CN1297482A - B组链球菌抗原 - Google Patents
B组链球菌抗原 Download PDFInfo
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- CN1297482A CN1297482A CN99805248A CN99805248A CN1297482A CN 1297482 A CN1297482 A CN 1297482A CN 99805248 A CN99805248 A CN 99805248A CN 99805248 A CN99805248 A CN 99805248A CN 1297482 A CN1297482 A CN 1297482A
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Abstract
本发明公开了B组链球菌(GBS)蛋白质和其编码多核苷酸。所述蛋白质是有抗原性的,所以是预防或治疗动物链球菌感染的有用疫苗成分。还公开了制备所述蛋白抗原的重组方法以及用于检测链球菌感染的诊断测试。
Description
发明领域
本发明涉及抗原，更特别地涉及用作治疗和/或预防的疫苗成分的B组链球菌(GBS)细菌病原体的蛋白质抗原。
发明背景
链球菌是革兰氏阳性细菌，它们按它们细胞表面上发现的A组至O组具体碳水化合物抗原来划分。链球菌组进一步通过具体类型荚膜多糖抗原来区分。对于B组链球菌(GBS)鉴定了几种血清型：Ⅰa,Ⅰb,Ⅱ,Ⅲ,Ⅳ,Ⅴ,Ⅵ,Ⅶh Ⅷ。GBS还包括已知为“C-蛋白质”(α,β,γ和δ)，其中一些已经被克隆出来。
尽管GBS是正常人阴道和结肠菌丛的共同成分，但是该病原体长期以来一直被认为是新生期脓毒症，脑膜炎，婴儿脑膜炎后遗症，产后子宫内膜炎以及牛奶场放牧人乳腺炎的主要病因。暴露给GBS的待产妇有产后感染的危险并且可能在孩子通过产道时将感染转移给她们的孩子。尽管微生物对抗生素敏感，但是由于新生期脓毒症和婴儿脑膜炎高的发生比例和快的发生速度而导致高的发病率和死亡率。
为了发现保护人们不受GBS感染的疫苗，研究转向特异型抗原。令人遗憾的是，证明了这些多糖在人体内具有不好的免疫原性并且限制于这些多糖发源的特殊血清型。此外，荚膜多糖激发T细胞独立响应，即没有IgG产生。结果荚膜多糖抗原不适合作为保护抗GBS感染的疫苗成分。
其它人观注C-蛋白β抗原，其被证明在小鼠和兔模型中有免疫原性。发现该性质不适合作为人疫苗，因为其与该亲和性相互作用和与人ⅠgA的Fc区的非免疫原性方式的不期望的性质。C-蛋白α抗原在负责大多数GBS介导的疾病的血清型GBS的Ⅲ型血清型中很少，因此作为疫苗成分几乎没有用途。
因此，对于可以用作预防和/或治疗GBS感染的疫苗成分的GBS抗原还有没有满足的要求。
发明概述
根据一方面，本发明提供编码与包括选自下面的一个序列的第二多肽至少70％一致性的多肽的分离的多核苷酸：
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，类似物或衍生物。
在其它方面，提供包括操作性连接了表达调控区的本发明多核苷酸的载体，以及用所述载体转染的宿主细胞，和产生多肽的方法，包括在适合表达的条件下培养所述宿主细胞。
在另一方面，提供本发明多核苷酸编码的新的多肽。
附图的简要描述
图1a是克隆1的DNA序列(SEQ ID NO:1)与相应的可读框的氨基酸序列；
图1b是氨基酸序列SEQ ID NO:2；
图1c是氨基酸序列SEQ ID NO:3；
图1d是氨基酸序列SEQ ID NO:4；
图1e是氨基酸序列SEQ ID NO:5；
图1f是氨基酸序列SEQ ID NO:6；
图2a是克隆2的DNA序列(SEQ ID NO:7)与相应的可读框的氨基酸序列；
图2b是氨基酸序列SEQ ID NO:8；
图2c是氨基酸序列SEQ ID NO:9；
图2d是氨基酸序列SEQ ID NO:10；
图2e是氨基酸序列SEQ ID NO:11；
图2f是氨基酸序列SEQ ID NO:12；
图3a是克隆3的DNA序列(SEQ ID NO:13)与相应的可读框的氨基酸序列；
图3b是氨基酸序列SEQ ID NO:14；
图3c是氨基酸序列SEQ ID NO:15；
图3d是氨基酸序列SEQ ID NO:16；
图3e是氨基酸序列SEQ ID NO:17；
图3f是氨基酸序列SEQ ID NO:18；
图3g是氨基酸序列SEQ ID NO:19；
图3h是氨基酸序列SEQ ID NO:20；
图3i是氨基酸序列SEQ ID NO:21；
图4a是克隆4的DNA序列(SEQ ID NO:22)与相应的可读框的氨基酸序列；
图4b是氨基酸序列SEQ ID NO:23；
图4c是氨基酸序列SEQ ID NO:24；
图4d是氨基酸序列SEQ ID NO:25；
