asn Leu Gln Glu Leu His Leu Ser Ser65 70 75 80Asn Gln Phe Thr Thr Leu Pro Lys Glu Ile Glu Gln Leu Gln Asn Leu
85 90 95Lys Ser Leu Asp Leu Trp Asp Asn Gln Leu Lys Thr Leu Pro Lys Glu
100 105 110Ile Gly Lys Leu Gln Asn Leu Lys Ser Leu Asp Leu Gly Ser Asn Gln
115 120 125Leu Thr Ile Leu Pro Lys Glu Ile Gly Gln Leu Gln Asn Leu Gln Lys
130 135 140Leu Asn Leu Trp Asn Asn Gln Leu Lys Thr Leu Pro Lys Glu Ile Gly145 150 155 160Gln Leu Gln Asn Leu Gln Lys Met Asn Leu Asp Lys Asn Arg Leu Asn
165 170 175Thr Leu Pro Asn Glu Ile Gly Gln Leu Gln Asn Leu Glu Ser Leu Tyr
180 185 190Leu Asn Tyr Asn Gln Leu Thr Ile Leu Pro Lys Glu Ile Gly Gln Leu
195 200 205Gln Asn Leu Glu Ser Leu Tyr Leu Asn Tyr Asn Gln Leu Thr Met Leu
210 215 220Pro Gln Glu Ile Gly Gln Leu Gln Asn Leu Glu Gly Leu Tyr Leu Lys225 230 235 240Tyr Asn Gln Leu Thr Thr Leu Pro Lys Glu Ile Gly Arg Leu Gln Asn
245 250 255Leu Lys Arg Leu Tyr Leu Lys Tyr Asn Gln Phe Ser Ser Lys Glu Lys
260 265 270Glu Lys Ile Gln Lys Leu Leu Pro Lys Tyr Pro Gly
275 280<210>51<211>936<212>DNA<213>钩端螺旋体(Leptospira interrogans)<400>51atgaactttc gcataacttt gaattactta caaaaaataa caattggtct tttgtttctc 60atgcatcttt cttgcgagat tcaagcagag gaatttgaac aacaagaaac ttatacggat 120ttaacaaaag cgctccaaaa tcctttgaag gttcgaacct tggatttgag tgctaaccga 180ttcaaaactc ttcctaagga aatcggaaaa ttgaaaaatt tacaagagtt gaatttgaat 240aagaaccaac tcacaactct tccacaagaa attggacaat tgaagaattt gaaatcgttg 300aatttgagtt ataaccaaat caaaactatt ccgaaagaaa ttgaaaaact acaaaagttg 360caatcgttgg gtttggataa taaccaactt acaactcttc cacaagaaat tggacaacta 420cagaacttgc aatcgttgga tttgagtact aaccgactca caactcttcc acaagaaatt 480ggacatctac agaatttgca agatttgtat ttggtttcta accaactcac aattcttcct 540aacgaaatcg gacaattaaa gaacttgcaa acgttgaatt tgcgtaataa ccgactcact 600actctttcta aagaaattga gcaactacag aatttgaaat cgttggattt gcgttctaac 660caacttacta cttttcctaa ggaaatcgag caattgaaga atttgcaagt tttggatttg 720ggttctaacc aactcactac tcttccagag ggaatcggac aattgaagaa cttgcaaaca 780ttggatttgg attctaacca acttacaact cttccacaag aaatcggaca actacagaat 840ttgcaagaat tgtttttaaa caacaatcaa ctctcatccc aagaaaaaaa aagaattcga 900aaacttcttc caaagtgcca aatttacttt gactaa 936<210>52<211>311<212>PRT<213>钩端螺旋体(Leptospira interrogans)<400>52Met Asn Phe Arg Ile Thr Leu Asn Tyr Leu Gln Lys Ile Thr Ile Gly1 5 10 15Leu Leu Phe Leu Met His Leu Ser Cys Glu Ile Gln Ala Glu Glu Phe
20 25 30Glu Gln Gln Glu Thr Tyr Thr Asp Leu Thr Lys Ala Leu Gln Asn Pro
35 40 45Leu Lys Val Arg Thr Leu Asp Leu Ser Ala Asn Arg Phe Lys Thr Leu
