專(zhuān)利名稱(chēng)：B組鏈球菌抗原的制作方法
技術(shù)領(lǐng)域：
本發(fā)明涉及抗原，更特別地涉及用作治療和/或預(yù)防的疫苗成分的B組鏈球菌(GBS)細(xì)菌病原體的蛋白質(zhì)抗原。
背景技術(shù)：
鏈球菌是革蘭氏陽(yáng)性細(xì)菌，它們按它們細(xì)胞表面上發(fā)現(xiàn)的A組至O組具體碳水化合物抗原來(lái)劃分。鏈球菌組進(jìn)一步通過(guò)具體類(lèi)型莢膜多糖抗原來(lái)區(qū)分。對(duì)于B組鏈球菌(GBS)鑒定了幾種血清型Ⅰa,Ⅰb,Ⅱ,Ⅲ,Ⅳ,Ⅴ,Ⅵ,Ⅶh Ⅷ。GBS還包括已知為“C-蛋白質(zhì)”(α,β,γ和δ)，其中一些已經(jīng)被克隆出來(lái)。
盡管GBS是正常人陰道和結(jié)腸菌叢的共同成分，但是該病原體長(zhǎng)期以來(lái)一直被認(rèn)為是新生期膿毒癥，腦膜炎，嬰兒腦膜炎后遺癥，產(chǎn)后子宮內(nèi)膜炎以及牛奶場(chǎng)放牧人乳腺炎的主要病因。暴露給GBS的待產(chǎn)婦有產(chǎn)后感染的危險(xiǎn)并且可能在孩子通過(guò)產(chǎn)道時(shí)將感染轉(zhuǎn)移給她們的孩子。盡管微生物對(duì)抗生素敏感，但是由于新生期膿毒癥和嬰兒腦膜炎高的發(fā)生比例和快的發(fā)生速度而導(dǎo)致高的發(fā)病率和死亡率。
為了發(fā)現(xiàn)保護(hù)人們不受GBS感染的疫苗，研究轉(zhuǎn)向特異型抗原。令人遺憾的是，證明了這些多糖在人體內(nèi)具有不好的免疫原性并且限制于這些多糖發(fā)源的特殊血清型。此外，莢膜多糖激發(fā)T細(xì)胞獨(dú)立響應(yīng)，即沒(méi)有IgG產(chǎn)生。結(jié)果莢膜多糖抗原不適合作為保護(hù)抗GBS感染的疫苗成分。
其它人觀注C-蛋白β抗原，其被證明在小鼠和兔模型中有免疫原性。發(fā)現(xiàn)該性質(zhì)不適合作為人疫苗，因?yàn)槠渑c該親和性相互作用和與人ⅠgA的Fc區(qū)的非免疫原性方式的不期望的性質(zhì)。C-蛋白α抗原在負(fù)責(zé)大多數(shù)GBS介導(dǎo)的疾病的血清型GBS的Ⅲ型血清型中很少，因此作為疫苗成分幾乎沒(méi)有用途。
因此，對(duì)于可以用作預(yù)防和/或治療GBS感染的疫苗成分的GBS抗原還有沒(méi)有滿(mǎn)足的要求。
發(fā)明概述根據(jù)一方面，本發(fā)明提供編碼與包括選自下面的一個(gè)序列的第二多肽至少70％一致性的多肽的分離的多核苷酸SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5,SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10,SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:15,SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19,SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:24,SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:29,SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:33, SEQ ID NO:34,SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:38, SEQ ID NO:39,SEQ ID NO:40, SEQ ID NO:41和 SEQ ID NO:44或者其片段，類(lèi)似物或衍生物。
在其它方面，提供包括操作性連接了表達(dá)調(diào)控區(qū)的本發(fā)明多核苷酸的載體，以及用所述載體轉(zhuǎn)染的宿主細(xì)胞，和產(chǎn)生多肽的方法，包括在適合表達(dá)的條件下培養(yǎng)所述宿主細(xì)胞。
在另一方面，提供本發(fā)明多核苷酸編碼的新的多肽。
附圖的簡(jiǎn)要描述

圖1a是克隆1的DNA序列(SEQ ID NO:1)與相應(yīng)的可讀框的氨基酸序列；圖1b是氨基酸序列SEQ ID NO:2；圖1c是氨基酸序列SEQ ID NO:3；圖1d是氨基酸序列SEQ ID NO:4；圖1e是氨基酸序列SEQ ID NO:5；圖1f是氨基酸序列SEQ ID NO:6；圖2a是克隆2的DNA序列(SEQ ID NO:7)與相應(yīng)的可讀框的氨基酸序列；圖2b是氨基酸序列SEQ ID NO:8；圖2c是氨基酸序列SEQ ID NO:9；圖2d是氨基酸序列SEQ ID NO:10；圖2e是氨基酸序列SEQ ID NO:11；圖2f是氨基酸序列SEQ ID NO:12；圖3a是克隆3的DNA序列(SEQ ID NO:13)與相應(yīng)的可讀框的氨基酸序列；
圖3b是氨基酸序列SEQ ID NO:14；圖3c是氨基酸序列SEQ ID NO:15；圖3d是氨基酸序列SEQ ID NO:16；圖3e是氨基酸序列SEQ ID NO:17；圖3f是氨基酸序列SEQ ID NO:18；圖3g是氨基酸序列SEQ ID NO:19；圖3h是氨基酸序列SEQ ID NO:20；圖3i是氨基酸序列SEQ ID NO:21；圖4a是克隆4的DNA序列(SEQ ID NO:22)與相應(yīng)的可讀框的氨基酸序列；圖4b是氨基酸序列SEQ ID NO:23；圖4c是氨基酸序列SEQ ID NO:24；圖4d是氨基酸序列SEQ ID NO:25；圖4e是氨基酸序列SEQ ID NO:26；圖5a是克隆5的DNA序列(SEQ ID NO:27)與相應(yīng)的可讀框的氨基酸序列；圖5b是氨基酸序列SEQ ID NO:28；圖5c是氨基酸序列SEQ ID NO:29；圖5d是氨基酸序列SEQ ID NO:30；圖5e是氨基酸序列SEQ ID NO:31；圖6a是克隆6的DNA序列(SEQ ID NO:32)與相應(yīng)的可讀框的氨基酸序列；圖6b是氨基酸序列SEQ ID NO:33；圖6c是氨基酸序列SEQ ID NO:34；圖6d是氨基酸序列SEQ ID NO:35；圖6e是氨基酸序列SEQ ID NO:36；圖7a是克隆7的DNA序列(SEQ ID NO:37)；圖7b是氨基酸序列SEQ ID NO:38；圖7c是氨基酸序列SEQ ID NO:39；圖7d是氨基酸序列SEQ ID NO:40；圖7e是氨基酸序列SEQ ID NO:41；圖8是包括信號(hào)序列的克隆7的一部分的DNA序列(SEQ ID NO:42)；圖9是不包括信號(hào)序列的克隆7的一部分的DNA序列(SEQ ID NO:43)；圖9a是氨基酸序列SEQ ID NO:44；圖10代表來(lái)自用相應(yīng)于SEQ ID NO:39的重組GBS蛋白質(zhì)免疫的CD-1小鼠血清的抗-GBS ELISA效價(jià)的分布。
本發(fā)明的詳細(xì)描述本發(fā)明涉及B組鏈球菌(GBS)的新的抗原多肽，特征在于是選自下面的氨基酸序列SEQ ID NO:2, SEQ ID NO:3SEQ ID NO:4, SEQ ID NO:5,SEQ ID NO:6, SEQ ID NO:8SEQ ID NO:9, SEQ ID NO:10,SEQ ID NO:11, SEQ ID NO:12 SEQ ID NO:14, SEQ ID NO:15,SEQ ID NO:16, SEQ ID NO:17 SEQ ID NO:18, SEQ ID NO:19,SEQ ID NO:20, SEQ ID NO:21 SEQ ID NO:23, SEQ ID NO:24,SEQ ID NO:25, SEQ ID NO:26 SEQ ID NO:28, SEQ ID NO:29,SEQ ID NO:30, SEQ ID NO:31 SEQ ID NO:33, SEQ ID NO:34,SEQ ID NO:35, SEQ ID NO:36 SEQ ID NO:38, SEQ ID NO:39,SEQ ID NO:40, SEQ ID NO:41和 SEQ ID NO:44或者其片段，類(lèi)似物或衍生物。
本發(fā)明的優(yōu)選實(shí)施方案包括SEQ ID NO:39和SEQ ID NO:44。
本發(fā)明的進(jìn)一步優(yōu)選實(shí)施方案包括SEQ ID NO39。
本發(fā)明的進(jìn)一步優(yōu)選實(shí)施方案包括SEQ ID NO:44。
如這里所使用的，本發(fā)明多肽的“片段”，“衍生物”或“類(lèi)似物”包括其中一個(gè)或多個(gè)氨基酸殘基被保守的或者不保守的氨基酸殘基(優(yōu)選保守的)取代并且可以是天然的或非天然的那些多肽。
本發(fā)明多肽的術(shù)語(yǔ)“片段”，“衍生物”或“類(lèi)似物”包括添加，缺失，取代氨基酸的修飾的多肽，前提是這些多肽保持誘導(dǎo)免疫應(yīng)答的能力。
術(shù)語(yǔ)“保守的氨基酸”是指一個(gè)或多個(gè)氨基酸被其它氨基酸取代，其中即使發(fā)生取代，給定抗原的抗原決定簇(包括其二級(jí)結(jié)構(gòu)和親水性質(zhì))也是完全或部分保守的。
例如，所述序列中一個(gè)或多個(gè)氨基酸殘基可以被另一個(gè)相似極性的作為功能等價(jià)物起作用的氨基酸取代，導(dǎo)致沉默改變。所述序列中氨基酸的取代物可以選自該氨基酸所屬于的種類(lèi)其它成員。例如，非極性(疏水性)氨基酸包括丙氨酸，亮氨酸，異亮氨酸，纈氨酸，脯氨酸，苯丙氨酸，色氨酸和蛋氨酸。極性中性氨基酸包括甘氨酸，絲氨酸，蘇氨酸，半胱氨酸，酪氨酸，天冬酰胺和谷氨酰胺。帶正電荷(堿性)氨基酸包括精氨酸，賴(lài)氨酸和組氨酸。帶負(fù)電荷(酸性)氨基酸包括天冬氨酸和谷氨酸。
優(yōu)選地，本發(fā)明多肽的衍生物和類(lèi)似物與圖中圖示說(shuō)明的那些序列或者其片段具有大約70％的一致性。也就是說(shuō)70％的殘基相同。更優(yōu)選地，多肽具有大于95％的同源性。在另一個(gè)優(yōu)選的實(shí)施方案中，本發(fā)明多肽的衍生物和類(lèi)似物具有少于大約20個(gè)氨基酸殘基的取代，修飾或缺失，更優(yōu)選地少于10個(gè)。優(yōu)選的取代物是本領(lǐng)域已知為保守的那些，即被取代的殘基具有相同的物理或化學(xué)性質(zhì)，例如疏水性，大小，電荷或者官能團(tuán)。
此外，在氨基酸區(qū)被發(fā)現(xiàn)是多態(tài)的情況下，可以預(yù)期改變一個(gè)或多個(gè)特殊氨基酸以更有效地模擬不同GBS株的不同表位。
還包括與改變多肽生物或藥學(xué)性質(zhì)的其它化合物，即聚乙二醇(PEG)融合以提高半壽期的多肽，易于純化的前導(dǎo)或分泌氨基酸序列；前序列原和前序列；和多糖。
此外，本發(fā)明的多肽可以通過(guò)末端-NH2?；饔?例如通過(guò)乙酰化作用，或者硫代乙二醇酸酰胺化作用，末端羧酸(carbosy)酰胺化作用，例如與氨或者甲胺)而被修飾，以提供連接或鍵合載體或其它分子的穩(wěn)定性，提高的疏水性。
還涉及所述多肽片段，類(lèi)似物和衍生物的雜聚和同聚多肽多聚體。這些聚合物形式包括，例如，與交聯(lián)劑例如親和素/生物素，戊二醛或二甲基超亞胺酯(dimethyl-superimidate)交聯(lián)的一種或多種多肽。這樣的聚合物形式還包括包含通過(guò)DNA重組技術(shù)產(chǎn)生的多順?lè)醋觤RNAs制備的兩個(gè)或多個(gè)串聯(lián)的或者倒位的連續(xù)序列的多肽。優(yōu)選地，本發(fā)明多肽的片段，類(lèi)似物或衍生物包括至少一個(gè)抗原區(qū)，即至少一個(gè)表位。
為了實(shí)現(xiàn)抗原聚合物(即合成的多聚體)的形成，可以使用具有二鹵代酰基，硝基芳基鹵化物等等的多肽，其中試劑對(duì)于硫基是特異性的。因此，不同肽的兩個(gè)巰基之間的連接可以是單鍵或者可以由至少兩個(gè)，典型地至少4個(gè)，不多于16個(gè)，但是通常不多于14個(gè)碳原子的連接基團(tuán)組成。
在具體的實(shí)施方案中，本發(fā)明的多肽片段，類(lèi)似物和衍生物不含有蛋氨酸(Met)起始?xì)埢?。?yōu)選地，多肽將不插入一個(gè)前導(dǎo)或分泌序列(信號(hào)序列)。根據(jù)已確定的分子生物學(xué)技術(shù)可以確定本發(fā)明多肽的信號(hào)部分。一般情況下，可以從GBS培養(yǎng)物分離所感興趣的多肽并且接著測(cè)序確定成熟蛋白的初始?xì)埢瑥亩_定成熟多肽的序列。
根據(jù)另一方面，提供了包括與藥學(xué)可接受載體稀釋劑或佐劑混合的一種或多種本發(fā)明GBS多肽的疫苗組合物。
合適的佐劑包括油，即弗氏完全佐劑或不完全佐劑；鹽，即AlK(SO4)2,AlNa(SO4)2,AlNH4(SO4)2,Al(OH)3,AlPO4，二氧化硅，高嶺土；皂草甙衍生物；碳多核苷酸，即多IC和多AU，還有解毒的霍亂毒素(CTB)和誘導(dǎo)粘膜免疫的大腸桿菌熱不穩(wěn)定毒素。優(yōu)選的佐劑包括QuilATM,AlhydrogelTM，和AdjuphosTM。本發(fā)明的疫苗可以通過(guò)注射，快速灌注，鼻咽吸收，皮膚吸收，非經(jīng)腸給藥，或者含化或者口服給藥。
本發(fā)明的疫苗組合物用于治療或預(yù)防鏈球菌感染和/或鏈球菌感染介導(dǎo)的疾病和癥狀，特別是A組鏈球菌(化膿鏈球菌)，B組鏈球菌(GBS或無(wú)乳鏈球菌)，停乳鏈球菌，乳房鏈球菌，諾氏鏈球菌以及金黃色葡萄球菌。關(guān)于鏈球菌的一般信息參見(jiàn)Manual of ClinicalMicrobiology,P.R.Murray等，(1995，第六版，ASM出版社，華盛頓)。更特別地是B組鏈球菌，無(wú)乳鏈球菌。在具體的實(shí)施方案中，對(duì)有GBS感染危險(xiǎn)的那些個(gè)體施用疫苗，所述個(gè)體是例如孕婦和嬰兒膿毒癥，腦膜炎和肺炎以及無(wú)免疫應(yīng)答的個(gè)體，例如糖尿病，肝病或癌癥的那些。疫苗也具有獸藥用途，例如用于治療上述細(xì)菌以及大腸桿菌介導(dǎo)的牛乳腺炎。
本發(fā)明的疫苗還可以用來(lái)制備用于治療或預(yù)防鏈球菌感染和/或鏈球菌感染介導(dǎo)的疾病和癥狀的藥物，所述鏈球菌特別是A組鏈球菌(化膿鏈球菌)，B組鏈球菌(GBS或無(wú)乳鏈球菌)，停乳鏈球菌，乳房鏈球菌，諾氏鏈球菌以及金黃色葡萄球菌。更特別地是B組鏈球菌，無(wú)乳鏈球菌。
疫苗組合物優(yōu)選以0.001至100μg/kg(抗原/體重)，更優(yōu)選地0.01至10μg/kg和最優(yōu)選地0.1至1μg/kg的單位劑量形式，免疫之間大約1至12周間隔免疫1至3次，更優(yōu)選地間隔1至6周。
根據(jù)另一方面，提供了編碼特征在于氨基酸序列選自下組的B組鏈球菌(GBS)多肽的多核苷酸SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:15SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO;23, SEQ ID NO:24SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:29SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:33, SEQ ID NO:34SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:38, SEQ ID NO:39SEQ ID NO:40, SEQ ID NO:41和 SEQ ID NO:44或者其片段，類(lèi)似物或衍生物。
優(yōu)選的多核苷酸是在圖1a(SEQ ID NO:1),2a(SEQ ID NO:7),3a(SEQ ID NO:13),4a(SEQ ID NO:22),5a(SEQ ID NO:27),6a(SEQ ID NO:32),7a(SEQ ID NO:37),8(SEQ ID NO:42)和9(SEQ ID NO:43)中圖示說(shuō)明的那些，它們相應(yīng)于編碼本發(fā)明多肽的可讀框。
優(yōu)選的多核苷酸是在圖1a(SEQ ID NO:1),2a(SEQ ID NO:7),3a(SEQ ID NO:13),4a(SEQ ID NO:22),5a(SEQ ID NO:27),6a(SEQ ID NO:32),7a(SEQ ID NO:37),8(SEQ ID NO:42)和9(SEQ ID NO:43)中圖示說(shuō)明的那些，和它們的片段，類(lèi)似物和衍生物。
更優(yōu)選的本發(fā)明的多核苷酸是圖7(SEQ ID NO:37),8(SEQ ID NO:42)和9(SEQ ID NO:43)中圖示說(shuō)明的那些。
最優(yōu)選的本發(fā)明的多核苷酸是圖8(SEQ ID NO:42)和9(SEQ ID NO:43)中圖示說(shuō)明的那些。
要理解圖中說(shuō)明的多核苷酸序列可以用也編碼本發(fā)明多肽的簡(jiǎn)并密碼子改變。
由于核苷酸編碼序列的簡(jiǎn)并性，本發(fā)明實(shí)施中也可以使用編碼本發(fā)明基本上相同的多肽的其它多核苷酸序列。這些包括但不限于編碼序列內(nèi)相同氨基酸殘基的不同密碼子的取代而改變從而產(chǎn)生沉默變化的核苷酸序列。
因此，本發(fā)明進(jìn)一步提供與上述多核苷酸序列雜交的多核苷酸(或者其互補(bǔ)序列)，序列之間具有50％，優(yōu)選地具有至少70％一致性。更優(yōu)選地，多核苷酸在嚴(yán)格條件下是可雜交的，即具有至少95％一致性，最優(yōu)選地大于97％一致性。
能在嚴(yán)格條件下雜交是指核苷酸分子在標(biāo)準(zhǔn)條件下，例如高溫和/或低鹽含量下，退火生成第二核酸序列的至少一個(gè)區(qū)(或者作為cDNA，或者作為基因組DNA)或者生成其互補(bǔ)鏈，其趨向于不利于非互補(bǔ)核苷酸序列的雜交。合適的方法描述于Maniatis T.等，分子克隆實(shí)驗(yàn)室手冊(cè)，冷泉港實(shí)驗(yàn)室，1982，其在這里引作參考。
再一方面，可以在DNA免疫方法中使用編碼本發(fā)明多肽的多核苷酸，或者其片段，類(lèi)似物或衍生物。即，它們可以被插入到載體中，該載體在注射后是可復(fù)制和可表達(dá)的，從而體內(nèi)產(chǎn)生抗原性多肽。例如于在真核細(xì)胞中起作用的CMV啟動(dòng)子控制下，多核苷酸可以插入到質(zhì)粒載體中。優(yōu)選地肌內(nèi)注射載體。
根據(jù)另一方面，提供了通過(guò)重組技術(shù)通過(guò)在宿主細(xì)胞中表達(dá)編碼所述多肽的多核苷酸并且回收表達(dá)的多肽產(chǎn)物而制備本發(fā)明多肽的方法?；蛘?，可以根據(jù)已確定的化學(xué)合成技術(shù)合成本發(fā)明多肽，即寡肽的液相或固相合成，連接寡肽產(chǎn)生全長(zhǎng)多肽(封閉連接)。
對(duì)于重組體生產(chǎn)，用編碼多肽的載體轉(zhuǎn)染宿主細(xì)胞，然后在經(jīng)改變適合激活啟動(dòng)子，選擇轉(zhuǎn)化物或者擴(kuò)增基因的營(yíng)養(yǎng)培養(yǎng)基中培養(yǎng)。合適的載體是在選擇的宿主內(nèi)可存活且可復(fù)制的那些載體，包括染色體的，非染色體的和合成的DNA序列，例如細(xì)菌質(zhì)粒，噬菌體DNA，桿狀病毒，酵母質(zhì)粒，質(zhì)粒和噬菌體DNA組合產(chǎn)生的載體。所述多肽序列可以使用限制酶在合適的位點(diǎn)插入載體中，這樣其操作連接包括啟動(dòng)子，核糖體結(jié)合位點(diǎn)(共有區(qū)或核糖體結(jié)合序列)，和任選地操縱基因(調(diào)控元件)的表達(dá)調(diào)控區(qū)。人們可以根據(jù)已確定的分子生物學(xué)原理(Sambrook等，分子克隆實(shí)驗(yàn)室手冊(cè)，第二版，冷泉港實(shí)驗(yàn)室，紐約，1989，其在這里引作參考)選擇適合給定宿主和載體的表達(dá)調(diào)控區(qū)的各成分。合適的啟動(dòng)子包括但不限于LTR或SV40啟動(dòng)子，大腸桿菌lac,tac或啟動(dòng)子和噬菌體λPL啟動(dòng)子。載體優(yōu)選插入復(fù)制起點(diǎn)以及選擇標(biāo)記，即氨芐青霉素抗性基因。合適的細(xì)菌載體包括pET,pQE70,pQE60,pQE-9,pbs,pD10phagescript,psiX174,pbluescriptSK,pbsks,pNH8A,pNH16a,pNH18A,pNH46A,ptrc99a,pKK223-3,pKK233-3,pDR540,pRIT5和真核載體pBlueBacⅢ,pWLNEO,pSV2CAT,pOG44,pXT1,pSG,pSVK3,pBPV,pMSG和pSVL。宿主細(xì)胞可以是細(xì)菌，即大腸桿菌，枯草芽孢桿菌，鏈霉菌屬；真菌，即黑曲霉，構(gòu)巢曲霉；酵母，即糖酵母屬或者真核的，即CHO,COS。
在培養(yǎng)基中表達(dá)多肽時(shí)，一般通過(guò)離心收集細(xì)胞，然后通過(guò)物理或化學(xué)方法破碎(如果表達(dá)的多肽沒(méi)有分泌到培養(yǎng)基中)，留下得到的粗提取物以分離感興趣的多肽。從培養(yǎng)基或溶解物中純化多肽可以根據(jù)多肽的性質(zhì)通過(guò)已確定的技術(shù)進(jìn)行，即利用硫酸銨或乙醇沉淀，酸提取，陰離子或陽(yáng)離子交換層析，磷酸纖維素層析，疏水作用層析，羥基磷灰石層析和凝集素層析?？梢詰?yīng)用HPLC實(shí)現(xiàn)最后的純化。
多肽可以在有或沒(méi)有前導(dǎo)或分泌序列下被表達(dá)。在前一種情況下，可以使用翻譯后處理去除前導(dǎo)序列(參見(jiàn)US4431739；4425437；和4338397，這里引作參考)或者可以化學(xué)去除前導(dǎo)序列接著純化表達(dá)的多肽。
根據(jù)又一方面，本發(fā)明的GBS多肽可以在診斷鏈球菌感染特別是GBS感染的診斷測(cè)定中使用。幾種診斷方法是可能的，例如檢測(cè)生物樣品中的鏈球菌生物，可以進(jìn)行下面的方法a)從患者獲得生物樣品；b)培養(yǎng)與本發(fā)明GBS多肽反應(yīng)的抗體或者其片段和生物樣品，形成混合物；和c)檢測(cè)混合物中特異性結(jié)合的抗體或者結(jié)合的片段，這表明鏈球菌的存在。
或者，可以如下進(jìn)行檢測(cè)含有或者懷疑含有所述抗體的生物樣品中對(duì)鏈球菌抗原特異性的抗體的方法a)從患者分離生物樣品；b)培養(yǎng)一種或多種本發(fā)明GBS多肽或者其片段和生物樣品，形成混合物；和c)檢測(cè)混合物中特異性結(jié)合的抗原或者結(jié)合的片段，這表明鏈球菌特異性抗體的存在。
本領(lǐng)域技術(shù)人員會(huì)理解可以以幾種形式進(jìn)行診斷測(cè)定，包括免疫測(cè)定，例如酶聯(lián)免疫吸附測(cè)定(ELISA)，放射免疫測(cè)定或者乳膠凝聚試驗(yàn)，特別是測(cè)定生物體內(nèi)是否存在對(duì)所述蛋白質(zhì)有特異性的抗體。
編碼本發(fā)明多肽的DNA序列也可以用來(lái)設(shè)計(jì)用于檢測(cè)懷疑含有鏈球菌的生物樣品中鏈球菌的存在。本發(fā)明的檢測(cè)方法包括a)從患者分離生物樣品；b)培養(yǎng)一種或多種具有編碼本發(fā)明多肽的DNA序列或者其片段的DNA探針和生物樣品，形成混合物；和c)檢測(cè)混合物中特異性結(jié)合的DNA探針，這表明鏈球菌的存在。
本發(fā)明的DNA探針也可以用來(lái)檢測(cè)樣品中的循環(huán)鏈球菌，即GBS核酸，例如使用聚合酶鏈反應(yīng)，作為診斷鏈球菌感染的方法。所述探針可以應(yīng)用常規(guī)技術(shù)合成和可以固定在固相上，或者可以用可檢測(cè)的標(biāo)記物標(biāo)記。用于本申請(qǐng)的優(yōu)選的DNA探針是具有與本發(fā)明GBS多肽的至少6個(gè)連續(xù)的核苷酸互補(bǔ)的序列的寡聚物。
檢測(cè)患者體內(nèi)鏈球菌的另一種診斷方法包括a)用可檢測(cè)的標(biāo)記物標(biāo)記與本發(fā)明多肽或者其片段反應(yīng)的抗體；b)對(duì)患者施用標(biāo)記的抗體或者標(biāo)記的片段；和c)檢測(cè)患者體內(nèi)特異性結(jié)合的標(biāo)記的抗體或者標(biāo)記的片段，這表明鏈球菌的存在。
本發(fā)明的再一方面是本發(fā)明的多肽作為用于產(chǎn)生用于診斷和特別是用于治療鏈球菌感染的特異性抗體的免疫原的用途?？梢允褂眠m當(dāng)?shù)暮Y選方法確定合適的抗體，例如通過(guò)在試驗(yàn)?zāi)Ｐ椭袦y(cè)定特定抗體對(duì)于抗鏈球菌感染的被動(dòng)免疫的能力。動(dòng)物模型的一個(gè)實(shí)施例是本中請(qǐng)實(shí)施例中描述的小鼠模型。所述抗體可以是整個(gè)抗體或者是其抗原結(jié)合片段，并且可以一般屬于任何免疫球蛋白種類(lèi)。所述抗體或片段可以是動(dòng)物來(lái)源的，特別是哺乳動(dòng)物源，更特別地是小鼠，大鼠或者人源。其可以是天然抗體或者其片段，或者如果期望，可以是重組體抗體或抗體片段。術(shù)語(yǔ)重組體抗體或抗體片段指應(yīng)用分子生物學(xué)技術(shù)產(chǎn)生的抗體或抗體片段。所述抗體或抗體片段可以是多克隆抗體，或者優(yōu)選地，單克隆抗體。其可以對(duì)多個(gè)與GBS多肽相關(guān)的表位是特異性的，但是優(yōu)選對(duì)于一個(gè)表位是特異性的。
實(shí)施例1致死量B組鏈球菌(GBS)感染的小鼠模型GBS感染的小鼠模型詳細(xì)描述于Lancefield等(J.Exp.Med.142:165-179,1975)。對(duì)GBS株C388/90(1990年從加拿大喔太華的Children’sHospital of Eastern Ontario腦膜炎患者的腦脊液獲得的臨床分離物)和NCS246(National Center for Streptococcus,ProvincialLaboratory of Public Health for Northern Alberta,Edmonton,加拿大)分別血清型測(cè)定為Ⅰa/c型和Ⅱ/R型。
為了提高它們的致病力，如前所述(Lancefield等J.Exp.Med.142:165-179,1975)連續(xù)在小鼠中傳代GBS株C388/90(血清型Ⅰa/c)和NCS246(血清型Ⅱ/R)。簡(jiǎn)要地說(shuō)，使用從感染小鼠的血液或脾獲得的在Todd-Hewitt肉湯中傳代培養(yǎng)物的系列稀釋物腹膜內(nèi)接種來(lái)監(jiān)測(cè)致病力的提高。最后傳代后，使用感染的血樣接種Todd-Hewitt肉湯.在37℃下7％CO2下培養(yǎng)2小時(shí)后，向培養(yǎng)物加入終濃度10％(v/v)的甘油。然后將培養(yǎng)物分成等份并且在-80℃下保存，用于GBS攻擊實(shí)驗(yàn)。測(cè)定這些冷凍的樣品中存在的GBS的cfu數(shù)。對(duì)于GBS株C388/90和NCS246分別測(cè)定100％殺死(LD100)18周齡小鼠所必須的細(xì)菌濃度是3.5×105和1.1×105，這相應(yīng)于肉湯菌株的致病力的顯著增加。事實(shí)上，這兩種菌株傳代之前記錄的LD100大于109cfu。
在細(xì)菌攻擊中，使用Todd-Hewitt肉湯將新解凍的致病GBS株等份調(diào)節(jié)到合適的細(xì)菌濃度并且對(duì)每一只雌性CD-1小鼠腹膜內(nèi)注射1毫升。用于被動(dòng)保護(hù)實(shí)驗(yàn)的小鼠是6-8周齡，而用于主動(dòng)保護(hù)實(shí)驗(yàn)的小鼠在受攻擊時(shí)大約18周齡。對(duì)所有的接種體證實(shí)菌落數(shù)。在攻擊后頭48小時(shí)每天4次觀察動(dòng)物的任何感染癥狀，接著下面的12天每天觀察。在該時(shí)期最后，從生存者獲得血樣并且在-20℃下冷凍。培養(yǎng)從受攻擊存活的每一只小鼠獲得脾以鑒定所有存留的GBS。
實(shí)施例2用甲醛殺死的GBS全細(xì)胞免疫和保護(hù)小鼠根據(jù)Lancefield等(J.Exp.Med.142:165-179,1975)描述的方法制備甲醛殺死的GBS全細(xì)胞。簡(jiǎn)要地說(shuō)，將GBS株在綿羊血瓊脂平板(Quelab Laboratories,Montreal，加拿大)上過(guò)夜培養(yǎng)物在PBS緩沖液(磷酸鹽緩沖鹽水，pH7.2)中清洗兩次，調(diào)節(jié)至大約3×109cfu/ml，并且在含有0.3％(v/v)甲醛的PBS中培養(yǎng)過(guò)夜。用PBS清洗殺死的GBS緩沖液并且在-80℃下冷凍保存。
以?xún)芍荛g隔，使用0.1毫升甲醛殺死的GBS株C388/900細(xì)胞(-6×107GBS)或者用于對(duì)照組的0.1毫升PBS，對(duì)6-8周齡CD-1雌性小鼠(Charles River,St-Constant,Quebec，加拿大)皮下注射三次。在免疫的前一天，向這些制劑中加入0.14毫克或0.21毫克Al終濃度的AlhydrogelTM(Superfos Biosetor,Frederikssund,Denmark)，并且在4℃攪拌下培養(yǎng)過(guò)夜。在免疫程序開(kāi)始前和在最后注射后兩周從每一只小鼠獲得血清樣品。血清在-20℃下冷凍。
第三次注射后1星期用1.5×104cfu的GBS株C388/90(Ⅰa/c)攻擊注射PBS的每一個(gè)對(duì)照組中的8只小鼠和用甲醛殺死的全細(xì)胞GBS株C388/90(Ⅰa/c)免疫的組。用甲醛殺死的GBS全細(xì)胞免疫的所有小鼠在同種攻擊中存活，而在攻擊后5天內(nèi)，注射PBS的8只小鼠只有4只存活。為了提高對(duì)照組中的死亡率，在細(xì)菌攻擊時(shí)必須根據(jù)小鼠年齡調(diào)節(jié)細(xì)菌懸浮液。在下面的攻擊實(shí)驗(yàn)中，當(dāng)小鼠大于15周齡時(shí)，細(xì)菌接種物的濃度提高到3.0×105和2.5×106cfu之間。