图4e是氨基酸序列SEQ ID NO:26；
图5a是克隆5的DNA序列(SEQ ID NO:27)与相应的可读框的氨基酸序列；
图5b是氨基酸序列SEQ ID NO:28；
图5c是氨基酸序列SEQ ID NO:29；
图5d是氨基酸序列SEQ ID NO:30；
图5e是氨基酸序列SEQ ID NO:31；
图6a是克隆6的DNA序列(SEQ ID NO:32)与相应的可读框的氨基酸序列；
图6b是氨基酸序列SEQ ID NO:33；
图6c是氨基酸序列SEQ ID NO:34；
图6d是氨基酸序列SEQ ID NO:35；
图6e是氨基酸序列SEQ ID NO:36；
图7a是克隆7的DNA序列(SEQ ID NO:37)；
图7b是氨基酸序列SEQ ID NO:38；
图7c是氨基酸序列SEQ ID NO:39；
图7d是氨基酸序列SEQ ID NO:40；
图7e是氨基酸序列SEQ ID NO:41；
图8是包括信号序列的克隆7的一部分的DNA序列(SEQ ID NO:42)；
图9是不包括信号序列的克隆7的一部分的DNA序列(SEQ ID NO:43)；
图9a是氨基酸序列SEQ ID NO:44；
图10代表来自用相应于SEQ ID NO:39的重组GBS蛋白质免疫的CD-1小鼠血清的抗-GBS ELISA效价的分布。
本发明的详细描述
本发明涉及B组链球菌(GBS)的新的抗原多肽，特征在于是选自下面的氨基酸序列：
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，类似物或衍生物。
本发明的优选实施方案包括SEQ ID NO:39和SEQ ID NO:44。
本发明的进一步优选实施方案包括SEQ ID NO：39。
本发明的进一步优选实施方案包括SEQ ID NO:44。
如这里所使用的，本发明多肽的“片段”，“衍生物”或“类似物”包括其中一个或多个氨基酸残基被保守的或者不保守的氨基酸残基(优选保守的)取代并且可以是天然的或非天然的那些多肽。
本发明多肽的术语“片段”，“衍生物”或“类似物”包括添加，缺失，取代氨基酸的修饰的多肽，前提是这些多肽保持诱导免疫应答的能力。
术语“保守的氨基酸”是指一个或多个氨基酸被其它氨基酸取代，其中即使发生取代，给定抗原的抗原决定簇(包括其二级结构和亲水性质)也是完全或部分保守的。
例如，所述序列中一个或多个氨基酸残基可以被另一个相似极性的作为功能等价物起作用的氨基酸取代，导致沉默改变。所述序列中氨基酸的取代物可以选自该氨基酸所属于的种类其它成员。例如，非极性(疏水性)氨基酸包括丙氨酸，亮氨酸，异亮氨酸，缬氨酸，脯氨酸，苯丙氨酸，色氨酸和蛋氨酸。极性中性氨基酸包括甘氨酸，丝氨酸，苏氨酸，半胱氨酸，酪氨酸，天冬酰胺和谷氨酰胺。带正电荷(碱性)氨基酸包括精氨酸，赖氨酸和组氨酸。带负电荷(酸性)氨基酸包括天冬氨酸和谷氨酸。
优选地，本发明多肽的衍生物和类似物与图中图示说明的那些序列或者其片段具有大约70％的一致性。也就是说70％的残基相同。更优选地，多肽具有大于95％的同源性。在另一个优选的实施方案中，本发明多肽的衍生物和类似物具有少于大约20个氨基酸残基的取代，修饰或缺失，更优选地少于10个。优选的取代物是本领域已知为保守的那些，即被取代的残基具有相同的物理或化学性质，例如疏水性，大小，电荷或者官能团。
此外，在氨基酸区被发现是多态的情况下，可以预期改变一个或多个特殊氨基酸以更有效地模拟不同GBS株的不同表位。
还包括与改变多肽生物或药学性质的其它化合物，即聚乙二醇(PEG)融合以提高半寿期的多肽，易于纯化的前导或分泌氨基酸序列；前序列原和前序列；和多糖。
此外，本发明的多肽可以通过末端-NH2酰化作用(例如通过乙酰化作用，或者硫代乙二醇酸酰胺化作用，末端羧酸(carbosy)酰胺化作用，例如与氨或者甲胺)而被修饰，以提供连接或键合载体或其它分子的稳定性，提高的疏水性。
还涉及所述多肽片段，类似物和衍生物的杂聚和同聚多肽多聚体。这些聚合物形式包括，例如，与交联剂例如亲和素/生物素，戊二醛或二甲基超亚胺酯(dimethyl-superimidate)交联的一种或多种多肽。这样的聚合物形式还包括包含通过DNA重组技术产生的多顺反子mRNAs制备的两个或多个串联的或者倒位的连续序列的多肽。优选地，本发明多肽的片段，类似物或衍生物包括至少一个抗原区，即至少一个表位。
为了实现抗原聚合物(即合成的多聚体)的形成，可以使用具有二卤代酰基，硝基芳基卤化物等等的多肽，其中试剂对于硫基是特异性的。因此，不同肽的两个巯基之间的连接可以是单键或者可以由至少两个，典型地至少4个，不多于16个，但是通常不多于14个碳原子的连接基团组成。