50 55 60Pro Lys Glu Ile Gly Lys Leu Lys Asn Leu Gln Glu Leu Asn Leu Asn65 70 75 80Lys Asn Gln Leu Thr Thr Leu Pro Gln Glu Ile Gly Gln Leu Lys Asn
85 90 95Leu Lys Ser Leu Asn Leu Ser Tyr Asn Gln Ile Lys Thr Ile Pro Lys
100 105 110Glu Ile Glu Lys Leu Gln Lys Leu Gln Ser Leu Gly Leu Asp Asn Asn
115 120 125Gln Leu Thr Thr Leu Pro Gln Glu Ile Gly Gln Leu Gln Asn Leu Gln
130 135 140Ser Leu Asp Leu Ser Thr Asn Arg Leu Thr Thr Leu Pro Gln Glu Ile145 150 155 160Gly His Leu Gln Asn Leu Gln Asp Leu Tyr Leu Val Ser Asn Gln Leu
165 170 175Thr Ile Leu Pro Asn Glu Ile Gly Gln Leu Lys Asn Leu Gln Thr Leu
180 185 190Asn Leu Arg Asn Asn Arg Leu Thr Thr Leu Ser Lys Glu Ile Glu Gln
195 200 205Leu Gln Asn Leu Lys Ser Leu Asp Leu Arg Ser Asn Gln Leu Thr Thr
210 215 220Phe Pro Lys Glu Ile Glu Gln Leu Lys Asn Leu Gln Val Leu Asp Leu225 230 235 240Gly Ser Asn Gln Leu Thr Thr Leu Pro Glu Gly Ile Gly Gln Leu Lys
245 250 255Asn Leu Gln Thr Leu Asp Leu Asp Ser Asn Gln Leu Thr Thr Leu Pro
260 265 270Gln Glu Ile Gly Gln Leu Gln Asn Leu Gln Glu Leu Phe Leu Asn Asn
275 280 285Asn Gln Leu Ser Ser Gln Glu Lys Lys Arg Ile Arg Lys Leu Leu Pro
290 295 300Lys Cys Gln Ile Tyr Phe Asp305 310<210>53<211>990<212>DNA<213>钩端螺旋体(Leptospira interrogans)<400>53atgaattttc gcataacttt gaattactta caaaaaataa taattggtct ttttctttct 60tgcaagattc aaacggaaaa agaagaacca ggaacttatt gggatttaac aaaagctctc 120caaaatcctt tggatgttcg aattttgaat ttgagtgaac aaaaactcac aactcttcct 180aaggaaatcg ggcaactaaa gaatctacaa gagttgaatt tgggtaataa ccaaattaca 240atacttccta aggagattgg acaattacag aatttgcaat tgttgggttt atattacaac 300caacttacaa ttcttcctaa ggaaatcgag caattgaaga acttgcaagc attatatttg 360ggtaataacc aaattacaat acttcctaag gagattagac aactacagaa tttgaaagtg 420ttgtttttga gtaataacca acttacaact cttcctaagg aaatcgagca attgaagaac 480ttgcaaactt tgtatttggg taataaccga cttacaactt ttccgaaaga aatcgaacaa 540ttgaagaatt tgcaattgtt gtatttatat gataaccaac tcacagttct tccacaagaa 600ataaaacaac tgaaaaattt acaattgttg gatttaagtt ataaccaact caaaactctt 660cctaaggaaa tcgagcaatt gaagaacttg caagagttga atttgggtta taaccaactc 720acagttcttc ctaaggaaat cgagcaattg aagaacttgc aaacattata tttgggttat 780aaccaactca ctactcttcc gaaagaaatt ggacaactac agaatttgaa agtgttgttt 840ttgaataata accaactcac tactcttccg aaagaaatcg gacaactgaa gaacttgcaa 900gaattgtatt taaacaacaa tcaactctca atcgaagaaa aagaaagaat ccaaaagctt 960attccaaagt gccaaatttc ctttgaataa 990<210>54<211>329<212>PRT<213>钩端螺旋体(Leptospira interrogans)<400>54Met Asn Phe Arg Ile Thr Leu Asn Tyr Leu Gln Lys Ile Ile Ile Gly1 5 10 15Leu Phe Leu Ser Cys Lys Ile Gln Thr Glu Lys Glu Glu Pro Gly Thr
20 25 30Tyr Trp Asp Leu Thr Lys Ala Leu Gln Asn Pro Leu Asp Val Arg Ile