表1用甲醛殺死的GBS全細(xì)胞免疫CD1小鼠并且接著同種攻擊(菌株C388/90(Ⅰa/c))和異種攻擊(菌株NCS246(Ⅱ/R))
1使用終濃度是0.14毫克或0.21毫克Al的alhydrogelTM；
2大約6×107cfu；3用調(diào)節(jié)到1.5×104cfu的含有GBS C388/90(Ⅰa/c)懸浮液的1毫升Todd-Hewitt培養(yǎng)基腹膜內(nèi)攻擊；4用調(diào)節(jié)到2.1×106cfu的含有GBS C388/90(Ⅰa/c)懸浮液的1毫升Todd-Hewitt培養(yǎng)基腹膜內(nèi)攻擊；5沒(méi)有做；6用調(diào)節(jié)到1.2×105cfu的含有GBS NCS246(Ⅱ/R)懸浮液的1毫升Todd-Hewitt培養(yǎng)基腹膜內(nèi)攻擊。
在另一項(xiàng)實(shí)驗(yàn)中，用PBS注射一組12只相應(yīng)于對(duì)照組的小鼠，而用甲醛殺死的GBSC388/90(Ⅰa/c)全細(xì)胞株免疫第二組12只小鼠。用2.1×106cfu的GBSC388/90(Ⅰa/c)株攻擊這兩組的每一個(gè)組的6只小鼠(表Ⅰ)。第一個(gè)攻擊實(shí)驗(yàn)中，所有用GBSC388/90(Ⅰa/c)株免疫的小鼠在同種攻擊中存活。注射PBS的6中小鼠中只有2只在感染中存活。
兩個(gè)組中的剩下的6只然后在一周后證明該抗原制劑是否帶來(lái)抗菌株NCS246(Ⅱ/R)的交叉保護(hù)性，其產(chǎn)生血清學(xué)特性被膜。用第二GBS菌株感染的小鼠沒(méi)有一只在感染中存活。后一種結(jié)果表明甲醛殺死的菌株C388/90誘導(dǎo)的保護(hù)性免疫應(yīng)答的大多數(shù)抗莢膜多糖，這只限于特殊血清型的菌株。這些結(jié)果清楚地表明該特定感染模型可以有效地用于研究接種帶來(lái)的保護(hù)作用。
實(shí)施例3用甲醛殺死的GBS全細(xì)胞免疫兔和小鼠的被動(dòng)免疫保護(hù)用甲醛殺死的GBS菌株C388/90(Ⅰa/c)細(xì)胞免疫新西蘭兔(2.5公斤，Charles River,St.Constant,Quebec，加拿大)，獲得超免疫血清。以三星期間隔用大約1.5×109cfu甲醛殺死的GBS菌株C388/90(Ⅰa/c)全細(xì)胞對(duì)該兔皮下注射三次，弗氏完全佐劑(GibcoBRL LifeTechnologies,Grand Island，紐約)用作第一次免疫的佐劑，弗氏不完全佐劑(GibcoBRL)用于下面的兩次注射。在免疫方案開(kāi)始之前和在最后注射后2星期取血清樣品。血清在-20℃下冷凍。
也評(píng)價(jià)了該特定兔超免疫血清對(duì)被動(dòng)保護(hù)小鼠抗GBS致命感染的能力。攻擊前18小時(shí)用15或25μL超免疫兔血清對(duì)小鼠腹膜內(nèi)注射，被保護(hù)的5只小鼠中有4只抗感染(80％)。相比之下，用PBS或用腦膜炎外膜制劑免疫的兔獲得的血清注射的對(duì)照組的小鼠存活率低于20％。該結(jié)果清楚地表明用殺死的GBS細(xì)胞免疫另一種動(dòng)物物種可以誘導(dǎo)產(chǎn)生可以被動(dòng)保護(hù)小鼠的抗體。該試劑也可以用來(lái)表征克隆。
表2用甲醛殺死的B組全鏈球菌(菌株C388/90(Ⅰa/c))抗原制劑免疫后獲得的兔血清帶來(lái)的對(duì)CD-1小鼠的被動(dòng)保護(hù)
1弗氏完全佐劑用于第一次免疫，弗氏不完全佐劑用于下面的兩次注射；2用含有調(diào)節(jié)到2×104cfu的GBS C388/90(Ⅰa/c)的1毫升Todd-Hewitt培養(yǎng)基腹膜內(nèi)攻擊。
實(shí)施例4 His.Tag-GBS融合蛋白的重組制備使用包含分別加入限制酶切位點(diǎn)BglⅡ(AGATCT)和HindⅢ(AAGCTT)的堿基延伸的寡聚物通過(guò)PCR(DNA Thermal Cycler GeneAmpPCT system2400 Perkin Elmer,San Jose,CA)從GBS菌株C388/90(Ⅰa/c)的基因組DNA擴(kuò)增GBS基因的編碼區(qū)。使用購(gòu)自Qiagen(Chatsworth,CA)的QiaexⅡ凝膠提取試劑盒用瓊脂糖凝膠純化PCR產(chǎn)物，用限制酶BglⅡ和HindⅢ(Pharmacia Canada Inc Baie d’Urfa，加拿大)消化，在乙醇沉淀之前用苯酚氯仿提取。用限制酶BglⅡ和HindⅢ消化包含硫氧還蛋白-His.Tag序列的pET-32b(+)載體(Novagen,Madison,WI)，用苯酚氯仿提取，然后乙醇沉淀之。BglⅡ-HindⅢ基因組DNA片段連接BglⅡ-HindⅢ pET-32b(+)載體，產(chǎn)生其基因在T7啟動(dòng)子控制下的硫氧還蛋白-His.Tag-GBS融合蛋白的編碼序列。根據(jù)Simanis的方法(Hanahan,D.DNA克隆，1985，D.M.Glover(編著)，pp.109-135)，將連接產(chǎn)物轉(zhuǎn)化到大腸桿菌菌株XLIBlue MRF’(Δ(mcrA)183A(mcrCB-hsdSMR-mrr)173 endA1 supE44 thi-1recAl gyrA96 relA1 lac(F’proAB lacIqZAM15Tn10(Tetr))c)(Stratagene,La Jolla,CA)中。使用Qiagen試劑盒(Qiagen,Chatsworth,CA)純化重組pET質(zhì)粒，并且通過(guò)DNA測(cè)序(Taq Dye,DeoxyTerminator Cycle Sequencing kit,ABI,Foster City,CA)證實(shí)所述DNA插入片段的核苷酸序列。通過(guò)電穿孔(Gene PulserⅡ apparatus,BIO-RAD Labs,Mississauga，加拿大)將重組pET質(zhì)粒轉(zhuǎn)化到大腸桿菌菌株AD494(DE3)(Δara-leu7697ΔlacX74 AphoA pvuⅡ phoRΔmalF3F’(lacI+(lacIq)pro)trxBKan(DE3))(Novagen,Madison,WI)k。在該大腸桿菌菌株中，其基因處于lac啟動(dòng)子控制下的T7 RNA聚合酶(存在于λDE3原噬菌體上)特異性識(shí)別控制所述融合蛋白表達(dá)的T7啟動(dòng)子，其中所述lac啟動(dòng)子由異丙基-β-D-硫代吡喃半乳糖(IPTG)誘導(dǎo)。
250rpm攪拌下在37℃下，在每毫升含有100μg氨芐青霉素(Sigma-Aldrich加拿大有限公司，Oakville，加拿大)的LB肉湯(蛋白胨10g/L，酵母提取物5g/L,NaCl 10g/L)中培養(yǎng)轉(zhuǎn)化體AD494(DE3)/rpET，直到A600達(dá)到0.6的值。為了誘導(dǎo)硫氧還蛋白-His.Tag-GBS融合蛋白的產(chǎn)生，在IPTG存在下以1mM終濃度將細(xì)胞再培養(yǎng)2小時(shí)。離心收集細(xì)菌細(xì)胞。
IPTG誘導(dǎo)2小時(shí)下AD494(DE3)/rpET32產(chǎn)生的重組融合蛋白部分以不溶的包函體獲得，通過(guò)分離不溶的團(tuán)聚體從內(nèi)源大腸桿菌蛋白質(zhì)將其純化(Gerlach,G.F.等1992,Infect.Immun.60:892)。從500毫升培養(yǎng)物誘導(dǎo)的細(xì)胞懸浮于20毫升25％蔗糖-50mM Tris-HCl緩沖液(pH8.0)并在-70℃下冷凍。通過(guò)加入5毫升250mM Tris-HCl緩沖液(pH8.0)中的溶菌酶溶液(10mg/ml)，接著在冰上溫育10-15分鐘，并且加入150毫升洗滌劑混合物(5份20mM Tris-HCl緩沖液(pH7.4)-300mM NaCl-2％脫氧膽酸-2％Nonidet P-40和4份100mM Tris-HCl緩沖液(pH8)-50mM EDTA-2％Triton X-100)，接著在冰上溫育5分鐘，實(shí)現(xiàn)解凍的懸浮液的溶解。超聲處理后，通過(guò)以35000xg離心30分鐘收集蛋白質(zhì)團(tuán)聚體并且保留溶解細(xì)胞級(jí)分的一個(gè)樣品。團(tuán)聚的蛋白質(zhì)溶解于6M鹽酸胍。使用在相應(yīng)的GBS菌株細(xì)菌攻擊中存活的注射了甲醛殺死的GBS菌株C388/90(Ⅰa/c)細(xì)胞的小鼠血清通過(guò)蛋白質(zhì)印跡分析表明溶解的和不溶的級(jí)分中都存在融合蛋白。
根據(jù)固定在His.Bind金屬螯合樹(shù)脂(Novagen，Madison,WI)上的與二價(jià)陽(yáng)離子(Ni2+)結(jié)合的His.Tag序列(6個(gè)連續(xù)的組氨酸殘基)的性質(zhì)通過(guò)親和層析進(jìn)行從IPTG-誘導(dǎo)的AD494(DE3)/rpET的溶解級(jí)分純化融合蛋白。使用的純化方法是pET system Manual，第六版(Novagen,Madison,WI)中描述的那些。簡(jiǎn)要地說(shuō)，將自IPTG誘導(dǎo)的100毫升培養(yǎng)物獲得的成粒的細(xì)胞懸浮于4毫升結(jié)合緩沖液(5mM咪唑-500mMNaCl-20mM Tris-HClpH7.9)，超聲，以39000xg離心20分鐘去除殘?jiān)?。過(guò)濾上清液(0.45μm孔徑的膜)并且沉積在結(jié)合緩沖液中平衡的His.Bind樹(shù)脂柱上。然后用10柱體積的結(jié)合緩沖液接著用6柱體積的洗滌緩沖液(20mM咪唑-500mMNaCl-20mM Tris-HClpH7.9)沖洗柱子。用洗脫緩沖液(1M咪唑-500mMNaCl-20mM Tris-HClpH7.9)洗脫硫氧還蛋白-His.Tag-GBS融合蛋白。通過(guò)在4℃下用3×1升PBS透析，從樣品中去除鹽和咪唑。
通過(guò)十二烷基硫酸鈉(SDS)-聚丙烯酰胺樹(shù)脂的考馬斯染色，用這些蛋白質(zhì)的系列稀釋物和牛血清白蛋白標(biāo)準(zhǔn)(Pierce ChemicalCo.Rockford,Ill)估計(jì)從大腸桿菌溶解或不溶細(xì)胞質(zhì)級(jí)分獲得的融合蛋白的量。
實(shí)施例5λPL啟動(dòng)子控制下GBS蛋白質(zhì)的重組制備GBS蛋白質(zhì)的DNA編碼區(qū)經(jīng)啟動(dòng)子λPL下游插入到翻譯載體pURV22中。該質(zhì)粒自p629(George等，1987,Bio/Technology5600)衍生，從p629去除I型單純皰疹病毒(HSV-Ⅰ)糖蛋白(gD-1)的一部分的編碼區(qū)，從質(zhì)粒載體pUC4K(Pharmacia Biotech Canada Inc.,Baie D’Urfe，加拿大)獲得的卡那霉素盒置換氨芐青霉素抗性基因。該載體包含λ噬菌體cI857溫度敏感阻抑蛋白基因盒，其中缺失了功能PR啟動(dòng)子。溫度從30-37℃提高至37-42℃范圍cI857阻抑蛋白的滅活導(dǎo)致λPL控制下的基因誘導(dǎo)。下游是BglⅡ限制位點(diǎn)(AGATCT)和ATGACTAAGGAGGTTAGATCTATG的核糖體結(jié)合位點(diǎn)cro控制該基因的翻譯。
根據(jù)供應(yīng)商說(shuō)明(Pharmacia Biotech Canada Inc.,Baie D’Urfe,加拿大；和New England Biolabs Ltd.,Mississauga，加拿大)使用限制酶和T4 DNA連接酶。根據(jù)Sambrook等所述(分子克隆實(shí)驗(yàn)室手冊(cè)，1989，冷泉港實(shí)驗(yàn)室出版社，N.Y.)進(jìn)行DNA片段的瓊脂糖凝膠電泳。根據(jù)Jayarao等(J.Clin.Microbiol.,1991,292774)中描述的方法制備GBS細(xì)菌的染色體DNA。使用DNA Thermal CyclerGeneAmp PCR系統(tǒng)2400(Perkin Elmer,San Jose,CA)進(jìn)行通過(guò)聚合酶鏈反應(yīng)(PCR)的DNA擴(kuò)增反應(yīng)。使用購(gòu)自Qiagen(Chatsworth,CA)的質(zhì)粒試劑盒純化用于DNA測(cè)序的質(zhì)粒。使用購(gòu)自Qiagen(Chatsworth,CA)的QiaexⅡ凝膠提取試劑盒從瓊脂糖凝膠純化DNA片段。通過(guò)Hanahan(DNA Clone,Glover(編著)pp.109-135,1985)描述的方法進(jìn)行質(zhì)粒轉(zhuǎn)化。使用寡核苷酸合成儀394型(Perkin-ElmerCorp.,Applied Biosystems Div,(ABI),Foster City,CA)合成的合成寡核苷酸進(jìn)行基因組DNA插入片段的測(cè)序。使用Taq DyeDeoxy Terminator Cycle Sequencing試劑盒(ABI,Foster City,CA)通過(guò)PCR進(jìn)行測(cè)序反應(yīng)，并且在自動(dòng)DNA測(cè)序儀373A(ABI,FosterCity,CA)上進(jìn)行DNA電泳。使用Sequencer 3.0程序(Gene CodesCorporation,Ann Arbor,MI)進(jìn)行DNA序列的組裝。應(yīng)用Gene Works2.45版程序(Intelligenetics,Inc.,Mountain ViewCA)進(jìn)行DNA序列分析和它們的預(yù)測(cè)的多肽的分析。
使用包含分別加入限制酶切位點(diǎn)BglⅡ(AGATCT)和XbaⅠ(TCTAGA)的堿基延伸的寡聚物通過(guò)PCR從GBS菌株C388/90(Ⅰa/c)的基因組DNA擴(kuò)增GBS基因的編碼區(qū)。使用購(gòu)自Qiagen(Chatsworth,CA)的QiaexⅡ凝膠提取試劑盒用瓊脂糖凝膠純化PCR產(chǎn)物，用限制酶BglⅡ和XbaⅠ消化，在乙醇沉淀之前用苯酚氯仿提取。用限制酶BglⅡ和XbaⅠ消化包含pURV22載體，用苯酚氯仿提取，然后乙醇沉淀之。BglⅡ-XbaI基因組DNA片段連接BglⅡ-XbaⅠ pURV22載體，其中GBS基因在λPL啟動(dòng)子控制下。根據(jù)上文Hanahan的方法，將連接產(chǎn)物轉(zhuǎn)化到大腸桿菌菌株XLI Blue MRF’(Δ(mcrA)183A(mcrCB-hsdSMR-mrr)173 endA1 supE44 thi-1 recA1 gyrA96 relA1 lac(F’proABlacIqZAM15Tn10(Tetr))c)(Stratagene,La Jolla,CA)中。通過(guò)將溶解的細(xì)胞在瓊脂糖凝膠上進(jìn)行電泳(Sambrook等，上文)分析來(lái)鑒定攜帶插入片段的帶有轉(zhuǎn)化體的質(zhì)粒。使用Qiagen試劑盒(Qiagen，Chatsworth,CA)純化重組pURV22質(zhì)粒，并且通過(guò)DNA測(cè)序證實(shí)所述DNA插入片段的核苷酸序列。
250rpm攪拌下在34℃下，在每毫升含有50μg卡那霉素的LB肉湯中培養(yǎng)轉(zhuǎn)化體XLI Blue MRF’/rpURV22，直到A600達(dá)到0.6的值。為了誘導(dǎo)融合蛋白的產(chǎn)生，將細(xì)菌細(xì)胞在39℃再培養(yǎng)4小時(shí)，再懸浮于樣品緩沖液中，沸騰10分鐘并且在-20℃下保存。
實(shí)施例6 CMV質(zhì)粒pCMV-GH中亞克隆GBS蛋白基因GBS蛋白DNA編碼區(qū)插入到人生長(zhǎng)激素(hGH)基因下游，其處于質(zhì)粒載體pCMV-GH中的巨細(xì)胞病毒(CMV)啟動(dòng)子的轉(zhuǎn)錄控制下(Tang等，Nature,1992,356:152)。CMV啟動(dòng)子在大腸桿菌細(xì)胞中是沒(méi)有功能的，但是施用真核細(xì)胞中的質(zhì)粒時(shí)是活性的。所述載體也插入氨芐青霉素抗性基因。
使用包含加入限制酶切位點(diǎn)BglⅡ(AGATCT)和HindⅢ(AAGCTT)的堿基延伸的寡聚物通過(guò)PCR從GBS菌株C388/90(Ⅰa/c)的基因組DNA擴(kuò)增所述基因的編碼區(qū)。使用購(gòu)自Qiagen(Chatsworth,CA)的QiaexⅡ凝膠提取試劑盒用瓊脂糖凝膠純化PCR產(chǎn)物，用限制酶BglⅡ和HindⅢ消化，在乙醇沉淀之前用苯酚氯仿提取。用限制酶BglⅡ和HindⅢ消化包含人生長(zhǎng)激素的pCMV-GH載體(StephenA.Johnston博士的實(shí)驗(yàn)室，德克薩斯大學(xué)，生物化學(xué)系，Dallas，德克薩斯州)，用苯酚氯仿提取，然后乙醇沉淀之。1.3-kb BglⅡ-HindⅢ基因組DNA片段連接BamHI-HindⅢ pCMV-GH載體，產(chǎn)生CMV啟動(dòng)子控制下的hGH-GBS融合蛋白。根據(jù)上文Hanahan的方法該連接產(chǎn)物轉(zhuǎn)化到大腸桿菌菌株DH5α(φ80 lacZ AM15 endAl recA1 hsdR17(rK-mK+)supE44 thi-1λ-gyrA1Δ(lacZYA-argF)U169)(GibcoBRL,Gaithersbury,MD)中。通過(guò)將溶解的細(xì)胞在瓊脂糖凝膠上進(jìn)行電泳(Sambrook等，上文)分析來(lái)鑒定攜帶插入片段的帶有轉(zhuǎn)化體的質(zhì)粒。使用Qiagen試劑盒(Qiagen，Chatsworth,CA)純化重組pCMV質(zhì)粒，并且通過(guò)DNA測(cè)序證實(shí)所述DNA插入片段的核苷酸序列。
實(shí)施例7 GBS蛋白質(zhì)對(duì)GBS攻擊的免疫活性以三星期間隔用0.1毫升下面的抗原制劑對(duì)6-8周齡的四組12只雌性CD-1小鼠(Charles River,St-Constant,Quebec，加拿大)皮下注射三次甲醛殺死的GBS菌株C388/90(6×107cfu),20μg來(lái)自不溶物(包函體)的硫氧還蛋白-His.Tag-GBS融合蛋白或者親和純化的(鎳柱)來(lái)自大腸桿菌細(xì)胞質(zhì)溶解級(jí)分的20μg融合蛋白，或者20μg親和純化的(鎳柱)硫氧還蛋白-His.Tag對(duì)照多肽。向每一抗原制劑中加入20μgQuilATM(Cedarlane Laboratories Ltd.Hornby，加拿大)作為佐劑。在免疫之前(PB)和在免疫過(guò)程中第20天(TB1)，41天(TB2)和54天(TB3)從每一只小鼠獲得血清樣品。血清在-20℃下冷凍。
每一次注射融合蛋白之后記錄ELISA效價(jià)的增加，表明好的初次應(yīng)答和第二次和第三次施用后特異性體液免疫應(yīng)答的加強(qiáng)。在免疫期最后，用從包函體獲得的20μg融合蛋白免疫的組相應(yīng)的ELISA效價(jià)的平均值是456145，相比之下，用得自大腸桿菌中可溶級(jí)分的蛋白質(zhì)免疫的小鼠組是290133。后一結(jié)果提示從包函體獲得的蛋白質(zhì)比可溶蛋白質(zhì)更具免疫原性。使用親和純化的硫氧還蛋白-His.Tag包被的平板在ELISA中的小鼠血清分析表明對(duì)于融合蛋白的硫氧還蛋白-His.Tag部分可忽略抗體效價(jià)。通過(guò)ELISA也試驗(yàn)了注射了重組融合蛋白的小鼠的血清對(duì)甲醛殺死的GBS菌株C388/90全細(xì)胞的反應(yīng)性。重組融合蛋白免疫誘導(dǎo)的抗體也識(shí)別GBS細(xì)胞上它們的特異性表位，表明它們的構(gòu)象足以接近天然的鏈球菌蛋白質(zhì)來(lái)誘導(dǎo)交叉反應(yīng)抗體。
為了證實(shí)免疫誘導(dǎo)的免疫應(yīng)答能否保護(hù)抗GBS感染，用3.5×105cfuGBS菌株C388/90(Ⅰa/c)和1.2×105cfu菌株NCS246(Ⅱ/R)攻擊小鼠，其結(jié)果分別在表3和4中說(shuō)明。用對(duì)照硫氧還蛋白-His.Tag肽免疫的小鼠都不能保護(hù)不受兩種GBS菌株的攻擊，而用甲醛殺死的GBS菌株C388/90全細(xì)胞免疫的那些小鼠只提供抗同種攻擊的保護(hù)。本發(fā)明硫氧還蛋白-His.Tag-GBS融合蛋白保護(hù)小鼠不受兩種GBS菌株的感染。這些小鼠的血和脾培養(yǎng)物表明不存在任何GBS。
表3在GBS菌株C388/90(Ⅰa/c)攻擊下存活試驗(yàn)1
1腹膜內(nèi)施用調(diào)節(jié)到3.5×105cfu的1毫升Todd-Hewitt培養(yǎng)基；2施用20μg；存活小鼠后腿麻痹；對(duì)血液和脾檢測(cè)GBS；3施用6×107cfu；4施用20μg。
表4在GBS菌株NCS246(Ⅱ/R)攻擊下存活試驗(yàn)1
1腹膜內(nèi)施用調(diào)節(jié)到1.2×105cfu的1毫升含有GBSNC246(Ⅱ/R)懸浮液的Todd-Hewitt培養(yǎng)基；2施用20μg；3施用6×107cfu；4一只小鼠在免疫期間死亡。
實(shí)施例8用重組GBS蛋白質(zhì)免疫賦與抗試驗(yàn)性GBS感染的保護(hù)性該實(shí)施例詳細(xì)說(shuō)明了通過(guò)用相應(yīng)于SEQ ID NO:39的重組蛋白免疫保護(hù)小鼠抗致命GBS感染的保護(hù)作用。
以三星期間隔，使用從攜帶包含相應(yīng)于SEQ ID NO:42的GBS基因的重組pURV22質(zhì)粒載體的大腸桿菌菌株BLR(Novagen)純化的20μg重組蛋白，在20μg QuilATM佐劑(Cedarlane Laboratories Ltd,Hornby，加拿大)存在下，對(duì)每組10只雌性CD-1小鼠(Charles River)皮下注射免疫三次，或者，作為對(duì)照，只使用PBS中的QuilATM注射。每一次免疫之前第1,22和43天和第三次注射之后第14天(57天)從眶下竇(orbital sinus)取血樣。一星期之后，用大約104至106CFU的各種強(qiáng)毒株GBS攻擊小鼠。在TSA/5％山羊血瓊脂平板上平板培養(yǎng)GBS攻擊種菌樣品以確定CFU和核實(shí)攻擊劑量。記錄14天的死亡情況，并且在攻擊后第14天，殺死存活的小鼠，并對(duì)血液和脾測(cè)定GBS微生物的存在。存活數(shù)據(jù)在表5中給出。
通過(guò)標(biāo)準(zhǔn)免疫測(cè)定法，對(duì)攻擊前血清分析與GBS反應(yīng)的抗體的存在。酶聯(lián)免疫吸附測(cè)定和免疫印跡分析表明用大腸桿菌中產(chǎn)生的重組GBS蛋白免疫誘發(fā)了與重組和天然GBS蛋白質(zhì)兩者都反應(yīng)的抗體。實(shí)施例9描述了對(duì)GBS應(yīng)答的抗體。
表5相應(yīng)于SEQ ID NO:39的重組GBS蛋白激發(fā)抗8種各異的GBS攻擊菌株的保護(hù)作用的能力
1使用每組10只小鼠的組，給出了感染存活的小鼠數(shù)目和死亡的小鼠的數(shù)目。應(yīng)用對(duì)于非參數(shù)分析的logrank測(cè)定，將相應(yīng)于重組GBS蛋白免疫的動(dòng)物的成活率曲線(xiàn)與相應(yīng)于模擬免疫的動(dòng)物的成活率曲線(xiàn)相比較。
2對(duì)NCS915-F-免疫的動(dòng)物的比較分析。
3在QuilATM佐劑存在下用甲醛殺死的GBS免疫動(dòng)物。
在攻擊后第14天，來(lái)自存活小鼠的所有血培養(yǎng)是陰性的。來(lái)自存活小鼠的所有脾培養(yǎng)是陰性的，除了實(shí)驗(yàn)MB-11的幾只小鼠。
實(shí)施例9用重組GBS蛋白質(zhì)接種激發(fā)對(duì)GBS的免疫應(yīng)答根據(jù)實(shí)施例8所述，用相應(yīng)于SEQ ID NO:39的重組GBS蛋白質(zhì)對(duì)10只雌性CD-1小鼠皮下免疫。為了評(píng)價(jià)對(duì)天然GBS蛋白的抗體反應(yīng)，通過(guò)ELISA，使用選自Ⅲ型NCS954菌株，Ib菌株ATCC12401，Ⅴ型菌株NCS535或Ⅵ型菌株NCS9842的甲醛殺死的GBS細(xì)胞包被的平板，對(duì)在每一次免疫之前和在第三次免疫之后14天收集的血樣血清測(cè)試抗體與GBS細(xì)胞的反應(yīng)性。對(duì)GBS細(xì)胞提取物和純化的重組抗原的蛋白質(zhì)印跡測(cè)定證實(shí)產(chǎn)生的抗體對(duì)GBS蛋白質(zhì)的特異性。圖10中顯示的結(jié)果清楚地證明動(dòng)物對(duì)用作免疫原的重組GBS蛋白質(zhì)強(qiáng)烈應(yīng)答，相應(yīng)的抗體效價(jià)中間值根據(jù)包被的抗原對(duì)于第三次免疫之后收集的血清來(lái)說(shuō)在12000和128000之間變化。當(dāng)在1∶100稀釋度下測(cè)定時(shí)，所有免疫前血清是負(fù)值。在一次注射重組GBS蛋白之后每一只動(dòng)物的血清中可檢測(cè)到與GBS反應(yīng)的抗體。
實(shí)施例10本發(fā)明GBS蛋白的抗原保守性使用特異于本發(fā)明的GBS蛋白的單克隆抗體(MAbs)來(lái)證明所有的GBS產(chǎn)生該表面抗原，并且其抗原性高度保守。
收集68個(gè)GBS分離物來(lái)評(píng)價(jià)GBS-特異性MAbs的反應(yīng)性。從加拿大Northern Alberta省立公共健康實(shí)驗(yàn)室，鏈球菌國(guó)家中心；魁北克大學(xué)中心醫(yī)院，Pavillon CHUL，魁北克，加拿大；美國(guó)典型培養(yǎng)物保藏中心，USA；加拿大魁北克Laboratoire de Sante Publique，美國(guó)西雅圖兒童醫(yī)院和醫(yī)藥中心獲得這些菌株。對(duì)下面菌株平行測(cè)試所有8個(gè)單克隆抗體血清型Ⅰa或Ⅰa/c的6個(gè)分離物，血清型Ⅰb的3個(gè)分離物，血清型Ⅱ的4個(gè)分離物，血清型Ⅳ的2個(gè)分離物，血清型V的2個(gè)分離物，血清型Ⅵ的2個(gè)分離物，血清型Ⅶ的2個(gè)分離物，血清型Ⅷ的1個(gè)分離物，沒(méi)有表征血清型的10個(gè)分離物和3個(gè)牛無(wú)菌鏈球菌菌株。Mab 3A2也與另外的GBS反應(yīng)血清型Ⅰa/c的9個(gè)分離物和血清型Ⅴ的10個(gè)分離物。37℃下5％CO2氣氛下在血瓊脂板上將菌株培養(yǎng)過(guò)夜。在-70℃下在含有20％(v/v)甘油的心浸液肉湯中保藏培養(yǎng)物。
為了獲得GBS蛋白質(zhì)特異性MAbs，以三星期間隔，用在20％QuilATM佐劑存在下的20μg純化的重組GBS蛋白質(zhì)(SEQ ID NO:44)三次免疫小鼠。從免疫小鼠回收的脾細(xì)胞與先前描述的非分泌SP2/0骨髓瘤細(xì)胞系(Hamel,J.等，1987,J.Med.Microbio1.23:163-170)融合產(chǎn)生雜交瘤細(xì)胞系。根據(jù)先前所述(Hamel,J.等，1987,J.Med.Microbiol.23:163-170)，使用甲醛滅活的GBS和純化的重組GBS蛋白質(zhì)(SEQ ID NO39或44)作為包被抗原，通過(guò)ELISA對(duì)雜交克隆上清液測(cè)試特異抗體產(chǎn)生。通過(guò)限制稀釋度克隆特異的雜化物，擴(kuò)展，并且在液氮中冷凍。實(shí)施例4和5中存在重組GBS蛋白質(zhì)的產(chǎn)生。通過(guò)使用不連續(xù)的Laemmli的緩沖體系根據(jù)生產(chǎn)商說(shuō)明通過(guò)電泳解離純化的重組GBS蛋白質(zhì)或甲醛滅活的GBS，然后轉(zhuǎn)移到硝基纖維素膜上用于蛋白質(zhì)免疫印跡測(cè)定，如先前所述(Martin等，1992,Infect.Immun.60:2718-2725)。
蛋白質(zhì)免疫印跡實(shí)驗(yàn)清楚地表明所有8個(gè)單克隆抗體識(shí)別相應(yīng)于純化的重組GBS蛋白質(zhì)(SEQ ID NO:39)的蛋白質(zhì)帶。這些單克隆抗體還與迄今為止試驗(yàn)的每一種GBS分離物中存在的蛋白質(zhì)帶反應(yīng)。表6中給出了這些GBS特異性單克隆抗體的反應(yīng)性。每一種單克隆抗體與所有46種GBS很好地反應(yīng)。另外，這些單克隆抗體也識(shí)別測(cè)試過(guò)的牛源的3個(gè)無(wú)菌鏈球菌菌株。MAb3A2還識(shí)別19種GBS；血清型Ⅰa/c的9個(gè)分離物和血清型Ⅴ的10個(gè)分離物。對(duì)其它MAbs沒(méi)有測(cè)定抗這些另外的菌株的性能。
這些結(jié)果證明迄今為止測(cè)定的所有65種GBS和牛源的3個(gè)無(wú)菌鏈球菌菌株都產(chǎn)生GBS蛋白質(zhì)(SEQ ID NO:39)。更重要的是，這些結(jié)果清楚地證明這8個(gè)GBS-特異性MAbs識(shí)別的表位分布廣泛并且在GBS中是保守的。這些結(jié)果還表明這些表位不限制于血清學(xué)相關(guān)的分離物，因?yàn)闇y(cè)試了所有已知GBS血清型，包括主要疾病引發(fā)組的代表。
結(jié)論是，該實(shí)施例中存在的數(shù)據(jù)清楚地證明所有的GBS都產(chǎn)生本發(fā)明的GBS蛋白質(zhì)并且其抗原性高度保守。
表6.根據(jù)蛋白質(zhì)印跡評(píng)價(jià)的8個(gè)GBS蛋白質(zhì)特異性單克隆抗體與不同的無(wú)菌鏈球菌菌株的反應(yīng)性
1MAb3A2識(shí)別血清型Ⅰa/c的9個(gè)另外的菌株和血清型Ⅴ的10個(gè)菌株。
2這些菌株沒(méi)有表征血清型。
序列表<110>BiOChem VaccinsRIOUX,ClémentDENIS,MartinBRODEUR,Bernard R.