在具体的实施方案中，本发明的多肽片段，类似物和衍生物不含有蛋氨酸(Met)起始残基。优选地，多肽将不插入一个前导或分泌序列(信号序列)。根据已确定的分子生物学技术可以确定本发明多肽的信号部分。一般情况下，可以从GBS培养物分离所感兴趣的多肽并且接着测序确定成熟蛋白的初始残基，从而确定成熟多肽的序列。
根据另一方面，提供了包括与药学可接受载体稀释剂或佐剂混合的一种或多种本发明GBS多肽的疫苗组合物。
合适的佐剂包括油，即弗氏完全佐剂或不完全佐剂；盐，即AlK(SO4)2,AlNa(SO4)2,AlNH4(SO4)2,Al(OH)3,AlPO4，二氧化硅，高岭土；皂草甙衍生物；碳多核苷酸，即多IC和多AU，还有解毒的霍乱毒素(CTB)和诱导粘膜免疫的大肠杆菌热不稳定毒素。优选的佐剂包括QuilATM,AlhydrogelTM，和AdjuphosTM。本发明的疫苗可以通过注射，快速灌注，鼻咽吸收，皮肤吸收，非经肠给药，或者含化或者口服给药。
本发明的疫苗组合物用于治疗或预防链球菌感染和/或链球菌感染介导的疾病和症状，特别是A组链球菌(化脓链球菌)，B组链球菌(GBS或无乳链球菌)，停乳链球菌，乳房链球菌，诺氏链球菌以及金黄色葡萄球菌。关于链球菌的一般信息参见Manual of ClinicalMicrobiology,P.R.Murray等，(1995，第六版，ASM出版社，华盛顿)。更特别地是B组链球菌，无乳链球菌。在具体的实施方案中，对有GBS感染危险的那些个体施用疫苗，所述个体是例如孕妇和婴儿脓毒症，脑膜炎和肺炎以及无免疫应答的个体，例如糖尿病，肝病或癌症的那些。疫苗也具有兽药用途，例如用于治疗上述细菌以及大肠杆菌介导的牛乳腺炎。
本发明的疫苗还可以用来制备用于治疗或预防链球菌感染和/或链球菌感染介导的疾病和症状的药物，所述链球菌特别是A组链球菌(化脓链球菌)，B组链球菌(GBS或无乳链球菌)，停乳链球菌，乳房链球菌，诺氏链球菌以及金黄色葡萄球菌。更特别地是B组链球菌，无乳链球菌。
疫苗组合物优选以0.001至100μg/kg(抗原/体重)，更优选地0.01至10μg/kg和最优选地0.1至1μg/kg的单位剂量形式，免疫之间大约1至12周间隔免疫1至3次，更优选地间隔1至6周。
根据另一方面，提供了编码特征在于氨基酸序列选自下组的B组链球菌(GBS)多肽的多核苷酸：
SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO: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，类似物或衍生物。
优选的多核苷酸是在图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)中图示说明的那些，它们相应于编码本发明多肽的可读框。
优选的多核苷酸是在图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)中图示说明的那些，和它们的片段，类似物和衍生物。
更优选的本发明的多核苷酸是图7(SEQ ID NO:37),8(SEQ ID NO:42)和9(SEQ ID NO:43)中图示说明的那些。
最优选的本发明的多核苷酸是图8(SEQ ID NO:42)和9(SEQ ID NO:43)中图示说明的那些。
要理解图中说明的多核苷酸序列可以用也编码本发明多肽的简并密码子改变。
由于核苷酸编码序列的简并性，本发明实施中也可以使用编码本发明基本上相同的多肽的其它多核苷酸序列。这些包括但不限于编码序列内相同氨基酸残基的不同密码子的取代而改变从而产生沉默变化的核苷酸序列。
因此，本发明进一步提供与上述多核苷酸序列杂交的多核苷酸(或者其互补序列)，序列之间具有50％，优选地具有至少70％一致性。更优选地，多核苷酸在严格条件下是可杂交的，即具有至少95％一致性，最优选地大于97％一致性。
能在严格条件下杂交是指核苷酸分子在标准条件下，例如高温和/或低盐含量下，退火生成第二核酸序列的至少一个区(或者作为cDNA，或者作为基因组DNA)或者生成其互补链，其趋向于不利于非互补核苷酸序列的杂交。合适的方法描述于Maniatis T.等，分子克隆：实验室手册，冷泉港实验室，1982，其在这里引作参考。
再一方面，可以在DNA免疫方法中使用编码本发明多肽的多核苷酸，或者其片段，类似物或衍生物。即，它们可以被插入到载体中，该载体在注射后是可复制和可表达的，从而体内产生抗原性多肽。例如于在真核细胞中起作用的CMV启动子控制下，多核苷酸可以插入到质粒载体中。优选地肌内注射载体。
根据另一方面，提供了通过重组技术通过在宿主细胞中表达编码所述多肽的多核苷酸并且回收表达的多肽产物而制备本发明多肽的方法。或者，可以根据已确定的化学合成技术合成本发明多肽，即寡肽的液相或固相合成，连接寡肽产生全长多肽(封闭连接)。
