鉤端螺旋體致病相關蛋白及其編碼序列的製作方法

2023-04-23 15:16:31

專利名稱：鉤端螺旋體致病相關蛋白及其編碼序列的製作方法
技術領域：
本發明屬於生物技術和醫學領域，具體地說，本發明涉及新的編碼鉤端螺旋體致病相關蛋白的多核苷酸，以及此多核苷酸編碼的多肽。本發明還涉及此多核苷酸和多肽的用途和製備。
背景技術：
鉤端螺旋體病是一種分布廣泛的人獸共患病，主要分布在中國南方以及東南亞、南亞等發展中國家和地區。在中國，本病1995年列入法定傳染病。1955-1993年，年平均發病率為十萬分之七，年平均死亡率佔發病人數的1.02％。
中國鉤端螺旋體病的的流行菌群以黃疸出血群和波摩那群為主。黃疸出血群有15個血清型，其中以賴型最為常見。
中國雖在該病的防治方面取得了巨大的成功，但對該病的致病免疫的機理、疫苗的高效保護等問題仍未解決。
鉤端螺旋體(Leptospira sp.)屬格蘭氏陰性菌。人類感染致病性鉤端螺旋體後有發熱、頭痛、肌肉酸痛、黃疸、肝腎功能損傷及體內各臟器的瀰漫性出血，嚴重者導致死亡。急性症狀患者常出現肺出血而暴死。
對於組織出血機制進行研究發現，人體內出血、凝血處於一個相互平衡的體系中。在正常情況下，參與凝血的因子有PAF(促進血小板凝集)，vWF(借導血小板凝集在血管內皮細胞破損後暴露出來的膠原蛋白上的一個蛋白)，凝血酶(促進血小板凝集)等因子，它們能促進血小板的凝集以及促進血小板堵塞在血管內皮組織破損處，而阻止出血。
然而目前為止，仍沒有公開過鉤端螺旋體致病相關蛋白。因此，本領域迫切需要開發新的鉤端螺旋體致病相關蛋白基因，以便為研究鉤端螺旋體病的致病機制及診斷提供了基礎。

發明內容
本發明目的是提供新的鉤端螺旋體致病相關蛋白以及其片段、類似物和衍生物。
本發明另一目的是提供編碼這些多肽的多核苷酸。
本發明另一目的是提供生產這些多肽的方法以及該多肽和編碼序列的用途。
在本發明的第一方面，提供新穎的分離出的致病相關蛋白多肽，它包含具有SEQ ID NO2、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胺基酸序列的多肽、或其保守性變異多肽、或其活性片段、或其活性衍生物。較佳地，該多肽是具有SEQ IDNO2、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胺基酸序列的多肽，更佳地該多肽不含信號肽序列。
在本發明的第二方面，提供分離的編碼上述多肽的多核苷酸，該多核苷酸包含一核苷酸序列，該核苷酸序列與選自下組的一種核苷酸序列有至少70％相同性(a)編碼上述鉤端螺旋體致病相關蛋白多肽的多核苷酸；和(b)與多核苷酸(a)互補的多核苷酸。較佳地，該多核苷酸編碼具有SEQ ID NO2、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所示胺基酸序列的多肽。更佳地，該多核苷酸的序列是選自下組的一種(a)具有SEQ ID NO1、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。
在本發明的第三方面，提供了含有上述多核苷酸的載體，以及被該載體轉化或轉導的宿主細胞或者被上述多核苷酸直接轉化或轉導的宿主細胞。
在本發明的第四方面，提供了製備鉤端螺旋體致病相關蛋白多肽的方法，該方法包含(a)在適合表達鉤端螺旋體致病相關蛋白的條件下，培養上述被轉化或轉導的宿主細胞；(b)從培養物中分離出鉤端螺旋體致病相關蛋白多肽。
在本發明的第五方面，提供了與上述的鉤端螺旋體致病相關蛋白多肽特異性結合的抗體。還提供了可用於檢測的核酸分子。
在本發明的第六方面，提供了檢測樣品中是否存在致病相關蛋白的方法，它包括將樣品與致病相關蛋白的特異性抗體接觸，觀察是否形成抗體複合物，形成了抗體複合物就表示樣品中存在致病相關蛋白。
在本發明的第七方面，提供了本發明多肽和編碼序列的用途。例如本發明的鉤端螺旋體致病相關蛋白的編碼序列或其片段，可被作為引物用於PCR擴增反應，或者作為探針用於雜交反應，或者用於製造基因晶片或微陣列。
本發明的其它方面由於本文的技術的公開，對本領域的技術人員而言是顯而易見的。


下列附圖用於說明本發明的具體實施方案，而不用於限定由權利要求書所界定的本發明範圍。
圖1A和B顯示了血小板激活因子乙醯水解酶PAF_ah和已知蛋白1WAB的三維結構。
圖2顯示了本發明血小板激活因子乙醯水解酶功能結構域。圖中的PD009782是乙醯水解酶PAF-AHγ亞基。
具體實施例方式
本發明人經過廣泛而深入的研究，從問號鉤端螺旋體(Leptospirainterrogans)黃疸出血群賴型賴株中分離純化和鑑定了一類新的致病相關蛋白。在此基礎上完成了本發明。
本發明人在鉤端螺旋體賴株基因組資料庫中鑑別出下列致病相關蛋白，包括(1)ColA(膠原酶，水解膠原蛋白)，(2)PAF-AH(血小板激活因子乙醯水解酶，抑制血小板凝集)，(3)vWF A結構域，以及(4)與凝血酶或者GPIb結合的蛋白。這些基因的產物均抑制血小板的凝集而破壞凝血系統，從而出現大出血現象。通過這四種因子的生物活性進行研究，有助於研究鉤端螺旋體病的致病機制。此外，本發明的蛋白還可作為篩選抗凝血的藥物靶點，開發抗凝血、抗血栓藥物，以及用於診斷出血性疾病。
在本發明中，術語「鉤端螺旋體致病相關蛋白多肽」或「鉤端螺旋體致病相關蛋白」可互換使用，都指具有鉤端螺旋體致病相關蛋白胺基酸序列(SEQ ID NO2、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。它們包括含有或不含起始甲硫氨酸的鉤端螺旋體致病相關蛋白，以及含有或不含有信號肽的成熟蛋白。
如本文所用，「分離的」是指物質從其原始環境中分離出來(如果是天然的物質，原始環境即是天然環境)。如活體細胞內的天然狀態下的多聚核苷酸和多肽是沒有分離純化的，但同樣的多聚核苷酸或多肽如從天然狀態中同存在的其他物質中分開，則為分離純化的。
如本文所用，「分離的致病相關蛋白或多肽」是指致病相關蛋白多肽基本上不含天然與其相關的其它蛋白、脂類、糖類或其它物質。本領域的技術人員能用標準的蛋白質純化技術純化致病相關蛋白。
本發明的多肽可以是重組多肽、天然多肽、合成多肽，優選重組多肽。本發明的多肽可以是天然純化的產物，或是化學合成的產物，或使用重組技術從原核或真核宿主(例如，細菌、酵母、高等植物、昆蟲和哺乳動物細胞)中產生。根據重組生產方案所用的宿主，本發明多肽可以是糖基化的，但優選是非糖基化的。
本發明還包括鉤端螺旋體致病相關蛋白的片段、衍生物和類似物。如本文所用，術語「片段」、「衍生物」和「類似物」是指基本上保持本發明的天然鉤端螺旋體致病相關蛋白相同的生物學功能或活性的多肽。本發明的多肽片段、衍生物或類似物可以是(i)有一個或多個保守或非保守性胺基酸殘基(優選保守性胺基酸殘基)被取代的多肽，而這樣的取代的胺基酸殘基可以是也可以不是由遺傳密碼編碼的，或(ii)在一個或多個胺基酸殘基中具有取代基團的多肽，或(iii)成熟多肽與另一個化合物(比如延長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)附加的胺基酸序列融合到此多肽序列而形成的多肽(如前導序列或分泌序列或用來純化此多肽的序列或蛋白原序列，或與抗原IgG片段的形成的融合蛋白)。根據本文的教導，這些片段、衍生物和類似物屬於本領域熟練技術人員公知的範圍。
在本發明中，術語「鉤端螺旋體致病相關蛋白多肽」指具有鉤端螺旋體致病相關蛋白活性或免疫原性的SEQ ID NO2、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。該術語還包括具有與鉤端螺旋體致病相關蛋白相同功能的、上述序列的變異形式。這些變異形式包括(但並不限於)若干個(通常為1-50個，較佳地1-30個，更佳地1-20個，最佳地1-10個)胺基酸的缺失、插入和/或取代，以及在C末端和/或N末端添加一個或數個(通常為10個以內，更佳地為5個以內)胺基酸。該術語還包括鉤端螺旋體致病相關蛋白的活性片段和活性衍生物。
該多肽的變異形式包括同源序列、保守性變異體、等位變異體、天然突變體、誘導突變體、在高或低的嚴緊度條件下能與鉤端螺旋體致病相關蛋白DNA雜交的DNA所編碼的蛋白、以及利用抗鉤端螺旋體致病相關蛋白多肽的抗血清獲得的多肽或蛋白。本發明還提供了其他多肽，如包含鉤端螺旋體致病相關蛋白多肽或其片段的融合蛋白。除了幾乎全長的多肽外，本發明還包括了鉤端螺旋體致病相關蛋白多肽的可溶性片段。通常，該片段具有鉤端螺旋體致病相關蛋白多肽序列的至少約10個連續胺基酸，通常至少約30個連續胺基酸，較佳地至少約50個連續胺基酸，更佳地至少約80個連續胺基酸，最佳地至少約100個連續胺基酸。這些片段也可用作免疫原以誘導針對鉤端螺旋體的免疫應答反應。
發明還提供鉤端螺旋體致病相關蛋白或多肽的類似物。這些類似物與天然鉤端螺旋體致病相關蛋白多肽的差別可以是胺基酸序列上的差異，也可以是不影響序列的修飾形式上的差異，或者兼而有之。這些多肽包括天然或誘導的遺傳變異體。誘導變異體可以通過各種技術得到，如通過輻射或暴露於誘變劑而產生隨機誘變，還可通過定點誘變法或其他已知分子生物學的技術。類似物還包括具有不同於天然L-胺基酸的殘基(如D-胺基酸)的類似物，以及具有非天然存在的或合成的胺基酸(如β、γ-胺基酸)的類似物。應理解，本發明的多肽並不限於上述例舉的代表性的多肽。
修飾(通常不改變一級結構)形式包括體內或體外的多肽的化學衍生形式如乙醯化或羧基化。修飾還包括糖基化，如那些在多肽的合成和加工中或進一步加工步驟中進行糖基化修飾而產生的多肽。修飾形式還包括具有磷酸化胺基酸殘基(如磷酸酪氨酸，磷酸絲氨酸，磷酸蘇氨酸)的序列。還包括被修飾從而提高了其抗蛋白水解性能或優化了溶解性能的多肽。
在本發明中，「鉤端螺旋體致病相關蛋白保守性變異多肽」指與SEQ ID NO2、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的胺基酸序列相比，有至多10個，較佳地至多8個，更佳地至多5個，最佳地至多3個胺基酸被性質相似或相近的胺基酸所替換而形成多肽。這些保守性變異多肽最好根據表1進行胺基酸替換而產生。
表1

本發明的多核苷酸可以是DNA形式或RNA形式。DNA形式包括cDNA、基因組DNA或人工合成的DNA。DNA可以是單鏈的或是雙鏈的。DNA可以是編碼鏈或非編碼鏈。以致病相關蛋白-膠原蛋白酶lep930-167為例，編碼成熟多肽的編碼區序列可以與SEQ ID NO1所示的編碼區序列相同或者是簡併的變異體。如本文所用，以lep930-167為例，「簡併的變異體」在本發明中是指編碼具有SEQ ID NO2的蛋白質，但與SEQ ID NO1所示的編碼區序列有差別的核酸序列。本發明的其他蛋白依此類推。本發明的核苷酸序列可以根據密碼子的對應關係，根據SEQ ID NO2、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的胺基酸推導出。
編碼成熟多肽的多核苷酸包括只編碼成熟多肽的編碼序列；成熟多肽的編碼序列和各種附加編碼序列；成熟多肽的編碼序列(和任選的附加編碼序列)以及非編碼序列。
術語「編碼多肽的多核苷酸」可以是包括編碼此多肽的多核苷酸，也可以是還包括附加編碼和/或非編碼序列的多核苷酸。
本發明還涉及上述多核苷酸的變異體，其編碼與本發明有相同的胺基酸序列的多肽或多肽的片段、類似物和衍生物。此多核苷酸的變異體可以是天然發生的等位變異體或非天然發生的變異體。這些核苷酸變異體包括取代變異體、缺失變異體和插入變異體。如本領域所知的，等位變異體是一個多核苷酸的替換形式，它可能是一個或多個核苷酸的取代、缺失或插入，但不會從實質上改變其編碼的多肽的功能。
本發明還涉及與上述的序列雜交且兩個序列之間具有至少50％，較佳地至少70％，更佳地至少80％相同性的多核苷酸。本發明特別涉及在嚴格條件下與本發明所述多核苷酸可雜交的多核苷酸。在本發明中，「嚴格條件」是指(1)在較低離子強度和較高溫度下的雜交和洗脫，如0.2×SSC，0.1％SDS，60℃；或(2)雜交時加有變性劑，如50％(v/v)甲醯胺，0.1％小牛血清/0.1％Ficoll，42℃等；或(3)僅在兩條序列之間的相同性至少在90％以上，更好是95％以上時才發生雜交。並且，可雜交的多核苷酸編碼的多肽與成熟的鉤端螺旋體致病相關蛋白有相同的生物學功能和活性。
本發明還涉及與上述的序列雜交的核酸片段。如本文所用，「核酸片段」的長度至少含15個核苷酸，較好是至少30個核苷酸，更好是至少50個核苷酸，最好是至少100個核苷酸以上。核酸片段可用於核酸的擴增技術(如PCR)以確定和/或分離編碼致病相關蛋白的多聚核苷酸。
本發明的鉤端螺旋體致病相關蛋白核苷酸全長序列或其片段通常可以用PCR擴增法、重組法或人工合成的方法獲得。對於PCR擴增法，可根據本發明所公開的有關核苷酸序列，尤其是開放閱讀框序列來設計引物，並用市售的DNA文庫或按本領域技術人員已知的常規方法所製備的DNA文庫作為模板，擴增而得有關序列。當序列較長時，常常需要進行兩次或多次PCR擴增，然後再將各次擴增出的片段按正確次序拼接在一起。
一旦獲得了有關的序列，就可以用重組法來大批量地獲得有關序列。這通常是將其克隆入載體，再轉入細胞，然後通過常規方法從增殖後的宿主細胞中分離得到有關序列。
此外，還可用人工合成的方法來合成有關序列，尤其是片段長度較短時。通常，通過先合成多個小片段，然後再進行連接可獲得序列很長的片段。
目前，已經可以完全通過化學合成來得到編碼本發明蛋白(或其片段，或其衍生物)的DNA序列。然後可將該DNA序列引入本領域中已知的各種現有的DNA分子(或如載體)和細胞中。此外，還可通過化學合成將突變引入本發明蛋白序列中。
本發明也涉及包含本發明的多核苷酸的載體，以及用本發明的載體或致病相關蛋白編碼序列經基因工程產生的宿主細胞，以及經重組技術產生本發明所述多肽的方法。
通過常規重組DNA技術(Science，1984；2241431)，可利用本發明的多聚核苷酸序列可用來表達或生產重組的致病相關蛋白多肽。一般來說有以下步驟(1).用本發明的編碼鉤端螺旋體致病相關蛋白多肽的多核苷酸(或變異體)，或用含有該多核苷酸的重組表達載體轉化或轉導合適的宿主細胞；(2).在合適的培養基中培養的宿主細胞；(3).從培養基或細胞中分離、純化蛋白質。
本發明中，鉤端螺旋體致病相關蛋白多核苷酸序列可插入到重組表達載體中。術語「重組表達載體」指本領域熟知的細菌質粒、噬菌體、酵母質粒、植物細胞病毒、哺乳動物細胞病毒如腺病毒、逆轉錄病毒或其他載體。在本發明中適用的載體包括但不限於；在細菌中表達的基於T7的表達載體；在哺乳動物細胞中表達的pMSXND表達載體和在昆蟲細胞中表達的來源於杆狀病毒的載體。總之，只要能在宿主體內複製和穩定，任何質粒和載體都可以用。表達載體的一個重要特徵是通常含有複製起點、啟動子、標記基因和翻譯控制元件。
本領域的技術人員熟知的方法能用於構建含鉤端螺旋體致病相關蛋白編碼DNA序列和合適的轉錄/翻譯控制信號的表達載體。這些方法包括體外重組DNA技術、DNA合成技術、體內重組技術等。所述的DNA序列可有效連接到表達載體中的適當啟動子上，以指導mRNA合成。表達載體還包括翻譯起始用的核糖體結合位點和轉錄終止子。
此外，表達載體優選地包含一個或多個選擇性標記基因，以提供用於選擇轉化的宿主細胞的表型性狀，如真核細胞培養用的二氫葉酸還原酶、新黴素抗性以及綠色螢光蛋白(GFP)，或用於大腸桿菌的四環素或氨苄青黴素抗性。
包含上述的適當DNA序列以及適當啟動子或者控制序列的載體，可以用於轉化適當的宿主細胞，以使其能夠表達蛋白質。
宿主細胞可以是原核細胞，如細菌細胞；或是低等真核細胞，如酵母細胞；或是高等真核細胞，如哺乳動物細胞。代表性例子有大腸桿菌，鏈黴菌屬；鼠傷寒沙門氏菌的細菌細胞；真菌細胞如酵母；植物細胞；果蠅S2或Sf9的昆蟲細胞；CHO的動物細胞等。
用重組DNA轉化宿主細胞可用本領域技術人員熟知的常規技術進行。當宿主為原核生物如大腸桿菌時，能吸收DNA的感受態細胞可在指數生長期後收穫，用CaCl2法處理，所用的步驟在本領域眾所周知。另一種方法是使用MgCl2。如果需要，轉化也可用電穿孔的方法進行。當宿主是真核生物，可選用如下的DNA轉染方法磷酸鈣共沉澱法，常規機械方法如顯微注射、電穿孔、脂質體包裝等。
獲得的轉化子可以用常規方法培養，表達本發明的基因所編碼的多肽。根據所用的宿主細胞，培養中所用的培養基可選自各種常規培養基。在適於宿主細胞生長的條件下進行培養。當宿主細胞生長到適當的細胞密度後，用合適的方法(如溫度轉換或化學誘導)誘導選擇的啟動子，將細胞再培養一段時間。
在上面的方法中的重組多肽可在細胞內、或在細胞膜上表達、或分泌到細胞外。如果需要，可利用其物理的、化學的和其它特性通過各種分離方法分離和純化重組的蛋白。這些方法是本領域技術人員所熟知的。這些方法的例子包括但並不限於常規的復性處理、用蛋白沉澱劑處理(鹽析方法)、離心、滲透破菌、超處理、超離心、分子篩層析(凝膠過濾)、吸附層析、離子交換層析、高效液相層析(HPLC)和其它各種液相層析技術及這些方法的結合。