HAMEL,JoséeCHARLEBOIS,IsabelleBOYER,Martine<120>新的B組鏈球菌抗原<130>12806-9PCT<150>60/075,425<151>1998-02-20<160>44<170>FastSEQ for window5 Version 3.0<210>1<211>4514<212>DNA<213>鏈球菌<220><221>CDS<222>(3)…(464)<221>CDS<222>(534)…(887)<223><221>CDS<222>(1024)…(1767)<221>CDS<222>(1841)…(4288)<221>CDS<222>(2735)…(4288)<400>1ta tct ggc aaa gag cca gct aat cgt ttt agt tgg gct aaa aat aaa47Ser Gly Lys Glu Pro Ala Asn Arg Phe Ser Trp Ala Lys Asn Lys1 5 10 15tta tta atc aat gga ttc att gca act cta gca gca act atc tta ttt 95Leu Leu Ile Asn Gly Phe Ile Ala Thr Leu Ala Ala Thr Ile Leu Phe20 25 30ttt gca gtt caa ttc ata ggt ctt aaa cca gat tac cct gga aaa acc 143Phe Ala Val Gln Phe Ile Gly Leu Lys Pro Asp Tyr Pro Gly Lys Thr35 40 45tac ttt att atc cta ttg aca gca tgg acc ttg atg gca tta gta act 191Tyr Phe Ile Ile Leu Leu Thr Ala Trp Thr Leu Met Ala Leu Val Thr50 55 60gct tta gtg gga tgg gat aat agg tat ggt tcc ttc ttg tcg tta tta 239Ala Leu Val Gly Trp Asp Ash Arg Tyr Gly Ser Phe Leu Ser Leu Leu65 70 75ata tta tta ttc cag ctt ggt tca agc gca gga act tac cca ata gaa 287Ile Leu Leu Phe Gln Leu Gly Ser Ser Ala Gly Thr Tyr Pro Ile Glu80 85 90 95ttg agt cct aag ttc ttt caa aca att caa cca ttt tta ccg atg act 335Leu Ser Pro Lys Phe Phe Gln Thr Ile Gln Pro Phe Leu Pro Met Thr100 105 110tac tct gtt tca gga tta aga gag acc atc tcg ttg acg gga gac gtt 383Tyr Ser Val Ser Gly Leu Arg Glu Thr Ile Ser Leu Thr Gly Asp Val115 120 125aac cat caa tgg aga atg cta gta atc ttt tta gta tca tcg atg ata 431Ash His Gln Trp Arg Met Leu Val Ile Phe Leu Val Ser Ser Met Ile130 135 140ctt gct ctt ctt att tat cgt aaa caa gaa gat taatagaaag tatctagtga 484Leu Ala Leu Leu Ile Tyr Arg Lys Gln Glu Asp145 150tagactaaca gtatgatatg gtatgtcaaa gtatttagga ggagaagat atg tct act 542Met Ser Thr155tta aca ata att att gca aca tta act gct ttg gaa cat ttt tat att 590Leu Thr Ile Ile Ile Ala Thr Leu Thr Ala Leu Glu His Phe Tyr Ile160 165 170atg tat ttg gag acg tta gcc acc cag tca aat atg act ggg aag att 638Met Tyr Leu Glu Thr Leu Ala Thr Gln Ser Asn Met Thr Gly Lys Ile175 180 185ttt agt atg tct aaa gaa gag ttg tca tat tta ccc gtt att aaa ctt 686Phe Ser Met Ser Lys Glu Glu Leu Ser Tyr Leu Pro Val Ile Lys Leu190 195 200 205ttt aag aat caa ggt gta tac aac ggc ttg att ggc cta ttc ctc ctt 734Phe Lys Asn Gln Gly Val Tyr Asn Gly Leu Ile Gly Leu Phe Leu Leu210 215 220tat ggg tta tat att tca cag aat caa gaa att gta gct gtt ttt tta 782Tyr Gly Leu Tyr Ile Ser Gln Asn Gln Glu Ile Val Ala Val Phe Leu225 230 235atc aat gta ttg cta gtt gct att tat ggt gct ttg aca gtt gat aaa 830Ile Ash Val Leu Leu Val Ala Ile Tyr Gly Ala Leu Thr Val Asp Lys240 245 250aaa atc tta tta aaa cag ggt ggt tta cct ata tta gct ctt tta aca 878Lys Ile Leu Leu Lys Gln Gly Gly Leu Pro Ile Leu Ala Leu Leu Thr255 260 265ttc tta ttt taatactact tagccgttcg atttagttga acggctttta 927Phe Leu Phe270gtaatcattt ttttctcata atacaggtag tttaagtaat ttgtctttaa aaatagtata 987atataactac gaattcaaag agaggtgact ttgatt atg act gag aac tgg tta1041Met Thr Glu ASn Trp Leu275cat act aaa gat ggt tca gat att tat tat cgt gtc gtt ggt caa ggt1089His Thr Lys Asp Gly Ser Asp Ile Tyr Tyr Arg Val Val Gly Gln Gly280 285 290caa ccg att gtt ttt tta cat ggc aat agc tta agt agt cgc tat ttt1137Gln Pro Ile Val Phe Leu His Gly Asn Ser Leu Ser Ser Arg Tyr Phe295 300 305 310gat aag caa ata gca tat ttt tct aag tat tac caa gtt att gtt atg1185Asp Lys Gln Ile Ala Tyr Phe Ser Lys Tyr Tyr Gln Val Ile Val Met315 320 325gat agt aga ggg cat ggc aaa agt cat gca aag cta aat acc att agt1233Asp Ser Arg Gly His Gly Lys Ser His Ala Lys Leu Asn Thr Ile Ser330 335 340ttc agg caa ata gca gtt gac tta aag gat atc tta gtt cat tta gag1281Phr Arg Gln Ile Ala Val Asp Leu Lys Asp Ile Leu Val His Leu Glu345 350 355att gat aaa gtt ata ttg gta ggc cat agc gat ggt gcc aat tta gct1329Ile Asp Lys Val Ile Leu Val Gly His Ser Asp Gly Ala Asn Leu Ala360 365 370tta gtt ttt caa acg atg ttt cca ggt atg gtt aga ggg ctt ttg ctt1377Leu Val Phe Gln Thr Met Phe Pro Gly Met Val Arg Gly Leu Leu Leu375 380 385 390aat tca ggg aac ctg act att cat ggt cag cga tgg tgg gat att ctt1425Asn Ser Gly ASn Leu Thr Ile His Gly Gln Arg Trp Trp Asp Ile Leu395 400 405tta gta agg att gcc tat aaa ttc ctt cac tat tta ggg aaa ctc ttt1473Leu Val Arg Ile Ala Tyr Lys Phe Leu His Tyr Leu Gly Lys Leu Phe410 415 420ccg tat atg agg caa aaa gct caa gtt att tcg ctt atg ttg gag gat1521Pro Tyr Met Arg Gln Lys Ala Gln Val Ile Ser Leu Met Leu Glu Asp425 430 435ttg aag att agt cca gct gat tta cag cat gtg tca act cct gta atg1569Leu Lys Ile Ser Pro Ala Asp Leu Gln His Val Ser Thr Pro Val Met440 445 450gtt ttg gtt gga aat aag gac ata att aag tta aat cat tct aag aaa1617Val Leu Val Gly Asn Lys Asp Ile Ile Lys Leu Asn His Ser Lys Lys455 460 465 470ctt gct tct tat ttt cca agg ggg gag ttt tat tct tta gtt ggc ttt1665Leu Ala Ser Tyr Phe Pro Arg Gly Glu Phe Tyr Ser Leu Val Gly Phe475 480 485ggg cat cac att att aag caa gat tcc cat gtt ttt aat att att gca1713Gly His His Ile Ile Lys Gln Asp Ser His Val Phe Asn Ile Ile Ala490 495 500aaa aag ttt atc aac gat acg ttg aaa gga gaa att gtt gaa aaa gct1761Lys Lys Phe Ile ASn Asp Thr Leu Lys Gly Glu Ile Val Glu Lys Ala505 510 515aat tga aaaagtcaaa tcactgactt ctgtgattaa aattgtattt tttatatctg 1817Asn *ttttagtgct tattattgtt gaa atg att cat ttg aaa cga act att tct gtt 1870Met Ile His Leu Lys Arg Thr Ile Ser Val520 525gag caa cta aag agt gtt ttt ggg caa tta tct cca atg aat ctt ttc1918Glu Gln Leu Lys Ser Val Phe Gly Gln Leu Ser Pro Met Asn Leu Phe530 535 540 545tta att atc ctt gtg ggg gtt atc gct gtc tta ccg aca acc gga tat1966Leu Ile Ile Leu Val Gly Val Ile Ala Val Leu Pro Thr Thr Gly Tyr550 555 560gac ttt gta ctg aat gga ctt tta cgt aca gat aaa agc aaa agg tat2014Asp Phe Val Leu Asn Gly Leu Leu Arg Thr Asp Lys Ser Lys Arg Tyr565 570 575att tta cag act agt tgg tgt atc aac act ttt aat aac ttg tca gga2062Ile Leu Gln Thr Ser Trp Cys Ile Asn Thr Phe ASn Asn Leu Ser Gly580 585 590ttc ggt ggc tta atc gat att ggg ttg cgc atg gct ttt tat ggt aaa2110Phe Gly Gly Leu Ile Asp Ile Gly Leu Arg Met Ala Phe Tyr Gly Lys595 600 605aaa ggt caa gag aag agt gac cta aga gaa gtg act cgt ttt tta ccc2158Lys Gly Gln Glu Lys Ser Asp Leu Arg Glu Val Thr Arg Phe Leu Pro610 615 620 625tat ctt att tct ggt ctg tca ttt att agt gtg att gcc tta atc atg2206Tyr Leu Ile Ser Gly Leu Ser Phe Ile Ser Val Ile Ala Leu Ile Met630 635 640agc cat att ttt cat gcc aaa gct agt gtt gat tac tat tat ttg gta2254Ser His Ile Phe His Ala Lys Ala Ser Val Asp Tyr Tyr Tyr Leu Val645 650 655tta att ggt gct agt atg tat ttt cct gtt att tat tgg att tct ggt2302Leu Ile Gly Ala Ser Met Tyr Phe Pro Val Ile Tyr Trp Ile Ser Gly660 665 670cat aaa gga agc cat tat ttc gga gat atg cca tct agt act cgt ata2350His Lys Gly Ser His Tyr Phe Gly Asp Met Pro Ser Ser Thr Arg Ile675 680 685aaa tta ggt gtt gtt tct ttt ttt gaa tgg gga tgt gcg gcc gca gca2398Lys Leu Gly Val Val Ser Phe Phe Glu Trp Gly Cys Ala Ala Ala Ala690 695 700 705ttt ata att atc ggt tat tta atg ggc att cat cta cca gtt tat aaa2446Phe Ile Ile Ile Gly Tyr Leu Met Gly Ile His Leu Pro Val Tyr Lys710 715 720att tta cca cta ttt tgt att ggt tgt gcc gtc ggg att gta tcc ctt2494Ile Leu Pro Leu Phe Cys Ile Gly Cys Ala Val Gly Ile Val Ser Leu725 730 735att ccc ggt gga tta gga agt ttt gaa tta gtt cta ttt aca ggg ttt2542Ile Pro Gly Gly Leu Gly Ser Phe Glu Leu Val Leu Phe Thr Gly Phe740 745 750gct gcc gag gga cta cct aaa gaa act gtg gtt gca tgg tta tta ctt2590Ala Ala Glu Gly Leu Pro Lys Glu Thr Val Val Ala Trp Leu Leu Leu755 760 765tat cgt tta gcc tac tat att att cca ttc ttt gca ggt atc tat ttc2638Tyr Arg Leu Ala Tyr Tyr Ile Ile Pro Phe Phe Ala Gly Ile Tyr Phe770 775 780 785ttt atc cat tat tta ggt agt caa ata aat caa cgt tat gaa aat gtc2686Phe Ile His Tyr Leu Gly Ser Gln Ile Asn Gln Arg Tyr Glu Asn Val790 795 800ccg aaa gag tta gta tca act gtt cta caa acc atg gtg agc cat ttg2734Pro Lys Glu Leu Val Ser Thr Val Leu Gln Thr Met Val Ser His Leu805 810 815atg cgt att tta ggt gca ttc tta ata ttt tca aca gca ttt ttt gaa2782Met Arg Ile Leu Gly Ala Phe Leu Ile Phe Ser Thr Ala Phe Phe Glu820 825 830aat att act tat att atg tgg ttg cag aag cta ggc ttg gac cca tta2830Asn Ile Thr Tyr Ile Met Trp Leu Gln Lys Leu Gly Leu Asp Pro Leu835 840 845caa gaa caa atg tta tgg cag ttt cca ggt tta ttg ctg ggg gtt tgt2878Gln Glu Gln Met Leu Trp Gln Phe Pro Gly Leu Leu Leu Gly Val Cys850 855 860 865ttt att ctc tta gct aga act att gat caa aaa gtg aaa aat gct ttt2926Phe Ile Leu Leu Ala Arg Thr Ile Asp Gln Lys Val Lys Asn Ala Phe870 875 880cca att gct att atc tgg att act ttg aca ttg ttt tat ctt aat tta2974Pro Ile Ala Ile Ile Trp Ile Thr Leu Thr Leu Phe Tyr Leu Asn Leu885 890 895ggt cat att agt tgg cga cra tct ttc tgg ttt att tta cta ttg tta3022Gly His Ile Ser Trp Arg Leu Ser Phe Trp Phe Ile Leu Leu Leu Leu900 905 910ggc tta tta gtc att aag cca act ctc tat aaa aaa caa ttt att tat3070Gly Leu Leu Val Ile Lys Pro Thr Leu Tyr Lys Lys Gln Phe Ile Tyr915 920 925agc tgg gaa gag cgt att aag gat gga atc att atc gtt agt tta atg3118Ser Trp Glu Glu Arg Ile Lys ASp Gly Ile Ile Ile Val Ser Leu Met930 935 940 945gga gtt cta ttt tat att gca gga cta cta ttc cct atc agg gct cat3166Gly Val Leu Phe Tyr Ile Ala Gly Leu Leu Phe Pro Ile Arg Ala His950 955 960att aca ggt ggt agt att gaa cgc ctg cat tat atc ata gca tgg gag3214Ile Thr Gly Gly Set Ile Glu Arg Leu His Tyr Ile Ile Ala Trp Glu965 970 975ccg ata gca ttg gct acg ttg att ctt act crc gtt tat tta tgt ttg3262Pro Ile Ala Leu Ala Thr Leu Ile Leu Thr Leu Val Tyr Leu Cys Leu980 985 990gtt aag att tta caa gga aaa tct tgt cag att ggt gat gtg ttc aat3310Val Lys Ile Leu Gln Gly Lys Ser Cys Gln Ile Gly Asp Val Phe Asn99510001005gtg gat cgt tat aaa aaa cta ctt caa gct tac ggt ggt tct tcg gat3358Val Asp Arg Tyr Lys Lys Leu Leu Gln Ala Tyr Gly Gly Ser Ser Asp1010 101510201025agc ggt tta gcc ttt tta aat gat aaa agg ctc tac tgg tac caa aaa3406Ser Gly Leu Ala Phe Leu Asn Asp Lys Arg Leu Tyr Trp Tyr Gln Lys103010351040aat gga gaa gat tgc gtt gcg ttc caa ttt gta att gtc aat aat aaa3454Asn Gly Glu Asp Cys Val Ala Phe Gln Phe Val Ile Val Asn Asn Lys104510501055tgt ctt att atg ggg gaa cca gcc ggt gat gac act tat att cgt gaa3502Cys Leu Ile Met Gly Glu Pro Ala Gly Asp Asp Thr Tyr Ile Arg Glu106010651070gct att gaa tcg ttt att gat gat gct gat aag cta gac tat gac ctt3550Ala Ile Glu Set Phe Ile Asp Asp Ala Asp Lys Leu Asp Tyr Asp Leu1075 10801085gtt ttt tac agt att gga cag aag ttg aca cta ctt tta cat gag tat3598Val Phe Tyr 5er Ile Gly Gln Lys Leu Thr Leu Leu Leu His Glu Tyr1090109511001105ggt ttt gac ttt atg aaa gtt ggt gag gat gct tta gtt aat tta gaa3646Gly Phe Asp Phe Met Lys Val Gly Glu Asp Ala Leu Val Asn Leu Glu111011151120acg ttt att ctt aaa ggg aat aag tac aaa cct ttc aga aat gcc cta3694Thr Phe Thr Ltu Lys Gly Asn Lys Tyr Lys Pro Phe Arg Asn Ala Leu112511301135aat aga gtt gaa aag gat ggt ttc tat ttc gaa gtt gta caa tcg cca3742Asn Arg Val Glu Lys Asp Gly Phe Tyr Phe Glu Val Val Gln Ser Pro114011451150cat agt caa gag cta cta aat agt ttg gaa gag att tct aat act tgg3790His Ser Gln Glu Leu Leu Asn Ser Leu Glu Glu Ile Ser Asn Thr Trp115511601165tta gaa gga cgc cct gaa aaa ggt ttc tca cta gga tat ttt aat aaa3838Leu Glu Gly Arg Pro Glu Lys Gly Phe Ser Leu Gly Tyr Phe Asn Lys1170117511801185gat tat ttc caa caa gcc cca ata gct ttg gta aaa aat gct gaa cac3886Asp Tyr Phe Gln Gln Ala Pro Ile Ala Leu Val Lys Asn Ala Glu His119011951200gaa gtt gct gct ttt gct aat att atg cca aac tat gaa aag agt att3934Glu Val Val Ala Phe Ala Asn Ile Met Pro Asn Tyr Glu Lys Ser Ile120512101215atc tct att gat tta atg cgt cac gat aaa cag aaa att ccg aat ggc3982Ile Ser Ile Asp Leu Met Arg His Asp Lys Gln Lys Ile Pro Asn Gly122012251230gtt atg gat ttc ctc ttt tta tca tta ttc tct tat tat caa gag aag4030Val Met Asp Phe Leu Phe Leu Ser Leu Phe Ser Tyr Tyr Gln Glu Lys123512401245gga tac cac tat ttt gat ttg ggg atg gca cct tta tca gga gtt ggt4078Gly Tyr His Tyr Phe Asp Leu Gly Met Ala Pro Leu Ser Gly Val Gly1250125512601265cgc gtt gaa aca agt ttt gct aaa gag aga atg gcg tat ctt gtc tat4126Arg Val Glu Thr Ser Phe Ala Lys Glu Arg Met Ala Tyr Leu Val Tyr127012751280cat ttc ggt agt cat trc tac tca ttt aat ggt tta cac aag tat aag4174His Phe Gly Ser His Phe Tyr Ser Phe Asn Gly Leu His Lys Tyr Lys128512901295aag aag ttt aca cca ttg tgg tcg gaa cgt tat att tct tgt tct cgt4222Lys Lys Phe Thr Pro Leu Trp Ser Glu Arg Tyr Ile Ser Cys Ser Arg130013051310tcg tcc tgg tta att tgt gct att tgt gcc cta tta atg gaa gat agt4270Ser Ser Trp Leu Ile Cys Ala Ile Cys Ala Leu Leu Met Glu Asp Ser131513201325aaa att aag att gtt aaa taagctttat ttggcaatta aaaagagcat 4318Lys Ile Lys Ile Val Lys13301335gtcatgcgac atgctctttt taaatcattt aataccattg attgcttgaa tctactttat 4378aatatgatgt gcttttaaat attgtttagc tactgtagct gctgatttat gctttacagc 4438tacttggtag ttcatttctt gcatttcttt ttcagtgata tgaccagcaa gtttattgag 4498agcttttttt acttga 4514<210>2<211>154<212>蛋白質(zhì)<213>鏈球菌<400>2Ser Gly Lys Glu Pro Ala Asn Arg Phe Ser Trp Ala Lys Asn Lys Leu1 5 10 15Leu Ile Asn Gly Phe Ile Ala Thr Leu Ala Ala Thr Ile Leu Phe Phe20 25 30Ala Val Gln Phe Ile Gly Leu Lys Pro Asp Tyr Pro Gly Lys Thr Tyr35 40 45Phe Ile Ile Leu Leu Thr Ala Trp Thr Leu Met Ala Leu Val Thr Ala50 55 60Leu Val Gly Trp Asp Asn Arg Tyr Gly Ser Phe Leu Ser Leu Leu Ile65 70 75 80Leu Leu Phe Gln Leu Gly Ser Ser Ala Gly Thr Tyr Pro Ile Glu Leu85 90 95Ser Pro Lys Phe Phe Gln Thr Ile Gln Pro Phe Leu Pro Met Thr Tyr100 105 110Ser Val Ser Gly Leu Arg Glu Thr Ile Ser Leu Thr Gly Asp Val Asn115 120 125His Gln Trp Arg Met Leu Val Ile Phe Leu Val Ser Ser Met Ile Leu130 135 140Ala Leu Leu Ile Tyr Arg Lys Gln Glu Asp145 150<210>3<211>118<212>蛋白質(zhì)<213>鏈球菌<400> 3Met Ser Thr Leu Thr Ile Ile Ile Ala Thr Leu Thr Ala Leu Glu His1 5 10 15Phe Tyr Ile Met Tyr Leu Glu Thr Leu Ala Thr Gln Ser Asn Met Thr20 25 30Gly Lys Ile Phe Ser Met Ser Lys Glu Glu Leu Ser Tyr Leu Pro Val35 40 45Ile Lys Leu Phe Lys Asn Gln Gly Val Tyr Asn Gly Leu Ile Gly Leu50 55 60Phe Leu Leu Tyr Gly Leu Tyr Ile Ser Gln Ash Gln Glu Ile Val Ala65 70 75 80Val Phe Leu Ile Asn Val Leu Leu Val Ala Ile Tyr Gly Ala Leu Thr85 90 95Val Asp Lys Lys Ile Leu Leu Lys Gln Gly Gly Leu Pro Ile Leu Ala100 105 110Leu Leu Thr Phe Leu Phe115<210>4<211>247<212>蛋白質(zhì)<213>鏈球菌<400>4Met Thr Glu Asn Trp Leu His Thr Lys Asp Gly Ser Asp Ile Tyr Tyr1 5 10 15Arg Val Val Gly Gln Gly Gln Pro Ile Val Phe Leu His Gly Asn Ser20 25 30Leu Ser Ser Arg Tyr Phe Asp Lys Gln Ile Ala Tyr Phe Ser Lys Tyr35 40 45Tyr Gln Val Ile Val Met Asp Ser Arg Gly His Gly Lys Ser His Ala50 55 60Lys Leu Asn Thr Ile Ser Phe Arg Gln Ile Ala Val Asp Leu Lys Asp65 70 75 80Ile Leu Val His Leu Glu Ile Asp Lys Val Ile Leu Val Gly His Ser85 90 95Asp Gly Ala Asn Leu Ala Leu Val Phe Gln Thr Met Phe Pro Gly Met100 105 110Val Arg Gly Leu Leu Leu Asn Ser Gly Asn Leu Thr Ile His Gly Gln115 120 125Arg Trp Trp Asp Ile Leu Leu Val Arg Ile Ala Tyr Lys Phe Leu His130 135 140Tyr Leu Gly Lys Leu Phe Pro Tyr Met Arg Gln Lys Ala Gln Val Ile145 150 155 160Ser Leu Met Leu Glu Asp Leu Lys Ile Ser Pro Ala Asp Leu Gln His165 170 175Val Ser Thr Pro Val Met Val Leu Val Gly Asn Lys Asp Ile Ile Lys180 185 190Leu Asn His Ser Lys Lys Leu Ala Ser Tyr Phe Pro Arg Gly Glu Phe195 200 205Tyr Ser Leu Val Gly Phe Gly His His Ile Ile Lys Gln Asp Ser His210 215 220Val Phe Asn Ile Ile Ala Lys Lys Phe Ile Asn Asp Thr Leu Lys Gly225 230 235 240Glu Ile Val Glu Lys Ala Asn245<210>5<211>816<212>蛋白質(zhì)<213>鏈球菌<400>5Met Ile His Leu Lys Arg Thr Ile Set Val Glu Gln Leu Lys Ser Val1 5 10 15Phe Gly Gln Leu Ser Pro Met Asn Leu Phe Leu Ile Ile Leu Val Gly20 25 30Val Ile Ala Val Leu Pro Thr Thr Gly Tyr Asp Phe Val Leu Asn Gly35 40 45Leu Leu Arg Thr Asp Lys Ser Lys Arg Tyr Ile Leu Gln Thr Ser Trp50 55 60Cys Ile Asn Thr Phe Asn Asn Leu Ser Gly Phe Gly Gly Leu Ile Asp65 70 75 80Ile Gly Leu Arg Met Ala Phe Tyr Gly Lys Lys Gly Gln Glu Lys Ser85 90 95Asp Leu Arg Glu Val Thr Arg Phe Leu Pro Tyr Leu Ile Ser Gly Leu100 105 110Ser Phe Ile Ser Val Ile Ala Leu Ile Met Ser His Ile Phe His Ala115 120 125Lys Ala Ser Val Asp Tyr Tyr Tyr Leu Val Leu Ile Gly Ala Ser Met130 135 140Tyr Phe Pro Val Ile Tyr Trp Ile Ser Gly His Lys Gly Ser His Tyr145 150 155 160Phe Gly Asp Met Pro Ser Ser Thr Arg Ile Lys Leu Gly Val Val Ser165 170 175Phe Phe Glu Trp Gly Cys Ala Ala Ala Ala Phe Ile Ile Ile Gly Tyr180 185 190Leu Met Gly Ile His Leu Pro Val Tyr Lys Ile Leu Pro Leu Phe Cys195 200 205Ile Gly Cys Ala Val Gly Ile Val Ser Leu Ile Pro Gly Gly Leu Gly210 215 220Ser Phe Glu Leu Val Leu Phe Thr Gly Phe Ala Ala Glu Gly Leu Pro225 230 235 240Lys Glu Thr Val Val Ala Trp Leu Leu Leu Tyr Arg Leu Ala Tyr Tyr245 250 255Ile Ile Pro Phe Phe Ala Gly Ile Tyr Phe Phe Ile His Tyr Leu Gly260 265 270Ser Gln Ile Asn Gln Arg Tyr Glu Asn Val Pro Lys Glu Leu Val Ser275 280 285Thr Val Leu Gln Thr Met Val Ser His Leu Met Arg Ile Leu Gly Ala290 295 300Phe Leu Ile Phe Ser Thr Ala Phe Phe Glu Asn Ile Thr Tyr Ile Met305 310 315 320Trp Leu Gln Lys Leu Gly Leu Asp Pro Leu Gln Glu Gln Met Leu Trp325 330 335Gln Phe Pro Gly Leu Leu Leu Gly Val Cys Phe Ile Leu Leu Ala Arg340 345 350Thr Ile Asp Gln Lys Val Lys Asn Ala Phe Pro Ile Ala Ile Ile Trp355 360 365Ile Thr Leu Thr Leu Phe Tyr Leu Asn Leu Gly His Ile Ser Trp Arg370 375 380Leu Ser Phe Trp Phe Ile Leu Leu Leu Leu Gly Leu Leu Val Ile Lys385 390 395 400Pro Thr Leu Tyr Lys Lys Gln Phe Ile Tyr Ser Trp Glu Glu Arg Ile405 410 415Lys Asp Gly Ile Ile Ile Val Ser Leu Met Gly Val Leu Phe Tyr Ile420 425 430Ala Gly Leu Leu Phe Pro Ile Arg Ala His Ile Thr Gly Gly Ser Ile435 440 445Glu Arg Leu His Tyr Ile Ile Ala Trp Glu Pro Ile Ala Leu Ala Thr450 455 460Leu Ile Leu Thr Leu Val Tyr Leu Cys Leu Val Lys Ile Leu Gln Gly465 470 475 480Lys Ser Cys Gln Ile Gly Asp Val Phe Asn Val Asp Arg Tyr Lys Lys485 490 495Leu Leu Gln Ala Tyr Gly Gly Ser Ser Asp Ser Gly Leu Ala Phe Leu500 505 510Asn Asp Lys Arg Leu Tyr Trp Tyr Gln Lya Asn Gly Glu Asp Cys Val515 520 525Ala Phe Gln Phe Val Ile Val Asn Asn Lys Cys Leu Ile Met Gly Glu530 535 540Pro Ala Gly Asp Asp Thr Tyr Ile Arg Glu Ala Ile Glu Ser Phe Ile545 550 555 560Asp Asp Ala Asp Lys Leu Asp Tyr Asp Leu Val Phe Tyr Ser Ile Gly565 570 575Gln Lys Leu Thr Leu Leu Leu His Glu Tyr Gly Phe Asp Phe Met Lys580 585 590Val Gly Glu Asp Ala Leu Val Asn Leu Glu Thr Phe Thr Leu Lys Gly595 600 605Asn Lys Tyr Lys Pro Phe Arg Asn Ala Leu Asn Arg Val Glu Lys Asp610 615 620Gly Phe Tyr Phe Glu Val Val Gln Ser Pro His Ser Gln Glu Leu Leu625 630 635 640Asn Ser Leu Glu Glu Ile Ser Asn Thr Trp Leu Glu Gly Arg Pro Glu645 650 655Lys Gly Phe Ser Leu Gly Tyr Phe Asn Lys Asp Tyr Phe Gln Gln Ala660 665 670Pro Ile Ala Leu Val Lys Asn Ala Glu His Glu Val Val Ala Phe Ala675 680 685Asn Ile Met Pro Asn Tyr Glu Lys Ser Ile Ile Ser Ile Asp Leu Met690 695 700Arg His Asp Lys Gln Lys Ile Pro Asn Gly Val Met Asp Phe Leu Phe705 710 715 720Leu Ser Leu Phe Ser Tyr Tyr Gln Glu Lys Gly Tyr His Tyr Phe Asp725 730 735Leu Gly Met Ala Pro Leu Ser Gly Val Gly Arg Val Glu Thr Ser Phe740 745 750Ala Lys Glu Arg Met Ala Tyr Leu Val Tyr His Phe Gly Ser His Phe755 760 765Tyr Ser Phe Asn Gly Leu His Lys Tyr Lys Lys Lys Phe Thr Pro Leu770 775 780Trp Ser Glu Arg Tyr Ile Ser Cys Ser Arg Ser Ser Trp Leu Ile Cys785 790 795 800Ala Ile Cys Ala Leu Leu Met Glu Asp Ser Lys Ile Lys Ile Val Lys805 810 815<210>6<211>518<212>蛋白質(zhì)<213>鏈球菌<400>6Met Arg Ile Leu Gly Ala Phe Leu Ile Phe Ser Thr Ala Phe Phe Glu1 5 10 15Asn Ile Thr Tyr Ile Met Trp Leu Gln Lys Leu Gly Leu Asp Pro Leu20 25 30Gln Glu Gln Met Leu Trp Gln Phe Pro Gly Leu Leu Leu Gly Val Cys35 40 45Phe Ile Leu Leu Ala Arg Thr Ile Asp Gln Lys Val Lys Asn Ala Phe50 55 60Pro Ile Ala Ile Ile Trp Ile Thr Leu Thr Leu Phe Tyr Leu Asn Leu65 70 75 80Gly His Ile Ser Trp Arg Leu Ser Phe Trp Phe Ile Leu Leu Leu Leu85 90 95Gly Leu Leu Val Ile Lys Pro Thr Leu Tyr Lys Lys Gln Phe Ile Tyr100 105 110Ser Trp Glu Glu Arg Ile Lys Asp Gly Ile Ile Ile Val Ser Leu Met115 120 125Gly Val Leu Phe Tyr Ile Ala Gly Leu Leu Phe Pro Ile Arg Ala His130 135 140Ile Thr Gly Gly Ser Ile Glu Arg Leu His Tyr Ile Ile Ala Trp Glu145 150 155 160Pro Ile Ala Leu Ala Thr Leu Ile Leu Thr Leu Val Tyr Leu Cys Leu165 170 175Val Lys Ile Leu Gln Gly Lys Ser Cys Gln Ile Gly Asp Val Phe Asn180 185 190Val Asp Arg Tyr Lys Lys Leu Leu Gln Ala Tyr Gly Gly Ser Ser Asp195 200 205Ser Gly Leu Ala Phe Leu Asn Asp Lys Arg Leu Tyr Trp Tyr Gln Lys210 215 220Asn Gly Glu Asp Cys Val Ala Phe Gln Phe Val Ile Val Asn Asn Lys225 230 235 240Cys Leu Ile Met Gly Glu Pro Ala Gly Asp Asp Thr Tyr Ile Arg Glu245 250 255Ala Ile Glu Ser Phe Ile Asp Asp Ala Asp Lys Leu Asp Tyr Asp Leu260 265 270Val Phe Tyr Ser Ile Gly Gln Lys Leu Thr Leu Leu Leu His Glu Tyr275 280 285Gly Phe Asp Phe Met Lys Val Gly Glu Asp Ala Leu Val Asn Leu Glu290 295 300Thr Phe Thr Leu Lys Gly Asn Lys Tyr Lys Pro Phe Arg Asn Ala Leu305 310 315 320Asn Arg Val Glu Lys Asp Gly Phe Tyr Phe Glu Val Val Gln Ser Pro325 330 335His Ser Gln Glu Leu Leu Asn Ser Leu Glu Glu Ile Ser Asn Thr Trp340 345 350Leu Glu Gly Arg Pro Glu Lys Gly Phe Ser Leu Gly Tyr Phe Asn Lys355 360 365Asp Tyr Phe Gln Gln Ala Pro Ile Ala Leu Val Lys Asn Ala Glu His370 375 380Glu Val Val Ala Phe Ala Asn Ile Met Pro Asn Tyr Glu Lys Ser Ile385 390 395 400Ile Ser Ile Asp Leu Met Arg His Asp Lys Gln Lys Ile Pro Asn Gly405 410 415Val Met Asp Phe Leu Phe Leu Ser Leu Phe Ser Tyr Tyr Gln Glu Lys420 425 430Gly Tyr His Tyr Phe Asp Leu Gly Met Ala Pro Leu Ser Gly Val Gly435 440 445Arg Val Glu Thr Ser Phe Ala Lys Glu Arg Met Ala Tyr Leu Val Tyr450 455 460His Phe Gly Ser His Phe Tyr Ser Phe Asn Gly Leu His Lys Tyr Lys465 470 475 480Lys Lys Phe Thr Pro Leu Trp Ser Glu Arg Tyr Ile Ser Cys Ser Arg485 490 495Ser Ser Trp Leu Ile Cys Ala Ile Cys Ala Leu Leu Met Glu Asp Ser500 505 510Lys Ile Lys Ile Val