对于重组体生产，用编码多肽的载体转染宿主细胞，然后在经改变适合激活启动子，选择转化物或者扩增基因的营养培养基中培养。合适的载体是在选择的宿主内可存活且可复制的那些载体，包括染色体的，非染色体的和合成的DNA序列，例如细菌质粒，噬菌体DNA，杆状病毒，酵母质粒，质粒和噬菌体DNA组合产生的载体。所述多肽序列可以使用限制酶在合适的位点插入载体中，这样其操作连接包括启动子，核糖体结合位点(共有区或核糖体结合序列)，和任选地操纵基因(调控元件)的表达调控区。人们可以根据已确定的分子生物学原理(Sambrook等，分子克隆：实验室手册，第二版，冷泉港实验室，纽约，1989，其在这里引作参考)选择适合给定宿主和载体的表达调控区的各成分。合适的启动子包括但不限于LTR或SV40启动子，大肠杆菌lac,tac或启动子和噬菌体λPL启动子。载体优选插入复制起点以及选择标记，即氨苄青霉素抗性基因。合适的细菌载体包括pET,pQE70,pQE60,pQE-9,pbs,pD10phagescript,psiX174,pbluescriptSK,pbsks,pNH8A,pNH16a,pNH18A,pNH46A,ptrc99a,pKK223-3,pKK233-3,pDR540,pRIT5和真核载体pBlueBacⅢ,pWLNEO,pSV2CAT,pOG44,pXT1,pSG,pSVK3,pBPV,pMSG和pSVL。宿主细胞可以是细菌，即大肠杆菌，枯草芽孢杆菌，链霉菌属；真菌，即黑曲霉，构巢曲霉；酵母，即糖酵母属或者真核的，即CHO,COS。
在培养基中表达多肽时，一般通过离心收集细胞，然后通过物理或化学方法破碎(如果表达的多肽没有分泌到培养基中)，留下得到的粗提取物以分离感兴趣的多肽。从培养基或溶解物中纯化多肽可以根据多肽的性质通过已确定的技术进行，即利用硫酸铵或乙醇沉淀，酸提取，阴离子或阳离子交换层析，磷酸纤维素层析，疏水作用层析，羟基磷灰石层析和凝集素层析。可以应用HPLC实现最后的纯化。
多肽可以在有或没有前导或分泌序列下被表达。在前一种情况下，可以使用翻译后处理去除前导序列(参见US4431739；4425437；和4338397，这里引作参考)或者可以化学去除前导序列接着纯化表达的多肽。
根据又一方面，本发明的GBS多肽可以在诊断链球菌感染特别是GBS感染的诊断测定中使用。几种诊断方法是可能的，例如检测生物样品中的链球菌生物，可以进行下面的方法：
a)从患者获得生物样品；
b)培养与本发明GBS多肽反应的抗体或者其片段和生物样品，形成混合物；和
c)检测混合物中特异性结合的抗体或者结合的片段，这表明链球菌的存在。
或者，可以如下进行检测含有或者怀疑含有所述抗体的生物样品中对链球菌抗原特异性的抗体的方法：
a)从患者分离生物样品；
b)培养一种或多种本发明GBS多肽或者其片段和生物样品，形成混合物；和
c)检测混合物中特异性结合的抗原或者结合的片段，这表明链球菌特异性抗体的存在。
本领域技术人员会理解可以以几种形式进行诊断测定，包括免疫测定，例如酶联免疫吸附测定(ELISA)，放射免疫测定或者乳胶凝聚试验，特别是测定生物体内是否存在对所述蛋白质有特异性的抗体。
编码本发明多肽的DNA序列也可以用来设计用于检测怀疑含有链球菌的生物样品中链球菌的存在。本发明的检测方法包括：
a)从患者分离生物样品；
b)培养一种或多种具有编码本发明多肽的DNA序列或者其片段的DNA探针和生物样品，形成混合物；和
c)检测混合物中特异性结合的DNA探针，这表明链球菌的存在。
本发明的DNA探针也可以用来检测样品中的循环链球菌，即GBS核酸，例如使用聚合酶链反应，作为诊断链球菌感染的方法。所述探针可以应用常规技术合成和可以固定在固相上，或者可以用可检测的标记物标记。用于本申请的优选的DNA探针是具有与本发明GBS多肽的至少6个连续的核苷酸互补的序列的寡聚物。
检测患者体内链球菌的另一种诊断方法包括：
a)用可检测的标记物标记与本发明多肽或者其片段反应的抗体；
b)对患者施用标记的抗体或者标记的片段；和
c)检测患者体内特异性结合的标记的抗体或者标记的片段，这表明链球菌的存在。
本发明的再一方面是本发明的多肽作为用于产生用于诊断和特别是用于治疗链球菌感染的特异性抗体的免疫原的用途。可以使用适当的筛选方法确定合适的抗体，例如通过在试验模型中测定特定抗体对于抗链球菌感染的被动免疫的能力。动物模型的一个实施例是本中请实施例中描述的小鼠模型。所述抗体可以是整个抗体或者是其抗原结合片段，并且可以一般属于任何免疫球蛋白种类。所述抗体或片段可以是动物来源的，特别是哺乳动物源，更特别地是小鼠，大鼠或者人源。其可以是天然抗体或者其片段，或者如果期望，可以是重组体抗体或抗体片段。术语重组体抗体或抗体片段指应用分子生物学技术产生的抗体或抗体片段。所述抗体或抗体片段可以是多克隆抗体，或者优选地，单克隆抗体。其可以对多个与GBS多肽相关的表位是特异性的，但是优选对于一个表位是特异性的。
实施例1致死量B组链球菌(GBS)感染的小鼠模型