另一方面，本發明還包括對鉤端螺旋體致病相關蛋白DNA或是其片段編碼的多肽具有特異性的多克隆抗體和單克隆抗體，尤其是單克隆抗體。這裡，「特異性」是指抗體能結合於鉤端螺旋體致病相關蛋白基因產物或片段。較佳地，指那些能與鉤端螺旋體致病相關蛋白基因產物或片段結合但不識別和結合於其它非相關抗原分子的抗體。本發明還包括那些能與修飾或未經修飾形式的鉤端螺旋體致病相關蛋白基因產物結合的抗體。
本發明不僅包括完整的單克隆或多克隆抗體，而且還包括具有免疫活性的抗體片段，如Fab′或(Fab)2片段；抗體重鏈；抗體輕鏈；或嵌合抗體。
本發明的抗體可以通過本領域內技術人員已知的各種技術進行製備。例如，純化的鉤端螺旋體致病相關蛋白基因產物或者其具有抗原性的片段，可被施用於動物以誘導多克隆抗體的產生。與之相似的，表達鉤端螺旋體致病相關蛋白或其具有抗原性的片段的細胞可用來免疫動物來生產抗體。本發明的抗體也可以是單克隆抗體。此類單克隆抗體可以利用雜交瘤技術來製備。
下面結合具體實施例，進一步闡述本發明。應理解，這些實施例僅用於說明本發明而不用於限制本發明的範圍。下列實施例中未註明具體條件的實驗方法，通常按照常規條件如Sambrook等人，分子克隆；實驗室手冊(New YorkColdSpring Harbor Laboratory Press，1989)中所述的條件，或按照製造廠商所建議的條件。
實施例1菌體培養1)培養基使用EMJH液體培養基，配製5×的EMJH培養基1000毫升，稱取BSA50g。無水醋酸鈉0.5g，丙酮酸鈉1g，硫酸亞鐵0.25g，1.5％的氯化鎂3.5ml，0.4％的硫酸鋅5ml，0.15的硫酸銅1ml，磷酸氫二鈉12.6g，磷酸二氫鉀1.5g，氯化鈉5g，氯化銨1.25g，0.05％的VB122ml，VB10.025g，調pH為7.2-7.6，定容至1000ml分別以0.7、0.45、0.2nm孔徑的硝酸纖維素膜過濾除菌。使用時，用無菌水其5倍稀釋。
2)培養條件30℃，100轉/分鐘搖動培養40小時。
3)菌株問號鉤端螺旋體黃疸出血群賴型賴株。
實施例2致病相關蛋白的核酸序列和胺基酸序列用常規方法抽提鉤端螺旋體DNA，構建文庫。測定基因組全序列。根據計算機分析結果，合成引物，以抽提的鉤端螺旋體DNA為模板，進行PCR擴增，獲得致病相關蛋白的ORF序列。再次測序驗證，與基因組序列相符。這些ORF編碼具有SEQID NO2、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所示胺基酸序列的多肽。本發明的8種致病相關蛋白的基本情況如下表所示。
表A致病相關蛋白的核酸序列和胺基酸序列


*信號肽一欄中給出的數字是信號肽的起始和終止胺基酸位置。
本發明的鉤端螺旋體致病相關蛋白中，Paf_ah與IWAB的胺基酸序列在一級結構上有41/128(32％)相同，70/128(54％)相似。在三維結構上也極其相似(圖1A和1B)。模擬PDB資料庫中的1WAB(已知的牛腦的PAF-AH)，發現鉤端螺旋體的PAF-AH的N-端Ser27，Asp171，His174與1WAB的Ser47，Asp192，His195的活性位點完全相同。
vWF A Domain LB54與LB55有較高的同源性。
膠原蛋白酶在一級結構上與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。
實施例3蛋白表達用常規基因工程方法將鉤端螺旋體致病相關蛋白基因克隆入融合表達載體pET28b(Novagene公司)，融合表達的受體菌選用大腸桿菌BL21(DE3)。常規條件發酵後，提取重組菌體蛋白，然後進行SDS-PAGE電泳，分子量與預測值基本相同。大腸桿菌中表達的致病相關蛋白(帶有His-tag的融合蛋白)，通過Ni-NTA柱分離純化，得到純的鉤端螺旋體致病相關蛋白(融合蛋白)。
實施例4製備抗體將實施例3中獲得的純化的、重組His-融合蛋白用來免疫動物以產生抗體，具體方法如下將融合蛋白用等量的完全Freund′s佐劑乳化。用200μg乳化過的蛋白，對家兔進行淋巴結注射。一個月後，用等量的非完全Freund′s佐劑乳化的同樣抗原，對家兔以200μg的劑量進行淋巴結注射以加強免疫。每隔14天進行一次加強免疫，至少進行三次。獲得的抗血清的特異反應活性用ELISA檢驗抗體滴度。結果發現，抗體可特異性地與本發明蛋白發生結合。
在本發明提及的所有文獻都在本申請中引用作為參考，就如同每一篇文獻被單獨引用作為參考那樣。此外應理解，在閱讀了本發明的上述講授內容之後，本領域技術人員可以對本發明作各種改動或修改，這些等價形式同樣落於本申請所附權利要求書所限定的範圍。
序列表110中國科學院上海生物工程研究中心120鉤端螺旋體致病相關蛋白及其編碼序列13002123216054170PatentIn version3.121012112667212DNA213鉤端螺旋體(Leptospira interrogans)4001atgataacga atataaagta taagctagca ataacgttaa gtgttcttgt aaccttttcg 60ttaggatgta atagccaaaa taatggctct aaaaatgatc ttagtttgtt tgcaacaatg 120tttgctcagg ctacacaaca aactacaatg agatcgacaa cagtaagcgt tagctcgtta 180aacgaactga taggttttaa acctattcaa gtagaaaacg agcaagactt tgacagaatc 240gcagatggaa aaaaaagaat tcttgttccg tcggttatga ctttcggacc aagtaattct 300tattcgaatg gaactgcaca aaatataact tcaaataatt gtaatggaag ttacgttgca 360cgtttaaaac cgaaagaatt cgtcagattc ttggaaacac atgatataaa ttgtgtggat 420aaatttgttt ggaattacga tcagtattcg gactatattt attcacaaga gaacatgctt 480gaagtagtca atcgactaaa cgatcttgct cctttttaca acggtaacaa cgatttgaac 540tttatacaat tgttcagaat gttttgggcc ggatattatg taaaacattc tcacccttcc 600ctcccatttg atacaaacca aatctctcag gctttagtta ctccaatgca gatttttgca 660agtagtgcac acttcttaga tggaaccaac gatgccggaa aagttttaga gttcttcttt 720acagtagcgg atagtacaaa aattgggcat acgatttatc 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Lys Ala Leu Thr Arg His Ser Asp Cys Ala Asn Leu Arg370 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 His405 410 415Thr Ala Leu Ser Ile Asp Glu Ile Gln Pro Leu Tyr His Ala Leu Lys420 425 430Gln Val Glu Ser Gln Phe Phe Arg Leu Ile Gly Ile His Ala Pro Val435 440 445Ser Gly Asp Thr Thr Asp Ser Ile Thr Met Tyr Val Tyr Lys Ser Arg450 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 Gln485 490 495Arg Thr Pro Ala Glu Ser Ile Tyr Thr Leu Glu Glu Leu Leu Arg His500 505 510Glu Tyr Ser His Tyr Leu Val Gly Arg Phe Ile Ile Pro Gly Leu Trp515 520 525Ser Gln Thr Ser Ala His Asp Asn Glu Arg Leu Thr Trp Phe Glu Glu530 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 Met565 570 575Asp Val Ser Lys Ile Leu Thr Ala Arg Tyr Gly Asp Phe Lys Phe Tyr580 585 590Arg Tyr Ser Gly Asn Phe Phe Asn Tyr Leu Tyr Thr Tyr Lys Lys Asp595 600 605Thr Leu Lys Asp Leu Ile Ser Ala Leu Arg Asp Ser Asn Ile Gln Thr610 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 Leu645 650 655Thr Asp Pro Ser Thr Pro Ala Pro Asp Leu Ala Asn Leu Ser Thr Asn660 665 670Asp Pro Lys Val Ile Arg Ser Phe Phe Arg Lys Thr Lys Thr Gly Leu675 680 685Lys Ala Lys Cys Thr Thr Ala Ala Phe Gly Leu Asn Ser Arg Phe Ser690 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 Thr725 730 735Leu Glu Ser Ser Gly Val Asn Asn Phe Gln Ser Met Asn Cys Arg Ile740 745 750Gly Glu Ile Arg Phe Asp Lys His Leu Asn Leu Phe Arg Pro Phe Thr755 760 765Ile Tyr Ser Cys Glu Gly Pro Leu Ala Tyr Arg Ser Arg Ile Ser Tyr770 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 Thr805 8l0 815Cys Asn Thr Pro Ile Val Thr Asn Ala Phe Ser Asn Thr Ala Thr Tyr820 825 830Glu Glu Met Tyr Gln Lys Leu Asn Gly Glu Leu Asn Glu Leu Lys Ser835 840 845Glu Val Tyr Thr Met Arg Pro Ser Phe Tyr Lys Lys Leu Asp Cys Asn850 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 Phe8852103211690212DNA213鉤端螺旋體(Leptospira interrogans)4003ttgattaaga aatcatatac ggacgattat tctagtccta attttgaatg ttggtctgga60tccggttttc 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 ctatctttaa6902104211229212PRT213鉤端螺旋體(Leptospira interrogans)4004Leu 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 Tyr20 25 30Lys Ser Ala Trp Ala Glu Leu Arg Asn Phe Tyr Lys Ile Glu Asn Gln35 40 45Lys Ile Lys Ser Ala Asn Ile Val Phe Val Gly Asp Ser Leu Ile Gln50 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 Glu85 90 95Glu Asp Val Leu Ser Leu Asn Pro Lys Val Ile Val Leu Glu Ile Gly100 105 110Gly Asn Asp Leu Ile Gln Gly Lys Cys Leu His Ile Thr Glu Thr Asn115 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 Tyr165 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 His195 200 205Val Asn Glu Lys Ala Tyr Arg Ala Trp Ile His Lys Leu Lys Pro Ile210 215 220Leu Leu Pro Tyr Leu2252105211963212DNA213鉤端螺旋體(Leptospira interrogans)4005ttggaaagga atctgggatc aaaaatgaat tttcaatatc cttacgcgct tttacttttg 60attccagttt ggatctgggc ttttatatat tataaaaaca aaatatactt aaaaaaaatg120gaaattcgtt ttccaggaag aagagaaagt actttttcca aatttgaaaa cttagggaaa180tttttaccga tacttcgccc gattgcgatt agtttgatga tactttctct ttccggacca240ggtaaaaaaa ttacctttct acccgacgaa aaagaaggag tagatatatt aatcgccttg300gacgtctcag gttccatgtc tagaagtaga gactttcttc cggaaactag attaggagtt360tctaaaaaac tattaaaaag atttatcgaa aaaagaaata atgatcgtct tggtttagtg420gtttttgcag gtgcggctta tttacaagct cctcttacag gtgatcgaga atctttgagc480gaaattttag aaaccataga agaagaaaca gtgacggaac aagggaccgc aatcggagac540gcgatcattc tttctacgta tcgattgagg aattctaagg ctcgttctaa agtgattgta600ctcatcaccg atggagtttc caatacgggt aaaatcgatc cagtaacagc gactgatctt660gcggaacaaa ttggagcgaa aatttattcc gtcggaattg gaaaagaaga cggttcttat720gaaatcaatt tcgaaatttt gcaagaactt tctgccaata ctggaggcag attttttagg780gcagaagatc cggaggaaat gaaagcggtt ctctcttcga tcgattctct tgaaaaagat840cctctgcaaa gtcctcccaa agaaataaga gaaactgaat atgaaatctg gctctataga900gccttagcgg ttttactttt agatttaata ttacgtgcgt tttttctgag atattatgta960tga 9632106211320212PRT213鉤端螺旋體(Leptospira interrogans)4006Met 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 Lys20 25 30Asn Lys Ile Tyr Leu Lys Lys Met Glu Ile Arg Phe Pro Gly Arg Arg35 40 45Glu Ser Thr Phe Ser Lys Phe Glu Asn Leu Gly Lys Phe Leu Pro Ile50 