Lys515<210>7<211>5126<212>DNA<213>鏈球菌<220><221>CDS<222>(1)…(687)<221>CDS<222>(701)…(2557)<221>CDS<222>(2566)…(3036)<221>CDS<222>(3106)…(4842)<221>CDS<222>(4850)…(5125)<400>7aat ttt gat atc gaa aca aca act ttt gag gca atg aaa aag cac gcg 48Asn Phe Asp Ile Glu Thr Thr Thr Phe Glu Ala Met Lys Lys His Ala1 5 10 15tca tta ttg gag aaa ata tct gtt gag cgt tct ttt att gaa ttt gat 96Ser Leu Leu Glu Lys Ile Ser Val Glu Arg Ser Phe Ile Glu Phe Asp20 25 30aaa ctt cta tta gca cct tat tgg cgt aaa gga atg ctg gca cta ata 144Lys Leu Leu Leu Ala Pro Tyr Trp Arg Lys Gly Met Leu Ala Leu Ile35 40 45gat agt cat gct ttt aat tat cta cca tgc tta aaa aat agg gaa tta 192Asp Ser His Ala Phe Asn Tyr Leu Pro Cys Leu Lys Asn Arg Glu Leu50 55 60caa tta agc gcc ttt ttg tcc cag tta gat aaa gat ttt tta ttt gag 240Gln Leu Ser Ala Phe Leu Ser Gln Leu Asp Lys Asp Phe Leu Phe Glu65 70 75 80aca rca gaa caa gct tgg gca tca ctc atc ttg agt atg gaa gtt gaa 288Thr Ser Glu Gln Ala Trp Ala Ser Leu Ile Leu Ser Met Glu Val Glu85 90 95cac aca aag act ttt tta aaa aaa tgg aag aca tca act cac ttt caa 336His Thr Lys Thr Phe Leu Lys Lys Trp Lys Thr Ser Thr His Phe Glnl00 105 110aaa gac gtt gag cat ata gtg gat gtt tat cgt att cgt gaa caa atg 384Lys Asp Val Glu His Ile Val Asp Val Tyr Arg Ile Arg Glu Gln Met115 120 125gga ttg gct aaa gaa cat ctt tat cgt tat gga aaa act ata ata aaa 432Gly Leu Ala Lys Glu His Leu Tyr Arg Tyr Gly Lys Thr Ile Ile Lys130 135 140caa gcg gaa ggt att cgc aaa gca aga ggc ttg atg gtt gat ttc gaa 480Gln Ala Glu Gly Ile Arg Lys Ala Arg Gly Leu Met Val Asp Phe Glu145 150 155 160aaa ata gaa caa cta gat agt gag tta gca atc cat gat agg cat gag 528Lys Ile Glu Gln Leu Asp Ser Glu Leu Ala Ile His Asp Arg His Glu165 170 175ata gtt gtc aat ggt ggc acc tta atc aag aaa tta gga ata aaa cct 576Ile Val Val Asn Gly Gly Thr Leu Ile Lys Lys Leu Gly Ile Lys Pro180 185 190ggt cca cag atg gga gat att atc tct caa att gaa tta gcc att gtt 624Gly Pro Gln Met Gly Asp Ile Ile Ser Gln Ile Glu Leu Ala Ile Val195 200 205tta gga caa ctg att aat gaa gaa gag gct att tta cat ttt gtt aag 672Leu Gly Gln Leu Ile Asn Glu Glu Glu Ala Ile Leu His Phe Val Lys210 215 220cag tac ttg atg gat tagagaggat tat atg agc gat ttt tta gta gat 721Gln Tyr Leu Met Asp Met Ser Asp Phe Leu Val Asp225 230 235gga ttg act aag tcg gtt ggt gat aag acg gtc ttt agt aat gtt tca 769Gly Leu Thr Lys Ser Val Gly Asp Lys Thr Val Phe Ser Asn Val Ser240 245 250rtt atc atc cat agt tta gac cgt att ggg att att ggt gtc aat gga 817Phe Ile Ile His Ser Leu Asp Arg Ile Gly Ile Ile Gly Val Asn Gly255 260 265act gga aag aca aca cta tta gat gtt att tcg ggt gaa tta ggt ttt 865Thr Gly Lys Thr Thr Leu Leu Asp Val Ile Ser Gly Glu Leu Gly Phe270 275 280gat ggt gat cgt tcc cct ttt tca tca gct aat gat tat aag att gct 913Asp Gly Asp Arg Ser Pro Phe Ser Ser Ala Asn Asp Tyr Lys Ile Ala285 290 295 300tat tta aaa caa gaa cca gac ttt gat gat tct cag aca att ttg gac 961Tyr Leu Lys Gln Glu Pro Asp Phe Asp Asp Ser Gln Thr Ile Leu Asp305 310 315acc gta ctt tct tct gac tta aga gag atg gct tta att aaa gaa tat1009Thr Val Leu Ser Ser Asp Leu Arg Glu Net Ala Leu Ile Lys Glu Tyr320 325 330gaa tta ttg ctt aat cac tac gaa gaa agt aag caa tca cgt cta gag1057Glu Leu Leu Leu Asn His Tyr Glu Glu Ser Lys Gln Ser Arg Leu Glu335 340 345aaa gta atg gca gaa atg gat tct tta gat gct tgg tct att gag agc1105Lys Val Met Ala Glu Met Asp Ser Leu Asp Ala Trp Ser Ile Glu Ser350 355 360gaa gtc aaa aca gta tta tcc aaa tta ggt att act gat ttg cag ttg1153Glu Val Lys Thr Val Leu Set Lys Leu Gly Ile Thr Asp Leu Gln Leu365 370 375 380tcg gtt ggt gaa tta tca gga gga tta cga aga cgt gtt caa tta gcg1201Set Val Gly Glu Leu Set Gly Gly Leu Arg Arg Arg Val Gln Leu Ala385 390 395caa gta tta tta aat gat gca gat tta ttg ctc tta gac gaa cct act1249Gln Val Leu Leu Asn Asp Ala Asp Leu Leu Leu Leu Asp Glu Pro Thr400 405 410aac cac tta gat att gac act att gca tgg tta acg aat ttt ttg aaa1297Asn His Leu Asp Ile Asp Thr Ile Ala Trp Leu Thr Asn Phe Leu Lys415 420 425aat agt aaa aag aca gtg ctt ttt ata act cat gat cgt tat ttt cta1345Asn Set Lys Lys Thr Val Leu Phe Ile Thr His Asp Arg Tyr Phe Leu430 435 440gac aat gtt gca aca cgt att ttt gaa tta gat aag gca cag att aca1393Asp Asn Val Ala Thr Arg Ile Phe Glu Leu Asp Lys Ala Gln Ile Thr445 450 455 460gaa tat caa ggc aat tat cag gat tat gtc cga ctt cgt gca gaa caa1441Glu Tyr Gln Gly Asn Tyr Gln Asp Tyr Val Arg Leu Arg Ala Glu Gln465 470 475gac gag cgt gat gct gct agt tta cat aaa aag aaa cag ctt tat aaa1489Asp Glu Arg Asp Ala Ala Ser Leu His Lys Lys Lys Gln Leu Tyr Lys480 485 490cag gaa cta gct tgg atg cgt act cag cca caa gct cgt gca acg aaa1537Gln Glu Leu Ala Trp Met Arg Thr Gln Pro Gln Ala Arg Ala Thr Lys495 500 505caa cag gct cgt att aat cgt ttt caa aat cta aaa aac gat tta cac1585Gln Gln Ala Arg Ile Asn Arg Phe Gln Asn Leu Lys Asn Asp Leu His510 515 520caa aca agc gat aca agc gat ttg gaa atg aca ttt gaa aca agt cga 1633Gln Thr Ser Asp Thr Ser Asp Leu Glu Met Thr Phe Glu Thr Ser Arg525 530 535 540att ggg aaa aag gtt att aat ttt gaa aat gtc tct ttt tct tac cca 1681Ile Gly Lys Lys Val Ile Asn Phe Glu Asn Val Ser Phe Ser Tyr Pro545 550 555gat aaa tct atc ttg aaa gac ttt aat ttg tta att caa aat aaa gac 1729Asp Lys Ser Ile Leu Lys Asp Phe Asn Leu Leu Ile Gln Asn Lys Asp560 565 570cgt att ggc atc gtt gga gat aat ggt gtt gga aag tca acc tta ctt 1777Arg Ile Gly Ile Val Gly Asp Asn Gly Val Gly Lys Ser Thr Leu Leu575 580 585aat tta att gtt caa gat tta cag ccg gat tcg ggt aat gtc tct att 1825Asn Leu Ile Val Gln Asp Leu Gln Pro Asp Ser Gly Asn Val Ser Ile590 595 600ggt gaa acg ata cgt gta ggt tac ttt tca caa caa ctt cat aat atg 1873Gly Glu Thr Ile Arg Val Gly Tyr Phe Ser Gln Gln Leu His Asn Met605 610 615 620gat ggc tca aaa cgt gtt att aat tat ttg caa gag gtt gca gat gag 1921Asp Gly Ser Lys Arg Val Ile Asn Tyr Leu Gln Glu Val Ala Asp Glu625 630 635gtt aaa act agt gtc ggt aca aca agt gtg aca gaa cta ttg gaa caa 1969Val Lys Thr Ser Val Gly Thr Thr Ser Val Thr Glu Leu Leu Glu Gln640 645 650ttt ctc ttt cca cgt tcg aca cat gga aca caa att gca aaa tta tca 2017Phe Leu Phe Pro Arg Ser Thr His Gly Thr Gln Ile Ala Lys Leu Ser655 660 665ggt ggt gag aaa aaa aga ctt tac ctt tta aaa atc ctg att gaa aag 2065Gly Gly Glu Lys Lys Arg Leu Tyr Leu Leu Lys Ile Leu Ile Glu Lys670 675 680cct aat gtg tta cta ctt gat gag ccg aca aat gac tta gat att gct 2113Pro Asn Val Leu Leu Leu Asp Glu Pro Thr Asn Asp Leu Asp Ile Ala685 690 695 700aca tta act gtt ctt gaa aat ttt tta caa ggc ttt ggt ggt cct gtg 2161Thr Leu Thr Val Leu Glu Asn Phe Leu Gln Gly Phe Gly Gly Pro Val705 710 715att aca gtt agt cac gat cgt tac ttt tta gat aaa gtg gct aat aaa 2209Ile Thr Val Ser His Asp Arg Tyr Phe Leu Asp Lys Val Ala Asn Lys720 725 730att att gcg ttt gaa gat aac gat atc cgt gaa ttt ttt ggt aat tat 2257Ile Ile Ala Phe Glu Asp Asn Asp Ile Arg Glu Phe Phe Gly Asn Tyr735 740 745act gat tat tta gat gaa aaa gca ttt aat gag caa aat aat gaa gtt 2305Thr Asp Tyr Leu Asp Glu Lys Ala Phe Asn Glu Gln Asn Asn Glu Val750 755 760atc agt aaa aaa gag agt acc aag aca agt cgt gaa aag caa agt cgt2353Ile Ser Lys Lys Glu Ser Thr Lys Thr Ser Arg Glu Lys Gln Ser Arg765 770 775 780aaa aga atg tct tac ttt gaa aaa caa gaa tgg gcg aca att gaa gac2401Lys Arg Met Ser Tyr Phe Glu Lys Gln Glu Trp Ala Thr Ile Glu Asp785 790 795gat att atg ata ttg gaa aat act atc act cgt ata gaa aat gat atg2449Asp Ile Met Ile Leu Glu Asn Thr Ile Thr Arg Ile Glu Asn Asp Met800 805 810caa aca tgt ggt agt gat ttt aca agg tta tct gat tta caa aag gaa2497Gln Thr Cys Gly Ser Asp Phe Thr Arg Leu Ser Asp Leu Gln Lys Glu815 820 825tta gat gca aaa aat gaa gca ctt cta gaa aag tat gac cgt tat gag2545Leu Asp Ala Lys Asn Glu Ala Leu Leu Glu Lys Tyr Asp Arg Tyr Glu830 835 840tac ctt agt gag ttagacac atg att atc cgt ccg att att aaa aat gat 2595Tyr Leu Ser Glu LeuAspThrMet Ile Ile Arg Pro Ile Ile Lys Asn Asp845850 855 860gac caa gca gtt gca caa tta att cga caa agt tta cgc gcc tat gat2643Asp Gln Ala Val Ala Gln Leu Ile Arg Gln Sar Leu Arg Ala Tyr Asp865 870 875tta gat aaa cct gat aca gca tat tca gac cct cac tta gat cat ttg2691Leu Asp Lys Pro Asp Thr Ala Tyr Ser Asp Pro His Leu Asp His Leu880 885 890acc tca tac tac gaa aaa ata gag aag tca gga ttc ttt gtc att gag2739Thr Ser Tyr Tyr Glu Lys Ile Glu Lys Ser Gly Phe Phe Val Ile Glu895 900 905gag aga gat gag att att ggc tgt ggc ggc ttt ggt ccg ctg aaa aat2787Glu Arg Asp Glu Ile Ile Gly Cys Gly Gly Phe Gly Pro Leu Lys Asn910 915 920 925cta att gca gag atg tag aag gtg tac att gca gaa cgt ttc cgt ggt2835Leu Ile Ala Glu Met Gln Lys Val Tyr Ile Ala Glu Arg Phe Arg Gly930 935 940aag ggg ctt gct act gat tta gtg aaa atg att gaa gta gaa gct cga2883Lys Gly Leu Ala Thr Asg Leu Val Lys Met Ile Glu Val Glu Ala Arg945 950 955aaa att ggg tat aga caa ctt tat tta gag aca gcc agt act ttg agt2931Lys Ile Gly Tyr Arg Gln Leu Tyr Leu Glu Thr Ala Ser Thr Leu Ser960 965 970agg gca act gcg gtt tat aag cat atg gga tat tgt gcc tta tcg caa2979Arg Ala Thr Ala Val Tyr Lys His Met Gly Tyr Cys Ala Leu Ser Gln975 980 985cca ata gca aat gat caa ggt cat aca gct atg gat att tgg atg att3027Pro Ile Ala Asn Asp Gln Gly His Thr Ala Met Asp Ile Trp Met Ile990 99510001005aaa gat tta taagttgaaa gtggattagt gaacatggat taattatttt3076Lys Asp Leugagataagag gaaagaaaag gagacatat atg gca tat att tgg tct tat ttg3129Met Ala Tyr Ile Trp Ser Tyr Leu10101015aaa agg tac ccc aat tgg tta tgg ctt gat tta cta gga gct atg ctt3177Lys Arg Tyr Pro Asn Trp Leu Trp Leu Asp Leu Leu Gly Ala Met Leu102010251030ttt gtg acg gtt atc cta gga atg ccc aca gcc tta gcg ggt atg att3225Phe Val Thr Val Ile Leu Gly Met Pro Thr Ala Leu Ala Gly Met Ile103510401045gat aat ggc gtt aca aaa ggt gat cgg act gga gtt tat ctg tgg acg3273Asp Asn Gly Val Thr Lys Gly Asp Arg Thr Gly Val Tyr Leu Trp Thr105010551060ttc atc atg ttt ata ttt gtt gta cta ggt att att ggg cgt att acg3321Phe Ile Met Phe Ile Phe Val Val Leu Gly Ile Ile Gly Arg Ile Thr1065107010751080atg gct tac gca tct agt cgc tta acg aca aca atg att aga gat atg3369Met Ala Tyr Ala Ser Ser Arg Leu Thr Thr Thr Met Ile Arg Asp Met108510901095cgt aat gat atg tat gct aag ctt caa gaa tac tcc cat cat gaa tat3417Arg Asn Asp Met Tyr Ala Lys Leu Gln Glu Tyr Ser His His Glu Tyr110011051110gaa cag ata ggt gta tct tca cta gtg aca cgt atg aca agc gat act3465Glu Gln Ile Gly Val Ser Ser Leu Val Thr Arg Met Thr Ser Asp Thr111511201125ttt gtt ttg atg caa ttt gct gaa atg tct tta cgt tta ggc cta gta3513Phe Val Leu Met Gln Phe Ala Glu Met Ser Leu Arg Leu Gly Leu Val113011351140act cct atg gta atg att ttt agc gtg gtt atg ata cta att acg agt3561Thr Pro Met Val Met Ile Phe Ser Val Val Met Ile Leu Ile Thr Ser1145115011551160cca tct ttg gct tgg ctt gta gcg gtt gcg atg cct ctt ttg gta gga3609Pro Ser Leu Ala Trp Leu Val Ala Val Ala Met Pro Leu Leu Val Gly116511701175gtc gtt tta tat gta gct ata aaa aca aaa cct tta tct gaa aga caa3657Val Val Leu Tyr Val Ala Ile Lys Thr Lys Pro Leu Ser Glu Arg Gln118011851190cag act atg ctt gat aaa atc aat caa tat gtt cgt gaa aat tta aca 3705Gln Thr Met Leu Asp Lys Ile Asn Gln Tyr Val Arg Glu Asn Leu Thr119512001205ggg tta cgc gtt gtt aga gcc ttt gca aga gag aat ttt caa tca caa 3753Gly Leu Arg Val Val Arg Ala Phe Ala Arg Glu Asn Phe Gln Ser Gln121012151220aaa ttt caa gtc gct aac caa cgt tac aca gat act tca act ggt ctt 3801Lys Phe Gln Val Ala Asn Gln Arg Tyr Thr Asp Thr Ser Thr Gly Leu1225123012351240ttt aaa tta aca ggg cta aca gaa cca ctt ttc gtt caa att att att 3849Phe Lys Leu Thr Gly Leu Thr Glu Pro Leu Phe Val Gln Ile Ile Ile124512501255gca atg att gtg gct atc gtt tgg ttt gct ttg gat ccc tta caa aga 3897Ala Met Ile Val Ala Ile Val Trp Phe Ala Leu Asp Pro Leu Gln Arg126012651270ggt gct att aaa ata ggg gat tta gtt gct ttt atc gaa tat agc ttc 3945Gly Ala Ile Lys Ile Gly Asp Leu Val Ala Phe Ile Glu Tyr Ser Phe127512801285cat gct ctc ttt tca ttt ttg cta ttt gcc aat ctt ttt act atg tat 3993His Ala Leu Phe Ser Phe Leu Leu Phe Ala Asn Leu Phe Thr Met Tyr129012951300cct cgt atg gtg gta tca agc cat cgt att aga gag gtg atg gat atg 4041Pro Arg Met Val Val Ser Ser His Arg Ile Arg Glu Val Met Asp Met1305131013151320cca atc tct atc aat cct aat gcc gaa ggt gtt acg gat acg aaa ctt 4089Pro Ile Ser Ile Asn Pro Asn Ala Glu Gly Val Thr Asp Thr Lys Leu132513301335aaa ggg cat tta gaa ttt gat aat gta aca ttc gct tat cca gga gaa 4137Lys Gly His Leu Glu Phe Asp Asn Val Thr Phe Ala Tyr Pro Gly Glu134013451350aca gag agt ccc gtt ttg cat gat att tct ttt aaa gct aag cct gga 4185Thr Glu Ser Pro Val Leu His Asp Ile Ser Phe Lys Ala Lys Pro Gly135513601365gaa aca att gct ttt att ggt tca aca ggt tca gga aaa tct tct ctt 4233Glu Thr Ile Ala Phe Ile Gly Ser Thr Gly Ser Gly Lys Ser Ser Leu137013751380gtt aat ttg att cca cgt ttt tat gat gtg aca ctt gga aaa atc tta 4281Val Asn Leu Ile Pro Arg Phe Tyr Asp Val Thr Leu Gly Lys Ile Leu1385139013951400gta gat gga gtt gat gta aga gat tat aac ctt aaa tca ctt cgc caa 4329Val Asp Gly Val Asp Val Arg Asp Tyr Asn Leu Lys Ser Leu Arg Gln140514101415aag att gga ttt atc ccc caa aaa gct ctt tta ttt aca ggg aca ata4377Lys Ile Gly Phe Ile Pro Gln Lys Ala Leu Leu Phe Thr Gly Thr Ile142014251430gga gag aat tta aaa tat gga aaa gct gat gct act att gat gat ctt4425Gly Glu Asn Leu Lys Tyr Gly Lys Ala Asp Ala Thr Ile Asp Asp Leu143514401445aga caa gcg gtt gat att tct caa gct aaa gag ttt att gag agt cac4473Arg Gln Ala Val Asp Ile Ser Gln Ala Lys Glu Phe Ile Glu Ser His145014551460caa gaa gcc ttt gaa acg cat tta gct gaa ggt ggg agc aat ctt tct4521Gln Glu Ala Phe Glu Thr His Leu Ala Glu Gly Gly Ser ASn Leu Ser1465147014751480ggg ggt caa aaa caa cgg tta tct att gct agg gct gtt gtt aaa gat4569Gly Gly Gln Lys Gln Arg Leu Ser Ile Ala Arg Ala Val Val Lys Asp148514901495cca gat tta tat att ttt gat gat tca ttt tct gct ctc gat tat aag4617Pro Asp Leu Tyr Ile Phe Asp Asp Ser Phe Ser Ala Leu Asp Tyr Lys150015051510aca gac gct act tta aga gcg cgt cta aaa gaa gta acc ggt gat tct4665Thr Asp Ala Thr Leu Arg Ala Arg Leu Lys Glu Val Thr Gly Asp Ser151515201525aca gtt ttg ata gtt gct caa agg gtg ggt acg att atg gat gct gat4713Thr Val Leu Ile Val Ala Gln Arg Val Gly Thr Ile Met Asp Ala Asp153015351540cag att att gtc ctt gat gaa ggc gaa att gtc ggt cgt ggt acc cac4761Gln Ile Ile Val Leu Asp Glu Gly Glu Ile Val Gly Arg Gly Thr His1545155015551560gct caa tta ata gaa aat aat gct att tat cgt gaa atc gct gag tca4809Ala Gln Leu Ile Glu Asn Asn Ala Ile Tyr Arg Glu Ile Ala Glu Ser156515701575caa ctg aag aac caa aac tta tca gaa gga gag tgattgt atg aga aaa4858Gln Leu Lys Asn Gln Asn Leu Ser Glu Gly GluMet Arg Lys15801585 1590aaa tct gtt ttt ttg aga tta tgg tct tac cta act cgc tac aaa gct4906Lys Ser Val Phe Leu Arg Leu Trp Ser Tyr Leu Thr Arg Tyr Lys Ala159516001605act ctt ttc tta gcg att ttt ttg aaa gtt tta tct agt ttt atg agt4954Thr Leu Phe Leu Ala Ile Phe Leu Lys Val Leu Ser Ser Phe Met Ser161016151620gtt ctg gag cct ttt att tta ggg tta gcg ata aca gag ttg act gct5002Val Leu Glu Pro Phe Ile Leu Gly Leu Ala Ile Thr Glu Leu Thr Ala162516301635aac ctt gtt gat atg gct aag gga gtt tct ggg gca gaa ttg aac gtt5050Asn Leu Val Asp Met Ala Lys Gly Val Ser Gly Ala Glu Leu Asn Val164016451650cct tat att gct ggt att ttg att att tat ttt ttc aga ggt gtt ttc5098Pro Tyr Ile Ala Gly Ile Leu Ile Ile Tyr Phe Phe Arg Gly Val Phe1655166016651670tat gaa tta ggt tct tat ggc tca aat t 5126Tyr Glu Leu Gly Ser Tyr Gly Ser Asn1675<220>8<211>229<212>蛋白質(zhì)<213>鏈球菌<400>8Asn Phe Asp Ile Glu Thr Thr Thr Phe Glu Ala Met Lys Lys His Ala1 5 10 15Ser Leu Leu Glu Lys Ile Ser Val Glu Arg Ser Phe Ile Glu Phe Asp20 25 30Lys Leu Leu Leu Ala Pro Tyr Trp Arg Lys Gly Met Leu Ala Leu Ile35 40 45Asp Ser His Ala Phe Asn Tyr Leu Pro Cys Leu Lys Asn Arg Glu Leu50 55 60Gln Leu Ser Ala Phe Leu Ser Gln Leu Asp Lys Asp Phe Leu Phe Glu65 70 75 80Thr Ser Glu Gln Ala Trp Ala Ser Leu Ile Leu Ser Met Glu Val Glu85 90 95His Thr Lys Thr Phe Leu Lys Lys Trp Lys Thr Ser Thr His Phe Gln100 105 110Lys Asp Val Glu His Ile Val Asp Val Tyr Arg Ile Arg Glu Gln Met115 120 125Gly Leu Ala Lys Glu His Leu Tyr Arg Tyr Gly Lys Thr Ile Ile Lys130 135 140Gln Ala Glu Gly Ile Arg Lys Ala Arg Gly Leu Met Val Asp Phe Glu145 150 155 160Lys Ile Glu Gln Leu Asp Ser Glu Leu Ala Ile His Asp Arg His Glu165 170 175Ile Val Val Asn Gly Gly Thr Leu Ile Lys Lys Leu Gly Ile Lys Pro180 185 190Gly Pro Gln Met Gly Asp Ile Ile Ser Gln Ile Glu Leu Ala Ile Val195 200 205Leu Gly Gln Leu Ile Asn Glu Glu Glu Ala Ile Leu His Phe Val Lys210 215 220Gln Tyr Leu Met Asp225<210>9<211>622<212>PRT<213>Streptococcus<400>9Met Ser Asp Phe Leu Val Asp Gly Leu Thr Lys Ser Val Gly Asp Lys1 5 10 15Thr Val Phe Ser Asn Val Ser Phe Ile Ile His Ser Leu Asp Arg Ile20 25 30Gly Ile Ile Gly Val Asn Gly Thr Gly Lys Thr Thr Leu Leu Asp Val35 40 45Ile Ser Gly Glu Leu Gly Phe Asp Gly Asp Arg Ser Pro Phe Ser Ser50 55 60Ala Asn Asp Tyr Lys Ile Ala Tyr Leu Lys Gln Glu Pro Asp Phe Asp65 70 75 80Asp Ser Gln Thr Ile Leu Asp Thr Val Leu Ser Ser Asp Leu Arg Glu85 90 95Met Ala Leu Ile Lys Glu Tyr Glu Leu Leu Leu Asn His Tyr Glu Glu100 105 110Ser Lys Gln Ser Arg Leu Glu Lys Val Met Ala Glu Met Asp Ser Leu115 120 125Asp Ala Trp Ser Ile Glu Ser Glu Val Lys Thr Val Leu Ser Lys Leu130 135 140Gly Ile Thr Asp Leu Gln Leu Ser Val Gly Glu Leu Ser Gly Gly Leu145 150 155 160Arg Arg Arg Val Gln Leu Ala Gln Val Leu Leu Asn Asp Ala Asp Leu165 170 175Leu Leu Leu Asp Glu Pro Thr Asn His Leu Asp Ile Asp Thr Ile Ala180 185 190Trp Leu Thr Asn Phe Leu Lys Asn Ser Lys Lys Thr Val Leu Phe Ile195 200 205Thr His Asp Arg Tyr Phe Leu Asp Asn Val Ala Thr Arg Ile Phe Glu210 215 220Leu Asp Lys Ala Gln Ile Thr Glu Tyr Gln Gly Asn Tyr Gln Asp Tyr225 230 235 240Val Arg Leu Arg Ala Glu Gln Asp Glu Arg Asp Ala Ala Ser Leu His245 250 255Lys Lys Lys Gln Leu Tyr Lys Gln Glu Leu Ala Trp Met Arg Thr Gln260 265 270Pro Gln Ala Arg Ala Thr Lys Gln Gln Ala Arg Ile Asn Arg Phe Gln275 280 285Asn Leu Lys Asn Asp Leu His Gln Thr Ser Asp Thr Ser Asp Leu Glu290 295 300Met Thr Phe Glu Thr Ser Arg Ile Gly Lys Lys Val Ile Asn Phe Glu305 310 315 320Asn Val Ser Phe Ser Tyr Pro Asp Lys Ser Ile Leu Lys Asp Phe Asn325 330 335Leu Leu Ile Gln Asn Lys Asp Arg Ile Gly Ile Val Gly Asp Asn Gly340 345 350Val Gly Lys Ser Thr Leu Leu Asn Leu Ile Val Gln Asp Leu Gln Pro355 360 365Asp Ser Gly Asn Val Ser Ile Gly Glu Thr Ile Arg Val Gly Tyr Phe370 375 380Ser Gln Gln Leu His Asn Met Asp Gly Ser Lys Arg Val Ile Asn Tyr385 390 395 400Leu Gln Glu Val Ala Asp Glu Val Lys Thr Ser Val Gly Thr Thr Ser405 410 415Val Thr Glu Leu Leu Glu Gln Phe Leu Phe Pro Arg Ser Thr His Gly420 425 430Thr Gln Ile Ala Lys Leu Ser Gly Gly Glu Lys Lys Arg Leu Tyr Leu435 440 445Leu Lys Ile Leu Ile Glu Lys Pro Asn Val Leu Leu Leu Asp Glu Pro450 455 460Thr Asn Asp Leu Asp Ile Ala Thr Leu Thr Val Leu Glu Asn Phe Leu465 470 475 480Gln Gly Phe Gly Gly Pro Val Ile Thr Val Ser His Asp Arg Tyr Phe485 490 495Leu Asp Lys Val Ala Asn Lys Ile Ile Ala Phe Glu Asp Asn Asp Ile500 505 510Arg Glu Phe Phe Gly Asn Tyr Thr Asp Tyr Leu Asp Glu Lys Ala Phe515 520 525Asn Glu Gln Asn Asn Glu Val Ile Ser Lys Lys Glu Ser Thr Lys Thr530 535 540Ser Arg Glu Lys Gln Ser Arg Lys Arg Met Ser Tyr Phe Glu Lys Gln545 550 555 560Glu Trp Ala Thr Ile Glu Asp Asp Ile Met Ile Leu Glu Asn Thr Ile565 570 575Thr Arg Ile Glu Asn Asp Met Gln Thr Cys Gly Ser Asp Phe Thr Arg580 585 590Leu Ser Asp Leu Gln Lys Glu Leu Asp Ala Lys Asn Glu Ala Leu Leu595 600 605Glu Lys Tyr Asp Arg Tyr Glu Tyr Leu Ser Glu Leu Asp Thr610 615 620<210>10<211>157<212>蛋白質(zhì)<213>鏈球菌<400>10Met Ile Ile Arg Pro Ile Ile Lys Asn Asp Asp Gln Ala Val Ala Gln1 5 10 15Leu Ile Arg Gln Ser Leu Arg Ala Tyr Asp Leu Asp Lys Pro Asp Thr20 25 30Ala Tyr Ser Asp Pro His Leu Asp His Leu Thr Ser Tyr Tyr Glu Lys35 40 45Ile Glu Lys Ser Gly Phe Phe Val Ile Glu Glu Arg Asp Glu Ile Ile50 55 60Gly Cys Gly Gly Phe Gly Pro Leu Lys Asn Leu Ile Ala Glu Met Gln65 70 75 80Lys Val Tyr Ile Ala Glu Arg Phe Arg Gly Lys Gly Leu Ala Thr Asp85 90 95Leu Val Lys Met Ile Glu Val Glu Ala Arg Lys Ile Gly Tyr Arg Gln100 105 110Leu Tyr Leu Glu Thr Ala Ser Thr Leu Ser Arg Ala Thr Ala Val Tyr115 120 125Lys His Met Gly Tyr Cys Ala Leu Ser Gln Pro Ile Ala Asn Asp Gln130 135 140Gly His Thr Ala Met Asp Ile Trp Met Ile Lys Asp Leu145150 155<210>11<211>579<212>蛋白質(zhì)<213>鏈球菌<400>11Met Ala Tyr Ile Trp Ser Tyr Leu Lys Arg Tyr Pro Asn Trp Leu Trp1 5 10 15Leu Asp Leu Leu Gly Ala Met Leu Phe Val Thr Val Ile Leu Gly Met20 25 30Pro Thr Ala Leu Ala Gly Met Ile Asp Asn Gly Val Thr Lys Gly Asp35 40 45Arg Thr Gly Val Tyr Leu Trp Thr Phe Ile Met Phe Ile Phe Val Val50 55 60Leu Gly Ile Ile Gly Arg Ile Thr Met Ala Tyr Ala Ser Ser Arg Leu65 70 75 80Thr Thr Thr Met Ile Arg Asp Met Arg Asn Asp Met Tyr Ala Lys Leu85 90 95Gln Glu Tyr Ser His His Glu Tyr Glu Gln Ile Gly Val Ser Ser Leu100 105 110Val Thr Arg Met Thr Ser Asp Thr Phe Val Leu Met Gln Phe Ala Glu115 120 125Met Ser Leu Arg Leu Gly Leu Val Thr Pro Met Val Met Ile Phe Ser130 135 140Val Val Met Ile Leu Ile Thr Ser Pro Ser Leu Ala Trp Leu Val Ala145 150 155 160Val Ala Met Pro Leu Leu Val Gly Val Val Leu Tyr Val Ala Ile Lys165 170 175Thr Lys Pro Leu Ser Glu Arg Gln Gln Thr Met Leu Asp Lys Ile Asn180 185 190Gln Tyr Val Arg Glu Asn Leu Thr Gly Leu Arg Val Val Arg Ala Phe195 200 205Ala Arg Glu Asn Phe Gln Ser Gln Lys Phe Gln Val Ala Asn Gln Arg210 215 220Tyr Thr Asp Thr Ser Thr Gly Leu Phe Lys Leu Thr Gly Leu Thr Glu225 230 235 240Pro Leu Phe Val Gln Ile Ile Ile Ala Met Ile Val Ala Ile Val Trp245 250 255Phe Ala Leu Asp Pro Leu Gln Arg Gly Ala Ile Lys Ile Gly Asp Leu260 265 270Val Ala Phe Ile Glu Tyr Ser Phe His Ala Leu Phe Ser Phe Leu Leu275 280 285Phe Ala Asn Leu Phe Thr Met Tyr Pro Arg Met Val Val Ser Ser His290 295 300Arg Ile Arg Glu Val Met Asp Met Pro Ile Ser Ile Asn Pro Asn Ala305 310 315 320Glu Gly Val Thr Asp Thr Lys Leu Lys Gly His Leu Glu Phe Asp Asn325 