GBS感染的小鼠模型详细描述于Lancefield等(J.Exp.Med.142:165-179,1975)。对GBS株C388/90(1990年从加拿大喔太华的Children’sHospital of Eastern Ontario脑膜炎患者的脑脊液获得的临床分离物)和NCS246(National Center for Streptococcus,ProvincialLaboratory of Public Health for Northern Alberta,Edmonton,加拿大)分别血清型测定为Ⅰa/c型和Ⅱ/R型。
为了提高它们的致病力，如前所述(Lancefield等J.Exp.Med.142:165-179,1975)连续在小鼠中传代GBS株C388/90(血清型Ⅰa/c)和NCS246(血清型Ⅱ/R)。简要地说，使用从感染小鼠的血液或脾获得的在Todd-Hewitt肉汤中传代培养物的系列稀释物腹膜内接种来监测致病力的提高。最后传代后，使用感染的血样接种Todd-Hewitt肉汤.在37℃下7％CO2下培养2小时后，向培养物加入终浓度10％(v/v)的甘油。然后将培养物分成等份并且在-80℃下保存，用于GBS攻击实验。测定这些冷冻的样品中存在的GBS的cfu数。对于GBS株C388/90和NCS246分别测定100％杀死(LD100)18周龄小鼠所必须的细菌浓度是3.5×105和1.1×105，这相应于肉汤菌株的致病力的显著增加。事实上，这两种菌株传代之前记录的LD100大于109cfu。
在细菌攻击中，使用Todd-Hewitt肉汤将新解冻的致病GBS株等份调节到合适的细菌浓度并且对每一只雌性CD-1小鼠腹膜内注射1毫升。用于被动保护实验的小鼠是6-8周龄，而用于主动保护实验的小鼠在受攻击时大约18周龄。对所有的接种体证实菌落数。在攻击后头48小时每天4次观察动物的任何感染症状，接着下面的12天每天观察。在该时期最后，从生存者获得血样并且在-20℃下冷冻。培养从受攻击存活的每一只小鼠获得脾以鉴定所有存留的GBS。
实施例2用甲醛杀死的GBS全细胞免疫和保护小鼠
根据Lancefield等(J.Exp.Med.142:165-179,1975)描述的方法制备甲醛杀死的GBS全细胞。简要地说，将GBS株在绵羊血琼脂平板(Quelab Laboratories,Montreal，加拿大)上过夜培养物在PBS缓冲液(磷酸盐缓冲盐水，pH7.2)中清洗两次，调节至大约3×109cfu/ml，并且在含有0.3％(v/v)甲醛的PBS中培养过夜。用PBS清洗杀死的GBS缓冲液并且在-80℃下冷冻保存。
以两周间隔，使用0.1毫升甲醛杀死的GBS株C388/900细胞(-6×107GBS)或者用于对照组的0.1毫升PBS，对6-8周龄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，向这些制剂中加入0.14毫克或0.21毫克Al终浓度的AlhydrogelTM(Superfos Biosetor,Frederikssund,Denmark)，并且在4℃搅拌下培养过夜。在免疫程序开始前和在最后注射后两周从每一只小鼠获得血清样品。血清在-20℃下冷冻。
第三次注射后1星期用1.5×104cfu的GBS株C388/90(Ⅰa/c)攻击注射PBS的每一个对照组中的8只小鼠和用甲醛杀死的全细胞GBS株C388/90(Ⅰa/c)免疫的组。用甲醛杀死的GBS全细胞免疫的所有小鼠在同种攻击中存活，而在攻击后5天内，注射PBS的8只小鼠只有4只存活。为了提高对照组中的死亡率，在细菌攻击时必须根据小鼠年龄调节细菌悬浮液。在下面的攻击实验中，当小鼠大于15周龄时，细菌接种物的浓度提高到3.0×105和2.5×106cfu之间。
表1用甲醛杀死的GBS全细胞免疫CD1小鼠并且接着同种攻击(菌株C388/90(Ⅰa/c))和异种攻击(菌株NCS246(Ⅱ/R))
| 用于免疫的抗原制剂 | 细菌攻击后14天存活小鼠的数目(存活百分率％) | |
| 同种攻击：菌株C388/90(Ia/c) | 异种攻击：菌株NCS246(Ⅱ/R) | |
| 第一次感染 | ||
| 甲醛杀死的GBSC388/90(Ia/c)2细胞株 | 8/8(100)3 | n.d.5 |
| 对照PBS | 4/8(50) | n.d. |
| 第二次感染 | ||
| 甲醛杀死的GBSC388/90(Ia/c)细胞株 | 6/6(100)4 | 0/6(0)6 |
| 对照PBS | 2/6(33) | 0/6(0) |
1使用终浓度是0.14毫克或0.21毫克Al的alhydrogelTM；
2大约6×107cfu；
3用调节到1.5×104cfu的含有GBS C388/90(Ⅰa/c)悬浮液的1毫升Todd-Hewitt培养基腹膜内攻击；