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 Ile85 90 95Leu Ile Ala Leu Asp Val Ser Gly Ser Met Ser Arg Ser Arg Asp Phe100 105 110Leu Pro Glu Thr Arg Leu Gly Val Ser Lys Lys Leu Leu Lys Arg Phe115 120 125Ile Glu Lys Arg Asn Asn Asp Arg Leu Gly Leu Val Val Phe Ala Gly130 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 Thr165 170 175Ala Ile Gly Asp Ala Ile Ile Leu Ser Thr Tyr Arg Leu Arg Asn Ser180 185 190Lys Ala Arg Ser Lys Val Ile Val Leu Ile Thr Asp Gly Val Ser Asn195 200 205Thr Gly Lys Ile Asp Pro Val Thr Ala Thr Asp Leu Ala Glu Gln Ile210 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 Gly245 250 255Arg Phe Phe Arg Ala Glu Asp Pro Glu Glu Met Lys Ala Val Leu Ser260 265 270Ser Ile Asp Ser Leu Glu Lys Asp Pro Leu Gln Ser Pro Pro Lys Glu275 280 285Ile Arg Glu Thr Glu Tyr Glu Ile Trp Leu Tyr Arg Ala Leu Ala Val290 295 300Leu Leu Leu Asp Leu Ile Leu Arg Ala Phe Phe Leu Arg Tyr Tyr Val305 310 315 32021072111044212DNA213鉤端螺旋體(Leptospira interrogans)4007atgaatcaag 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 atga10442108211347212PRT213鉤端螺旋體(Leptospira interrogans)4008Met 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 Ser20 25 30Lys Ile Arg Glu Trp Lys Ile Phe Tyr Pro Gly Leu Glu Arg Ser Ser35 40 45Ser Phe Pro Asp Val Arg Trp Val Leu Val Arg Ile Leu Leu Ile Phe50 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 Leu85 90 95Val Asp Val Ser Leu Ser Met Gln Ala Ile Asp Ser Ser Pro Thr Arg100 105 110Leu Ala Lys Phe Lys Glu Val Leu Leu Arg Met Leu Pro Ser Leu Ser115 120 125Gly Asn Arg Phe Gly Met Ile Val Phe Ala Gly Ser Pro Phe Leu Tyr130 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 Ala165 170 175Phe Thr Lys Ala Glu Ala Leu Leu Arg Ser Glu Lys Val Phe Arg Asn180 185 190Arg Ile Leu Ile Leu Val Thr Asp Gly Glu Asp Gln Asn Asp Pro Gln195 200 205Ala Ile Ser Phe Pro Ala Ser Phe Gln Val Trp Ala Ala Gly Thr Glu210 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 Gly245 250 255Ile Ile His Ser Lys Met Asn Arg Thr Phe Leu Lys Asn Leu Ala Asp260 265 270Lys Asn Asp Gly Asn Phe Tyr Ser Leu Asp Ser Asn Pro Pro Asp Ser275 280 285Thr Ile Leu Gln Lys Glu Ile Phe Ser Leu Glu Glu Asn Leu Tyr Ser290 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 Leu325 330 335Leu Phe Pro Lys Arg Lys Ser Val Ser Val Ile340 34521092112826212DNA213鉤端螺旋體(Leptospira interrogans)4009atgatcaaag 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 282621010211941212PRT213鉤端螺旋體(Leptospira interrogans)40010Met 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 Gln20 25 30Leu Asn Gly Ile Ala Gln Lys Val Lys Phe Lys Gly Leu Lys Asp Met35 40 45Ala Asn Thr Phe Met Glu Ser Ile Ala Lys Lys Lys Gly Leu Arg Lys50 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 Phe85 90 95Gly Pro Asp Met Lys Pro Met Ile Lys Asp Glu Asp Gly Lys Ile Lys100 105 110Asp Asp Leu Pro Lys Pro Asn Ser Lys Asp Asp Ser Asp Leu Ala Asn115 120 125Thr Ser Val Glu Glu Trp Lys Leu Met Lys Lys Gln Ile Arg Glu Ile130 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 Leu165 170 175Met Thr His Ile Ala Lys Thr Leu Leu Trp Trp Val Tyr Phe Pro Asp180 185 190Gln Asn Gln Ser Ile Asn Val Phe Arg Leu Thr Glu Glu Lys Asp Tyr195 200 205Ala Asp Ile His Asp Asn Ser Leu Asn Leu Gln Gly Gly Ala Tyr Ile210 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 Ile245 250 255Ser Arg Pro Val Tyr Val Leu Ser Gly Glu Asp Lys Asn Lys Gln Glu260 265 270Ile Pro Gly Phe Thr Asn Gln Lys Val Lys Ala Glu Gln Leu Val Phe275 280 285Gly Leu Glu Lys Met Gly Trp Ser Arg Gly Ser Ala Gly Asp Gly Gly290 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 Gly325 330 335Gln Asp Leu Asp Leu Glu Glu Ala Tyr Phe Val Lys Gly Leu Arg Glu340 345 350Pro Ser Phe Ser Val Gly Lys Glu Thr Lys Leu Thr Leu Glu Glu Ile355 360 365Asn Pro Val Ala Phe Ser Glu Thr Leu His Ser Leu Leu Gln Val Ser370 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 Val405 4l0 415Phe Asp Glu Val Asp Tyr Thr Glu Ile Tyr Asn Gly Phe Ser Thr Ala420 425 430Trp Lys Lys Phe Leu Ser Asn Asp His Gln Ile Thr Arg Ser Lys Asn435 440 445His Lys Ser Ile Ser Lys Ile Thr Lys Met Tyr Ile Arg Ser Ser Asp450 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 Leu485 490 495Lys Asn Leu Lys Lys Ile Glu Leu Gln Asn Trp Asn Leu Lys Asp Leu500 505 510Asn Val Leu Asn Ser Cys Thr Asn Leu Glu Lys Val Glu Leu Val Asp515 520 525Ile Gln Gly Phe Glu Thr Asp Phe Asp Cys Ser Gly Leu Leu Asn Glu530 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 Leu565 570 575Arg Asp Cys Lys Leu Ser Glu Val Pro Glu Ser Ile Gly Asn Leu Lys580 585 590Arg Leu Ile Asn Leu Tyr Leu Asp Lys Asn Gln Leu Thr Thr Leu Pro595 600 605Thr Ser Leu Gly Thr Leu Glu Gln Leu Thr Gln Leu His Ile Asp Ser610 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 Glu645 650 655Ile Gly Asn Leu Thr Ser Leu Glu Asp Leu Asn Leu His Asp Asn Gln660 665 670Leu Ser Ser Leu Pro Thr Thr Ile Gln Asn Leu Ser Ser Leu Thr Lys675 680 685Ile Gly Leu Ser Lys Asn Lys Phe Ser Glu Phe Pro Glu Pro Ile Leu690 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 Asp725 730 735Ile Lys Glu Thr Trp Ile Glu Ser Leu Pro Gln Ser Ile Gln Asn Leu740 745 750Thr Gln Leu Glu Thr Ile Tyr Leu Pro Lys Ala Lys Phe Arg Asn Leu755 760 765Pr0 Asp Phe Leu Ala Asn Met Glu Ser Leu Lys Lys Ile Lys Phe Glu770 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 Asp805 810 815Lys Ile Thr Asn Leu Phe Thr Thr Lys Gly Glu Asp Phe Leu Lys Leu820 825 830Asn Gln Trp Glu Val Lys Leu Lys Ile Ser Glu Ser Tyr Tyr Arg Lys835 840 845Ala Glu Ala Phe Ala Thr Val Val Tyr Ala Leu Ala Thr Ile Leu Lys850 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 Thr885 890 895Gly Gln Lys Glu Pro Met Leu Gln Ser Ile Arg Lys Ser Val Lys Leu900 905 910Gly Thr Lys Ala Asp Glu Phe Thr Lys Glu Lys Asp Phe Ala Ser Tyr915 920 925Leu Lys Asp Pro Asp Phe Leu Glu Ala Ile Arg Lys Asn930 935 9402101l211504212DNA213鉤端螺旋體(Leptospira interrogans)40011gtggttggaa agaaaagcgg agaaattttt atgaagtgcc actttatatt tgttagcatt 60ggattgatct tgtctttgta ctgtagtctg aaaatgacaa aaacgtatca taatttgcag120gatgctcttg ataacccttt ggacgtttta attcttgaga tgagtatgaa tacgcgtata180agtatgggtt tgcatgaact cgaatctctt ccaagggtaa ttgggttatt tcaaaattta240gaaaaattga atttagatgg taatcaactt acgtctcttc cgaaagaaat tggacaactt300caaaatttat tcgaattaaa tctacaagac aacaaactca agactcttcc gaaagaaatc360gagcaacttc aaaatttaca agtgctgcgg ttgtattcga actctttttc attaaaagaa420aaacagaaaa ttcaagagct acttcctaac tgtgaaatcg attttgaaag tgaaggtaaa480agcgaatctt ctttgactga atga 50421012211167212PRT213鉤端螺旋體(Leptospira interrogans)40012Met 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 Met20 25 30Thr Lys Thr Tyr His Asn Leu Gln Asp Ala Leu Asp Asn Pro Leu Asp35 40 45Val Leu Ile Leu Glu Met Ser Met Asn Thr Arg Ile Ser Met Gly Leu50 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 Glu85 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 Val115 120 125Leu Arg Leu Tyr Ser Asn Ser Phe Ser Leu Lys Glu Lys Gln Lys Ile130 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 Glu165210132111272212DNA213鉤端螺旋體(Leptospira interrogans)40013atgaattata gatttacaaa catcctaaag gttataattt tcttttgttt tattttcgaa60atacaatcag 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 127221014211423212PRT213鉤端螺旋體(Leptospira interrogans)40014Met 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 Asn20 25 30Leu Ala Lys Ala Leu Gln Asn Pro Ala Asp Val Arg