330 335Val Thr Phe Ala Tyr Pro Gly Glu Thr Glu Ser Pro Val Leu His Asp340 345 350Ile Ser Phe Lys Ala Lys Pro Gly Glu Thr Ile Ala Phe Ile Gly Ser355 360 365Thr Gly Ser Gly Lys Ser Ser Leu Val Asn Leu Ile Pro Arg Phe Tyr370 375 380Asp Val Thr Leu Gly Lys Ile Leu Val Asp Gly Val Asp Val Arg Asp385 390 395 400Tyr Asn Leu Lys Ser Leu Arg Gln Lys Ile Gly Phe Ile Pro Gln Lys405 410 415Ala Leu Leu Phe Thr Gly Thr Ile Gly Glu Asn Leu Lys Tyr Gly Lys420 425 430Ala Asp Ala Thr Ile Asp Asp Leu Arg Gln Ala Val Asp Ile Ser Gln435 440 445Ala Lys Glu Phe Ile Glu Ser His Gln Glu Ala Phe Glu Thr His Leu450 455 460Ala Glu Gly Gly Ser Asn Leu Ser Gly Gly Gln Lys Gln Arg Leu Ser465 470 475 480Ile Ala Arg Ala Val Val Lys Asp Pro Asp Leu Tyr Ile Phe Asp Asp485 490 495Ser Phe Ser Ala Leu Asp Tyr Lys Thr Asp Ala Thr Leu Arg Ala Arg500 505 510Leu Lys Glu Val Thr Gly Asp Ser Thr Val Leu Ile Val Ala Gln Arg515 520 525Val Gly Thr Ile Met Asp Ala Asp Gln Ile Ile Val Leu Asp Glu Gly530 535 540Glu Ile Val Gly Arg Gly Thr His Ala Gln Leu Ile Glu Asn Asn Ala545 550 555 560Ile Tyr Arg Glu Ile Ala Glu Ser Gln Leu Lys Asn Gln Asn Leu Ser565 570 575Glu Gly Glu<210>12<211>92<212>蛋白質(zhì)<213>鏈球菌<400>12Met Arg Lys Lys Ser Val Phe Leu Arg Leu Trp Ser Tyr Leu Thr Arg1 5 10 15Tyr Lys Ala Thr Leu Phe Leu Ala Ile Phe Leu Lys Val Leu Ser Ser20 25 30Phe Met Ser Val Leu Glu Pro Phe Ile Leu Gly Leu Ala Ile Thr Glu35 40 45Leu Thr Ala Asn Leu Val Asp Met Ala Lys Gly Val Ser Gly Ala Glu50 55 60Leu Asn Val Pro Tyr Ile Ala Gly Ile Leu Ile Ile Tyr Phe Phe Arg65 70 75 80Gly Val Phe Tyr Glu Leu Gly Ser Tyr Gly Ser Asn85 90<210>13<211>5215<212>DNA<213>鏈球菌<220><221>CDS<222>(3)…(122)<221>CDS<222>(133)…(2511)<221>CDS<222>(367)…(2511)<221>CDS<222>(2946)(2716)<223>互補(bǔ)鏈<221>CDS<222>(3252)…(2995)<223>互補(bǔ)鏈<221>CDS<222>(3676)…(3299)<223>互補(bǔ)鏈<221>CDS<222>(4124)…(3837)<223>互補(bǔ)鏈<221>CDS<222>(5214)…(4351)<223>互補(bǔ)鏈<400>13aa ttt gga agt gct cta tca aca gtt gaa gta aag gag att att agt47Phe Gly Ser Ala Leu Ser Thr Val Glu Val Lys Glu Ile Ile Ser1 5 10 15gaa gaa aac ata tgg tta tat cgg ctc agt tgc tgc cat ttt act agc 95Glu Glu Asn Ile Trp Leu Tyr Arg Leu Ser Cys Cys His Phe Thr Ser20 25 30tac tca tat tgg aag tta cca act tgg taagcatcat atg ggt cta gca 144Tyr Ser Tyr Trp Lys Leu Pro Thr TrpMet Gly Leu Ala35 40aca aag gac aat cag att gcc tat att gat gac agc aaa ggt aag gca 192Thr Lys Asp Asn Gln Ile Ala Tyr Ile Asp Asp Ser Lys Gly Lys Ala45 50 55 60aaa gcc cct aaa aca aac aaa acg atg gat caa atc agt gct gaa gaa 240Lys Ala Pro Lys Thr Asn Lys Thr Met Asp Gln Ile Ser Ala Glu Glu65 70 75ggc atc tct gct gaa cag atc gta gtc aaa att act gac caa ggc tat 288Gly Ile Ser Ala Glu Gln Ile Val Val Lys Ile Thr Asp Gln Gly Tyr80 85 90gtg acc tca cac ggt gac cat tat cat ttt tac aat ggg aaa gtt cct 336Val Thr Ser His Gly Asp His Tyr His Phe Tyr Asn Gly Lys Val Pro95 100 105tat gat gcg att att agt gaa gag ttg ttg atg acg gat cct aat tac 384Tyr Asp Ala Ile Ile Ser Glu Glu Leu Leu Met Thr Asp Pro Asn Tyr110 115 120cgt ttt aaa caa tca gac gtt atc aat gaa atc tta gac ggt tac gtt 432Arg Phe Lys Gln Ser Asp Val Ile Asn Glu Ile Leu Asp Gly Tyr Val125 130 135 140att aaa gtc aat ggc aac tat tat gtt tac ctc aag cca ggt agt aag 480Ile Lys Val Asn Gly Asn Tyr Tyr Val Tyr Leu Lys Pro Gly Ser Lys145 150 155cgc aaa aac att cga acc aaa caa caa att gct gag caa gta gcc aaa 528Arg Lys Asn Ile Arg Thr Lys Gln Gln Ile Ala Glu Gln Val Ala Lys160 165 170gga act aaa gaa gct aaa gaa aaa ggt tta gct caa gtg gcc cat ctc 576Gly Thr Lys Glu Ala Lys Glu Lys Gly Leu Ala Gln Val Ala His Leu175 180 185agt aaa gaa gaa gtt gcg gca gtc aat gaa gca aaa aga caa gga cgc 624Ser Lys Glu Glu Val Ala Ala Val Asn Glu Ala Lys Arg Gln Gly Arg190 195 200tat act aca gac gat ggc tat att ttt agt ccg aca gat atc att gat 672Tyr Thr Thr Asp Asp Gly Tyr Ile Phe Ser Pro Thr Asp Ile Ile Asp205 210 215 220gat tta gga gat gct tat tta gta cct cat ggt aat cac tat cat tat 720Asp Leu Gly Asp Ala Tyr Leu Val Pro His Gly Asn His Tyr His Tyr225 230 235att cct aaa aag gat ttg tct cca agt gag cta gct gct gca caa gcc 768Ile Pro Lys Lys Asp Leu Ser Pro Ser Glu Leu Ala Ala Ala Gln Ala240 245 250tac tgg agt caa aaa caa ggt cga ggt gct aga ccg tct gat tac cgc 816Tyr Trp Ser Gln Lys Gln Gly Arg Gly Ala Arg Pro Ser Asp Tyr Arg255 260 265ccg aca cca gcc cca ggt cgt agg aaa gcc cca att cct gat gtg acg 864Pro Thr Pro Ala Pro Gly Arg Arg Lys Ala Pro Ile Pro Asp Val Thr270 275 280cct aac cct gga caa ggt cat cag cca gat aac ggt ggc tat cat cca 912Pro Asn Pro Gly Gln Gly His Gln Pro Asp Asn Gly Gly Tyr His Pro285 290 295 300gcg cct cct agg cca aat gat gcg tca caa aac aaa cac caa aga gat 960Ala Pro Pro Arg Pro Asn Asp Ala Ser Gln Asn Lys His Gln Arg Asp305 310 315gag ttt aaa gga aaa acc ttt aag gaa ctt tta gat caa cta cac cgt 1008Glu Phe Lys Gly Lys Thr Phe Lys Glu Leu Leu Asp Gln Leu His Arg320 325 330ctt gat ttg aaa tac cgt cat gtg gaa gaa gat ggg ttg att ttt gaa 1056Leu Asp Leu Lys Tyr Arg His Val Glu Glu Asp Gly Leu Ile Phe Glu335 340 345ccg act caa gtg atc aaa tca aac gct ttt ggg tat gtg gtg cct cat 1104Pro Thr Gln Val Ile Lys Ser Asn Ala Phe Gly Tyr Val Val Pro His350 355 360gga gat cat tat cat att atc cca aga agt cag tta tca cct ctt gaa 1152Gly Asp His Tyr His Ile Ile Pro Arg Ser Gln Leu Ser Pro Leu Glu365 370 375 380atg gaa tta gca gat cga tac tta gct ggc caa act gag gac aat gac 1200Met Glu Leu Ala Asp Arg Tyr Leu Ala Gly Gln Thr Glu Asp Asn Asp385 390 395tca ggt tca gag cac tca aaa cca tca gat aaa gaa gtg aca cat acc 1248Ser Gly Ser Glu His Ser Lys Pro Ser Asp Lys Glu Val Thr His Thr400 405 410ttt ctt ggt cat cgc atc aaa gct tac gga aaa ggc tta gat ggt aaa 1296Phe Leu Gly His Arg Ile Lys Ala Tyr Gly Lys Gly Leu Asp Gly Lys415 420 425cca tat gat acg agt gat gct tat gtt ttt agt aaa gaa tcc att cat 1344Pro Tyr Asp Thr Ser Asp Ala Tyr Val Phe Ser Lys Glu Ser Ile His430 435 440tca gtg gat aaa tca gga gtt aca gct aaa cac gga gat cat ttc cac 1392Ser Val Asp Lys Ser Gly Val Thr Ala Lys His Gly Asp His Phe His445 450 455 460tat ata gga ttt gga gaa ctt gaa caa tat gag ttg gat gag gtc gct 1440Tyr Ile Gly Phe Gly Glu Leu Glu Gln Tyr Glu Leu Asp Glu Val Ala465 470 475aac tgg gtg aaa gca aaa ggt caa gct gat gag ctt gct gct gct ttg 1488Asn Trp Val Lys Ala Lys Gly Gln Ala Asp Glu Leu Ala Ala Ala Leu480 485 490gat cag gaa caa ggc aaa gaa aaa cca ctc ttt gac act aaa aaa gtg 1536Asp Gln Glu Gln Gly Lys Glu Lys Pro Leu Phe Asp Thr Lys Lys Val495 500 505agt cgc aaa gta aca aaa gat ggt aaa gtg ggc tat atg atg cca aaa 1584Ser Arg Lys Val Thr Lys Asp Gly Lys Val Gly Tyr Met Met Pro Lys510 515 520gat ggt aag gac tat ttc tat gct cgt gat caa ctt gat ttg act cag 1632Asp Gly Lys Asp Tyr Phe Tyr Ala Arg Asp Gln Leu Asp Leu Thr Gln525 530 535 540att gcc ttt gcc gaa caa gaa cra atg ctt aaa gat aag aag cat tac 1680Ile Ala Phe Ala Glu Gln Glu Leu Met Leu Lys Asp Lys Lys His Tyr545 550 555cgt tat gac att gtt gac aca ggt att gag cca cga ctt gct gta gat 1728Arg Tyr Asp Ile Val Asp Thr Gly Ile Glu Pro Arg Leu Ala Val Asp560 565 570gtg tca agt ctg ccg atg cat gct ggt aat gct act tac gat act gga 1776Val Ser Ser Leu Pro Met His Ala Gly Asn Ala Thr Tyr Asp Thr Gly575 580 585agt tcg ttt gtt arc cca cat att gat cat atc cat gtc gtt ccg tat 1824Ser Ser Phe Val Ile Pro His Ile Asp His Ile His Val Val Pro Tyr590 595 600tca tgg ttg acg cgc gat cag att gca aca gtc aag tat gtg atg caa 1872Ser Trp Leu Thr Arg Asp Gln Ile Ala Thr Val Lys Tyr Val Met Gln605 610 615 620cac ccc gaa gtt cgt ccg gat gta tgg tct aag cca ggg cat gaa gag 1920His Pro Glu Val Arg Pro Asp Val Trp Ser Lys Pro Gly His Glu Glu625 630 635tca ggt tcg gtc att cca aat gtt acg cct ctt gat aaa cgt gct ggt 1968Ser Gly Ser Val Ile Pro Asn Val Thr Pro Leu Asp Lys Arg Ala Gly640 645 650atg cca aac tgg caa att arc cat tct gct gaa gaa gtt caa aaa gcc 2016Met Pro Asn Trp Gln Ile Ile His Ser Ala Glu Glu Val Gln Lys Ala655 660 665cta gca gaa ggt cgt ttt gca aca cca gac ggc tat att ttc gat cca 2064Leu Ala Glu Gly Arg Phe Ala Thr Pro Asp Gly Tyr Ile Phe Asp Pro670 675 680cga gat gtt ttg gcc aaa gaa act ttt gta tgg aaa gat ggc tcc ttt 2112Arg Asp Val Leu Ala Lys Glu Thr Phe Val Trp Lys Asp Gly Ser Phe685 690 695 700agc atc cca aga gca gat ggc agt tca ttg aga acc att aat aaa tct 2160Ser Ile Pro Arg Ala Asp Gly Ser Ser Leu Arg Thr Ile Asn Lys Ser705 710 715gat cta tcc caa gct gag tgg caa caa gct caa gag tta ttg gca aag 2208Asp Leu Ser Gln Ala Glu Trp Gln Gln Ala Gln Glu Leu Leu Ala Lys720 725 730aaa aat act ggt gat gct act gat acg gat aaa ccc aaa gaa aag caa 2256Lys Asn Thr Gly Asp Ala Thr Asp Thr Asp Lys Pro Lys Glu Lys Gln735 740 745cag gca gat aag agc aat gaa aac caa cag cca agt gaa gcc agt aaa 2304Gln Ala Asp Lys Ser Asn Glu Asn Gln Gln Pro Ser Glu Ala Ser Lys750 755 760gaa gaa aaa gaa tca gat gac ttt ata gac agt tta cca gac tat ggt 2352Glu Glu Lys Glu Ser Asp Asp Phe Ile Asp Ser Leu Pro Asp Tyr Gly765 770 775 780cta gat aga gca acc cta gaa gat cat atc aat caa tta gca caa aaa 2400Leu Asp Arg Ala Thr Leu Glu Asp His Ile Asn Gln Leu Ala Gln Lys785 790 795gct aat atc gat cct aag tat ctc att ttc caa cca gaa ggt gtc caa 2448Ala Asn Ile Asp Pro Lys Tyr Leu Ile Phe Gln Pro Glu Gly Val Gln800 805 810ttt tat aat aaa aat ggt gaa ttg gta act tat gat atc aag aca ctt2496Phe Tyr Asn Lys Asn Gly Glu Leu Val Thr Tyr Asp Ile Lys Thr Leu815 820 825caa caa ata aac cct taaccaaaag aagatctcat tgttaaagca ctgctttgtc 2551Gln Gln Ile Asn Pro830aaagcaagtt acggtgattt tgaagtcatt ctatgtaacg agtagtgata aaagttggat 2611aatagcggtt ttcttttgca aagaaatggt atccatgtta gaatagtaaa aaaagaggag 2671gattcttgga ctaatgtcaa ataagtagac agaaaactgt gttattttattgcgt2726taaaataatt ttcttctttc tgattagggg ttagtcctag attagccgta tgtgggttgt 2786aattgttata aaaattctca atgtattcaa agcagtctaa ttgaacctgt ttgatatttt 2846gataatgttt tcggttgatt tgtctatgct ttaaatactt gaaaaatgct tcagttacgg 2906cattatcata aggatatcca ggattagaaa aagaatgcat gatattggca ctgcacccta 2966atagtgagac gcaagaaaaa cacttttaggcaatcagtt ttctgtactg tacaggcgac3025tggtcgttta atctctgttg aattctagtt tcattataaa atgtaatgta atttttaaca 3085atatttgtta tactatcttt gttgtatttt ctcctattat ggaaataaaa ggtttcagtc 3145tttaggacgg tgtgaaacca ttcaatacag gcattatctg caggtgttcc ttttcgagac 3205attgagcgga taatgtcttt ttccgtgcaa gcctggtagt aagccataga agtatacact 3265gagccttggt cactgtgtaa gattgctcct ttatttaggcaatt ttaactgatt 3319aagggtgtct agtacaaaat ccgtgtcctg acaatctgag atagtgtaag ctataatttc 3379tcggttatag agattcataa ttgatgagag atacaattta cagttaccga aatataggta 3439ggtaatatct gttacgagct tttccttagg cttatcggca tggaaatccc gactcaattt 3499attatctgtt aaataataag ctttacccaa attgggaact ttcttggtac gtgtccgaca 3559aagccagcca ttatttttca tgatacgata gactttcttt gtattaacag tcaatccgtg 3619gatttttttg agcaatcgtg taatggtacg atagccataa ataaagtgat tctccataca 3679gagctgttca attaattcaa taaggtcatc tttttttgcg gcttctcata ctcctttttc 3739caacggtaat aggtcgaccg cttgacctta aaacagtcta gaatgaaaac tatcgggtag 3799ttgtttttat agtcttccac aagcttgata agacttactttatcgatt tccttatcaa 3857gcctcgatac ttttttaaga ggtcaacctg taattgtaat tgttccactt cagacagatg 3917ttccaagcct ttaccgtagg tatattgctt gccaacacct tgatgaaaac gataaagctc 3977ctcgttttcg taccatttca tccaagtata gatttgacta ttatttttga tgcctaaagt 4037ctccataata actctgttag acttgcctgc tttcttcata tcgatgcaag ccagcttagt 4097ttcccatgaa tatgcttttt taaccataat aaaacattcc tgtttctagt ttactaaatt 4157tcaacaggag tgtttttctt ttgtctcatt ttagggattc agtgcctatt gttgtcatca 4217attatttttc taaattcccc ggacttaaat tgtgaccctt ggtcggaatg aaagagaagt 4277gttccttcaa tctttctttt attaagtgaa aaggcaacac ttttctgtac aacatttata 4337aagtgttttt ctaggcaattaatc ttttagtcat tggtgtttgg tagttgagac 4391taccatgaat gcggtggtaa ttccaccaat gaacatagtc tttagtctta agagctagtt 4451cttccagcaa ttgaaaggtt tcttgataaa caaattcaat tttgaaagca cgatacgtac 4511tttcagctac ggcattgtca taaggataac cagcctgact aagcgaacgt gtgattccaa 4571aggcttccaa tatttcatca attaactgat tatcaaactc tttgccacga tctgaatgga 4631acatcttgac tttggtcagg gcgtaaggga tgctttgtat ggcttgctta acgagttcag 4691cggtcttgtg ccaaccaaga gacaggccga tgatttcacg gttgtatagg tcaatgatga 4751ggcaaacata agcccaacga ttgcctacac gaacataggt taagtcagtg actaaggctt 4811gtagtggtct ttcttgctta aattgcctgt ctaagtggtt gggaataggg gcttcattct 4871tgcctctaga atgtggtttg aaggtggctt tctgataaac agaaaccaaa ttgagtcgct 4931tcataatgcg tcgaatccga cgacgtgaaa gtgtgatacc ttcgttattc aagcatattt 4991tgatttttct ggatccgtat ctagactcgc tatcgagaaa aattctttta atagtttctt 5051caaactccgt ttcagatact gactccacgg cttgatagta ataacttgag tgtggcatat 5111tcagccagcg acacatcttt gaaatgctgt atttatcctt attagcagtg attatttccc 5171tttttgtgcc ataatcaccg ctgcttgctt taggatatct aatt 5215<210>14<211>40<212>蛋白質(zhì)<213>鏈球菌<400>14Phe Gly Ser Ala Leu Ser Thr Val Glu Val Lys Glu Ile Ile Ser Glu1 5 10 15Glu Asn Ile Trp Leu Tyr Arg Leu Ser Cys Cys His Phe Thr Ser Tyr20 25 30Ser Tyr Trp Lys Leu Pro Thr Trp35 40<210>15<211>793<212>蛋白質(zhì)<213>鏈球菌<400>15Met Gly Leu Ala Thr Lys Asp Asn Gln Ile Ala Tyr Ile Asp Asp Ser1 5 10 15Lys Gly Lys Ala Lys Ala Pro Lys Thr Asn Lys Thr Met Asp Gln Ile20 25 30Ser Ala Glu Glu Gly Ile Ser Ala Glu Gln Ile Val Val Lys Ile Thr35 40 45Asp Gln Gly Tyr Val Thr Ser His Gly Asp His Tyr His Phe Tyr Asn50 55 60Gly Lys Val Pro Tyr Asp Ala Ile Ile Ser Glu Glu Leu Leu Met Thr65 70 75 80Asp Pro Asn Tyr Arg Phe Lys Gln Ser Asp Val Ile Asn Glu Ile Leu85 90 95Asp Gly Tyr Val Ile Lys Val Asn Gly Asn Tyr Tyr Val Tyr Leu Lys100 105 110Pro Gly Ser Lys Arg Lys Asn Ile Arg Thr Lys Gln Gln Ile Ala Glu115 120 125Gln Val Ala Lys Gly Thr Lys Glu Ala Lys Glu Lys Gly Leu Ala Gln130 135 140Val Ala His Leu Ser Lys Glu Glu Val Ala Ala Val Asn Glu Ala Lys145 150 155 160Arg Gln Gly Arg Tyr Thr Thr Asp Asp Gly Tyr Ile Phe Ser Pro Thr165 170 175Asp Ile Ile Asp Asp Leu Gly Asp Ala Tyr Leu Val Pro His Gly Asn180 185 190His Tyr His Tyr Ile Pro Lys Lys Asp Leu Ser Pro Ser Glu Leu Ala195 200 205Ala Ala Gln Ala Tyr Trp Ser Gln Lys Gln Gly Arg Gly Ala Arg Pro210 215 220Ser Asp Tyr Arg Pro Thr Pro Ala Pro Gly Arg Arg Lys Ala Pro Ile225 230 235 240Pro Asp Val Thr Pro Asn Pro Gly Gln Gly His Gln Pro Asp Asn Gly245 250 255Gly Tyr His Pro Ala Pro Pro Arg Pro Asn Asp Ala Ser Gln Asn Lys260 265 270His Gln Arg Asp Glu Phe Lys Gly Lys Thr Phe Lys Glu Leu Leu Asp275 280 285Gln Leu His Arg Leu Asp Leu Lys Tyr Arg His Val Glu Glu Asp Gly290 295 300Leu Ile Phe Glu Pro Thr Gln Val Ile Lys Ser Asn Ala Phe Gly Tyr305 310 315 320Val Val Pro His Gly Asp His Tyr His Ile Ile Pro Arg Ser Gln Leu325 330 335Ser Pro Leu Glu Met Glu Leu Ala Asp Arg Tyr Leu Ala Gly Gln Thr340 345 350Glu Asp Asn Asp Ser Gly Ser Glu His Ser Lys Pro Ser Asp Lys Glu355 360 365Val Thr His Thr Phe Leu Gly His Arg Ile Lys Ala Tyr Gly Lys Gly370 375 380Leu Asp Gly Lys Pro Tyr Asp Thr Ser Asp Ala Tyr Val Phe Ser Lys385 390 395 400Glu Ser Ile His Ser Val Asp Lys Ser Gly Val Thr Ala Lys His Gly405 410 415Asp His Phe His Tyr Ile Gly Phe Gly Glu Leu Glu Gln Tyr Glu Leu420 425 430Asp Glu Val Ala Asn Trp Val Lys Ala Lys Gly Gln Ala Asp Glu Leu435 440 445Ala Ala Ala Leu Asp Gln Glu Gln Gly Lys Glu Lys Pro Leu Phe Asp450 455 460Thr Lys Lys Val Ser Arg Lys Val Thr Lys Asp Gly Lys Val Gly Tyr465 470 475 480Met Met Pro Lys Asp Gly Lys Asp Tyr Phe Tyr Ala Arg Asp Gln Leu485 490 495Asp Leu Thr Gln Ile Ala Phe Ala Glu Gln Glu Leu Met Leu Lys Asp500 505 510Lys Lys His Tyr Arg Tyr Asp Ile Val Asp Thr Gly Ile Glu Pro Arg515 520 525Leu Ala Val Asp Val Ser Ser Leu Pro Met His Ala Gly Asn Ala Thr530 535 540Tyr Asp Thr Gly Ser Ser Phe Val Ile Pro His Ile Asp His Ile His545 550 555 560Val Val Pro Tyr Ser Trp Leu Thr Arg Asp Gln Ile Ala Thr Val Lys565 570 575Tyr Val Met Gln His Pro Glu Val Arg Pro Asp Val Trp Ser Lys Pro580 585 590Gly His Glu Glu Ser Gly Ser Val Ile Pro Asn Val Thr Pro Leu Asp595 600 605Lys Arg Ala Gly Met Pro Asn Trp Gln Ile Ile His Ser Ala Glu Glu610 615 620Val Gln Lys Ala Leu Ala Glu Gly Arg Phe Ala Thr Pro Asp Gly Tyr625 630 635 640Ile Phe Asp Pro Arg Asp Val Leu Ala Lya Glu Thr Phe Val Trp Lys645 650 655Asp Gly Ser Phe Ser Ile Pro Arg Ala Asp Gly Ser Ser Leu Arg Thr660 665 670Ile Asn Lys Ser Asp Leu Ser Gln Ala Glu Trp Gln Gln Ala Gln Glu675 680 685Leu Leu Ala Lys Lys Asn Thr Gly Asp Ala Thr Asp Thr Asp Lys Pro690 695 700Lys Glu Lys Gln Gln Ala Asp Lys Ser Asn Glu Asn Gln Gln Pro Ser705 710 715 720Glu Ala Ser Lys Glu Glu Lys Glu Ser Asp Asp Phe Ile Asp Ser Leu725 730 735Pro Asp Tyr Gly Leu Asp Arg Ala Thr Leu Glu Asp His Ile Asn Gln740 745 750Leu Ala Gln Lys Ala Asn Ile Asp Pro Lys Tyr Leu Ile Phe Gln Pro755 760 765Glu Gly Val Gln Phe Tyr Asn Lys Asn Gly Glu Leu Val Thr Tyr Asp770 775 780Ile Lys Thr Leu Gln Gln Ile Asn Pro785 790<210>16<211>715<212>蛋白質(zhì)<213>鏈球菌<400>16Met Thr Asp Pro Asn Tyr Arg Phe Lys Gln Ser Asp Val Ile Asn Glu1 5 10 15I1e Leu Asp Gly Tyr Val Ile Lys Val Asn Gly Asn Tyr Tyr Val Tyr20 25 30Leu Lys Pro Gly Ser Lys Arg Lys Asn Ile Arg Thr Lys Gln Gln Ile35 40 45Ala Glu Gln Val Ala Lys Gly Thr Lys Glu Ala Lys Glu Lys Gly Leu50 55 60Ala Gln Val Ala His Leu Ser Lys Glu Glu Val Ala Ala Val Asn Glu65 70 75 80Ala Lys Arg Gln Gly Arg Tyr Thr Thr Asp Asp Gly Tyr Ile Phe Ser85 90 95Pro Thr Asp Ile Ile Asp Asp Leu Gly Asp Ala Tyr Leu Val Pro His100 105 110Gly Asn His Tyr His Tyr Ile Pro Lys Lys Asp Leu Ser Pro Ser Glu115 120 125Leu Ala Ala Ala Gln Ala Tyr Trp Ser Gln Lys Gln Gly Arg Gly Ala130 135 140Arg Pro Ser Asp Tyr Arg Pro Thr Pro Ala Pro Gly Arg Arg Lys Ala145 150 155 160Pro Ile Pro Asp Val Thr Pro Asn Pro Gly Gln Gly His Gln Pro Asp165 170 175Asn Gly Gly Tyr His Pro Ala Pro Pro Arg Pro Asn Asp Ala Ser Gln180 185 190Asn Lys His Gln Arg Asp Glu Phe Lys Gly Lys Thr Phe Lys Glu Leu195 200 205Leu Asp Gln Leu His Arg Leu Asp Leu Lys Tyr Arg His Val Glu Glu210 215 220Asp Gly Leu Ile Phe Glu Pro Thr Gln Val Ile Lys Ser Asn Ala Phe225 230 235 240Gly Tyr Val Val Pro His Gly Asp His Tyr His Ile Ile Pro Arg Ser245 250 255Gln Leu Ser Pro Leu Glu Met Glu Leu Ala Asp Arg Tyr Leu Ala Gly260 265 270Gln Thr Glu Asp Asn Asp Ser Gly Ser Glu His Ser Lys Pro Ser Asp275 280 285Lys Glu Val Thr His Thr Phe Leu Gly His Arg Ile Lys Ala Tyr Gly290 295 300Lys Gly Leu Asp Gly Lys Pro Tyr Asp Thr Ser Asp Ala Tyr Val Phe305 310 315 320Ser Lys Glu Ser Ile His Ser Val Asp Lys Ser Gly Val Thr Ala Lys325 330 335His Gly Asp His Phe His Tyr Ile Gly Phe Gly Glu Leu Glu Gln Tyr340 345 350Glu Leu Asp Glu Val Ala Asn Trp Val Lys Ala Lys Gly Gln Ala Asp355 360 365Glu Leu Ala Ala Ala Leu Asp Gln Glu Gln Gly Lys Glu Lys Pro Leu370 375 380Phe Asp Thr Lys Lys Val Ser Arg Lys Val Thr Lys Asp Gly Lys Val385 390 395 400Gly Tyr Met Met Pro Lys Asp Gly Lys Asp Tyr Phe Tyr Ala Arg Asp405 410 415Gln Leu Asp Leu Thr Gln Ile Ala Phe Ala Glu Gln Glu Leu Met Leu420 425 430Lys Asp Lys Lys His Tyr Arg Tyr Asp Ile Val Asp Thr Gly Ile Glu435 440 445Pro Arg Leu Ala Val Asp Val Ser Ser Leu Pro Met His Ala Gly Asn450 455 460Ala Thr Tyr Asp Thr Gly Ser Ser Phe Val Ile Pro His Ile Asp His465 470 475 480Ile His Val Val Pro Tyr Ser Trp Leu Thr Arg Asp Gln Ile Ala Thr485 490 495Val Lys Tyr Val Met Gln His Pro Glu Val Arg Pro Asp Val Trp Ser500 505 510Lys Pro Gly His Glu Glu Ser Gly Ser Val Ile Pro Asn Val Thr Pro515 520 525Leu Asp Lys Arg Ala Gly Met Pro Asn Trp Gln Ile Ile His Ser Ala530 535 540Glu Glu Val Gln Lys Ala Leu Ala Glu Gly Arg Phe Ala Thr Pro Asp545 550 555 560Gly Tyr Ile Phe Asp Pro Arg Asp Val Leu Ala Lys Glu Thr Phe Val565 570 575Trp Lys Asp Gly Ser Phe Ser Ile Pro Arg Ala Asp Gly Ser Ser Leu580 585 590Arg Thr Ile Asn Lys Ser Asp Leu Ser Gln Ala Glu Trp Gln Gln Ala595 600 605Gln Glu Leu Leu Ala Lys Lys Asn Thr Gly Asp Ala Thr Asp Thr Asp610 615 620Lys Pro Lys Glu Lys Gln Gln Ala Asp Lys Ser Asn Glu Asn Gln Gln625 630 635 640Pro Ser Glu Ala Ser Lys Glu Glu Lys Glu Ser Asp Asp Phe Ile Asp645 650 655Ser Leu Pro Asp Tyr Gly Leu Asp Arg Ala Thr Leu Glu Asp His Ile660 665 670Asn Gln Leu Ala Gln Lys Ala Asn Ile Asp Pro Lys Tyr Leu Ile Phe675 680 685Gln Pro Glu Gly Val Gln Phe Tyr Asn Lys Asn Gly Glu Leu Val Thr690 695 700Tyr Asp Ile Lys Thr Leu Gln Gln Ile Asn Pro705 710 715<210>17<211>77<212>蛋白質(zhì)<213>鏈球菌<400>17Met His Ser Phe Ser Asn Pro Gly Tyr Pro Tyr Asp Asn Ala Val Thr1 5 10 15Glu Ala Phe Phe Lys Tyr Leu Lys His Arg Gln Ile Asn Arg Lys His20 25 30Tyr Gln Asn Ile Lys Gln Val Gln Leu Asp Cys Phe Glu Tyr Ile Glu35 40 45Asn Phe Tyr Asn Asn Tyr Asn Pro His Thr Ala Asn Leu Gly Leu Thr50 55 60Pro Asn Gln Lys Glu Glu Asn Tyr Phe Asn Ala Ile Lys65 70 75<210>18<211>86<212>蛋白質(zhì)<213>鏈球菌<400>18Met Ala Tyr Tyr Gln Ala Cys Thr Glu Lys Asp Ile Ile Arg Ser Met1 5 10 15Ser Arg Lys Gly Thr Pro Ala Asp Asn Ala Cys Ile Glu Trp Phe His20 25 30Thr Val Leu Lys Thr Glu Thr Phe Tyr Phe His Asn Arg Arg Lys Tyr35 40 45Asn Lys Asp Ser Ile Thr Asn Ile Val Lys Asn Tyr Ile Thr Phe Tyr50 55 60Asn Glu Thr Arg Ile Gln Gln Arg Leu Asn Asp Gln Ser Pro Val Gln65 70 75 80Tyr Arg Lys Leu Ile Ala85<210>19<211>126<212>蛋白質(zhì)<213>鏈球菌<400>19Met Glu Asn His Phe Ile Tyr Gly Tyr Arg Thr Ile Thr Arg Leu Leu1 5 10 15Lys Lys Ile His Gly Leu Thr Val Asn Thr Lys Lys Val Tyr Arg Ile20 25 30Met Lys Asn Asn Gly Trp Leu Cys Arg Thr Arg Thr Lys Lys Val Pro35 40 45Asn Leu Gly Lys Ala Tyr Tyr Leu Thr Asp Asn Lys Leu Ser Arg Asp50 55 60Phe His Ala Asp Lys Pro Lys Glu Lys Leu Val Thr Asp Ile Thr Tyr65 70 75 80Leu Tyr Phe Gly Asn Cys Lys Leu Tyr Leu Ser Ser Ile Met Asn Leu85 90 95Tyr Asn Arg Glu Ile Ile Ala Tyr Thr Ile Ser Asp Cys Gln Asp Thr100 105 110Asp Phe Val Leu Asp Thr Leu Asn Gln Leu Lys Leu Pro Lys115 120 125<210>20<211>96<212>蛋白質(zhì)<213>鏈球菌<400>20Met Val Lys Lys Ala Tyr Ser Trp Glu Thr Lys Leu Ala Cys Ile Asp1 5 10 15Met Lys Lys Ala Gly Lys Ser Asn Arg Val Ile Met Glu Thr Leu Gly20 25 30Ile Lys Asn Asn Ser Gln Ile Tyr Thr Trp Met Lys Trp Tyr Glu Asn35 40 45Glu Glu Leu Tyr Arg Phe His Gln Gly Val Gly Lys Gln Tyr Thr Tyr50 55 60Gly hys Gly Leu