4用调节到2.1×106cfu的含有GBS C388/90(Ⅰa/c)悬浮液的1毫升Todd-Hewitt培养基腹膜内攻击；
5没有做；
6用调节到1.2×105cfu的含有GBS NCS246(Ⅱ/R)悬浮液的1毫升Todd-Hewitt培养基腹膜内攻击。
在另一项实验中，用PBS注射一组12只相应于对照组的小鼠，而用甲醛杀死的GBSC388/90(Ⅰa/c)全细胞株免疫第二组12只小鼠。用2.1×106cfu的GBSC388/90(Ⅰa/c)株攻击这两组的每一个组的6只小鼠(表Ⅰ)。第一个攻击实验中，所有用GBSC388/90(Ⅰa/c)株免疫的小鼠在同种攻击中存活。注射PBS的6中小鼠中只有2只在感染中存活。
两个组中的剩下的6只然后在一周后证明该抗原制剂是否带来抗菌株NCS246(Ⅱ/R)的交叉保护性，其产生血清学特性被膜。用第二GBS菌株感染的小鼠没有一只在感染中存活。后一种结果表明甲醛杀死的菌株C388/90诱导的保护性免疫应答的大多数抗荚膜多糖，这只限于特殊血清型的菌株。这些结果清楚地表明该特定感染模型可以有效地用于研究接种带来的保护作用。
实施例3用甲醛杀死的GBS全细胞免疫兔和小鼠的被动免疫保护
用甲醛杀死的GBS菌株C388/90(Ⅰa/c)细胞免疫新西兰兔(2.5公斤，Charles River,St.Constant,Quebec，加拿大)，获得超免疫血清。以三星期间隔用大约1.5×109cfu甲醛杀死的GBS菌株C388/90(Ⅰa/c)全细胞对该兔皮下注射三次，弗氏完全佐剂(GibcoBRL LifeTechnologies,Grand Island，纽约)用作第一次免疫的佐剂，弗氏不完全佐剂(GibcoBRL)用于下面的两次注射。在免疫方案开始之前和在最后注射后2星期取血清样品。血清在-20℃下冷冻。
也评价了该特定兔超免疫血清对被动保护小鼠抗GBS致命感染的能力。攻击前18小时用15或25μL超免疫兔血清对小鼠腹膜内注射，被保护的5只小鼠中有4只抗感染(80％)。相比之下，用PBS或用脑膜炎外膜制剂免疫的兔获得的血清注射的对照组的小鼠存活率低于20％。该结果清楚地表明用杀死的GBS细胞免疫另一种动物物种可以诱导产生可以被动保护小鼠的抗体。该试剂也可以用来表征克隆。
表2用甲醛杀死的B组全链球菌(菌株C388/90(Ⅰa/c))抗原制剂免疫后获得的兔血清带来的对CD-1小鼠的被动保护
| 组 | GBS菌株C388/90(Ia/c)2细菌攻击后14天存活小鼠数 | 存活率(％) |
| 兔超免疫血清2-25μl | 4/5 | 80 |
| 兔超免疫血清2-15μl | 4/5 | 80 |
| 对照兔血清-25μl | 1/5 | 20 |
| 对照PBS | 1/10 | 10 |
1弗氏完全佐剂用于第一次免疫，弗氏不完全佐剂用于下面的两次注射；
2用含有调节到2×104cfu的GBS C388/90(Ⅰa/c)的1毫升Todd-Hewitt培养基腹膜内攻击。
实施例4 His.Tag-GBS融合蛋白的重组制备
使用包含分别加入限制酶切位点BglⅡ(AGATCT)和HindⅢ(AAGCTT)的碱基延伸的寡聚物通过PCR(DNA Thermal Cycler GeneAmpPCT system2400 Perkin Elmer,San Jose,CA)从GBS菌株C388/90(Ⅰa/c)的基因组DNA扩增GBS基因的编码区。使用购自Qiagen(Chatsworth,CA)的QiaexⅡ凝胶提取试剂盒用琼脂糖凝胶纯化PCR产物，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消化包含硫氧还蛋白-His.Tag序列的pET-32b(+)载体(Novagen,Madison,WI)，用苯酚：氯仿提取，然后乙醇沉淀之。BglⅡ-HindⅢ基因组DNA片段连接BglⅡ-HindⅢ pET-32b(+)载体，产生其基因在T7启动子控制下的硫氧还蛋白-His.Tag-GBS融合蛋白的编码序列。根据Simanis的方法(Hanahan,D.DNA克隆，1985，D.M.Glover(编著)，pp.109-135)，将连接产物转化到大肠杆菌菌株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试剂盒(Qiagen,Chatsworth,CA)纯化重组pET质粒，并且通过DNA测序(Taq Dye,DeoxyTerminator Cycle Sequencing kit,ABI,Foster City,CA)证实所述DNA插入片段的核苷酸序列。通过电穿孔(Gene PulserⅡ apparatus,BIO-RAD Labs,Mississauga，加拿大)将重组pET质粒转化到大肠杆菌菌株AD494(DE3)(Δara-leu7697ΔlacX74 AphoA pvuⅡ phoRΔmalF3F’(lacI+(lacIq)pro)trxB：：Kan(DE3))(Novagen,Madison,WI)k。在该大肠杆菌菌株中，其基因处于lac启动子控制下的T7 RNA聚合酶(存在于λDE3原噬菌体上)特异性识别控制所述融合蛋白表达的T7启动子，其中所述lac启动子由异丙基-β-D-硫代吡喃半乳糖(IPTG)诱导。