Asn Leu Asp Leu35 40 45Ser Phe Gln Gly Leu Lys Thr Leu Pro Asn Lys Ile Gly Gln Leu Lys50 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 Asn85 90 95Asn Lys Leu Thr Val Leu Pro Lys Glu Ile Gly Gln Leu Gln Asn Leu100 105 110Gln Glu Leu Ser Leu His Ser Asn Glu Leu Val Asn Leu Pro Lys Glu115 120 125Ile Gly Gln Phe Lys Asn Leu Gln Lys Leu Asn Leu Asp Asn Asn Lys130 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 Glu165 170 175Gln Leu Lys Ser Leu Lys Asn Leu Asp Leu Asn His Asn Glu Leu Thr180 185 190Thr Val Ser Lys Glu Val Met Leu Leu Glu Thr Leu Glu Asn Leu Asp195 200 205Leu Arg Ser Asn Lys Leu Lys Thr Ile Pro Lys Glu Ile Arg Gln Leu210 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 Gly245 250 255Glu Asn Arg Phe Gln Ile Leu Pro Val Glu Ile Leu Glu Leu Lys Asn260 265 270Leu Leu Glu Leu Asn Leu Tyr Tyr Asn Gln Leu Val Glu Phe Pro Lys275 280 285Glu Val Gly Gln Leu Lys Ser Leu Lys Tyr Leu Ser Leu Tyr His Asn290 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 Ile325 330 335Leu Gln Leu Lys Asn Leu Glu Trp Leu Ser Leu Ser Asn Asn Lys Leu340 345 350Asn Ala Leu Pro Lys Glu Ile Gly Gln Leu Lys Lys Leu Gln Arg Leu355 360 365Glu Leu Gly Asn Asn Gln Leu Thr Thr Leu Pro Lys Glu Ile Glu Gln370 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 Asp405 410 415Phe Glu Gly Gly Gly Gly Glu42021015211369212DNA213鉤端螺旋體(Leptospira interrogans)40015atgaaaatca tgaatttaca aaaaatagga accttaatct ttctttgttt tttgagccaa 60ctcaaagccg aagagaaagg tcactatcaa aatctaacca aggctcttaa aaatccaacg120gatgtccaaa ctttagattt aagtaataat caactaataa ctcttccaaa agaaattgga180caattgaaag agttagaatg gctaagttta agcaagaatc aactcaaaac tcttcctaaa240gaaatcgaaa ctttgaaaaa gttagaagaa ttatttttag atgatatacc cgttttaaag300tctcaagaaa aaaagattca gaaattactt cccaaggcgc aaattgactt tatcgaaata360aaaaagtaa36921016211122212PRT213鉤端螺旋體(Leptospira interrogans)40016Met 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 Leu20 25 30Thr Lys Ala Leu Lys Asn Pro Thr Asp Val Gln Thr Leu Asp Leu Ser35 40 45Asn Asn Gln Leu Ile Thr Leu Pro Lys Glu Ile Gly Gln Leu Lys Glu50 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 Ile85 90 95Pro Val Leu Lys Ser Gln Glu Lys Lys Ile Gln Lys Leu Leu Pro Lys100 105 110Ala Gln Ile Asp Phe Ile Glu Ile Lys Lys115 12021017211654212DNA213鉤端螺旋體(Leptospira interrogans)40017atgaatttcc tatctttaaa aaaaatcttt aagctctatt tctttctttg ggtttcgttt 60tttgggatcg attgtaaaaa aaacgcggta gagattttag aagaagcaaa aagaaaatcg120gaatcgattc aaatcttaga tttaggaatg caaaagttaa cctcgattcc agaaggagtt180tgctcttttc cgaatttgac tcagctggat ttacgtttga acagtttgaa ttctcttcct240ggttggattg gagcttgtaa aaatctggaa caaattaacc ttttcggaaa tgatctaaat300acggttccat cttctttttc taaattaaag aatttgaaag tattactctt gggtaataac360gattttactt ttctaccttc ggaacttttg tttctgcctt tgcttaaaac cctgtatttg420gatcaaaaca agcttacgtt gactgaaacg gatgttgaaa ttctagcttc tctttccagt480ttggaagaat tggatcttaa tttgaactct ggaatcaaag tgcttccttc taactataat540aaattgaaaa atctgactca tttaaaaaga ttgaatatta aaaaaacttc attaaagggg600gaggatgctg ataaattaca agcagttctc ccaaatacaa gaatagacta ttga 65421018211217212PRT213鉤端螺旋體(Leptospira interrogans)40018Met 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 Ile20 25 30Leu Glu Glu Ala Lys Arg Lys Ser Glu Ser Ile Gln Ile Leu Asp Leu35 40 45Gly Met Gln Lys Leu Thr Ser Ile Pro Glu Gly Val Cys Ser Phe Pro50 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 Gly85 90 95Asn Asp Leu Asn Thr Val Pro Ser Ser Phe Ser Lys Leu Lys Asn Leu100 105 110Lys Val Leu Leu Leu Gly Asn Asn Asp Phe Thr Phe Leu Pro Ser Glu115 120 125Leu Leu Phe Leu Pro Leu Leu Lys Thr Leu Tyr Leu Asp Gln Asn Lys130 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 Pro165 170 175Ser Asn Tyr Asn Lys Leu Lys Asn Leu Thr His Leu Lys Arg Leu Asn180 185 190Ile Lys Lys Thr Ser Leu Lys Gly Glu Asp Ala Asp Lys Leu Gln Ala195 200 205Val Leu Pro Asn Thr Arg Ile Asp Tyr210 21521019211531212DNA213鉤端螺旋體(Leptospira interrogans)40019gtgtggatgt cttcttatgt ttcggcattc agaacttgga acaaaaagcg cgatcgctta 60aagtattgta aaattacttc tttcattgaa aaaaatattt cgctgaaaca acgcgtatcc120ttttttgttc attgcaataa aattttgatt tggaaaatcc gtcgcatgaa cttttttaca180acacagattc gttttaaacg aatcggaatg gttcttttga tttctatctt ttttttcttc240aacgaggtag aagcacaaga gcaacttgtg gaagtaaaag cttacacaaa tttaatagag300gctttccaaa attcttcgga tgttcgtatt ctgcatttga acggtcaagg atttaaaaat360cttccgagac aaatcggaaa tctacaaaat ttaacagagt tgaatttagg gagcaattca420cttacgacag ttcctaaaga aatcggtcgg ctacgcaact tacaaagttt gtggttgttt480ggaaactggc tggcaacctt cctgaagaag tcggggaact tcacaattta g 53121020211176212PRT213鉤端螺旋體(Leptospira interrogans)40020Met 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 Asn20 25 30Ile Ser Leu Lys Gln Arg Val Ser Phe Phe Val His Cys Asn Lys Ile35 40 45Leu Ile Trp Lys Ile Arg Arg Met Asn Phe Phe Thr Thr Gln Ile Arg50 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 Thr85 90 95Asn Leu Ile Glu Ala Phe Gln Asn Ser Ser Asp Val Arg Ile Leu His100 105 110Leu Asn Gly Gln Gly Phe Lys Asn Leu Pro Arg Gln Ile Gly Asn Leu115 120 125Gln Asn Leu Thr Glu Leu Asn Leu Gly Ser Asn Ser Leu Thr Thr Val130 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 Ile165 170 17521021211300212DNA213鉤端螺旋體(Leptospira interrogans)40021ttgaatctat ctgacaatcg tcccactttc accgttcttc caaaagaaat cggagaacta 60cggaatttaa aagagttgaa tttagagaac aattcactta cgacaattcc taaagaaatc120ggaaaactac aaaatttaaa atggttgcgt ttagggagca attcacttac gacgcttcca180aaagaaatcg gaaaactacg gaatttaaaa gagttggatt tgagttcgaa ttctctttcg240gtcaaagaga aaaagagaat tcgaaagtta cttccaaaat gttctatcat ctatttttga3002102221199212PRT213鉤端螺旋體(Leptospira interrogans)40022Met 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 Ser20 25 30Leu Thr Thr Ile Pro Lys Glu Ile Gly Lys Leu Gln Asn Leu Lys Trp35 40 45Leu Arg Leu Gly Ser Asn Ser Leu Thr Thr Leu Pro Lys Glu Ile Gly50 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 Ile85 90 95Ile Tyr Phe21023211282212DNA213鉤端螺旋體(Leptospira interrogans)40023atggatcgcc gttctgggct caaaaaggca catttttcaa gtgttccgat aaatgagtct 60ttttacttgc aaaaagtatg tttttctgat agagaaaaat tttccgaacc ttaccgcctc120ccaaaaatag agaaaactca acacaacgct ctttatgaat cgtgttgtgg agtgggaact180aagttttaca gaagattttg tcgtaattcc gacagattta tattgagatc aaatacttgt240ggggttggtg atgaatttgt agaaactact gcattttatt aa 2822102421193212PRT213鉤端螺旋體(Leptospira interrogans)40024Met 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 Glu20 25 30Lys Phe Ser Glu Pro Tyr Arg Leu Pro Lys Ile Glu Lys Thr Gln His35 40 45Asn Ala Leu Tyr Glu Ser Cys Cys Gly Val Gly Thr Lys Phe Tyr Arg50 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 Tyr85 90210252111581212DNA213鉤端螺旋體(Leptospira interrogans)40025atgaaggatt attctgtaat tctttctagt ttatcgatta tgaaaataca atacacggct60ttaaatagga 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 a158121026211526212PRT213鉤端螺旋體(Leptospira interrogans)40026Met 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 Arg20 25 30Arg His Val Arg Ile Phe Leu Thr Ile Val Phe Gly Leu Phe Ser Val35 40 45Gln Leu Gln Ala Asn Asp Gly Tyr Glu Thr Phe Thr Asn Leu Lys Glu50 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 Glu85 90 95Leu Glu Tyr Leu Glu Ile Leu Lys Leu Glu Glu Asn Arg Ile Thr Thr100 105 110Leu Pro Arg Glu Ile Asn Lys Leu Lys Asn Leu Lys Glu Leu Tyr Leu115 120 125Asn Gly Asn Lys Leu Thr Ile Val Pro Lys Glu Ile Trp Glu