Glu His Leu Ser Glu Val Glu Gln Leu Gln Leu Gln65 70 75 80Val Asp Leu Leu Lys Lys Tyr Arg Gly Leu Ile Arg Lys ger Ile Lys85 90 95<210>21<211>288<212>蛋白質(zhì)<213>鏈球菌<400>21Ile Arg Tyr Pro Lys Ala Ser Ser Gly Asp Tyr Gly Thr Lys Arg Glu1 5 10 15Ile Ile Thr Ala Asn Lys Asp Lys Tyr Ser Ile Ser Lys Met Cys Arg20 25 30Trp Leu Asn Met Pro His Ser Ser Tyr Tyr Tyr Gln Ala Val Glu Ser35 40 45Val Ser Glu Thr Glu Phe Glu Glu Thr Ile Lys Arg Ile Phe Leu Asp50 55 60Ser Glu Ser Arg Tyr Gly Ser Arg Lys Ile Lys Ile Cys Leu Asn Asn65 70 75 80Glu Gly Ile Thr Leu Ser Arg Arg Arg Ile Arg Arg Ile Met Lys Arg85 90 95Leu Asn Leu Val Ser Val Tyr Gln Lys Ala Thr Phe Lys Pro His Ser100105 110Arg Gly Lys Asn Glu Ala Pro Ile Pro Asn His Leu Asp Arg Gln Phe115 120 125Lys Gln Glu Arg Pro Leu Gln Ala Leu Val Thr Asp Leu Thr Tyr Val130 135 140Arg Val Gly Asn Arg Trp Ala Tyr Val Cys Leu Ile Ile Asp Leu Tyr145 150 155 160Asn Arg Glu Ile Ile Gly Leu Ser Leu Gly Trp His Lys Thr Ala Glu165 170 175Leu Val Lys Gln Ala Ile Gln Ser Ile Pro Tyr Ala Leu Thr Lys Val180 185 190Lys Met Phe His Ser Asp Arg Gly Lys Glu Phe Asp Asn Gln Leu Ile195 200 205Asp Glu Ile Leu Glu Ala Phe Gly Ile Thr Arg Ser Leu Ser Gln Ala210 215 220Gly Tyr Pro Tyr Asp Asn Ala Val Ala Glu Ser Thr Tyr Arg Ala Phe225 230 235 240Lys Ile Glu Phe Val Tyr Gln Glu Thr Phe Gln Leu Leu Glu Glu Leu245 250 255Ala Leu Lys Thr Lys Asp Tyr Val His Trp Trp Asn Tyr His Arg Ile260 265 270His Gly Ser Leu Asn Tyr Gln Thr Pro Met Thr Lys Arg Leu Ile Ala275 280 285<210>22<211>5058<212>DNA<213>鏈球菌<220><221>CDS<222>(1)…(663)<221>CDS<222>(763)…(1344)<221>CDS<222>(1362)…(1739)<221>CDS<222>(2266)…(5058)<400>22aat ttg aaa gca gaa tta tct gta gaa gat gag caa tat aca gca aca 48Asn Leu Lys Ala Glu Leu Ser Val Glu Asp Glu Gln Tyr Thr Ala Thr1 5 10 15gtt tat ggt aaa tct gct cat ggt tca aca cca caa gaa ggt gtt aat 96Val Tyr Gly Lys Ser Ala His Gly Ser Thr Pro Gln Glu Gly Val Asn20 25 30ggg gcg act tat tta gct ctt tat cta agt caa ttt gat ttt gaa ggt 144Gly Ala Thr Tyr Leu Ala Leu Tyr Leu Ser Gln Phe Asp Phe Glu Gly35 40 45cct gct cgt gct ttc tta gat gtt aca gcc aac att att cac gaa gac 192Pro Ala Arg Ala Phe Leu Asp Val Thr Ala Asn Ile Ile His Glu Asp50 55 60ttc tca ggt gaa aaa ctt gga gta gct tat gaa gat gac tgt atg gga 240Phe Ser Gly Glu Lys Leu Gly Val Ala Tyr Glu Asp Asp Cys Met Gly65 70 75 80cca ttg agc atg aat gca ggt gtc ttc cag ttt gat gaa act aat gat 288Pro Leu Set Met Asn Ala Gly Val Phe Gln Phe Asp Glu Thr Asn Asp85 90 95gat aat act atc gct ctt aat ttc cgt tac cca caa ggg aca gat gct 336Asp Asn Thr Ile Ala Leu Asn Phe Arg Tyr Pro Gln Gly Thr Asp Ala100 105 110aaa act atc caa act aag ctt gag aaa ctt aac gga gtt gaa aaa gtg 384Lys Thr Ile Gln Thr Lys Leu Glu Lys Leu Asn Gly Val Glu Lys Val115 120 125act ctt tct gac cat gaa cac aca cca cac tat gta cct atg gac gat 432Thr Leu Ser Asp His Glu His Thr Pro His Tyr Val Pro Met Asp Asp130 135 140gaa tta gta tca acc tta cta gct gtc tat gaa aag caa act ggt ctt 480Glu Leu Val Ser Thr Leu Leu Ala Val Tyr Glu Lys Gln Thr Gly Leu145 150 155 160aaa gga cac gaa cag gtt att ggt ggt ggg aca ttt ggt cgc tta ctt 528Lys Gly His Glu Gln Val Ile Gly Gly Gly Thr Phe Gly Arg Leu Leu165 170 175gaa cgg ggt gtt gca tac ggt gcc atg ttc cca gga gat gaa aac act 576Glu Arg Gly Val Ala Tyr Gly Ala Met Phe Pro Gly Asp Glu ASn Thr180 185 190atg cat caa gct aat gag tac atg cct tta gaa aat att ttc cgt tcg 624Met His Gln Ala Asn Glu Tyr Met Pro Leu Glu Asn Ile Phe Arg Ser195 200 205gct gct atc tac gca gaa gct atc tat gaa tta atc aaa taaaataatc 673Ala Ala Ile Tyr Ala Glu Ala Ile Tyr Glu Leu Ile Lys210 215 220cttaaactaa atatgtgatc aatgataaag ggtggtgaag acatgaaagt gtctttgcct733cttttcataa ggttagattt ggagacttt atg act gac ttg gaa aaa att att 786Met Thr Asp Leu Glu Lys Ile Ile225aaa gca ata aaa agt gat tca cag aat caa aat tat aca gaa aat ggt 834Lys Ala Ile Lys Ser Asp Ser Gln Asn Gln Asn Tyr Thr Glu Asn Gly230 235 240 245att gat cct ttg ttt gct gct cct aaa aca gct agg atc aat att gtt 882Ile Asp Pro Leu Phe Ala Ala Pro Lys Thr Ala Arg Ile Asn Ile Val250 255 260ggc caa gca cct ggt tta aaa act caa gaa gca aga ctc tat tgg aaa 930Gly Gln Ala Pro Gly Leu Lys Thr Gln Glu Ala Arg Leu Tyr Trp Lys265 270 275gat aaa tct gga gat cgt cta cgc cag tgg ctt gga gtt gat gaa gag 978Asp Lys Ser Gly Asp Arg Leu Arg Gln Trp Leu Gly Val Asp Glu Glu280 285 290aca ttt tac cat tct gga aaa ttt gct gtt tta cct tta gat ttt tat 1026Thr Phe Tyr His Ser Gly Lys Phe Ala Val Leu Pro Leu Asp Phe Tyr295 300 305tac cca ggc aaa gga aaa tca gga gat tta ccc cct aga aaa ggt ttt 1074Tyr Pro Gly Lys Gly Lys Ser Gly Asp Leu Pro Pro Arq Lys Gly Phe310 315 320 325gcg gag aaa tgg cac cct ctt att tta aaa gaa atg cct aat gtt caa 1122Ala Glu Lys Trp His Pro Leu Ile Leu Lys Glu Met Pro Asn Val Gln330 335 340ttg acc ttg cta gtt ggt cag tat gct cag aaa tat tat ctt gga agc 1170Leu Thr Leu Leu Val Gly Gln Tyr Ala Gln Lys Tyr Tyr Leu Gly Ser345 350 355tcc gca cat aaa aat cta aca gaa aca gtt aaa gct tac aaa gac tat1218Ser Ala His Lys Asn Leu Thr Glu Thr Val Lys Ala Tyr Lys Asp Tyr360 365 370cta ccc gat tat tta ccc ctg gtt cac cca tca ccg cga aat caa att1266Leu Pro Asp Tyr Leu Pro Leu Val His Pro Ser Pro Arg Asn Gln Ile375 380 385tgg cta aag aag aat cca tgg ttt gaa aaa gat cta atc gtt gat tta1314Trp Leu Lys Lys Asn Pro Trp Phe Glu Lys Asp Leu Ile Val Asp Leu390 395 400 405caa aag ata gta gca gat att tta aaa gat taaggatagg agttggt atg 1364Gln Lys Ile Val Ala Asp Ile Leu Lys Asp Met410 415aga gat aat cat cta cac acg tat ttt tcc tat gat tgt caa acg gca1412Arg Asp Asn His Leu His Thr Tyr Phe Ser Tyr Asp Cys Gln Thr Ala420 425 430ttt gag gac tat att aat ggt ttt aca ggt gaa ttt atc acg aca gaa1460Phe Glu Asp Tyr Ile Asn Gly Phe Thr Gly Glu Phe Ile Thr Thr Glu435 440 445cat ttt gat tta tca aat cct tac acc ggt caa gac gat gtt cct gat1508His Phe Asp Leu Ser Asn Pro Tyr Thr Gly Gln Asp Asp Val Pro Asp450 455 460tat agt gct tat tgt caa aaa ata gat tat ctt aat cag aaa tat gga1556Tyr Ser Ala Tyr Cys Gln Lys Ile Asp Tyr Leu Asn Gln Lys Tyr Gly465 470 475 480aat cga ttt aaa aaa gga att gaa atc ggt tat ttt aaa gat agg gaa1604Asn Arg Phe Lys Lys Gly Ile Glu Ile Gly Tyr Phe Lys Asp Arg Glu485 490 495tca gat att tta gat tat tta aaa aat aaa gaa ttt gat tta aaa cta1652Ser Asp Ile Leu Asp Tyr Leu Lys Asn Lys Glu Phe Asp Leu Lys Leu500 505 510ttg tca atc cat cat aat ggt agg tat gat tat ctg caa gaa gaa gct1700Leu Ser Ile His His Asn Gly Arg Tyr Asp Tyr Leu Gln Glu Glu Ala515 520 525ctg aaa gta cca aca aag gga gct ttt agc aga tta ctt taatcgtatg 1749Leu Lys Val Pro Thr Lys Gly Ala Phe Ser Arg Leu Leu530 535 540gaatttgcca taggccgtgt ggaagcgcac gttttagctc actttgatta tggttttcgt 1809aagttaaact tagatgtaga agatttaaaa ccgtttgaaa cgcaattgaa gcgcattttc 1869ataaagatgt tatctaaggg gttagctttt gaactaaata ccaaatccct ttatctatat 1929gggaatgaaa aactttatcg ctatgcttta gagatactca aacagcttgg ttgtaaacaa 1989tactctatag gctctgacgg tcatattcct gaacattttt gttatgaatt tgatagactt 2049caaggtctgc taaaggacta tcaaattgat gaaaatcatt tgatatgagg aaatttttga 2109taaaaaagct aggcaatatt gcttagcttt tttgtaatgc tattgatagt tttagtgaaa 2169atttcaaaaa aataaagaaa tcatttactt gttgcaagcg cttgcgtaaa ttgttatgat 2229tttattggta acaattcatt aaaaaaggag aatgat atg aaa aga aaa gac tta2283Met Lys Arg Lys Asp Leu545ttt ggt gat aaa caa act caa tac acg att aga aag tta agt gtt gga2331Phe Gly Asp Lys Gln Thr Gln Tyr Thr Ile Arg Lys Leu Ser Val Gly550 555 560gta gct tca gtt aca aca ggg gta tgt att ttt ctt cat agt cca cag2379Val Ala Ser Val Thr Thr Gly Val Cys Ile Phe Leu His Ser Pro Gln565 570 575gta ttt gct gaa gaa gta agt gtt tct cct gca act aca gcg att gca2427Val Phe Ala Glu Glu Val Ser Val Ser Pro Ala Thr Thr Ala Ile Ala580 585 590 595gag tcg aat att aat cag gtt gac aac caa caa tct act aat tta aaa2475Glu Ser Asn Ile Asn Gln Val Asp Asn Gln Gln Ser Thr Asn Leu Lys600 605 610gat gac ara aac tca aac tct gag acg gtt gtg aca ccc tca gat atg2523Asp Asp Ile Asn Ser Asn Ser Glu Thr Val Val Thr Pro Ser Asp Met615 620 625ccg gat acc aag caa tta gta tca gat gaa act gac act caa aag gga2571Pro Asp Thr Lys Gln Leu Val Ser Asp Glu Thr Asp Thr Gln Lys Gly630 635 640gtg aca gag ccg gat aag gcg aca agc ctg ctt gaa gaa aat aaa ggt2619Val Thr Glu Pro Asp Lys Ala Thr Ser Leu Leu Glu Glu Asn Lys Gly645 650 655cct gtt tca gat aaa aat acc tta gat tta aaa gta gca cca tct aca2667Pro Val Ser Asp Lys Asn Thr Leu Asp Leu Lys Val Ala Pro ger Thr660 665 670 675ttg caa aat act ccc gac aaa act tct caa gct ata ggt gct cca agc2715Leu Gln Asn Thr Pro Asp Lys Thr Ser Gln Ala Ile Gly Ala Pro Ser680 685 690cct acc ttg aaa gta gct aat caa gct cca cgg att gaa aat ggt tac2763Pro Thr Leu Lys Val Ala Asn Gln Ala Pro Arg Ile Glu Asn Gly Tyr695 700 705ttt agg cta cat ctt aaa gaa ttg cct caa ggt cat cct gta gaa agc2811Phe Arg Leu His Leu Lys Glu Leu Pro Gln Gly His Pro Val Glu Ser710 715 720act gga ctt tgg ata tgg gga gat gtt gat caa ccg tct agt aat tgg2859Thr Gly Leu Trp Ile Trp Gly Asp Val Asp Gln Pro Ser Ser Asn Trp725 730 735cca aat ggt gct att cct atg act gat gct aag aaa gat gat tac ggt2907Pro Asn Gly Ala Ile Pro Met Thr Asp Ala Lys Lys Asp Asp Tyr Gly740 745 750 755tat tat gtt gat ttt aaa tta tct gaa aaa caa cga aaa caa ata tct2955Tyr Tyr Val Asp Phe Lys Leu Ser Glu Lys Gln Arg Lys Gln Ile Ser760 765 770ttt tta att aat aac aaa gca ggg aca aat tta agc ggc gat cat cat3003Phe Leu Ile Asn Asn Lys Ala Gly Thr Asn Leu Ser Gly Asp His His775 780 785att cca tta tta cga cct gag atg aac caa gtt tgg att gat gaa aag3051Ile Pro Leu Leu Arg Pro Glu Met Asn Gln Val Trp Ile Asp Glu Lys790 795 800tac ggt ata cat act tat caa ccc ctc aaa gaa ggg tat gtc cgt att3099Tyr Gly Ile His Thr Tyr Gln Pro Leu Lys Glu Gly Tyr Val Arg Ile805 810 815aac tat ttg agt tcc tcu agt aac tat gac cac tta tca gca tgg ctc3147Asn Tyr Leu Ser Ser Ser Ser Asn Tyr Asp His Leu Ser Ala Trp Leu820 825 830 835ttt aaa gat gtt gca acc ccy tca aca act tgg cca gat ggt agt aat3195Phe Lys Asp Val Ala Thr Xaa Ser Thr Thr Trp Pro Asp Gly Ser Asn840 845 850ttt gtg aat caa gga cta tat gga agg tat att gat gta tca cta aaa3243Phe Val Asn Gln Gly Leu Tyr Gly Arg Tyr Ile Asp Val Ser Leu Lys855 860 865act aac gcc aaa gag att ggt ttt cta atc tta gat gaa agt aag aca3291Thr Asn Ala Lys Glu Ile Gly Phe Leu Ile Leu Asp Glu Ser Lys Thr870 875 880gga gat gca gtg aaa gtt caa ccc aac gac tat gtt ttt aga gat tta3339Gly Asp Ala Val Lys Val Gln Pro Asn Asp Tyr Val Phe Arg Asp Leu885 890 895gct aac cat aac caa att ttt gta aaa gat aag gat cca aag gtt tat3387Ala Asn His Asn Gln Ile Phe Val Lys Asp Lys Asp Pro Lys Val Tyr900 905 910 915aat aat cct tat tac att gat caa gtg cag cta aag gat gcc caa caa3435Asn Asn Pro Tyr Tyr Ile Asp Gln Val Gln Leu Lys Asp Ala Gln Gln920 925 930att gat tta aca agt att caa gca agt ttt aca act cta gat ggg gta3483Ile Asp Leu Thr Ser Ile Gln Ala Ser Phe Thr Thr Leu Asp Gly Val935 940 945gat aaa act gaa att tta aaa gaa ttg aaa gtg act gat aaa aat caa3531Asp Lys Thr Glu Ile Leu Lys Glu Leu Lys Val Thr Asp Lys Asn Gln950 955 960aat gct ata caa att tct gat atc act ctc gat act agt aaa tct ctt3579Asn Ala Ile Gln Ile Ser Asp Ile Thr Leu Asp Thr Ser Lya Ser Leu965 970 975tta ata atc aaa ggc gac ttt aat cct aaa caa ggt cat ttc aac ata 3627Leu Ile Ile Lys Gly Asp Phe Asn Pro Lys Gln Gly His Phe Asn Ile980 985 990 995tct tat aat ggt aac aat gtc atg aca agg caa tct tgg gaa ttt aaa 3675Ser Tyr Asn Gly Asn Asn Val Met Thr Arg Gln Ser Trp Glu Phe Lys100010051010gac caa ctt tat gct tat agt gga aat tta ggt gca gtt ctc aat caa 3723Asp Gln Leu Tyr Ala Tyr Ser Gly Asn Leu Gly Ala Val Leu Asn Gln101510201025gat ggt tca aaa gtt gaa gcc agc ctc tgg tca ccg agt gct gat agt 3771Asp Gly Ser Lys Val Glu Ala Ser Leu Trp Ser Pro Ser Ala Asp Ser103010351040gtc act atg att att tat gac aaa gat aac caa aac agg gtt gta gcg 3819Val Thr Met Ile Iie Tyr Asp Lys Asp Asn Gln Asn Arg Val Val Ala104510501055act acc ccc ctt gtg aaa aat aat aaa ggt gtt tgg cag acg ata ctt 3867Thr Thr Pro Leu Val Lys Asn Asn Lys Gly Val Trp Gln Thr Ile Leu1060106510701075gat act aaa tta ggt att aaa aac tat act ggt tac tat tat ctt tac 3915Asp Thr Lys Leu Gly Ile Lys Asn Tyr Thr Gly Tyr Tyr Tyr Leu Tyr108010851090gaa ara aaa aga ggt aag gat aag gtt aag att tta gat cct tat gca 3963Glu Ile Lys Arg Gly Lys Asp Lys Val Lys Ile Leu Asp Pro Tyr Ala109511001105aag tca tta gca gag tgg gat agt aat act gtt aat gat gat att aaa 4011Lys Ser Leu Ala Glu Trp Asp Ser Asn Thr Val Asn Asp Asp Ile Lys111011151120acg gct aaa gca gct ttt gta aat cca agt caa ctt gga cct caa aat 4059Thr Ala Lys Ala Ala Phe Val Asn Pro Ser Gln Leu Gly Pro Gln Asn112511301135tta agt ttt gct aaa att gct aat ttt aaa gga aga caa gat gct gtt 4107Leu Ser Phe Ala Lys Ile Ala Asn Phe Lys Gly Arg Gln Asp Ala Val1140114511501155ata tac gaa gca cat gta aga gac ttc act tct gat cga tct ttg gat 4155Ile Tyr Glu Ala His Val Arg Asp Phe Thr Ser Asp Arg Ser Leu Asp116011651170gga aaa tta aaa aat caa ttt ggt acc ttt gca gcc ttt tca gag aaa 4203Gly Lys Leu Lys Asn Gln Phe Gly Thr Phe Ala Ala Phe Ser Glu Lys117511801185cta gat tat tta cag aaa tta gga gtt aca cac att cag ctt tta ccg 4251Leu Asp Tyr Leu Gln Lys Leu Gly Val Thr His Ile Gln Leu Leu Pro119011951200gta ttg agt tat ttt tat gtt aat gaa atg gat aag tca cgc tca aca 4299Val Leu Ser Tyr Phe Tyr Val Asn Glu Met Asp Lys Ser Arg Ser Thr120512101215gct tac act tcc tca gac aat aat tac aat tgg ggc tat gac cca cag 4347Ala Tyr Thr Ser Ser Asp Asn Asn Tyr Asn Trp Gly Tyr Asp Pro Gln1220122512301235agc tat ttt gct ctt tct ggg atg tat tca gag aaa cca aaa gat cca 4395Ser Tyr Phe Ala Leu Ser Gly Met Tyr Ser Glu Lys Pro Lys Asp Pro124012451250tca gca cgt atc gcc gaa tta aaa caa tta ata cat gat att cat aaa 4443Ser Ala Arg Ile Ala Glu Leu Lys Gln Leu Ile His Asp Ile His Lys125512601265cgt ggc atg ggg gtt ata ctt gat gtc gtc tat aat cac act gca aaa 4491Arg Gly Met Gly Val Ile Leu Asp Val Val Tyr Asn His Thr Ala Lys127012751280act tat ctc ttt gag gat ata gaa cct aat tat tat cac ttt atg aat 4539Thr Tyr Leu Phe Glu Asp Ile Glu Pro Asn Tyr Tyr His Phe Met Asn128512901295gaa gat ggt tca cca aga gaa agt ttt gga ggg gga cgt tta gga acc 4587Glu Asp Gly Ser Pro Arg Glu Ser Phe Gly Gly Gly Arg Leu Gly Thr1300130513101315act cat gca atg agt cgt cgt gtt ttg gtt gat tcc att aaa tat ctt 4635Thr His Ala Met Ser Arg Arg Val Leu Val Asp Ser Ile Lys Tyr Leu132013251330aca agt gaa ttt aaa gtt gat ggt ttc cgt ttt gat atg atg gga gat 4683Thr Ser Glu Phe Lys Val Asp Gly Phe Arg Phe Asp Met Met Gly Asp133513401345cat gat gcg gct gcg att gaa tta gct tat aaa gaa gct aaa gct att 4731His Asp Ala Ala Ala Ile Glu Leu Ala Tyr Lys Glu Ala Lys Ala Ile135013551360aat cct aat atg att atg att ggt gag ggc tgg aga aca ttc caa ggc 4779Asn Pro Asn Met Ile Met Ile Gly Glu Gly Trp Arg Thr Phe Gln Gly136513701375gat caa ggt cag ccg gtt aaa cca gct gac caa gat tgg atg aag tca 4827Asp Gln Gly Gln Pro Val Lys Pro Ala Asp Gln Asp Trp Met Lys Ser1380138513901395acc gat aca gtt ggc gtc ttt tca gat gat att cgt aat agc ttg aaa 4875Thr Asp Thr Val Gly Val Phe Ser Asp Asp Ile Arg Asn Ser Leu Lys140014051410tct ggt ttt cca aat gaa ggt act cca gct ttc atc aca ggt ggc cca 4923Ser Gly Phe Pro Asn Glu Gly Thr Pro Ala Phe Ile Thr Gly Gly Pro141514201425caa tct tta caa ggt att ttt aaa aat atc aaa gca caa cct ggg aat 4971Gln Ser Leu Gln Gly Ile Phe Lys Asn Ile Lys Ala Gln Pro Gly Asn143014351440ttt gaa gca gat tcg cca gga gat gtg gtg cag tat att gct gca cat 5019phe Glu Ala Asp Ser Pro Gly Asp Val Val Gln Tyr Ile Ala Ala His144514501455gat aac ctt acc ttg cat gat gtg att gca aaa tca att 5058Asp Asn Leu Thr Leu His Asp Val Ile Ala Lys Ser Ile146014651470<210>23<211>221<212>蛋白質(zhì)<213>鏈球菌<400>23Asn Leu Lys Ala Glu Leu Ser Val Glu Asp Glu Gln Tyr Thr Ala Thr1 5 10 15Val Tyr Gly Lys Ser Ala His Gly Ser Thr Pro Gln Glu Gly Val Asn20 25 30Gly Ala Thr Tyr Leu Ala Leu Tyr Leu Ser Gln Phe Asp Phe Glu Gly35 40 45Pro Ala Arg Ala Phe Leu Asp Val Thr Ala Asn Ile Ile His Glu Asp50 55 60Phe Ser Gly Glu Lys Leu Gly Val Ala Tyr Glu Asp Asp Cys Met Gly65 70 75 80Pro Leu Ser Met Asn Ala Gly Val Phe Gln Phe Asp Glu Thr Asn Asp85 90 95Asp Asn Thr Ile Ala Leu Asn Phe Arg Tyr Pro Gln Gly Thr Asp Ala100 105 110Lys Thr Ile Gln Thr Lys Leu Glu Lys Leu Asn Gly Val Glu Lys Val115 120 125Thr Leu Ser Asp His Glu His Thr Pro His Tyr Val Pro Met Asp Asp130 135 140Glu Leu Val Ser Thr Leu Leu Ala Val Tyr Glu Lys Gln Thr Gly Leu145 150 155 160Lys Gly His Glu Gln Val Ile Gly Gly Gly Thr Phe Gly Arg Leu Leu165 170 175Glu Arg Gly Val Ala Tyr Gly Ala Met Phe Pro Gly Asp Glu Asn Thr180 185 190Met His Gln Ala Asn Glu Tyr Met Pro Leu Glu Asn Ile Phe Arg Ser195 200 205Ala Ala Ile Tyr Ala Glu Ala Ile Tyr Glu Leu Ile Lys210 215 220<210>24<211>194<212>蛋白質(zhì)<213>鏈球菌<400>24Met Thr Asp Leu Glu Lys Ile Ile Lys Ala Ile Lys Ser Asp Ser Gln1 5 10 15Asn Gln Asn Tyr Thr Glu Asn Gly Ile Asp Pro Leu Phe Ala Ala Pro20 25 30Lys Thr Ala Arg Ile Asn Ile Val Gly Gln Ala Pro Gly Leu Lys Thr35 40 45Gln Glu Ala Arg Leu Tyr Trp Lys Asp Lys Ser Gly Asp Arg Leu Arg50 55 60Gln Trp Leu Gly Val Asp Glu Glu Thr Phe Tyr His Ser Gly Lys Phe65 70 75 80Ala Val Leu Pro Leu Asp Phe Tyr Tyr Pro Gly Lys Gly Lys Ser Gly85 90 95Asp Leu Pro Pro Arg Lys Gly Phe Ala Glu Lys Trp His Pro Leu Ile100 105 110Leu Lys Glu Met Pro Asn Val Gln Leu Thr Leu Leu Val Gly Gln Tyr115 120 125Ala Gln Lys Tyr Tyr Leu Gly Ser Ser Ala His Lys Asn Leu Thr Glu130 135 140Thr Val Lys Ala Tyr Lys Asp Tyr Leu Pro Asp Tyr Leu Pro Leu Val145 150 155 160His Pro Ser Pro Arg Asn Gln Ile Trp Leu Lys Lys Asn Pro Trp Phe165 170 175Glu Lys Asp Leu Ile Val Asp Leu Gln Lys Ile Val Ala Asp Ile Leu180 185 190Lys Asp<210>25<211>126<212>蛋白質(zhì)<213>鏈球菌<400>25Met Arg Asp Asn His Leu His Thr Tyr Phe Ser Tyr Asp Cys Gln Thr1 5 10 15Ala Phe Glu Asp Tyr Ile Asn Gly Phe Thr Gly Glu Phe Ile Thr Thr20 25 30Glu His Phe Asp Leu Ser Asn Pro Tyr Thr Gly Gln Asp Asp Val Pro35 40 45Asp Tyr Ser Ala Tyr Cys Gln Lys Ile Asp Tyr Leu Asn Gln Lys Tyr50 55 60Gly Asn Arg Phe Lys Lys Gly Ile Glu Ile Gly Tyr Phe Lys Asp Arg65 70 75 80Glu Ser Asp Ile Leu Asp Tyr Leu Lys Asn Lys Glu Phe Asp Leu Lys85 90 95Leu Leu Ser Ile His His Asn Gly Arg Tyr Asp Tyr Leu Gln Glu Glu100 105 110Ala Leu Lys Val Pro Thr Lys Gly Ala Phe Ser Arg Leu Leu115 120 125<210>26<211>931<212>蛋白質(zhì)<213>鏈球菌<400>26Met Lys Arg Lys Asp Leu Phe Gly Asp Lys Gln Thr Gln Tyr Thr Ile1 5 10 15Arg Lys Leu Ser Val Gly Val Ala Ser Val Thr Thr Gly Val Cys Ile20 25 30Phe Leu His Ser Pro Gln Val Phe Ala Glu Glu Val Ser Val Ser Pro35 40 45Ala Thr Thr Ala Ile Ala Glu Ser Asn Ile Asn Gln Val Asp Asn Gln50 55 60Gln Ser Thr Asn Leu Lys Asp Asp Ile Asn Ser Asn Ser Glu Thr Val65 70 75 80Val Thr Pro Ser Asp Met Pro Asp Thr Lys Gln Leu Val Ser Asp Glu85 90 95Thr Asp Thr Gln Lys Gly Val Thr Glu Pro Asp Lys Ala Thr Ser Leu100 105 110Leu Glu Glu Asn Lys Gly Pro Val Ser Asp Lys Asn Thr Leu Asp Leu115 120 125Lys Val Ala Pro Ser Thr Leu Gln Asn Thr Pro Asp Lys Thr Ser Gln130 135 140Ala Ile Gly Ala Pro Ser Pro Thr Leu Lys Val Ala Asn Gln Ala Pro145 150 155 160Arg Ile Glu Asn Gly Tyr Phe Arg Leu His Leu Lys Glu Leu Pro Gln165 170 175Gly His Pro Val Glu Ser Thr Gly Leu Trp Ile Trp Gly Asp Val Asp180 185 190Gln Pro Ser Ser Asn Trp Pro Asn Gly Ala Ile Pro Met Thr Asp Ala195 200 205Lys Lys Asp Asp Tyr Gly Tyr Tyr Val Asp Phe Lys Leu Ser Glu Lys210 215 220Gln Arg Lys Gln Ile Ser Phe Leu Ile Asn Asn Lys Ala Gly Thr Asn225 230 235 240Leu Ser Gly Asp His His Ile Pro Leu Leu Arg Pro Glu Met Asn Gln245 250 255Val Trp Ile Asp Glu Lys Tyr Gly Ile His Thr Tyr Gln Pro Leu Lys260 265 270Glu Gly Tyr Val Arg Ile Asn Tyr Leu Ser Ser Ser Ser Asn Tyr Asp275 280 285His Leu Ser Ala Trp Leu Phe Lys Asp Val Ala Thr Xaa Ser Thr Thr290 295 300Trp Pro Asp Gly Ser Asn Phe Val Asn Gln Gly Leu Tyr Gly Arg Tyr305 310 315 320Ile Asp Val Ser Leu Lys Thr Asn Ala Lys Glu Ile Gly Phe Leu Ile325 330 335Leu Asp Glu Ser Lys Thr Gly Asp Ala Val Lys Val Gln Pro Asn Asp340 345 350Tyr Val Phe Arg Asp Leu Ala Asn His Asn Gln Ile Phe Val Lys Asp355 360 365Lys Asp Pro Lys Val Tyr Asn Asn Pro Tyr Tyr Ile Asp Gln Val Gln370 375 380Leu Lys Asp Ala Gln Gln Ile Asp Leu Thr Ser Ile Gln Ala Ser Phe385 390 395 400Thr Thr Leu Asp Gly Val Asp Lys Thr Glu Ile Leu Lys Glu Leu Lys405 410 415Val Thr Asp Lys Asn Gln Asn Ala Ile Gln Ile Ser Asp Ile Thr Leu420 425 430Asp Thr Ser Lys Ser Leu Leu Ile Ile Lys Gly Asp Phe Asn Pro Lys435 440 445Gln Gly His Phe Asn Ile Ser Tyr Asn Gly Asn Asn Val Met Thr Arg450 455 460Gln Ser Trp Glu Phe Lys Asp Gln Leu Tyr Ala Tyr Ser Gly Asn Leu465 470 475 480Gly Ala Val Leu Asn Gln Asp Gly Ser Lys Val Glu Ala Ser Leu Trp485 490 495Ser Pro Ser Ala Asp Ser Val Thr Met Ile Ile Tyr Asp Lys Asp Asn500 505 510Gln Asn Arg Val Val Ala Thr Thr Pro Leu Val Lys Asn Asn Lys Gly515 520 525Val Trp Gln Thr Ile Leu Asp Thr Lys Leu Gly Ile Lys Asn Tyr Thr530 535 540Gly Tyr Tyr Tyr Leu Tyr Glu Ile Lys Arg Gly Lys Asp Lys Val Lys545 550 555 560Ile Leu Asp Pro Tyr Ala Lys Ser Leu Ala Glu Trp Asp Ser Asn Thr565 570 575Val Asn Asp Asp Ile Lys Thr Ala Lys Ala Ala Phe Val Asn Pro Ser580 585 590Gln Leu Gly Pro Gln Asn Leu Ser Phe Ala Lys Ile Ala Asn Phe Lys595 600 605Gly Arg Gln Asp Ala Val Ile Tyr Glu Ala His Val Arg Asp Phe Thr610 615 620Ser Asp Arg Ser Leu Asp Gly Lys Leu Lys Asn Gln Phe Gly Thr Phe625 630 635 640Ala Ala Phe Ser Glu Lys Leu Asp Tyr Leu Gln Lys Leu Gly Val Thr645 650 655His Ile Gln Leu Leu Pro Val Leu Ser Tyr Phe Tyr Val Asn Glu Met660 665 670Asp Lys Ser Arg Ser Thr Ala Tyr Thr Ser Ser Asp Asn Asn Tyr Asn675 680 685Trp Gly Tyr Asp Pro Gln Ser Tyr Phe Ala Leu Ser Gly Met Tyr Ser690 695 700Glu Lys Pro Lys Asp Pro Ser Ala Arg Ile Ala Glu Leu Lys Gln Leu705 710 715 720Ile His Asp Ile His Lys Arg Gly Met Gly Val Ile Leu Asp Val Val725 730 