250rpm搅拌下在37℃下，在每毫升含有100μg氨苄青霉素(Sigma-Aldrich加拿大有限公司，Oakville，加拿大)的LB肉汤(蛋白胨10g/L，酵母提取物5g/L,NaCl 10g/L)中培养转化体AD494(DE3)/rpET，直到A600达到0.6的值。为了诱导硫氧还蛋白-His.Tag-GBS融合蛋白的产生，在IPTG存在下以1mM终浓度将细胞再培养2小时。离心收集细菌细胞。
IPTG诱导2小时下AD494(DE3)/rpET32产生的重组融合蛋白部分以不溶的包函体获得，通过分离不溶的团聚体从内源大肠杆菌蛋白质将其纯化(Gerlach,G.F.等1992,Infect.Immun.60:892)。从500毫升培养物诱导的细胞悬浮于20毫升25％蔗糖-50mM Tris-HCl缓冲液(pH8.0)并在-70℃下冷冻。通过加入5毫升250mM Tris-HCl缓冲液(pH8.0)中的溶菌酶溶液(10mg/ml)，接着在冰上温育10-15分钟，并且加入150毫升洗涤剂混合物(5份20mM Tris-HCl缓冲液(pH7.4)-300mM NaCl-2％脱氧胆酸-2％Nonidet P-40和4份100mM Tris-HCl缓冲液(pH8)-50mM EDTA-2％Triton X-100)，接着在冰上温育5分钟，实现解冻的悬浮液的溶解。超声处理后，通过以35000xg离心30分钟收集蛋白质团聚体并且保留溶解细胞级分的一个样品。团聚的蛋白质溶解于6M盐酸胍。使用在相应的GBS菌株细菌攻击中存活的注射了甲醛杀死的GBS菌株C388/90(Ⅰa/c)细胞的小鼠血清通过蛋白质印迹分析表明溶解的和不溶的级分中都存在融合蛋白。
根据固定在His.Bind金属螯合树脂(Novagen，Madison,WI)上的与二价阳离子(Ni2+)结合的His.Tag序列(6个连续的组氨酸残基)的性质通过亲和层析进行从IPTG-诱导的AD494(DE3)/rpET的溶解级分纯化融合蛋白。使用的纯化方法是pET system Manual，第六版(Novagen,Madison,WI)中描述的那些。简要地说，将自IPTG诱导的100毫升培养物获得的成粒的细胞悬浮于4毫升结合缓冲液(5mM咪唑-500mMNaCl-20mM Tris-HClpH7.9)，超声，以39000xg离心20分钟去除残渣。过滤上清液(0.45μm孔径的膜)并且沉积在结合缓冲液中平衡的His.Bind树脂柱上。然后用10柱体积的结合缓冲液接着用6柱体积的洗涤缓冲液(20mM咪唑-500mMNaCl-20mM Tris-HClpH7.9)冲洗柱子。用洗脱缓冲液(1M咪唑-500mMNaCl-20mM Tris-HClpH7.9)洗脱硫氧还蛋白-His.Tag-GBS融合蛋白。通过在4℃下用3×1升PBS透析，从样品中去除盐和咪唑。
通过十二烷基硫酸钠(SDS)-聚丙烯酰胺树脂的考马斯染色，用这些蛋白质的系列稀释物和牛血清白蛋白标准(Pierce ChemicalCo.Rockford,Ill)估计从大肠杆菌溶解或不溶细胞质级分获得的融合蛋白的量。
实施例5λPL启动子控制下GBS蛋白质的重组制备
GBS蛋白质的DNA编码区经启动子λPL下游插入到翻译载体pURV22中。该质粒自p629(George等，1987,Bio/Technology5：600)衍生，从p629去除I型单纯疱疹病毒(HSV-Ⅰ)糖蛋白(gD-1)的一部分的编码区，从质粒载体pUC4K(Pharmacia Biotech Canada Inc.,Baie D’Urfe，加拿大)获得的卡那霉素盒置换氨苄青霉素抗性基因。该载体包含λ噬菌体cI857温度敏感阻抑蛋白基因盒，其中缺失了功能PR启动子。温度从30-37℃提高至37-42℃范围cI857阻抑蛋白的灭活导致λPL控制下的基因诱导。下游是BglⅡ限制位点(AGATCT)和ATG：ACTAAGGAGGTTAGATCTATG的核糖体结合位点cro控制该基因的翻译。