Leu Glu130 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 Gly165 170 175Asn Arg Leu Val Thr Leu Pro Gly Glu Ile Gly Glu Leu Lys Leu Leu180 185 190Glu Glu Leu Asn Leu Glu Asn Asn Arg Ile Lys Ile Leu Pro Asn Glu195 200 205Ile Gly Ala Leu Glu Asn Leu Trp Ile Phe Asn Leu Ser Gly Asn Lys210 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 Glu245 250 255Lys Leu Gln Asp Leu Glu Val Leu Asn Leu Leu Ile Asn Pro Leu Leu260 265 270Ser Lys Glu Arg Lys Lys Ile Gln Ala Leu Leu Pro Asp Cys Asp Ile275 280 285Asp Leu Arg Asp Val Glu Glu Gly Glu Thr Tyr Arg Asn Leu Asn Leu290 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 Arg325 330 335Ser Leu Ser Leu Tyr Asp Thr Ser Leu Val Ala Leu Pro Lys Glu Ile340 345 350Val Arg Leu Lys His Leu Glu His Leu Ser Leu Gly Leu Asn Gln Leu355 360 365Lys Ser Leu Pro Lys Glu Ile Gly Leu Leu Arg Asn Leu Arg Ser Leu370 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 Lys405 410 415Ile Phe Pro Lys Glu Ile Trp Glu Leu Lys Lys Leu Val Ile Leu Asn420 425 430Val Asn Thr Asn Gln Leu Asp Ala Leu Pro Glu Lys Ile Gly Arg Leu435 440 445Lys Gly Leu Gln Met Leu Asp Leu Ser His Asn Arg Leu Thr Thr Leu450 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 Asn485 490 495Leu Arg Lys Leu Thr Leu Tyr Glu Asn Pro Ile Pro Pro Gln Glu Leu500 505 510Asp Lys Ile Arg Lys Leu Leu Pro Asn Cys Glu Ile Arg Phe515 520 52521027211234212DNA213鉤端螺旋體(Leptospira interrogans)40027ttggatttaa atgataatca atttacagtt cttcctaagg aaattggaaa actgaaaaaa 60ttacaaacat tggatttacg gaataaccaa ctcacaaccc ttcctaccga aatcggacaa 120ctacaaaatt tacaatggtt gtatttacag aataatcaac tttcactcaa agaacaagaa 180aggattcgaa agttacttcc actaaaatgt aaaataattt ttgaagttta ttag2342102821177212PRT213鉤端螺旋體(Leptospira interrogans)40028Met 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 Thr20 25 30Thr Leu Pro Thr Glu Ile Gly Gln Leu Gln Asn Leu Gln Trp Leu Tyr35 40 45Leu Gln Asn Asn Gln Leu Ser Leu Lys Glu Gln Glu Arg Ile Arg Lys50 55 60Leu Leu Pro Leu Lys Cys Lys Ile Ile Phe Glu Val Tyr65 70 75210292111497212DNA213鉤端螺旋體(Leptospira interrogans)40029atgaacttcc gcataacgtt aatttactta caaaaaataa caattggtct tttgcttctc60attactcttt 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 149721030211498212PRT213鉤端螺旋體(Leptospira interrogans)40030Met 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 Val20 25 30Glu Pro Gly Thr Tyr Arg Asp Leu Thr Lys Ala Leu Gln Asn Pro Leu35 40 45Asp Val Arg Val Leu Asn Leu Ser Gly Glu Lys Leu Thr Ala Leu Pro50 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 Leu85 90 95Gln Glu Leu Asp Leu Arg Asp Asn Gln Leu Ala Thr Phe Pro Ala Val100 105 110Ile Val Glu Leu Gln Lys Leu Glu Ser Leu Asp Leu Ser Glu Asn Arg115 120 125Leu Ile Ile Leu Pro Asn Glu Ile Gly Arg Leu Gln Asn Leu Gln Asp130 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 Thr165 170 175Ala Leu Pro Lys Glu Ile Gly Gln Leu Lys Asn Leu Gln Thr Leu Asp180 185 190Leu Gln Asp Asn Gln Phe Thr Ile Leu Pro Lys Glu Ile Gly Gln Leu195 200 205Gln Asn Leu Gln Thr Leu Asn Leu Ser Asp Asn Gln Leu Ala Thr Leu210 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 Asn245 250 255Leu Gln Met Leu Cys Ser Pro Glu Asn Arg Leu Thr Ala Leu Pro Lys260 265 270Glu Met Gly Gln Leu Gln Asn Leu Gln Thr Leu Asn Leu Val Asn Asn275 280 285Arg Leu Thr Val Phe Pro Lys Glu Ile Gly Gln Leu Gln Asn Leu Gln290 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 Ala325 330 335Glu Asn Gly Val Tyr Arg Asn Leu Asn Leu Ala Gln Glu Glu Pro Leu340 345 350Lys Val Phe Glu Leu Ser Leu Glu Tyr Lys Asp Phe Ser Gln Ser Phe355 360 365Pro Lys Val Ile Leu Lys Phe Arg Asn Leu Arg Gly Leu Asn Leu Tyr370 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 Ser405 410 415Glu Ile Gly Gln Leu Lys Asn Leu Glu Ala Leu Asn Leu Glu Ala Asn420 425 430Glu Leu Glu Arg Leu Pro Lys Glu Ile Gly Gln Leu Arg Asn Leu Gln435 440 445Lys Leu Ser Leu His Gln Asn Thr Leu Lys Ile Phe Pro Ala Glu Ile450 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 Leu485 490 495Asn Leu210312111287212DNA213鉤端螺旋體(Leptospira interrogans)40031atgaacttcc 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 128721032211428212PRT213鉤端螺旋體(Leptospira interrogans)40032Met 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 Val20 25 30Lys Pro Gly Thr Tyr Met Asp Leu Thr Lys Ala Leu Gln Asn Pro Leu35 40 45Asn Val Arg Val Leu Asp Leu Ser Gly Gln Asn Phe Thr Thr Leu Pro50 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 Leu85 90 95Gln Glu Leu Asn Leu Ser Ser Asn Gln Leu Thr Ile Leu Pro Lys Glu100 105 110Ile Gly Lys Leu Glu Asn Leu Gln Arg Leu Asp Leu Tyr Asp Asn Arg115 120 125Leu Thr Ile Leu Pro Ile Glu Ile Gly Lys Leu Gln Asn Leu Gln Thr130 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 Thr165 170 175Thr Leu Pro Gln Glu Ile Gly Gln Leu Gln Asn Leu Gln Thr Leu Asn180 185 190Leu Lys Ser Asn Gln Leu Thr Thr Leu Phe Lys Glu Ile Glu Gln Leu195 200 205Lys Asn Leu Gln Thr Leu Asn Leu Ser Asp Asn Gln Leu Thr Thr Leu210 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 Asn245 250 255Leu His Thr Leu Asn Leu Ser Asp Asn Gln Leu Thr Thr Leu Pro Ile260 265 270Glu Ile Gly Lys Leu Gln Asn Leu His Thr Leu Asn Leu Ser Gly Asn275 280 285Gln Leu Thr Thr Leu Ser Ile Glu Ile Gly Lys Leu Gln Asn Leu Gln290 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 Leu340 345 350Asn Leu Trp Asn Asn Gln Leu Thr Ala Leu Pro Ile Glu Ile Gly Gln355 360 365Leu Gln Asn Leu Gln Thr Leu Ser Leu Tyr Lys Asn Arg Leu Met Thr370 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 Lys405 410 415Leu Leu Pro Asn Cys Lys Ile Tyr Phe Gly Asp Asn420 425210332111902212DNA213鉤端螺旋體(Leptospira interrogans)40033atgaacttcc gcataacgtt gatttactta caaaagatta caattggtct tttgtttctc60attactcttt 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 190221034211633212PRT213鉤端螺旋體(Leptospira interrogans)40034Met 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 Pro20 25 30Glu Asp Glu Pro Gly Thr Tyr Arg Asp Leu Thr Glu Ala Leu Gln Asn35 40 45Pro Leu Asn Val Arg Val Leu Asn Leu Ser Gly Gln Asn Phe Thr Thr50 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 Gln85 90 95Lys Leu Glu Ser Leu Asp Leu Ser Glu Asn Arg Leu Val Met Leu Pro100 105 110Asn Glu Ile Gly Arg Leu Gln Asn Leu Gln Glu Leu Gly Leu Tyr Lys115 120 125Asn Lys Leu Ile Thr Phe Pro Lys Glu Ile Gly Gln Leu Gln Asn Leu130 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 Arg165 170 175Leu Thr Val Leu Pro Lys Glu Ile Gly Gln Leu Gln Asn Leu Gln Thr180 185 190Leu Asn Leu Gln Asp Asn Gln Leu Ala Thr Leu Pro Val Glu Ile Gly195 200 205Gln Leu Gln Asn Leu Gln Thr Leu Gly Leu Ser Glu Asn Gln Leu Thr210 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 Leu245 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 Gly275 280 285Arg Asn Gln Leu Thr Thr Phe Pro Lys Glu Ile Gly Gln Leu Lys Asn290 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 Asn325 330 335Gln Leu Ala Thr Leu Pro Ala Glu Ile Gly Gln Leu Lys Lys Leu Gln340 345 350Asp Leu Ser Leu Gly Arg Asn Gln Leu Thr Thr Leu Pro Lys Glu Ile355 360 365Gly Gln Leu Lys Asn Leu Tyr Asn Leu Asp Leu Gly Thr Asn Gln Leu370 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 Gln405 410 415Leu Glu Asn Leu Gln Glu Leu Asp Leu Trp Asn Asn Arg Leu Thr Ala420 425 430Leu Pro Lys Glu Ile Gly Gln Leu Lys Asn Leu Glu Asn Leu Glu Leu435 440 445Ser Glu Asn Gln Leu Thr Thr Phe Pro Lys