35 40 45Leu Asn Leu Ser Glu Gln Lys Leu Thr Thr Leu Pro Lys Glu Ile Gly
50 55 60Gln Leu Lys Asn Leu Gln Glu Leu Asn Leu Gly Asn Asn Gln Ile Thr65 70 75 80Ile Leu Pro Lys Glu Ile Gly Gln Leu Gln Asn Leu Gln Leu Leu Gly
85 90 95Leu Tyr Tyr Asn Gln Leu Thr Ile Leu Pro Lys Glu Ile Glu Gln Leu
100 105 110Lys Asn Leu Gln Ala Leu Tyr Leu Gly Asn Asn Gln Ile Thr Ile Leu
115 120 125Pro Lys Glu Ile Arg Gln Leu Gln Asn Leu Lys Val Leu Phe Leu Ser
130 135 140Asn Asn Gln Leu Thr Thr Leu Pro Lys Glu Ile Glu Gln Leu Lys Asn145 150 155 160Leu Gln Thr Leu Tyr Leu Gly Asn Asn Arg Leu Thr Thr Phe Pro Lys
165 170 175Glu Ile Glu Gln Leu Lys Asn Leu Gln Leu Leu Tyr Leu Tyr Asp Asn
180 185 190Gln Leu Thr Val Leu Pro Gln Glu Ile Lys Gln Leu Lys Asn Leu Gln
195 200 205Leu Leu Asp Leu Ser Tyr Asn Gln Leu Lys Thr Leu Pro Lys Glu Ile
210 215 220Glu Gln Leu Lys Asn Leu Gln Glu Leu Asn Leu Gly Tyr Asn Gln Leu225 230 235 240Thr Val Leu Pro Lys Glu Ile Glu Gln Leu Lys Asn Leu Gln Thr Leu
245 250 255Tyr Leu Gly Tyr Asn Gln Leu Thr Thr Leu Pro Lys Glu Ile Gly Gln
260 265 270Leu Gln Asn Leu Lys Val Leu Phe Leu Asn Asn Asn Gln Leu Thr Thr
275 280 285Leu Pro Lys Glu Ile Gly Gln Leu Lys Asn Leu Gln Glu Leu Tyr Leu
290 295 300Asn Asn Asn Gln Leu Ser Ile Glu Glu Lys Glu Arg Ile Gln Lys Leu305 310 315 320Ile Pro Lys Cys Gln Ile Ser Phe Glu
325
Claims (9)
1.一种分离的钩端螺旋体致病相关蛋白多肽，其特征在于，它包含：具有选自SEQ ID NO：2、4、6、8、10、12、14、16、18、20、22、24、26、28、30、32、34、36、38、40、42、44、46、48、50、52、54的氨基酸序列的多肽、或其保守性变异多肽、或其活性片段、或其活性衍生物。
2.如权利要求1所述的多肽，其特征在于，该多肽是具有SEQ ID NO：2、4、6、8、10、12、14、16、18、20、22、24、26、28、30、32、34、36、38、40、42、44、46、48、50、52、54的氨基酸序列的多肽。
3.一种分离的多核苷酸，其特征在于，它包含一核苷酸序列，该核苷酸序列与选自下组的一种核苷酸序列有至少70％相同性：
(a)编码如权利要求1和2所述多肽的多核苷酸；
(b)与多核苷酸(a)互补的多核苷酸。
4.如权利要求3所述的多核苷酸，其特征在于，该多核苷酸编码具有SEQ IDNO：2、4、6、8、10、12、14、16、18、20、22、24、26、28、30、32、34、36、38、40、42、44、46、48、50、52、54所示氨基酸序列的多肽。
5.如权利要求3所述的多核苷酸，其特征在于，该多核苷酸的序列选自下组：
(a)具有SEQ ID NO：1、3、5、7、9、11、13、15、17、19、21、23、25、
27、29、31、33、35、37、39、41、43、45、47、49、51、53。
6.一种载体，其特征在于，它含有权利要求3所述的多核苷酸。
7.一种遗传工程化的宿主细胞，其特征在于，它含有权利要求6所述的载体。
8.一种钩端螺旋体致病相关蛋白多肽的制备方法，其特征在于，该方法包含：
(a)在适合表达钩端螺旋体致病相关蛋白的条件下，培养权利要求7所述的宿主细胞；
(b)从培养物中分离出钩端螺旋体致病相关蛋白多肽。
9.一种能与权利要求1所述的钩端螺旋体致病相关蛋白特异性结合的抗体。
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| CN111518823A (zh) * | 2020-04-24 | 2020-08-11 | 杭州医学院 | 一种钩端螺旋体黏附蛋白rvWFA3-1的制备方法和应用 |
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| CN111518823A (zh) * | 2020-04-24 | 2020-08-11 | 杭州医学院 | 一种钩端螺旋体黏附蛋白rvWFA3-1的制备方法和应用 |
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