735Tyr Asn His Thr Ala Lys Thr Tyr Leu Phe Glu Asp Ile Glu Pro Asn740 745 750Tyr Tyr His Phe Met Asn Glu Asp Gly Ser Pro Arg Glu Ser Phe Gly755 760 765Gly Gly Arg Leu Gly Thr Thr His Ala Met Ser Arg Arg Val Leu Val770 775 780Asp Ser Ile Lys Tyr Leu Thr Ser Glu Phe Lys Val Asp Gly Phe Arg785 790 795 800Phe Asp Met Met Gly Asp His Asp Ala Ala Ala Ile Glu Leu Ala Tyr805 810 815Lys Glu Ala Lys Ala Ile Asn Pro Asn Met Ile Met Ile Gly Glu Gly820 825 830Trp Arg Thr Phe Gln Gly Asp Gln Gly Gln Pro Val Lys Pro Ala Asp835 840 845Gln Asp Trp Met Lys Ser Thr Asp Thr Val Gly Val Phe Ser Asp Asp850 855 860Ile Arg Asn Ser Leu Lys Ser Gly Phe Pro Asn Glu Gly Thr Pro Ala865 870 875 880Phe Ile Thr Gly Gly Pro Gln Ser Leu Gln Gly Ile Phe Lys Asn Ile885 890 895Lys Ala Gln Pro Gly Asn Phe Glu Ala Asp Ser Pro Gly Asp Val Val900 905 910Gln Tyr Ile Ala Ala His Asp Asn Leu Thr Leu His Asp Val Ile Ala915 920 925Lys Ser Ile930<210>27<211>5607<212>DNA<213>鏈球菌<220><221>CDS<222>(2)…(301)<400>27a att caa agt ttg aca gaa ggt caa ctt cgt tct gat atc cct gag ttc49Ile Gln Ser Leu Thr Glu Gly Gln Leu Arg Ser Asp Ile Pro Glu Phe1 5 10 15cgt gct ggt gat act gta cgt gtt cac gct aaa gtt gtt gaa ggt act 97Arg Ala Gly Asp Thr Val Arg Val His Ala Lys Val Val Glu Gly Thr20 25 30cgc gaa cgt att cag atc ttt gaa ggt gtt gtt atc tca cgt aaa ggt 145Arg Glu Arg Ile Gln Ile Phe Glu Gly Val Val Ile Ser Arg Lys Gly35 40 45caa gga atc tca gaa atg tac aca gta cgt aaa att tct ggt ggt atc 193Gln Gly Ile Ser Glu Met Tyr Thr Val Arg Lys Ile Ser Gly Gly Ile50 55 60ggt gta gag cgt aca ttc cca att cac act cct cgt gtt gat aaa atc 241Gly Val Glu Arg Thr Phe Pro Ile His Thr Pro Arg Val Asp Lys Ile65 70 75 80gaa gtt gtt cgt tat ggt aaa gta cgt cgt gct aaa ctt tac tac tta 289Glu Val Val Arg Tyr Gly Lys Val Arg Arg Ala Lys Leu Tyr Tyr Leu85 90 95cgc gca ttg caa ggtaaagctg cacgtattaa agaaatccgt cgttaatttt 341Arg Ala Leu Gln100gatgatcaga ttttaaaaat gcttggttgt ttgaggatag taactatgtt ttaaaactgg 401acaaccaaga cgtaaaaaat ctgcctgtgg gcagtttttt tactaggtcc ccttagttca 461atggatataa caactccctc ctaaggagta attgctggtt cgattccggc aggggacata 521ttcattgcat gtaaatagcg gtttagagct attttgcccc aaatttctct gattaagttt 581atcgttccta tctttttgtt cttgtaattg atgtgcgtaa acttctaaag tgatatttaa 641attctcgtga tctaaaactt gagagatgga aattagatag cttgcaaatg tatgcctgag 701agagtgcact cgtacctcgc gaccagttat ttttcggata gttttattga ctgcattatt 761tgaaagtttg tcgaataatc tgtcgttttt attttttgta aattcatgca aaaaaaataa 821tgtatcattg tcaattggta tatttctgat actacttttg ttttttgttg gcaggtatct 881ttggttgaaa tgataatccc aagttttatt aattgataaa tatttgttag tgtaatcaat 941atcattaact gttaaaccta aacattcagc gaagcgcatg ccagttttag cgatgaggta 1001taacgctgca cacgattgat gttgtgattt ttctttacaa atttttatca agcgtaagta 1061ttcattggtt tcaagaaatt ttatctctat ttacgcccct tattttttgc tttaacctta 1121gtgaataaac aaaaattttt ttctatatat ccctcgtgaa cagccatgga tacgcaggct 1181tttacatgta tgttaaaacg ctttactgta tcttgcacat gcgtttgact ataatgattt 1241atgacttgtt gatatttagt ggaagtaata ttgcaaagta atatatttcc tattatatgt 1301ttatacgana ttcgatattc ccacccgttg tcgcgtttac ggaaatacgc cattgatata 1361ctccacatta gctaaagaac agggtgttca aggctacctt gatggaaaag gctctcttag 1421agatatttgt aaatggtatg atatctcaag tcgctctgtt ctccaaaagt ggataaaacg 1481gtatactagt ggtgaagact tgaaagccac tagtagagga tatagccgta tgaaacaagg 1541aaggcaagcc acatttgaag aacgtgtaga gattgttaac tacaccattg cccatgggaa 1601agactatcaa gcagctattg agaagtttgg tgtttcctac caacaaattt attcttgggt 1661gcgtaagctt gagaagaatg gctcacaagg tttggttgat agacgtgtga aagggttgga 1721gagtaggcct gatttaaccg agattgagca actttaactc aagattaaac aattggagga 1781acgtaatcgt ctcttagaaa tcgaggttag tttactaaaa aagttagaag acatcaaacg 1841aggaaacaga cggtaagact aggtaagcat ttagcggagt tccaagtaat caagaattat 1901tacgatgagg aatctaatgt gcctattcag gccttatgcc aactcttgaa ggggtctcgt 1961tcaggctatt acaagtggct caatcgtcaa aaaacagatt ttgagacaaa aaatacaaag 2021ctaatggcta aaatcaagga acttcgtaga ctctacaatg gtatcttagg ttatcgccgt 2081atgacaacat ttattaatcg tcaacttggg acaacttaaa acaagaaacg gattcgttga 2141ttgatgaaca ttctggggat tagttcagtc attcgtcgtg ttagccatgc ttgtacaaaa 2201gctggtgaca gattttacga agaaaatatt cttaatcgtg aatttacagc cacagctcat 2261aaccagaaat ggtgcacaga tgtcacctat cttcaatacg gtctgggagc taaagcttat 2321ctcagtgcga ttaaagacct gtataacggt tctattatcg cttatgagat tagtcacaac 2381aatgaaatcc acttgttatg aagaccatta aaaaggggct agagctcaat ccaggagcca 2441cacctatcat ccatagcgat tgaggtagtc aatatacttc caaagaatac cgttatatca 2501tacaacaagc tggtctgacc ttatccatgt cccggattgg caaatgtatt gataatgcac 2561caactgaaag tttctttggg tttttcaaga ctgagtctta ccaccttaag aaatacaact 2621cttatgatga gttggtcaat gatgtggcac gttatatcga attctacaac acacaacgtt 2681atcaatcaaa attaaacaac ctgactcctc tagaattcag gaatcaggtt gcataactta 2741tcttttatta tttgactgtc tacttgacag ggagccgttc agattgctta acctttctaa 2801atttgctaaa atagctacaa gaaaacgagc catttaatgc ttatttctta tactgtcttg 2861cctcacgctc tcctcgacca aaaattgagc gtgaggcttt ttgtttcatt aaacgatgat 2921atttccatat tcatcagttt gttttccgag agccattaaa gcttcgataa ggtcgataat 2981tccaggaata aaggtaatac taaaaataat atataaaaaa acctggccta tttttcctgc 3041gtaaaattta tgcgctccaa tgccgcccaa aagaacgtta ataaaacata aactactatg 3101ttagcataag actttatttt tacaactgaa tttcatataa atggattaga gtaagggata 3161aaagaaatta gcatagctct tttgaaaata aaaaaattaa tataatatgg aaaaaatttt 3221atttcataaa cgtttcataa aaggtatgta atctagtatt taggcaacac tattttgtca 3281ctggtgtcta gtaacttata gattgataat tttactagta aacgtaattc ttcgctttaa 3341gagttaaatg tctatttatt gtaagctaaa ttgggaggtg aacttatgta aaattagata 3401ggtactgtca agtacgggat gattattgaa acagccagta tgcatcataa aatctgtatt 3461gcttaataac tatttcctta accagacatc agttcattgt ttatcatcgc taccctaagt 3521ctagtttttt caatagagca ttaggtagtt tttgataata aaactatata aacatgagaa 3581ttagatttcg tattgcattc ttcataatga gttatttgag attttccttt gaataaatag 3641atacgaaatt cagtaacttc atatataaac ggctctatca ttgagatagt ttgtcaaatg 3701aagaaatttt taatggaaat agttttaaaa acattagttg taggcgatgt aaaaatatta 3761atccagtgga tgcaatagtt gcggagtaaa aatagagagg agtaattagg aagtgataaa 3821aaatgctata gcatatatta ccagaaaaaa aaatagaaca cttattatat ttgctatttt 3881aacaattgtt ctttcttgct tgtattcatg tttaacaata atgaaatcaa gtaatgaaat 3941agaaaaggct ttatatgaaa gttctaattc ttcaatatca attacaaaaa aagatggtaa 4001atattttaat attaatcaat ttaagaatat tgaaaaaata aaagaggttg aagaaaaaat 4061atttcaatat gatggattag caaaattgaa agatcttaaa gtagttagtg gtgagcaaag 4121tataaataga gaagatttat ctgacgaatt taaaaatgtt gtttcactag aagctacaag 4181taatactaaa agaaatcttt tatttagtag tggagtattt agttttaaag aaggaaaaaa 4241tatagaagaa aatgataaga attcaattct tgttcatgaa gaatttgcta aacaaaacaa 4301actaaaattg ggtgatgaaa ttgatcttga attactagat acggaaaaaa gtggaaaaat 4361aaaaagtcat aaatttaaaa ttataggaat cttttctggt aaaaaacagg aaacatatac 4421aggattatca tctgatttta gcgaaaatat ggtttttgta gattattcaa ctagccaaga 4481aatattaaat aaatcagaga ataatagaat tgcaaataaa attttaatgt attctggtag 4541tttagaatct acagagcttg ccttaaacaa attgaaagac tttaaaattg ataagtcaaa 4601gtattctatt aagaaagata ataaagcatt cgaagagtct ttagagtcag tgagtggaat 4661aaaacatata attaaaataa tgacttattc gattatgtta ggtggaatag ttgttctttc 4721attaatcttg attctatggt taagagaaag aatttatgaa ataggtatat ttttatctat 4781tggaacaact aagatacaaa ttataaggca atttatattt gagttaatat tcatatcaat 4841accaagtata atatcctcct tatttttagg gaatctacta ttaaaagtaa ttgtagaagg 4901atttattaac tcagagaact caatgatttt cggtggaagt ttaataaata aaagcagttt 4961tatgttaaac ataacaacac ttgcagaaag ttatttaata ttaataagta ttattgtttt 5021atcagttgta atggcctctt cattaatatt atttaagaaa ccacaagaaa tattatcaaa 5081aataagttag gagcaaataa tggatatatt agaaataaag aatgtaaatt acagttacgc 5141aaattctaaa gaaaaagttt tgtcaggagt aaatcaaaaa tttgaacttg gaaagtttta 5201tgcgatagta gggaagtcag gaacaggaaa atccacactt ctttccttac ttgcaggact 5261tgataaagtt caaacaggaa aaatcttgtt taagaatgaa gatatagaaa agaaaggata 5321tagtaatcac agaaaaaata atatatcttt ggtatttcaa aattataatt taatagatta 5381tttatcgccg attgaaaata ttagactagt aaataaatca gtagatgaga gtatcttgtt 5441cgaattaggt ttagataaaa aacaaataaa aagaaatgtt atgaaattat ctggtggtca 5501gcaacaaagg gtagctattg ctagggcact ggtatcagat gccccaataa tactagctga 5561tgagcctacc ggtaacctag acagtgttac tgctggagaa ataatt 5607<210>28<211>111<212>蛋白質(zhì)<213>鏈球菌<400>28Ile Gln Ser Leu Thr Glu Gly Gln Leu Arg Ser Asp Ile Pro Glu Phe1 5 10 15Arg Ala Gly Asp Thr Val Arg Val His Ala Lys Val Val Glu Gly Thr20 25 30Arg Glu Arg Ile Gln Ile Phe Glu Gly Val Val Ile Ser Arg Lys Gly35 40 45Gln Gly Ile Ser Glu Met Tyr Thr Val Arg Lys Ile Ser Gly Gly Ile50 55 60Gly Val Glu Arg Thr Phe Pro Ile His Thr Pro Arg Val Asp Lys Ile65 70 75 80Glu Val Val Arg Tyr Gly Lys Val Arg Arg Ala Lys Leu Tyr Tyr Leu85 90 95Arg Ala Leu Gln Gly Lys Ala Ala Arg Ile Lys Glu Ile Arg Arg100 105 110<210>29<211>173<212>蛋白質(zhì)<213>鏈球菌<400>29Met Arg Phe Ala Glu Cys Leu Gly Leu Thr Val Asn Asp Ile Asp Tyr1 5 10 15Thr Asn Lys Tyr Leu Ser Ile Asn Lys Thr Trp Asp Tyr His Phe Asn20 25 30Gln Arg Tyr Leu Pro Thr Lys Asn Lys Ser Ser Ile Arg Asn Ile Pro35 40 45Ile Asp Asn Asp Thr Leu Phe Phe Leu His Glu Phe Thr Lys Asn Lys
50 55 60Asn Asp Arg Leu Phe Asp Lys Leu Ser Asn Asn Ala Val Asn Lys Thr65 70 75 80Ile Arg Lys Ile Thr Gly Arg Glu Val Arg Val His Ser Leu Arg His85 90 95Thr Phe Ala Ser Tyr Leu Ile Ser Ile Ser Gln Val Leu Asp His Glu100 105 110Asn Leu Asn Ile Thr Leu Glu Val Tyr Ala His Gln Leu Gln Glu Gln115 120 125Lys Asp Arg Asn Asp Lys Leu Asn Gln Arg Asn Leu Gly Gln Asn Ser130 135 140Ser Lys Pro Leu Phe Thr Cys Asn Glu Tyr Val Pro Cys Arg Asn Arg145 150 155 160Thr Ser Asn Tyr Ser Leu Gly Gly Ser Cys Tyr Ile His165 170<210>30<211>389<212>蛋白質(zhì)<213>鏈球菌<400>30Met Lys Ser Ser Asn Glu Ile Glu Lys Ala Leu Tyr Glu Ser Ser Asn1 5 10 15Ser Ser Ile Ser Ile Thr Lys Lys Asp Gly Lys Tyr Phe Asn Ile Asn20 25 30Gln Phe Lys Asn Ile Glu Lys Ile Lys Glu Val Glu Glu Lys Ile Phe35 40 45Gln Tyr Asp Gly Leu Ala Lys Leu Lys Asp Leu Lys Val Val Ser Gly50 55 60Glu Gln Ser Ile Asn Arg Glu Asp Leu Ser Asp Glu Phe Lys Asn Val65 70 75 80Val Ser Leu Glu Ala Thr Ser Asn Thr Lys Arg Asn Leu Leu Phe Ser85 90 95Ser Gly Val Phe Ser Phe Lys Glu Gly Lys Asn Ile Glu Glu Asn Asp100 105 110Lys Asn Ser Ile Leu Val His Glu Glu Phe Ala Lys Gln Asn Lys Leu115 120 125Lys Leu Gly Asp Glu Ile Asp Leu Glu Leu Leu Asp Thr Glu Lys Ser130 135 140Gly Lys Ile Lys Ser His Lys Phe Lys Ile Ile Gly Ile Phe Ser Gly145 150 155 160Lys Lys Gln Glu Thr Tyr Thr Gly Leu Ser Ser Asp Phe Ser Glu Asn165 170 175Met Val Phe Val Asp Tyr Ser Thr Ser Gln Glu Ile Leu Asn Lys Ser180 185 190Glu Asn Asn Arg Ile Ala Asn Lys Ile Leu Met Tyr Ser Gly Ser Leu195 200 205Glu Ser Thr Glu Leu Ala Leu Asn Lys Leu Lys Asp Phe Lys Ile Asp210 215 220Lys Ser Lys Tyr Ser Ile Lys Lys Asp Asn Lys Ala Phe Glu Glu Ser225 230 235 240Leu Glu Ser Va1 Ser Gly Ile Lys His Ile Ile Lys Ile Met Thr Tyr245 250 255Ser Ile Met Leu Gly Gly Ile Val Val Leu Ser Leu Ile Leu Ile Leu260 265 270Trp Leu Arg Glu Arg Ile Tyr Glu Ile Gly Ile Phe Leu Ser Ile Gly275 280 285Thr Thr Lys Ile Gln Ile Ile Arg Gln Phe Ile Phe Glu Leu Ile Phe290 295 300Ile Sar Ile Pro Ser Ile Ile Ser Ser Leu Phe Leu Gly Asn Leu Leu305 310 315 320Leu Lys Val Ile Val Glu Gly Phe Ile Asn Ser Glu Asn Ser Met Ile325 330 335Phe Gly Gly Ser Leu Ile Asn Lys Ser Ser Phe Met Leu Asn Ile Thr340 345 350Thr Leu Ala Glu Ser Tyr Leu Ile Leu Ile Ser Ile Ile Val Leu Ser355 360 365Val Val Met Ala Ser Ser Leu Ile Leu Phe Lys Lys Pro Gln Glu Ile370 375 380Leu Ser Lys Ile Ser385<210>31<211>169<212>蛋白質(zhì)<213>鏈球菌<400>31Met Asp Ile Leu Glu Ile Lys Asn Val Asn Tyr Ser Tyr Ala Asn Ser1 5 10 15Lys Glu Lys Val Leu Ser Gly Val Asn Gln Lys Phe Glu Leu Gly Lys20 25 30Phe Tyr Ala Ile Val Gly Lys Ser Gly Thr Gly Lys Ser Thr Leu Leu35 40 45Ser Leu Leu Ala Gly Leu Asp Lys Val Gln Thr Gly Lys Ile Leu Phe50 55 60Lys Asn Glu Asp Ile Glu Lys Lys Gly Tyr Ser Asn His Arg Lys Asn65 70 75 80Asn Ile Ser Leu Val Phe Gln Asn Tyr Asn Leu Ile Asp Tyr Leu Ser85 90 95Pro Ile Glu Asn Ile Arg Leu Val Asn Lys Ser Val Asp Glu Ser Ile100 105 110Leu Phe Glu Leu Gly Leu Asp Lys Lys Gln Ile Lys Arg Asn Val Met115 120 125Lys Leu Ser Gly Gly Gln Gln Gln Arg Val Ala Ile Ala Arg Ala Leu130 135 140Val Ser Asp Ala Pro Ile Ile Leu Ala Asp Glu Pro Thr Gly Asn Leu145 150 155 160Asp Ser Val Thr Ala Gly Glu Ile Ile165<210>32<211>4171<212>DNA<213>鏈球菌<400>32catatgacaa tatttttcaa agtctacatc acttactcgc ctgtcgtgga aaatctggca 60atacattaat cgaccaatta gttgctgatg gtttacttca tgcagataat cactaccatt 120ttttcaatgg gaagtctctg gccactttca atactaacca attgattcgc gaagttgtct 180atgttgaaat atccttagat actatgtcta gtggtgaaca tgatttagta aaagttaaca 240ttatcagacc cactaccgag catactatcc ccacgatgat gacagctagc ccctatcatc 300aaggtatcaa tgatcctgcc gcagaccaaa aaacatacca aatggagggt gcgctagcag 360ttaaacagcc taaacacata caagttgaca caaaaccatt taaagaagaa gtaaaacatc 420cttcaaaatt acccatcagc cctgcaactg aaagcttcac acacattgac agttatagtc 480tcaatgacta ttttctttct cgtggttttg ctaatatata cgtttcaggt gtgggtactg 540ctggctctac gggtttcatg accagtgggg attaccaaca aatacaaagc tttaaagcag 600tcattgattg gttaaatggt aaggttactg cattcacaag tcataaacga gataaacaag 660tcaaggctga ttggtcaaac ggccttgtag caaccacagg taaatcttat ctcggtacca 720tgtcaactgg tttagcaaca actggcgttg aggggctgaa agtcattatc gctgaagccg 780caatctccac atggtatgat tattatcgag aaaatgggct tgtgtgtagt ccaggcggct 840accccggtga agatttagac gttttaacag aattaacata ctcacgaaac ctcttagctg 900gtgattacat caaaaacaac gattgctatc aagcattgtt aaatgaacaa tcaaaagcaa 960ttgaccgtca aagtggggat tacaaccaat actggcatga ccgtaattac ctaactcacg 1020tcaataatgt caaaagtcga gtagtttaca ctcatggact acaggattgg aatgttaagc 1080caagacatgt ctacaaagtt ttcaatgcat tgcctcaaac catcaaaaaa cacctttttt 1140tacatcaagg tcaacatgtg tatatgcata attggcagtc gattgatttt cgtgaaagca 1200tgaatgcctt actaagccaa gaactacttg gcattgacaa tcatttccaa ttagaagagg 1260tcatttggca agataatact actgagcaaa cttggcaagt tttagatgct ttcggaggaa 1320accatcaaga gcaaattggt ttaggtgata gtaaaaaact tattgataac cattatgaca 1380aagaagcctt tgatacttat tgtaaagact tcaatgtgtt caaaaatgat cttttcaagg 1440gaaataataa aaccaatcaa atcactatta atcttcctct aaagaaaaat tatctcctga 1500atggacagtg caaactccat ctacgtgtta aaactagtga caaaaaggcc attttatcag 1560cccaaatctt agactatggt cctaaaaaac gattcaaaga tacaccaacc atcaaattct 1620taaacagcct tgataatggt aaaaattttg ccagagaagc tttacgtgaa ctcccgttta 1680ctaaagatca ttatcgtgtc atcagtaaag gtgtcttgaa ccttcaaaat cgtacagact 1740tacttacaat tgaggctatc gagccagaac aatggtttga tatcgagttt agcctccaac 1800caagtatata tcaattgagt aaaggtgata atctaaggat tatcctttat acaactgatt 1860ttgaacatac cattcgagat aatgctagtt actctataac agtagatttg agtcaatctt 1920atttaactat cccaactaat caaggaaatt aacttatgaa acttcttact aaagaacggt 1980ttgatgattc tcaacacttt tggtaccaga tcaatttatt acaagagagt aacttcggag 2040cagtttttga ccatgataat aaaaacattc cacaggttgt tgcaactatt gttgatgatt 2100tacaaggttc cggaagttcg aatcatttct ggtattttgg caatactact gatacttcca 2160tccttatgat tgctcattta aatcgaaaat tctatattca ggttaattta aaggactttg 2220actttgcact caatttaata gctataaata attggaagag tctcctccaa actcaacttg 2280aagctctaaa cgatacccta gcaatatttc aataaataag gtagaatgga gtgacaaagc 2340aacgcgaggg agactgatta atgtcatctt attggaataa ctatcctgaa cttaaaaaaa 2400atattgatga aaccaatcaa ctaattcaag aaagaataca ggtcagaaat aaagatattg 2460aagcggcgct aagccaactc acagctgcgg gaggaaaaca gctcagacca gcattctttt 2520accttttttc tcaacttggt aataaggaga atcaagatac tcagcaacta aagaaaatcg 2580ctgcttcttt agaaatcctt cacgttgcta cattaatcca tgatgatgtc attgatgact 2640caccactaag acgtggaaat atgaccattc aaagcaagtt tggcaaagac atcgcagttt 2700atactgggga tttacttttc acagtctttt tcgatcttat tttagaatct atgactgata 2760caccatttat gaggattaat gcaaaatcta tgcgtaaaat tctcatggga gaattggacc 2820agatgcacct tcgttacaat caacaacaag gtatccatca ctatttacgt gcgatttcag 2880gtaagacagc cgaactcttt aaattagcta gcaaagaagg agcttacttt ggtggtgcag 2940agaaggaggt tgttcgtcta gcaggccata tcggctttaa cattggtatg acattccaaa 3000ttttggatga tatcctggat tatactgcag ataaaaaaac atttaataag cctgtcttag 3060aggatttaac acaaggcgtt tacagccttc ctctacttct tgccattgaa gaaaatcctg 3120atattttcaa acctatttta gataaaaaaa cagatatggc tactgaagac atggaaaaaa 31e0ttgcttatct cgtcgtttcc catagaggtg ttgacaaagc tcgccatcta gctcgtaaat 3240ttactgagaa agctattagt gacataaata agctacccca gaactctgca aaaaaacagt 3300tgctacaatt aactaattac cttttaaaac gcaaaattta aataataaaa aaacattcca 3360caatgctaga aaagcagtta gggaatgttt ttttattatc atttatttat cgcacctatc 3420aatcatcata gatcaccatc atcagcggct ttcagctgac ggtaacgttg actactttga 3480gacaattctt gaggagaacc ttccaactct aattgcccat tttctataaa taagatacga 3540tcagcatgtt caataccttt taagtgatgt gtaatccaaa ctaaggtctt accttccaat 3600tctttcataa atacccttag taaggcttgt tcagtaatag gatcaagtcc aacagttggc 3660tcatctaaga taacaattgg gacatctttt agtaagattc tagccaaagc aattctatgc 3720ctttcgccac ctgaaaacct aagtccagct tcatcaacca ttgtatagag accatctgat 3780aaatcagtga ccatctcttt caatccaact cgttcaagaa ctttccatac atcttcttca 3840ctagcatctt ggtttccaat gcgaatgtta tttagcaggg ttgtattaaa aaggtagggc 3900gcttgttgta tcactccaat atagttagaa atgcaatcac caactattga aacatcagca 3960ccgcctaggg taatcttccc ttgacttgct ttcaagtcgc cacgaagtag actagctaag 4020gtactcttgc cagaaccact ccgccctaaa atagcaattt tttctccttc tttaatatcc 4080aaatctaaat gatgcaaaac ccatttctct tgtggcttat actggaaact taaattcttg 4140acggaaaaat catatggctt attaggcaat t 4171<210>33<211>649<212>蛋白質(zhì)<213>鏈球菌<400>33Tyr Asp Asn Ile Phe Gln Ser Leu His His Leu Leu Ala Cys Arg Gly1 5 10 15Lys Ser Gly Asn Thr Leu Ile Asp Gln Leu Val Ala Asp Gly Leu Leu20 25 30His Ala Asp Asn His Tyr His Phe Phe Asn Gly Lys Ser Leu Ala Thr35 40 45Phe Asn Thr Asn Gln Leu Ile Arg Glu Val Val Tyr Val Glu Ile Ser50 55 60Leu Asp Thr Met Ser Ser Gly Glu His Asp Leu Val Lys Val Asn Ile65 70 75 80Ile Arg Pro Thr Thr Glu His Thr Ile Pro Thr Met Met Thr Ala Ser85 90 95Pro Tyr His Gln Gly Ile Asn Asp Pro Ala Ala Asp Gln Lys Thr Tyr100 105 110Gln Met Glu Gly Ala Leu Ala Val Lys Gln Pro Lys His Ile Gln Val115 120 125Asp Thr Lys Pro Phe Lys Glu Glu Val Lys His Pro Ser Lys Leu Pro130 135 140Ile Ser Pro Ala Thr Glu Ser Phe Thr His Ile Asp Ser Tyr Ser Leu145 150 155 160Asn Asp Tyr Phe Leu Ser Arg Gly Phe Ala Asn Ile Tyr Val Ser Gly165 170 175Val Gly Thr Ala Gly Ser Thr Gly Phe Met Thr Ser Gly Asp Tyr Gln180 185 190Gln Ile Gln Ser Phe Lys Ala Val Ile Asp Trp Leu Asn Gly Lys Val195 200 205Thr Ala Phe Thr Ser His Lys Arg Asp Lys Gln Val Lys Ala Asp Trp210 215 220Ser Asn Gly Leu Val Ala Thr Thr Gly Lys Ser Tyr Leu Gly Thr Met225 230 235 240Ser Thr Gly Leu Ala Thr Thr Gly Val Glu Gly Leu Lys Val Ile Ile245 250 255Ala Glu Ala Ala Ile Ser Thr Trp Tyr Asp Tyr Tyr Arg Glu Asn Gly260 265 270Leu Val Cys Ser Pro Gly Gly Tyr Pro Gly Glu Asp Leu Asp Val Leu275 280 285Thr Glu Leu Thr Tyr Ser Arg Asn Leu Leu Ala Gly Asp Tyr Ile Lys290 295 300Asn Asn Asp Cys Tyr Gln Ala Leu Leu Asn Glu Gln Ser Lys Ala Ile305 310 315 320Asp Arg Gln Ser Gly Asp Tyr Asn Gln Tyr Trp His Asp Arg Asn Tyr325 330 335Leu Thr His Val Asn Asn Val Lys Ser Arg Val Val Tyr Thr His Gly340 345 350Leu Gln Asp Trp Asn Val Lys Pro Arg His Val Tyr Lys Val Phe Asn355 360 365Ala Leu Pro Gln Thr Ile Lys Lys His Leu Phe Leu His Gln Gly Gln370 375 380His Val Tyr Met His Asn Trp Gln Ser Ile Asp Phe Arg Glu Ser Met385 390 395 400Asn Ala Leu Leu Ser Gln Glu Leu Leu Gly Ile Asp Asn His Phe Gln405 410 415Leu Glu Glu Val Ile Trp Gln Asp Asn Thr Thr Glu Gln Thr Trp Gln420 425 430Val Leu Asp Ala Phe Gly Gly Asn His Gln Glu Gln Ile Gly Leu Gly435 440 445Asp Ser Lys Lys Leu Ile Asp Asn His Tyr Asp Lys Glu Ala Phe Asp450 455 460Thr Tyr Cys Lys Asp Phe Asn Val Phe Lys Asn Asp Leu Phe Lys Gly465 470 475 480Asn Asn Lys Thr Asn Gln Ile Thr Ile Asn Leu Pro Leu Lys Lys Asn485 490 495Tyr Leu Leu Asn Gly Gln Cys Lys Leu His Leu Arg Val Lys Thr Ser500 505 510Asp Lys Lys Ala Ile Leu Ser Ala Gln Ile Leu Asp Tyr Gly Pro Lys515 520 525Lys Arg Phe Lys Asp Thr Pro Thr Ile Lys Phe Leu Asn Ser Leu Asp530 535 540Asn Gly Lys Asn Phe Ala Arg Glu Ala Leu Arg Glu Leu Pro Phe Thr545 550 555 560Lys Asp His Tyr Arg Val Ile Ser Lys Gly Val LeuAsn Leu Gln Asn565 570575Arg Thr Asp Leu Leu Thr Ile Glu Ala Ile Glu Pro Glu Gln Trp Phe580 585 590Asp Ile Glu Phe Ser Leu Gln Pro Ser Ile Tyr Gln Leu Ser Lys Gly595 600 605Asp Asn Leu Arg Ile Ile Leu Tyr Thr Thr Asp Phe Glu His Thr Ile610 615 620Arg Asp Asn Ala Ser Tyr Ser Ile Thr Val Asp Leu Ser Gln Ser Tyr625 630 635 640Leu Thr Ile Pro Thr Asn Gln Gly Asn645<210>34<211>119<212>蛋白質(zhì)<213>鏈球菌<400>34Met Lys Leu Leu Thr Lys Glu Arg Phe Asp Asp Ser Gln His Phe Trp1 5 10 15Tyr Gln Ile Asn Leu Leu Gln Glu Ser Asn Phe Gly Ala Val Phe Asp20 25 30His Asp Asn Lys Asn Ile Pro Gln Val Val Ala Thr Ile Val Asp Asp35 40 45Leu Gln Gly Ser Gly Ser Ser Asn His Phe Trp Tyr Phe Gly Asn Thr50 55 60Thr Asp Thr Ser Ile Leu Met Ile Ala His Leu Asn Arg Lys Phe Tyr65 70 75 80Ile Gln Val Asn Leu Lys Asp Phe Asp Phe Ala Leu Asn Leu lle Ala85 90 95Ile Asn Asn Trp Lys Ser Leu Leu Gln Thr Gln Leu Glu Ala Leu Asn100 105 110Asp Thr Leu Ala Ile Phe Gln115<210>35<211>326<212>蛋白質(zhì)<213>鏈球菌<400>35Met Ser Ser Tyr Trp Asn Asn Tyr Pro Glu Leu Lys Lys Asn lle Asp1 5 10 15Glu Thr Asn Gln Leu Ile Gln Glu Arg Ile Gln Val Arg Asn Lys Asp20 25 30lle Glu Ala Ala Leu Ser Gln Leu Thr Ala Ala Gly Gly Lys Gln Leu35 40 45Arg Pro Ala Phe Phe Tyr Leu Phe Ser Gln Leu Gly Asn Lys Glu Asn50 55 60Gln Asp Thr Gln Gln Leu Lys Lys Ile Ala Ala Ser Leu Glu Ile Leu65 70 75 80His Val Ala Thr Leu Ile His Asp Asp Val Ile Asp Asp Ser Pro Leu85 90 95Arg Arg Gly Asn Met Thr Ile Gln Ser Lys Phe Gly Lys Asp Ile Ala100 105 110Val Tyr Thr Gly Asp Leu Leu Phe Thr Val Phe Phe Asp Leu Ile Leu115 120 125Glu Ser Met Thr Asp Thr Pro Phe Met Arg Ile Asn Ala Lys Ser Met130 135 140Arg Lys Ile Leu Met Gly Glu Leu Asp Gln Met His Leu Arg Tyr Asn145 150 155 160Gln Gln Gln G1y Ile His His Tyr Leu Arg Ala Ile Ser Gly Lys Thr165 170 175Ala Glu Leu Phe Lys Leu Ala Ser Lys Glu Gly Ala Tyr Phe Gly Gly180 185 190Ala Glu Lys Glu Val Val Arg Leu Ala Gly