Glu Ile Gly Gln Leu Lys450 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 Tyr485 490 495Asn Arg Leu Val Ile Leu Pro Lys Glu Ile Gly Gln Leu Lys Asn Leu500 505 510Gln Met Leu Asp Leu Cys Tyr Asn Gln Phe Lys Thr Val Ser Lys Glu515 520 525Ile Gly Gln Leu Lys Asn Leu Leu Gln Leu Asn Leu Ser Tyr Asn Gln530 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 Gly565 570 575Gln Leu Lys Asn Leu Tyr Asn Leu Gly Leu Gly Thr Asn Gln Leu Thr580 585 590Thr Leu Pro Lys Glu Ile Gly Gln Leu Lys Lys Leu Gln Trp Leu Leu595 600 605Leu Ser Val Asn Pro Ile Leu Pro Gln Glu Lys Glu Arg Ile Arg Lys610 615 620Leu Leu Pro Asn Cys Glu Ile Arg Phe625 63021035211870212DNA213鉤端螺旋體(Leptospira interrogans)40035atgacgtgct 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 87021036211289212PRT213鉤端螺旋體(Leptospira interrogans)40036Met 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 Ile20 25 30Tyr Leu Gln Lys Ile Thr Ile Phe Leu Leu Phe Leu Ile Tyr Leu Ser35 40 45Cys Glu Ile Gln Ala Glu Ala Pro Glu Asp Glu Pro Gly Thr Tyr Arg50 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 Leu85 90 95Lys Asn Leu Gln Met Leu Asp Leu Cys Tyr Asn Gln Phe Lys Thr Val100 105 110Pro Lys Glu Ile Glu Gln Leu Lys Asn Leu Gln Met Leu Asp Leu Cys115 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 Asn165 170 175Gln Leu Ile Thr Phe Pro Lys Glu Ile Gly Lys Leu Glu Asn Leu Gln180 185 190Val Leu Asn Leu Gly Ser Asn Arg Leu Lys Thr Leu Pro Lys Gly Ile195 200 205Glu Gln Leu Lys Asn Leu Gln Thr Leu Tyr Leu Asn Tyr Asn Gln Leu210 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 Gln245 250 255Leu Gln Asn Leu Arg Lys Leu Thr Leu Tyr Glu Asn Pro Ile Pro Pro260 265 270Gln Glu Leu Asp Lys Ile Arg Lys Leu Leu Pro Asn Cys Glu Ile Arg275 280 285Phe21037211867212DNA213鉤端螺旋體(Leptospira interrogans)40037atgattttta 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 86721038211288212PRT213鉤端螺旋體(Leptospira interrogans)40038Met 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 Lys20 25 30Pro Gly Glu Tyr Thr Asp Leu Glu Glu Ala Phe Lys Asn Pro Lys Asp35 40 45Val Leu Val Leu Asp Tyr Arg Asp Asn Glu Glu Asn Pro Leu Lys Thr50 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 Lys85 90 95Asn Leu Gln Val Leu Ser Leu Asn Gly Asn Arg Leu Glu Thr Ile Pro100 105 110Lys Glu Ile Gly Asn Leu Lys Asn Leu Lys Glu Leu Ser Ile Glu Trp115 120 125Asn Lys Leu Gln Thr Leu Pro Lys Glu Ile Gly Asn Leu Lys Asn Leu130 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 Glu165 170 175Leu Thr Lys Leu Pro Gln Glu Ile Lys Asn Leu Glu Ser Leu Leu Glu180 185 190Ile Tyr Leu Tyr Asp Asn Gln Phe Thr Thr Leu Pro Lys Glu Ile Gly195 200 205Asn Leu Lys Asn Leu Arg Asn Leu Val Leu Gly Arg Asn Gln Leu Ile210 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 Leu245 250 255Lys Gln Leu Ala Arg Leu Ser Leu Lys Gly Asn Gln Phe Pro Ser Glu260 265 270Glu Lys Glu Arg Ile Gln Arg Leu Leu Pro Lys Cys Asn Ile Ser Phe275 280 285210392111281212DNA213鉤端螺旋體(Leptospira interrogans)40039atgaatttcc aatccataaa cattcgttta aataaaagac tcatgattct tttgattctg60atctgttttt cctgcaaact acaagcccaa tctaacgaag tgcaaacata ttatcgtaat120ataacagagg ttcttaaaaa tccacagaat gttcgaattc tgaatttgag cggtagcaaa180ctcacaaccc ttccgggaga aatcggaaaa ctgcaaaatt tacaactgtt gaatttagat240gataaccaac ttatagctct tccgaaagaa attggaaaac ttcaaaattt acaacaattg300catttgagta agaaccaact catggcgctt ccagaagaaa tcggacaact tcagaattta360caaaagttga aattgtatga aaaccagctt acggccattc ccaaagagat tggacaacta420caaaatttac aagagctgaa tttggcacat aaccaactcg caactctacc agaagacatc480gaacaattac aaagattaca aacattgtat ttgggtcata accaatttaa ctctattctc540aaagagattg gacaattaca aaatttagaa tcattaggtt tggatcataa tcaacttaac600gtccttccca aagaaattgg acaacttcga aatttagaat cattaggttt ggatcataac660caacttaacg tccttcccaa agaaattgga caacttcaaa atttacaaat attacatttg720cgtaataacc aactcacaac tcttcccaaa gaaatcgggc agcttcagaa tttacaaaag780ttacttttaa ataaaaacaa actcacaact cttcccaaag aaattggtca gcttcagaat840ttacaaaagt tgaaattgta tgaaaaccaa cttacaaccc ttccgaaaga gattggacaa900ctacaaaatt tacaagagtt ggatttagat ggtaaccaac tcacaactct accagaaaac960atcggacaat 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 128121040211426212PRT213鉤端螺旋體(Leptospira interrogans)40040Met 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 Asn20 25 30Glu Val Gln Thr Tyr Tyr Arg Asn Ile Thr Glu Val Leu Lys Asn Pro35 40 45Gln Asn Val Arg Ile Leu Asn Leu Ser Gly Ser Lys Leu Thr Thr Leu50 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 Asn85 90 95Leu Gln Gln Leu His Leu Ser Lys Asn Gln Leu Met Ala Leu Pro Glu100 105 110Glu Ile Gly Gln Leu Gln Asn Leu Gln Lys Leu Lys Leu Tyr Glu Asn115 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 Phe165 170 175Asn Ser Ile Leu Lys Glu Ile Gly Gln Leu Gln Asn Leu Glu Ser Leu180 185 190Gly Leu Asp His Asn Gln Leu Asn Val Leu Pro Lys Glu Ile Gly Gln195 200 205Leu Arg Asn Leu Glu Ser Leu Gly Leu Asp His Asn Gln Leu Asn Val210 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 Gln245 250 255Asn Leu Gln Lys Leu Leu Leu Asn Lys Asn Lys Leu Thr Thr Leu Pro260 265 270Lys Glu Ile Gly Gln Leu Gln Asn Leu Gln Lys Leu Lys Leu Tyr Glu275 280 285Asn Gln Leu Thr Thr Leu Pro Lys Glu Ile Gly Gln Leu Gln Asn Leu290 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 Gln325 330 335Leu Asn Phe Leu Pro Asn Lys Val Glu Gln Leu Gln Asn Leu Glu Ser340 345 350Leu Asp Leu Glu His Asn Gln Leu Asn Ala Leu Pro Lys Glu Ile Gly355 360 365Lys Leu Gln Lys Leu Gln Thr Leu Asn Leu Lys Tyr Asn Gln Leu Ala370 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 Lys405 410 415Leu Leu Pro Gln Cys Asn Ile Tyr Phe Glu420 425210412111227212DNA213鉤端螺旋體(Leptospira interrogans)40041atgaaaccga atttaatacg acaacgctgt tttcaaaaag tggaaatcct ttttttaata 60tttgtatgtt cccttacgca attccatgcg gaagaaaatc acacaacgaa agaaggtctt120tatacaaatt taaccgaggc tcttaaaaat ccgaatgagg ttcggatttt agatttaagt180agaagcaaac tcaagattct tccgaaagaa atcgggcaac tgcaaaattt acaaatcctg240aattcagaaa ataatcaact cacgaccctt cccaaagaaa tcggaaaatt gcagaacttg300caagagttac atctgcagaa taaccaacta acaacccttc cagaagaaat agggcagtta360cagaatttaa aagtattaca tttgaacaat aatcaactca ctactcttcc agaagaaatc420ggaaaactac aaaatctaca agagttgaat ttatttgtta atcgacttaa catccttcca480aaggaaatcg gacgactaca aaatttacaa gagttatatt tgtctcttaa tcggcttaca540atccttccag aagaaattgg acaattagag agtttacgaa agttgagttt gggtggtaag600aataagccat tcacgattct tccaaaagaa atcacgcagc ttcagaattt acaagagttg660catttgaaat ttaaccggct cacggtcctt cccaaagaaa ttggacaact tcaaaatttg720cggatattag atttgtatca aaaccgactc acgattcttc cgaaagaaat tggacaactc780aaaaatttac tagtattgga tttgtctggt aaccaactta caattcttcc gaaagaaatc840acacaacttc agaatttaca agaattgaat ttggaataca atcgattcga agcttttcca900aaagaaatca cacagtttca gaatttacaa gtgttggatt tgtatcaaaa ccgactcacg960actcttccaa 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 122721042211408212PRT213鉤端螺旋體(Leptospira interrogans)40042Met 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 Glu20 25 30Asn His Thr Thr Lys Glu Gly Leu Tyr Thr Asn Leu Thr Glu Ala Leu35 40 45Lys Asn Pro Asn Glu Val Arg Ile Leu Asp Leu Ser Arg Ser Lys Leu50 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 Lys85 90 95Leu Gln Asn Leu Gln Glu Leu His Leu Gln Asn Asn Gln Leu Thr Thr100 105 110Leu Pro Glu Glu Ile Gly Gln Leu Gln Asn Leu Lys Val Leu His Leu115 120 125Asn Asn Asn Gln Leu Thr Thr Leu Pro Glu Glu Ile Gly Lys Leu Gln130 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 Leu180 185 190Arg Lys Leu Ser Leu Gly Gly Lys Asn Lys Pro Phe Thr Ile Leu Pro195 200 205Lys Glu Ile Thr Gln Leu Gln Asn Leu Gln Glu Leu His Leu Lys Phe210 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 Glu245 250 255Ile Gly Gln Leu Lys Asn Leu Leu Val Leu Asp Leu Ser Gly Asn Gln260 265 270Leu Thr Ile Leu Pro Lys Glu Ile Thr Gln Leu Gln Asn Leu Gln Glu275 280 285Leu Asn Leu Glu Tyr Asn Arg Phe Glu Ala Phe Pro Lys Glu Ile Thr290 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 His325 330 335Leu Ser Arg Asn Gln Leu Thr Thr Leu Pro Lys Glu Ile Gly Arg Leu340 345 350Gln Lys Leu Glu Ser Leu Gly Leu Asp His Asn Gln Leu Ala Thr Leu355 360 365Pro Glu Glu Ile Lys Gln Leu Lys Asn Leu Lys Lys Leu Tyr Leu His370 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 Glu405210432112058212DNA213鉤端螺旋體(Leptospira interrogans)40043atgaaacact 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 tcgaaacgtc600cctgtagtcg atttttctaa ttttattaaa aatcaactca aacagaaacc agaaaaaatc660cgggaagtat tggaagacga agacttttct ctgataaaag acatactgtg taaactaacg720caggaagtaa ttatacaaac tttaaaagca gaagttttta aacaaatccc aaaacaacaa780ccgtattttc taatgttctc ctattaccac gatgaagatg cttctgtatt tttcgattcg840aacgcaaagg aacaaaaaaa tttatactcc atactcaaaa tctcaaaatc ttctctattt900aaatattttt taagcgggga gcaatcttta atcatttcta atgtttcaga aatagatctt960accgatataa 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 205821044211685212PRT213鉤端螺旋體(Leptospira interrogans)40044Met 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 Thr20 25 30Gly Thr Ser Gly Gln Thr Gln Ile Lys Thr Phe Asp Asp Lys Glu Ile35 40 45Cys Leu Lys Glu Ala Lys Lys Leu Leu Ser Glu Lys Leu Lys Lys Gly50 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 Lys85 90 95Ser Lys Ile Asp Lys Pro Ala Ser Leu Thr Ser Glu Thr Phe His Gln100 105 110Phe Leu Lys Val Arg Ile Gln Asp Ile Ser Ser Ser Lys Tyr Ser Lys115 120 125Ile Leu Lys Asn Thr Asp Trp Glu Asn Leu Ala Gly Asp Ile Phe Asp130 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 Cys165 170 175Ile Glu Phe Asp Tyr Asp Leu Glu Ser Asn Asp Pro Lys Asn Ser Met180 185 190Glu Glu Gly Ala Ile Arg Asn Val Pro Val Val Asp Phe Ser Asn Phe195 200 205Ile Lys Asn Gln Leu Lys Gln Lys Pro Glu Lys Ile Arg Glu Val Leu210 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 Ile245 250 255Pro Lys Gln Gln Pro Tyr Phe Leu Met Phe Ser Tyr Tyr His Asp Glu260 265 270Asp Ala Ser Val Phe Phe Asp Ser Asn Ala Lys Glu Gln Lys Asn Leu275 280 285Tyr Ser Ile Leu Lys Ile Ser Lys Ser Ser Leu Phe Lys Tyr Phe Leu290 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 Asn325 330 335Cys Glu Asn Ile Ile Ser Leu Gln Ser Leu Ser Glu Leu Lys Asn Ile340 345 350Gln Ser Ile Ser Ile Lys Gly Ala Lys Leu Ser Glu Phe Pro Asp Phe355 360 365Leu Leu Asn Leu Pro Ser Leu Lys Ser Leu Tyr Leu Thr Asn Ser Asn370 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 Val405 410 415Phe Gln Leu Pro Gln Leu Lys Lys Leu Leu Phe Met Asp Asn Gln Leu420 425 430Thr Glu Leu Pro Asp Arg Leu Ala Asp Leu Lys Phe Leu Arg Asn Leu435 440 445Asn Leu Ser Gly Asn Lys Ile Thr Gln Ile Ser Asn Leu Thr Lys Glu450 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 Trp485 490 495Gly Asn Glu Leu Glu Thr Ile Ser Pro Glu Ile Phe Asn Leu Lys Asn500 505 510Leu Thr Arg Ile Asp Thr Thr Lys Asn Lys Ile Ser Ser Phe Pro Asn515 520 525Ile Gly Ile Thr Leu Glu Ser Val Thr Asn Leu Ser Leu Ser Asn Asn530 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 Leu565 570 575Phe Lys Asn Phe Gln Lys Leu Glu Thr Leu Ala Leu Ser Asn Asn Arg580 585 590Leu Ser Asn Leu Pro Lys Ser Ile Ser Gln Leu Glu Ser Leu Lys Asn595 600 605Ile Tyr Leu Lys Asn Asn Gln Phe Ile Gln Ile Pro Glu Ile Leu Lys610 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 Lys645 650 655Ile Gly Asn Asn Pro Ile Ala Gln Asn Pro Glu Ser Val Glu Thr Lys660 665 670Met Lys Glu Ile Asn Ser Lys Val Val Leu Tyr Leu Ser675 680 685210452111134212DNA213鉤端螺旋體(Leptospira interrogans)40045atgaacttcc gcataacgtt aattcactta caaaaaatta caattggtct tttgtttctc 60attcatcttt cttacgagat tcaagcagaa gaaccaggaa cttatagaga tttaacaaaa120gcgctccaaa atcctttgga ggttcgagtt ttggatttga gtcgtcaaga actcaaaacc180cttcctatag aaatcggaaa attgaagaat ctgcaaaggt tgtatttaca ttataaccaa240cttacagttc ttccacaaga aatagaacaa ctgaaaaatt tacaattgtt gtatttgcgt300tctaaccgac tcactactct tcctaacgaa atcgagcaat tgaagaactt gcaagttttg360gatttgggtt ctaaccaact tacaattctt ccacaagaaa tagaacagct gaaaaattta420caattgttgt atttgcattc taaccgactc actactcttt ctaaagacat tgaacaacta480cagaatttga aatcgttgga tttgagtaat aaccaactta caactcttcc taacgaaatc540gagcaattga aaaacttgaa atcgttgtat ttgagtgaga accaattcgc aacttttcct600aaagaaattg gacaactaca gaatttaaaa gtgttgtttt tgaataataa ccaaatcaca660atacttccca 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 ataa113421046211377212PRT213鉤端螺旋體(Leptospira interrogans)40046Met 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 Pro20 25 30Gly Thr Tyr Arg Asp Leu Thr Lys Ala Leu Gln Asn Pro Leu Glu Val35 40 45Arg Val Leu Asp Leu Ser Arg Gln Glu Leu Lys Thr Leu Pro Ile Glu50 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 Leu85 90 95Leu Tyr Leu Arg Ser Asn Arg Leu Thr Thr Leu Pro Asn Glu Ile Glu100 105 110Gln Leu Lys Asn Leu Gln Val Leu Asp Leu Gly Ser Asn Gln Leu Thr115 120 125Ile Leu Pro Gln Glu Ile Glu Gln Leu Lys Asn Leu Gln Leu Leu Tyr130 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 Leu165 170 175Pro Asn Glu Ile Glu Gln Leu Lys Asn Leu Lys Ser Leu Tyr Leu Ser180 185 190Glu Asn Gln Phe Ala Thr Phe Pro Lys Glu Ile Gly Gln Leu Gln Asn195 200 205Leu Lys Val Leu Phe Leu Asn Asn Asn Gln Ile Thr Ile Leu Pro Asn210 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 Gln245 250 255Thr Leu Asp Leu Ser Tyr Asn Gln Leu Thr Ile Leu Pro Lys Glu Val260 265 270Gly Gln Leu Glu Asn Leu Gln Thr Leu Asp Leu Arg Asn Asn Gln Leu275 280 285Lys Thr Leu Pro Lys Glu Ile Glu Gln Leu Lys Asn Leu Gln Thr Leu290 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 Thr325 330 335Leu Pro Asn Glu Ile Glu Gln Leu Lys Asn Leu Gln Thr Leu Tyr Leu340 345 350Asn Asn Asn Gln Phe Ser Ser Gln Glu Lys Lys Arg Ile Arg Lys Leu355 360 365Leu Pro Lys Cys Gln Ile Ser Phe Glu370 375210472111359212DNA213鉤端螺旋體(Leptospira interrogans)40047atgaacttcc 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 135921048211452212PRT213鉤端螺旋體(Leptospira interrogans)40048Met 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 Ser20 25 30Glu Ser Gly Thr Tyr Thr Asp Leu Ala Lys Thr Leu Gln Asn Pro Leu35 40 45Lys Val Arg Thr Leu Asp Leu Arg Tyr Gln Lys Leu Thr Thr Leu Pro50 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 Leu85 90 95Gln Glu Leu Asp Leu Ser Phe Asn Ser Leu Thr Thr Leu Pro Lys Glu100 105 110Val Gly Gln Leu Glu Asn Leu Gln Arg Leu Asp Leu His Gln Asn Arg115 120 125Leu Ala Thr Leu Pro Met Glu Ile Gly Gln Leu Lys Asn Leu Gln Glu130 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 Thr165 170 175Thr Leu Pro Lys Glu Ile Gly Gln Leu Gln Asn Leu Lys Thr Leu Asn180 185 190Leu Ile Val Thr Gln Leu Thr Thr Leu Pro Lys Glu Ile Gly Glu Leu195 200 205Gln Asn Leu Lys Thr Leu Asn Leu Leu Asp Asn Gln Leu Thr Thr Leu210 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 Asn245 250 255Leu Gln Trp Leu Asp Leu His Gln Asn Gln Leu Thr Thr Leu Pro