His Ile Gly Phe Asn Ile195 200 205Gly Met Thr Phe Gln Ile Leu Asp Asp Ile Leu Asp Tyr Thr Ala Asp210 215 220Lys Lys Thr Phe Asn Lys Pro Val Leu Glu Asp Leu Thr Gln Gly Val225 230 235 240Tyr Ser Leu Pro Leu Leu Leu Ala Ile Glu Glu Asn Pro Asp Ile Phe245 250 255Lys Pro Ile Leu Asp Lys Lys Thr Asp Met Ala Thr Glu Asp Met Glu260 265 270Lys Ile Ala Tyr Leu Val Val Ser His Arg Gly Val Asp Lys Ala Arg275 280 285His Leu Ala Arg Lys Phe Thr Glu Lys Ala Ile Ser Asp Ile Asn Lys290 295 300Leu Pro Gln Asn Ser Ala Lys Lys Gln Leu Leu Gln Leu Thr Asn Tyr305 310 315 320Leu Leu Lys Arg Lys Ile325<210>36<211>247<212>蛋白質(zhì)<213>鏈球菌<400>36Leu Pro Asn Lys Pro Tyr Asp Phe Ser Val Lys Asn Leu Ser Phe Gln1 5 10 15Tyr Lys Pro Gln Glu Lys Trp Val Leu His His Leu Asp Leu Asp Ile20 25 30Lys Glu Gly Glu Lys Ile Ala Ile Leu Gly Arg Ser Gly Ser Gly Lys35 40 45Ser Thr Leu Ala Ser Leu Leu Arg Gly Asp Leu Lys Ala Ser Gln Gly50 55 60Lys Ile Thr Leu Gly Gly Ala Asp Val Ser Ile Val Gly Asp Cys Ile65 70 75 80Ser Asn Tyr Ile Gly Val Ile Gln Gln Ala Pro Tyr Leu Phe Asn Thr85 90 95Thr Leu Leu Asn Asn Ile Arg Ile Gly Asn Gln Asp Ala Ser Glu Glu100 105 110Asp Val Trp Lys Val Leu Glu Arg Val Gly Leu Lys Glu Met Val Thr115 120 125Asp Leu Ser Asp Gly Leu Tyr Thr Met Val Asp Glu Ala Gly Leu Arg130 135 140Phe Ser Gly Gly Glu Arg His Arg Ile Ala Leu Ala Arg Ile Leu Leu145 150 155 160Lys Asp Val Pro Ile Val Ile Leu Asp Glu Pro Thr Val Gly Leu Asp165 170 175Pro Ile Thr Glu Gln Ala Leu Leu Arg Val Phe Met Lys Glu Leu Glu180 185 190Gly Lys Thr Leu Val Trp Ile Thr His His Leu Lys Gly Ile Glu His195 200 205Ala Asp Arg Ile Leu Phe Ile Glu Asn Gly Gln Leu Glu Leu Glu Gly210 215 220Ser Pro Gln Glu Leu Ser Gln Ser Ser Gln Arg Tyr Arg Gln Leu Lys225 230 235 240Ala Ala Asp Asp Gly Asp Leu245<210>37<211>3480<212>DNA<213>鏈球菌<400>37aattctattt ggaggttttt cttgaataaa tggttagtta aggcaagttc cttagttgtt50ttaggtggta tggttttatc tgcgggttcc cgagttttag cggatactta tgtccgtcca 120attgataatg gtagaattac aacaggtttc aatggttatc ctggacattg tggggtggat 180tatgctgttc cgactggaac gattattagg gcagtggcag atggtactgt gaaatttgca 240ggagctggag ccaacttttc ttggatgaca gacttagcag gaaattgtgt catgattcaa 300catgcggatg gaatgcatag tggttacgct catatgtcac gtgtggtggc taggactggg 360gaaaaagtca aacaaggaga tatcatcggt tacgtaggag caactggtat ggcgacggga 420cctcaccttc attttgaatt tttaccagct aaccctaatt ttcaaaatgg tttccatgga 480cgtatcaatc caacgtcact aattgctaac gttgcgacct ttagtggaaa aacgcaagca 540tcagctccaa gcattaagcc attacaatca gctcctgtac agaatcaatc tagtaaatta 600aaagtgtatc gagtagacga attacaaaag gttaatggtg tttggttagt caaaaataac 660accctaacgc cgactgggtt tgattggaac gataatggta taccagcatc agaaattgat 720gaggttgatg ctaatggtaa tttgacagct gaccaggttc ttcaaaaagg tggttacttt 780atctttaatc ctaaaactct taagactgta gaaaaaccca tccaaggaac agctggttta 840acttgggcta agacacgctt tgctaatggt agttcagttt ggcttcgcgt tgacaacagt 900caagaactgc tttacaaata gtttgaggta ttgattcatt gttttaaatg acagttttgt 960tactaactaa gtacaatttc tttaaaccgt ctgaaaataa ttttatagtc cagtaaagtg 1020tgatattata gtctcggact aataaaaagg aaataggaat tgaagcaatg aaaatgaata 1080aaaaggtact attgacatcg acaatggcag cttcgctatt atcagtcgca agtgttcaag 1140cacaagaaac agatacgacg tggacagcac gtactgtttc agaggtaaag gctgatttgg 1200taaagcaaga caataaatca tcatatactg tgaaatatgg tgatacacta agcgttattt 1260cagaagcaat gtcaattgat atgaatgtct tagcaaaaat taataacatt gcagatatca 1320atcttattta tcctgagaca acactgacag taacttacga tcagaagagt catactgcca 1380cttcaatgaa aatagaaaca ccagcaacaa atgctgctgg tcaaacaaca gctactgtgg 1440atttgaaaac caatcaagtt tctgttgcag accaaaaagt ttctctcaat acaatttcgg 1500aaggtatgac accagaagca gcaacaacga ttgtttcgcc aatgaagaca tattcttctg 1560cgccagcttt gaaatcaaaa gaagtattag cacaagagca agctgttagt caagcagcag 1620ctaatgaaca ggtatcaaca gctcctgtga agtcgattac ttcagaagtt ccagcagcta 1680aagaggaagt taaaccaact cagacgtcag tcagtcagtc aacaacagta tcaccagctt 1740ctgttgccgc tgaaacacca gctccagtag ctaaagtagc accggtaaga actgtagcag 1800cccctagagt ggcaagtgtt aaagtagtca ctcctaaagt agaaactggt gcatcaccag 1860agcatgtatc agctccagca gttcctgtga ctacgacttc aacagctaca gacagtaagt 1920tacaagcgac tgaagttaag agcgttccgg tagcacaaaa agctccaaca gcaacaccgg 1980tagcacaacc agcttcaaca acaaatgcag tagctgcaca tcctgaaaat gcagggctcc 2040aacctcatgt tgcagcttat aaagaaaaag tagcgtcaac ttatggagtt aatgaattca 2100gtacataccg tgcaggtgat ccaggtgatc atggtaaagg tttagcagtc gactttattg 2160taggtaaaaa ccaagcactt ggtaatgaag ttgcacagta ctctacacaa aatatggcag 2220caaataacat ttcatatgtt atctggcaac aaaagtttta ctcaaataca aatagtattt 2280atggacctgc taatacttgg aatgcaatgc cagatcgtgg tggcgttact gccaaccatt 2340atgaccatgt tcacgtatca tttaacaaat aatataaaaa aggaagctat ttggcttctt 2400ttttatatgc cttgaataga ctttcaaggt tcttatctaa tttttattaa attgaggaga 2460ttaagctata agtctgaaac tactttcacg ttaaccgtga ctaaatcaaa acgctaaaac 2520taaaatctaa gtctgtaaag attattgaaa acgctttaaa aacagatata ataaggtttg 2580tagatatcta aaattaaaaa agataaggaa gtgagaatat gccacatcta agtaaagaag 2640cttttaaaaa gcaaataaaa aatggcatta ttgtgtcatg tcaagctttg cctggggagc 2700ctctttatac tgaaagtgga ggtgttatgc ctcttttagc tttggcagct caagaagcag 2760gagcggttgg tataagagcc aatagtgtcc gcgacattaa ggaaattcaa gaagttacta 2820atttacctat catcggcatt attaaacgtg aatatcctcc acaagaacca tttatcactg 2880ctacgatgac agaggtggat caattagcta gtttagatat tgcagtaata gccttagatt 2940gtacatttag agagcgtcat gatggtttga gtgtagctga gtttattcaa aagataaaag 3000ggaaatatcc tgaacagttg ctaatggctg atataagtac ttttgaagaa ggtaaaaatg 3060cttttgaagc aggagttgat tttgtgggta caactctatc tggatacaca gattacagcc 3120gccaagaaga aggaccggat atagaactcc ttaataagct ttgtcaagcc ggtatagatg 3180tgattgcgga aggtaaaatt catactccta agcaagctaa tgaaattaat catataggtg 3240ttgcaggaat tgtagttggt ggtgctatca ctagaccaaa agaaatagcg gagcgtttca 3300tctcaggact tagttaaaag tgttactcaa aaatcaaaat caaaataaaa aaggggaata 3360gttatgagta tcaaaaaaag tgtgattggt ttttgcctcg gagctgcagc attatcaatg 3420tttgcttgtg tagacagtag tcaatctgtt atggctgccg agaaggataa agtcgaaatt 3480<210>38<211>306<212>蛋白質(zhì)<213>鏈球菌<400>38Asn Ser Ile Trp Arg Phe Phe Leu Asn Lys Trp Leu Val Lys Ala Ser1 5 10 15Ser Leu Val Val Leu Gly Gly Met Val Leu Ser Ala Gly Ser Arg Val20 25 30Leu Ala Asp Thr Tyr Val Arg Pro Ile Asp Asn Gly Arg Ile Thr Thr35 40 45Gly Phe Asn Gly Tyr Pro Gly His Cys Gly Val Asp Tyr Ala Val Pro50 55 60Thr Gly Thr Ile Ile Arg Ala Val Ala Asp Gly Thr Val Lys Phe Ala65 70 75 80Gly Ala Gly Ala Asn Phe Ser Trp Met Thr Asp Leu Ala Gly Asn Cys85 90 95Val Met Ile Gln His Ala Asp Gly Met His Ser Gly Tyr Ala His Met100 105 110Ser Arg Val Val Ala Arg Thr Gly Glu Lys Val Lys Gln Gly Asp Ile115 120 125Ile Gly Tyr Val Gly Ala Thr Gly Met Ala Thr Gly Pro His Leu His130 135 140Phe Glu Phe Leu Pro Ala Asn Pro Asn Phe Gln Asn Gly Phe His Gly145 150 155 160Arg Ile Asn Pro Thr Ser Leu Ile Ala Asn Val Ala Thr Phe Ser Gly165 170 175Lys Thr Gln Ala Ser Ala Pro Ser Ile Lys Pro Leu Gln Ser Ala Pro180 185 190Val Gln Asn Gln Ser Ser Lys Leu Lys Val Tyr Arg Val Asp Glu Leu195 200 205Gln Lys Val Asn Gly Val Trp Leu Val Lys Asn Asn Thr Leu Thr Pro210 215 220Thr Gly Phe Asp Trp Asn Asp Asn Gly Ile Pro Ala Ser Glu Ile Asp225 230 235 240Glu Val Asp Ala Asn Gly Asn Leu Thr Ala Asp Gln Val Leu Gln Lys245 250 255Gly Gly Tyr Phe Ile Phe Asn Pro Lys Thr Leu Lys Thr Val Glu Lys260 265 270Pro Ile Gln Gly Thr Ala Gly Leu Thr Trp Ala Lys Thr Arg Phe Ala275 280 285Asn Gly Ser Ser Val Trp Leu Arg Val Asp Asn Ser Gln Glu Leu Leu290 295 300Tyr Lys305<210>39<211>434<212>蛋白質(zhì)<213>鏈球菌<400>39Met Lys Met Asn Lys Lys Val Leu Leu Thr Ser Thr Met Ala Ala Ser1 5 10 15Leu Leu Ser Val Ala Ser Val Gln Ala Gln Glu Thr Asp Thr Thr Trp20 25 30Thr Ala Arg Thr Val Ser Glu Val Lys Ala Asp Leu Val Lys Gln Asp35 40 45Asn Lys Ser Ser Tyr Thr Val Lys Tyr Gly Asp Thr Leu Ser Val Ile
50 55 60Ser Glu Ala Met Ser Ile Asp Met Asn Val Leu Ala Lys Ile Asn Asn65 70 75 80Ile Ala Asp Ile Asn Leu Ile Tyr Pro Glu Thr Thr Leu Thr Val Thr85 90 95Tyr Asp Gln Lys Ser His Thr Ala Thr Ser Met Lys Ile Glu Thr Pro100 105 110Ala Thr Asn Ala Ala Gly Gln Thr Thr Ala Thr Val Asp Leu Lys Thr115 120 125Asn Gln Val Ser Val Ala Asp Gln Lys Val Ser Leu Asn Thr Ile Ser130 135 140Glu Gly Met Thr Pro Glu Ala Ala Thr Thr Ile Val Ser Pro Met Lys145 150 155 160Thr Tyr Ser Ser Ala Pro Ala Leu Lys Ser Lys Glu Val Leu Ala Gln165 170 175Glu Gln Ala Val Ser Gln Ala Ala Ala Asn Glu Gln Val Ser Thr Ala180 185 190Pro Val Lys Ser Ile Thr Ser Glu Val Pro Ala Ala Lys Glu Glu Val195 200 205Lys Pro Thr Gln Thr Ser Val Ser Gln Ser Thr Thr Val Ser Pro Ala210 215 220Ser Val Ala Ala Glu Thr Pro Ala Pro Val Ala Lys Val Ala Pro Val225 230 235 240Arg Thr Val Ala Ala Pro Arg Val Ala Ser Val Lys Val Val Thr Pro245 250 255Lys Val Glu Thr Gly Ala Ser Pro Glu His Val Ser Ala Pro Ala Val260 265 270Pro Val Thr Thr Thr Ser Thr Ala Thr Asp Ser Lys Leu Gln Ala Thr275 280 285Glu Val Lys Ser Val Pro Val Ala Gln Lys Ala Pro Thr Ala Thr Pro290 295 300Val Ala Gln Pro Ala Ser Thr Thr Asn Ala Val Ala Ala His Pro Glu305 310 315 320Asn Ala Gly Leu Gln Pro His Val Ala Ala Tyr Lys Glu Lys Val Ala325 330 335Ser Thr Tyr Gly Val Asn Glu Phe Ser Thr Tyr Arg Ala Gly Asp Pro340 345 350Gly Asp His Gly Lys Gly Leu Ala Val Asp Phe Ile Val Gly Lys Asn355 360 365Gln Ala Leu Gly Asn Glu Val Ala Gln Tyr Ser Thr Gln Asn Met Ala370 375 380Ala Asn Asn Ile Ser Tyr Val Ile Trp Gln Gln Lys Phe Tyr Ser Asn385 390 395 400Thr Asn Ser Ile Tyr Gly Pro Ala Asn Thr Trp Asn Ala Met Pro Asp405 410 415Arg Gly Gly Val Thr Ala Asn His Tyr Asp His Val His Val Ser Phe420 425 430Asn Lys<210>40<211>232<212>蛋白質(zhì)<213>鏈球菌<400>40Met Pro His Leu Ser Lys Glu Ala Phe Lys Lys Gln Ile Lys Asn Gly1 5 10 15Ile Ile Val Ser Cys Gln Ala Leu Pro Gly Glu Pro Leu Tyr Thr Glu20 25 30Ser Gly Gly Val Met Pro Leu Leu Ala Leu Ala Ala Gln Glu Ala Gly35 40 45Ala Val Gly Ile Arg Ala Asn Ser Val Arg Asp Ile Lys Glu Ile Gln50 55 60Glu Val Thr Asn Leu Pro Ile Ile Gly Ile Ile Lys Arg Glu Tyr Pro65 70 75 80Pro Gln Glu Pro Phe Ile Thr Ala Thr Met Thr Glu Val Asp Gln Leu85 90 95Ala Ser Leu Asp Ile Ala Val Ile Ala Leu Asp Cys Thr Leu Arg Glu100 105 110Arg His Asp Gly Leu Ser Val Ala Glu Phe Ile Gln Lys Ile Lys Gly115 120 125Lys Tyr Pro Glu Gln Leu Leu Met Ala Asp Ile Ser Thr Phe Glu Glu130 135 140Gly Lys Asn Ala Phe Glu Ala Gly Val Asp Phe Val Gly Thr Thr Leu145 150 155 160Ser Gly Tyr Thr Asp Tyr Xaa Arg Gln Glu Glu Gly Pro Asp Ile Glu165 170 175Leu Leu Asn Lys Leu Cys Gln Ala Gly Ile Asp Val Ile Ala Glu Gly180 185 190Lys Ile His Thr Pro Lys Gln Ala Asn Glu Ile Asn His Ile Gly Val195 200 205Ala Gly Ile Val Val Gly Gly Ala Ile Thr Arg Pro Lys Glu Ile Ala210 215 220Glu Arg Phe Ile Ser Gly Leu Ser225 230<210>41<211>39<212>蛋白質(zhì)<213>鏈球菌<400>41Met Ser Ile Lys Lys Ser Val Ile Gly Phe Cys Leu Gly Ala Ala Ala1 5 10 15Leu Ser Met Phe Ala Cys Val Asp Ser Ser Gln Ser Val Met Ala Ala20 25 30Glu Lys Asp Lys Val Glu Ile35<210>42<211>1305<212>DNA<213>鏈球菌<400>42atgaaaatga ataaaaaggt actattgaca tcgacaatgg cagcttcgct attatcagtc60gcaagtgttc aagcacaaga aacagatacg acgtggacag cacgtactgt ttcagaggta 120aaggctgatt tggtaaagca agacaataaa tcatcatata ctgtgaaata tggtgataca 180ctaagcgtta tttcagaagc aatgtcaatt gatatgaatg tcttagcaaa aattaataac 240attgcagata tcaatcttat ttatcctgag acaacactga cagtaactta cgatcagaag 300agtcatactg ccacttcaat gaaaatagaa acaccagcaa caaatgctgc tggtcaaaca 360acagctactg tggatttgaa aaccaatcaa gtttctgttg cagaccaaaa agtttctctc 420aatacaattt cggaaggtat gacaccagaa gcagcaacaa cgattgtttc gccaatgaag 480acatattctt ctgcgccagc tttgaaatca aaagaagtat tagcacaaga gcaagctgtt 540agtcaagcag cagctaatga acaggtatca acagctcctg tgaagtcgat tacttcagaa 600gttccagcag ctaaagagga agttaaacca actcagacgt cagtcagtca gtcaacaaca 660gtatcaccag cttctgttgc cgctgaaaca ccagctccag tagctaaagt agcaccggta 720agaactgtag cagcccctag agtggcaagt gttaaagtag tcactcctaa agtagaaact 780ggtgcatcac cagagcatgt atcagctcca gcagttcctg tgactacgac ttcaacagct 840acagacagta agttacaagc gactgaagtt aagagcgttc cggtagcaca aaaagctcca 900acagcaacac cggtagcaca accagcttca acaacaaatg cagtagctgc acatcctgaa 960aatgcagggc tccaacctca tgttgcagct tataaagaaa aagtagcgtc aacttatgga 1020gttaatgaat tcagtacata ccgtgcaggt gatccaggtg atcatggtaa aggtttagca 1080gtcgacttta ttgtaggtaa aaaccaagca cttggtaatg aagttgcaca gtactctaca 1140caaaatatgg cagcaaataa catttcatat gttatctggc aacaaaagtt ttactcaaat 1200acaaatagta tttatggacc tgctaatact tggaatgcaa tgccagatcg tggtggcgtt 1260actgccaacc attatgacca tgttcacgta tcatttaaca aataa 1305<2l0>43<211>1230<212>DNA<213>鏈球菌<400>43caagaaacag atacgacgtg gacagcacgt actgtttcag aggtaaaggc tgatttggta60aagcaagaca ataaatcatc atatactgtg aaatatggtg atacactaag cgttatttca 120gaagcaatgt caattgatat gaatgtctta gcaaaaatta ataacattgc agatatcaat 180cttatttatc ctgagacaac actgacagta acttacgatc agaagagtca tactgccact 240tcaatgaaaa tagaaacacc agcaacaaat gctgctggtc aaacaacagc tactgtggat 300ttgaaaacca atcaagtttc tgttgcagac caaaaagttt ctctcaatac aatttcggaa 360ggtatgacac cagaagcagc aacaacgatt gtttcgccaa tgaagacata ttcttctgcg 420ccagctttga aatcaaaaga agtattagca caagagcaag ctgttagtca agcagcagct 480aatgaacagg tatcaacagc tcctgtgaag tcgattactt cagaagttcc agcagctaaa 540gaggaagtta aaccaactca gacgtcagtc agtcagtcaa caacagtatc accagcttct 600gttgccgctg aaacaccagc tccagtagct aaagtagcac cggtaagaac tgtagcagcc 660cctagagtgg caagtgttaa agtagtcact cctaaagtag aaactggtgc atcaccagag 720catgtatcag ctccagcagt tcctgtgact acgacttcaa cagctacaga cagtaagtta 780caagcgactg aagttaagag cgttccggta gcacaaaaag ctccaacagc aacaccggta 840gcacaaccag cttcaacaac aaatgcagta gctgcacatc ctgaaaatgc agggctccaa 900cctcatgttg cagcttataa agaaaaagta gcgtcaactt atggagttaa tgaattcagt 960acataccgtg caggtgatcc aggtgatcat ggtaaaggtt tagcagtcga ctttattgta 1020ggtaaaaacc aagcacttgg taatgaagtt gcacagtact ctacacaaaa tatggcagca 1080aataacattt catatgttat ctggcaacaa aagttttact caaatacaaa tagtatttat 1140ggacctgcta atacttggaa tgcaatgcca gatcgtggtg gcgttactgc caaccattat 1200gaccatgttc acgtatcatt taacaaataa 1230<210>44<211>409<212>蛋白質(zhì)<213>鏈球菌<400>44Gln Glu Thr Asp Thr Thr Trp Thr Ala Arg Thr Val Ser Glu Val Lys1 5 10 15Ala Asp Leu Val Lys Gln Asp Asn Lys Ser Ser Tyr Thr Val Lys Tyr20 25 30Gly Asp Thr Leu Ser Val Ile Ser Glu Ala Met Ser Ile Asp Met Asn
35 40 45Val Leu Ala Lys Ile Asn Asn Ile Ala Asp Ile Asn Leu Ile Tyr Pro50 55 60Glu Thr Thr Leu Thr Val Thr Tyr Asp Gln Lys Ser His Thr Ala Thr65 70 75 80Ser Met Lys Ile Glu Thr Pro Ala Thr Asn Ala Ala Gly Gln Thr Thr85 90 95Ala Thr Val Asp Leu Lys Thr Asn Gln Val Ser Val Ala Asp Gln Lys100 105 110Val Ser Leu Asn Thr Ile Ser Glu Gly Met Thr Pro Glu Ala Ala Thr115 120 125Thr Ile Val Ser Pro Met Lys Thr Tyr Ser Ser Ala Pro Ala Leu Lys130 135 140Ser Lys Glu Val Leu Ala Gln Glu Gln Ala Val Ser Gln Ala Ala Ala145 150 155 160Asn Glu Gln Val Ser Thr Ala Pro Val Lys Ser Ile Thr Ser Glu Val165 170 175Pro Ala Ala Lys Glu Glu Val Lys Pro Thr Gln Thr Ser Val Ser Gln180 185 190Ser Thr Thr Val Ser Pro Ala Ser Val Ala Ala Glu Thr Pro Ala Pro195 200 205Val Ala Lys Val Ala Pro Val Arg Thr Val Ala Ala Pro Arg Val Ala210 215 220Ser Val Lys Val Val Thr Pro Lys Val Glu Thr Gly Ala Ser Pro Glu225 230 235 240His Val Ser Ala Pro Ala Val Pro Val Thr Thr Thr Ser Thr Ala Thr245 250 255Asp Ser Lys Leu Gln Ala Thr Glu Val Lys Ser Val Pro Val Ala Gln260 265 270Lys Ala Pro Thr Ala Thr Pro Val Ala Gln Pro Ala Ser Thr Thr Asn275 280 285Ala Val Ala Ala His Pro Glu Asn Ala Gly Leu Gln Pro His Val Ala290 295 300Ala Tyr Lys Glu Lys Val Ala Ser Thr Tyr Gly Val Asn Glu Phe Ser305 310 315 320Thr Tyr Arg Ala Gly Asp Pro Gly Asp His Gly Lys Gly Leu Ala Val325 330 335Asp Phe Ile Val Gly Lys Asn Gln Ala Leu Gly Asn Glu Val Ala Gln340 345 350Tyr Ser Thr Gln Asn Met Ala Ala Asn Asn Ile Ser Tyr Val Ile Trp355 360 365Gln Gln Lys Phe Tyr Ser Asn Thr Asn Ser Ile Tyr Gly Pro Ala Asn370 375 380Thr Trp Asn Ala Met Pro Asp Arg Gly Gly Val Thr Ala Asn His Tyr385 390 395 400Asp His Val His Val Ser Phe Asn Lys40權(quán)利要求
1．分離的多核苷酸，其編碼的多肽與選自含有如下之序列的第二多肽有至少70％一致性SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5,SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10,SEQ ID NO:11,SEQ ID NO:12 SEQ ID NO:14 SEQ ID NO:1S,SEQ ID NO:16,SEQ ID NO:17,SEQ ID NO:18,SEQ ID NO:19,SEQ ID NO:20,SEQ ID NO:21,SEQ ID NO:23,SEQ ID NO:24,SEQ ID NO:25,SEQ ID NO:26,SEQ ID NO:28,SEQ ID NO:29,SEQ ID NO:30,SEQ ID NO:31,SEQ ID NO:33,SEQ ID NO:34,SEQ ID NO:35,SEQ ID NO:36,SEQ ID NO:38,SEQ ID NO:39,SEQ ID NO:40,SEQ ID NO:41和SEQ ID NO:44或者其片段，類(lèi)似物或衍生物。
2．根據(jù)權(quán)利要求1的多核苷酸，其中所述多核苷酸編碼的多肽與第二多肽有至少95％的一致性。
3．分離的多核苷酸，其所編碼的多肽能產(chǎn)生對(duì)含有選自如下的序列的多肽有特異性結(jié)合性的抗體SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5,SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10,SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:14 SEQ ID NO:15,SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19,SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:24,SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:29,SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:35, SEQ ID NO:34,SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:38, SEQ ID NO:39,SEQ ID NO:40, SEQ ID NO:41和 SEQ ID NO:44或者其片段，類(lèi)似物或衍生物。
4．與權(quán)利要求1的多核苷酸互補(bǔ)的分離的多核苷酸。
5．與權(quán)利要求3的多核苷酸互補(bǔ)的分離的多核苷酸。
6．權(quán)利要求1的多核苷酸，其中所述多核苷酸是DNA。
7．權(quán)利要求3的多核苷酸，其中所述多核苷酸是DNA。
8．權(quán)利要求1的多核苷酸，其中所述多核苷酸是RNA。
9．權(quán)利要求3的多核苷酸，其中所述多核苷酸是RNA。
10．在嚴(yán)格條件下與含有選自下組之序列的第二多核苷酸雜交的多核苷酸SEQ ID NO:1,SEQ ID NO:7,SEQ ID NO:13,SEQ IDNO:22,SEQ ID NO:27,SEQ ID NO:32,SEQ ID NO:37,SEQ ID NO:42和SEQ ID NO:43或者其片段，類(lèi)似物或衍生物。
11．在嚴(yán)格條件下與具有選自下面的一個(gè)序列的第二多核苷酸雜交的多核苷酸SEQ ID NO:37,SEQ ID NO:42和SEQ ID NO:43。
12．根據(jù)權(quán)利要求11的多核苷酸，其在嚴(yán)格條件下與具有SEQ IDNO:37序列的第二多核苷酸雜交。
13．根據(jù)權(quán)利要求11的多核苷酸，其在嚴(yán)格條件下與具有SEQ IDNO:42序列的第二多核苷酸雜交。
14．根據(jù)權(quán)利要求11的多核苷酸，其在嚴(yán)格條件下與具有SEQ IDNO:43序列的第二多核苷酸雜交。
15．根據(jù)權(quán)利要求10的多核苷酸，其中所述多核苷酸具有與所述第二多核苷酸至少95％的互補(bǔ)性。
16．根據(jù)權(quán)利要求11的多核苷酸，其中所述多核苷酸具有與所述第二多核苷酸至少95％的互補(bǔ)性。
17．包含權(quán)利要求1的多核苷酸的載體，其中所述多核苷酸與表達(dá)調(diào)控區(qū)操作連接。
18．包含權(quán)利要求3的多核苷酸的載體，其中所述多核苷酸與表達(dá)調(diào)控區(qū)操作連接。
19．用權(quán)利要求17的載體轉(zhuǎn)染的宿主細(xì)胞。
20．用權(quán)利要求18的載體轉(zhuǎn)染的宿主細(xì)胞。
21．制備多肽的方法，包括在適合表達(dá)所述多肽的條件下培養(yǎng)權(quán)利要求19的宿主細(xì)胞。
22．制備多肽的方法，包括在適合表達(dá)所述多肽的條件下培養(yǎng)權(quán)利要求20的宿主細(xì)胞。
23．與含有選自下組之序列的第二多肽有至少70％一致性的分離的多肽SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5,SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10,SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:15,SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19,SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:24,SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:29,SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:33, SEQ ID NO:34,SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:38, SEQ ID NO:39,SEQ ID NO:40, SEQ ID NO:41 和SEQ ID NO:44或者其片段，類(lèi)似物或衍生物。
24．具有SEQ ID NO39序列的權(quán)利要求23的分離的多肽。
25．具有SEQ ID NO44序列的權(quán)利要求23的分離的多肽。
26．能產(chǎn)生對(duì)于含有選自下組之序列的第二多肽有結(jié)合特異性的抗體的分離的多肽SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5,SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10,SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:15,SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19,SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:24,SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:29,SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:33, SEQ ID NO:34,SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:38, SEQ ID NO:39,SEQ ID NO:40, SEQ ID NO:41和 SEQ ID NO:44或者其片段，類(lèi)似物或衍生物。
27．具有SEQ ID NO:39序列的權(quán)利要求26的分離的多肽。
28．具有SEQ ID NO:44序列的權(quán)利要求26的分離的多肽。
29．具有選自下組之氨基酸序列的分離的多肽SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5,SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10,SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:15,SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19,SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:24,SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:29,SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:33, SEQ ID NO:34,SEQ ID NO:35, SEQ ID NO:36, SEQ ID NO:38, SEQ ID NO:39,SEQ ID NO:40 和 SEQ ID NO:41或者其片段，類(lèi)似物或衍生物。
30．具有SEQ ID NO:39的氨基酸序列的權(quán)利要求29的分離的多肽。
31．具有SEQ ID NO:44的氨基酸序列的分離的多肽。
32．權(quán)利要求29-31任一項(xiàng)的分離的多肽，其中N-末端Met殘基缺失。
33．權(quán)利要求29-30任一項(xiàng)的分離的多肽，其中分泌氨基酸序列缺失。
34．疫苗組合物，包括根據(jù)權(quán)利要求23-31任一項(xiàng)的一種多肽和藥學(xué)可接受載體，稀釋劑或佐劑。
35．疫苗組合物，包括根據(jù)權(quán)利要求32的一種多肽和藥學(xué)可接受載體，稀釋劑或佐劑。
36．疫苗組合物，包括根據(jù)權(quán)利要求33的一種多肽和藥學(xué)可接受載體，稀釋劑或佐劑。
37．一種治療或預(yù)防懷疑感染鏈球菌的動(dòng)物鏈球菌感染的方法，包括對(duì)所述動(dòng)物施用治療或預(yù)防量的權(quán)利要求34的組合物。
38．一種治療或預(yù)防懷疑感染鏈球菌的動(dòng)物鏈球菌感染的方法，包括對(duì)所述動(dòng)物施用治療或預(yù)防量的權(quán)利要求35的組合物。
39．一種治療或預(yù)防懷疑感染鏈球菌的動(dòng)物鏈球菌感染的方法，包括對(duì)所述動(dòng)物施用治療或預(yù)防量的權(quán)利要求36的組合物。
40．根據(jù)權(quán)利要求37-39任一項(xiàng)的方法，其中所述動(dòng)物是牛。
41．根據(jù)權(quán)利要求37-39任一項(xiàng)的方法，其中所述動(dòng)物是人。
42．根據(jù)權(quán)利要求37-39任一項(xiàng)的方法，其中所述細(xì)菌感染選自A組鏈球菌和B組鏈球菌。
43．根據(jù)權(quán)利要求42的方法，其中所述細(xì)菌感染是B組鏈球菌。
44．根據(jù)權(quán)利要求34的一種疫苗組合物治療或預(yù)防懷疑或者感染了鏈球菌的動(dòng)物鏈球菌感染的用途，包括對(duì)所述動(dòng)物施用治療或預(yù)防量的該組合物。
45．根據(jù)權(quán)利要求35-36任一項(xiàng)的一種疫苗組合物治療或預(yù)防懷疑或者感染了鏈球菌的動(dòng)物鏈球菌感染的用途，包括對(duì)所述動(dòng)物施用治療或預(yù)防量的該組合物。
46．根據(jù)權(quán)利要求23-31任一項(xiàng)的一種疫苗組合物制備用于治療或預(yù)防懷疑感染了鏈球菌的動(dòng)物鏈球菌感染的疫苗的用途，所述治療或預(yù)防包括對(duì)所述動(dòng)物施用治療或預(yù)防量的該組合物。
47．根據(jù)權(quán)利要求32的一種疫苗組合物在制備用于治療或預(yù)防懷疑感染了鏈球菌的動(dòng)物鏈球菌感染的疫苗中的用途，所述治療或預(yù)防包括對(duì)所述動(dòng)物施用治療或預(yù)防量的該組合物。
48．根據(jù)權(quán)利要求33的一種疫苗組合物在制備用于治療或預(yù)防懷疑感染了鏈球菌的動(dòng)物鏈球菌感染的疫苗中的用途，所述治療或預(yù)防包括對(duì)所述動(dòng)物施用治療或預(yù)防量的該組合物。
全文摘要
本發(fā)明公開(kāi)了B組鏈球菌(GBS)蛋白質(zhì)和其編碼多核苷酸。所述蛋白質(zhì)是有抗原性的,所以是預(yù)防或治療動(dòng)物鏈球菌感染的有用疫苗成分。還公開(kāi)了制備所述蛋白抗原的重組方法以及用于檢測(cè)鏈球菌感染的診斷測(cè)試。
文檔編號(hào)C12N1/15GK1297482SQ99805248
公開(kāi)日2001年5月30日 申請(qǐng)日期1999年2月17日 優(yōu)先權(quán)日1998年2月20日
發(fā)明者B·R·布羅多伊爾, C·里奧克斯, M·波耶爾, I·查勒波伊斯, J·哈梅爾, D·馬丁 申請(qǐng)人:生化制藥有限公司