Lys260 265 270Glu Ile Gly Gln Leu Gln Asn Leu Gln Arg Leu Asp Leu His Gln Asn275 280 285Gln Leu Thr Thr Leu Pro Lys Glu Ile Gly Gln Leu Gln Asn Leu Gln290 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 Leu325 330 335Thr Thr Leu Pro Lys Glu Val Leu Arg Leu Gln Ser Leu Gln Val Leu340 345 350Ala Leu Gly Ser Asn Arg Leu Ser Thr Leu Pro Lys Glu Ile Gly Gln355 360 365Leu Gln Asn Leu Gln Val Leu Gly Leu Ile Ser Asn Gln Leu Thr Thr370 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 Lys405 410 415Asn Leu Gln Glu Leu His Leu Tyr Leu Asn Pro Leu Ser Ser Lys Glu420 425 430Lys Lys Arg Ile Arg Arg Leu Leu Pro Lys Cys Glu Ile His Phe Glu435 440 445Glu Tyr His Ile45021049211855212DNA213鉤端螺旋體(Leptospira interrogans)40049atgaaatttc gcttaacttt gaattactta caaaatataa caattggcct tctgtttctc 60attcatcttt cttgcaagat tcaaacggaa aaagtagaac cagtaactta tagggattta120acaaaagcgc tccaaaatcc tttgggtgtt cgaattttga atttgagtcg tcaaaaactc180aaaacttttc ctaaagaaat tgggcaattg aagaacttgc aagagttgca tttgagtagt240aaccaattca ctactcttcc taaagaaatc gaacaactac aaaatttgaa atcattggat300ttatgggata atcaacttaa aacccttcct aaggaaatcg gaaaactaca gaacttgaaa360tcgttggatt tgggtagcaa ccaactcaca atccttccaa aggaaatcgg acaactacag420aatttgcaaa aattgaattt atggaataat caacttaaaa ctcttcctaa ggaaattgga480caattacaga acttgcaaaa aatgaattta gataaaaaca ggctcaatac acttcctaat540gaaattggac aactacagaa tttggaatcg ttgtatttga attataacca actcactata600cttccaaaag aaattggaca actacagaat ttggaatcgt tgtatttgaa ttataaccaa660ctcacaatgc ttccacaaga aattggacaa ctacagaatt tggaagggtt gtatctgaaa720tataatcaac tcacaaccct tcctaaagaa attggacgac tacagaattt gaaaaggttg780tatctgaaat ataatcaatt ctcatccaaa gaaaaagaaa aaatccaaaa gcttcttcca840aagtatccag ggtaa 85521050211284212PRT213鉤端螺旋體(Leptospira interrogans)40050Met 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 Val20 25 30Glu Pro Val Thr Tyr Arg Asp Leu Thr Lys Ala Leu Gln Asn Pro Leu35 40 45Gly Val Arg Ile Leu Asn Leu Ser Arg Gln Lys Leu Lys Thr Phe Pro50 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 Leu85 90 95Lys Ser Leu Asp Leu Trp Asp Asn Gln Leu Lys Thr Leu Pro Lys Glu100 105 110Ile Gly Lys Leu Gln Asn Leu Lys Ser Leu Asp Leu Gly Ser Asn Gln115 120 125Leu Thr Ile Leu Pro Lys Glu Ile Gly Gln Leu Gln Asn Leu Gln Lys130 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 Asn165 170 175Thr Leu Pro Asn Glu Ile Gly Gln Leu Gln Asn Leu Glu Ser Leu Tyr180 185 190Leu Asn Tyr Asn Gln Leu Thr Ile Leu Pro Lys Glu Ile Gly Gln Leu195 200 205Gln Asn Leu Glu Ser Leu Tyr Leu Asn Tyr Asn Gln Leu Thr Met Leu210 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 Asn245 250 255Leu Lys Arg Leu Tyr Leu Lys Tyr Asn Gln Phe Ser Ser Lys Glu Lys260 265 270Glu Lys Ile Gln Lys Leu Leu Pro Lys Tyr Pro Gly275 28021051211936212DNA213鉤端螺旋體(Leptospira interrogans)40051atgaactttc gcataacttt gaattactta caaaaaataa caattggtct tttgtttctc 60atgcatcttt cttgcgagat tcaagcagag gaatttgaac aacaagaaac ttatacggat120ttaacaaaag cgctccaaaa tcctttgaag gttcgaacct tggatttgag tgctaaccga180ttcaaaactc ttcctaagga aatcggaaaa ttgaaaaatt tacaagagtt gaatttgaat240aagaaccaac tcacaactct tccacaagaa attggacaat tgaagaattt gaaatcgttg300aatttgagtt ataaccaaat caaaactatt ccgaaagaaa ttgaaaaact acaaaagttg360caatcgttgg gtttggataa taaccaactt acaactcttc cacaagaaat tggacaacta420cagaacttgc aatcgttgga tttgagtact aaccgactca caactcttcc acaagaaatt480ggacatctac agaatttgca agatttgtat ttggtttcta accaactcac aattcttcct540aacgaaatcg gacaattaaa gaacttgcaa acgttgaatt tgcgtaataa ccgactcact600actctttcta aagaaattga gcaactacag aatttgaaat cgttggattt gcgttctaac660caacttacta cttttcctaa ggaaatcgag caattgaaga atttgcaagt tttggatttg720ggttctaacc aactcactac tcttccagag ggaatcggac aattgaagaa cttgcaaaca780ttggatttgg attctaacca acttacaact cttccacaag aaatcggaca actacagaat840ttgcaagaat tgtttttaaa caacaatcaa ctctcatccc aagaaaaaaa aagaattcga900aaacttcttc caaagtgcca aatttacttt gactaa 93621052211311212PRT213鉤端螺旋體(Leptospira interrogans)40052Met 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 Phe20 25 30Glu Gln Gln Glu Thr Tyr Thr Asp Leu Thr Lys Ala Leu Gln Asn Pro35 40 45Leu Lys Val Arg Thr Leu Asp Leu Ser Ala Asn Arg Phe Lys Thr Leu50 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 Asn85 90 95Leu Lys Ser Leu Asn Leu Ser Tyr Asn Gln Ile Lys Thr Ile Pro Lys100 105 110Glu Ile Glu Lys Leu Gln Lys Leu Gln Ser Leu Gly Leu Asp Asn Asn115 120 125Gln Leu Thr Thr Leu Pro Gln Glu Ile Gly Gln Leu Gln Asn Leu Gln130 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 Leu165 170 175Thr Ile Leu Pro Asn Glu Ile Gly Gln Leu Lys Asn Leu Gln Thr Leu180 185 190Asn Leu Arg Asn Asn Arg Leu Thr Thr Leu Ser Lys Glu Ile Glu Gln195 200 205Leu Gln Asn Leu Lys Ser Leu Asp Leu Arg Ser Asn Gln Leu Thr Thr210 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 Lys245 250 255Asn Leu Gln Thr Leu Asp Leu Asp Ser Asn Gln Leu Thr Thr Leu Pro260 265 270Gln Glu Ile Gly Gln Leu Gln Asn Leu Gln Glu Leu Phe Leu Asn Asn275 280 285Asn Gln Leu Ser Ser Gln Glu Lys Lys Arg Ile Arg Lys Leu Leu Pro290 295 300Lys Cys Gln Ile Tyr Phe Asp305 31021053211990212DNA213鉤端螺旋體(Leptospira interrogans)40053atgaattttc 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 99021054211329212PRT213鉤端螺旋體(Leptospira interrogans)40054Met 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 Thr20 25 30Tyr Trp Asp Leu Thr Lys Ala Leu Gln Asn Pro Leu Asp Val Arg Ile35 40 45Leu Asn Leu Ser Glu Gln Lys Leu Thr Thr Leu Pro Lys Glu Ile Gly50 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 Gly85 90 95Leu Tyr Tyr Asn Gln Leu Thr Ile Leu Pro Lys Glu Ile Glu Gln Leu100 105 110Lys Asn Leu Gln Ala Leu Tyr Leu Gly Asn Asn Gln Ile Thr Ile Leu115 120 125Pro Lys Glu Ile Arg Gln Leu Gln Asn Leu Lys Val Leu Phe Leu Ser130 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 Lys165 170 175Glu Ile Glu Gln Leu Lys Asn Leu Gln Leu Leu Tyr Leu Tyr Asp Asn180 185 190Gln Leu Thr Val Leu Pro Gln Glu Ile Lys Gln Leu Lys Asn Leu Gln195 200 205Leu Leu Asp Leu Ser Tyr Asn Gln Leu Lys Thr Leu Pro Lys Glu Ile210 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 Leu245 250 255Tyr Leu Gly Tyr Asn Gln Leu Thr Thr Leu Pro Lys Glu Ile Gly Gln260 265 270Leu Gln Asn Leu Lys Val Leu Phe Leu Asn Asn Asn Gln Leu Thr Thr275 280 285Leu Pro Lys Glu Ile Gly Gln Leu Lys Asn Leu Gln Glu Leu Tyr Leu290 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 Glu32權利要求
1.一種分離的鉤端螺旋體致病相關蛋白多肽，其特徵在於，它包含具有選自SEQ ID NO2、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的胺基酸序列的多肽、或其保守性變異多肽、或其活性片段、或其活性衍生物。
2.如權利要求1所述的多肽，其特徵在於，該多肽是具有SEQ ID NO2、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的胺基酸序列的多肽。
3.一種分離的多核苷酸，其特徵在於，它包含一核苷酸序列，該核苷酸序列與選自下組的一種核苷酸序列有至少70％相同性(a)編碼如權利要求1和2所述多肽的多核苷酸；(b)與多核苷酸(a)互補的多核苷酸。
4.如權利要求3所述的多核苷酸，其特徵在於，該多核苷酸編碼具有SEQ IDNO2、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所示胺基酸序列的多肽。
5.如權利要求3所述的多核苷酸，其特徵在於，該多核苷酸的序列選自下組(a)具有SEQ ID NO1、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。
6.一種載體，其特徵在於，它含有權利要求3所述的多核苷酸。
7.一種遺傳工程化的宿主細胞，其特徵在於，它含有權利要求6所述的載體。
8.一種鉤端螺旋體致病相關蛋白多肽的製備方法，其特徵在於，該方法包含(a)在適合表達鉤端螺旋體致病相關蛋白的條件下，培養權利要求7所述的宿主細胞；(b)從培養物中分離出鉤端螺旋體致病相關蛋白多肽。
9.一種能與權利要求1所述的鉤端螺旋體致病相關蛋白特異性結合的抗體。
全文摘要
本發明提供了一類新的鉤端螺旋體致病相關蛋白，編碼致病相關蛋白的多核苷酸和經重組技術產生這類致病相關蛋白的方法。本發明還公開了這類致病相關蛋白及其編碼序列的用途。
文檔編號C07K16/18GK1443773SQ0211099
公開日2003年9月24日 申請日期2002年3月11日 優先權日2002年3月11日
發明者任雙喜, 傅剛, 曾嶸, 蔣秀高, 張怡軒, 趙國屏, 陳竺, 繆有剛, 徐海, 郭小奎 申請人:中國科學院上海生物工程研究中心