根据供应商说明(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连接酶。根据Sambrook等所述(分子克隆：实验室手册，1989，冷泉港实验室出版社，N.Y.)进行DNA片段的琼脂糖凝胶电泳。根据Jayarao等(J.Clin.Microbiol.,1991,29：2774)中描述的方法制备GBS细菌的染色体DNA。使用DNA Thermal CyclerGeneAmp PCR系统2400(Perkin Elmer,San Jose,CA)进行通过聚合酶链反应(PCR)的DNA扩增反应。使用购自Qiagen(Chatsworth,CA)的质粒试剂盒纯化用于DNA测序的质粒。使用购自Qiagen(Chatsworth,CA)的QiaexⅡ凝胶提取试剂盒从琼脂糖凝胶纯化DNA片段。通过Hanahan(DNA Clone,Glover(编著)pp.109-135,1985)描述的方法进行质粒转化。使用寡核苷酸合成仪394型(Perkin-ElmerCorp.,Applied Biosystems Div,(ABI),Foster City,CA)合成的合成寡核苷酸进行基因组DNA插入片段的测序。使用Taq DyeDeoxy Terminator Cycle Sequencing试剂盒(ABI,Foster City,CA)通过PCR进行测序反应，并且在自动DNA测序仪373A(ABI,FosterCity,CA)上进行DNA电泳。使用Sequencer 3.0程序(Gene CodesCorporation,Ann Arbor,MI)进行DNA序列的组装。应用Gene Works2.45版程序(Intelligenetics,Inc.,Mountain ViewCA)进行DNA序列分析和它们的预测的多肽的分析。
使用包含分别加入限制酶切位点BglⅡ(AGATCT)和XbaⅠ(TCTAGA)的碱基延伸的寡聚物通过PCR从GBS菌株C388/90(Ⅰa/c)的基因组DNA扩增GBS基因的编码区。使用购自Qiagen(Chatsworth,CA)的QiaexⅡ凝胶提取试剂盒用琼脂糖凝胶纯化PCR产物，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载体，用苯酚：氯仿提取，然后乙醇沉淀之。BglⅡ-XbaI基因组DNA片段连接BglⅡ-XbaⅠ pURV22载体，其中GBS基因在λPL启动子控制下。根据上文Hanahan的方法，将连接产物转化到大肠杆菌菌株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。通过将溶解的细胞在琼脂糖凝胶上进行电泳(Sambrook等，上文)分析来鉴定携带插入片段的带有转化体的质粒。使用Qiagen试剂盒(Qiagen，Chatsworth,CA)纯化重组pURV22质粒，并且通过DNA测序证实所述DNA插入片段的核苷酸序列。
250rpm搅拌下在34℃下，在每毫升含有50μg卡那霉素的LB肉汤中培养转化体XLI Blue MRF’/rpURV22，直到A600达到0.6的值。为了诱导融合蛋白的产生，将细菌细胞在39℃再培养4小时，再悬浮于样品缓冲液中，沸腾10分钟并且在-20℃下保存。
实施例6 CMV质粒pCMV-GH中亚克隆GBS蛋白基因
GBS蛋白DNA编码区插入到人生长激素(hGH)基因下游，其处于质粒载体pCMV-GH中的巨细胞病毒(CMV)启动子的转录控制下(Tang等，Nature,1992,356:152)。CMV启动子在大肠杆菌细胞中是没有功能的，但是施用真核细胞中的质粒时是活性的。所述载体也插入氨苄青霉素抗性基因。
使用包含加入限制酶切位点BglⅡ(AGATCT)和HindⅢ(AAGCTT)的碱基延伸的寡聚物通过PCR从GBS菌株C388/90(Ⅰa/c)的基因组DNA扩增所述基因的编码区。使用购自Qiagen(Chatsworth,CA)的QiaexⅡ凝胶提取试剂盒用琼脂糖凝胶纯化PCR产物，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消化包含人生长激素的pCMV-GH载体(StephenA.Johnston博士的实验室，德克萨斯大学，生物化学系，Dallas，德克萨斯州)，用苯酚：氯仿提取，然后乙醇沉淀之。1.3-kb BglⅡ-HindⅢ基因组DNA片段连接BamHI-HindⅢ pCMV-GH载体，产生CMV启动子控制下的hGH-GBS融合蛋白。根据上文Hanahan的方法该连接产物转化到大肠杆菌菌株DH5α(φ80 lacZ AM15 endAl recA1 hsdR17(rK-mK+)supE44 thi-1λ-gyrA1Δ(lacZYA-argF)U169)(GibcoBRL,Gaithersbury,MD)中。通过将溶解的细胞在琼脂糖凝胶上进行电泳(Sambrook等，上文)分析来鉴定携带插入片段的带有转化体的质粒。使用Qiagen试剂盒(Qiagen，Chatsworth,CA)纯化重组pCMV质粒，并且通过DNA测序证实所述DNA插入片段的核苷酸序列。
实施例7 GBS蛋白质对GBS攻击的免疫活性
以三星期间隔用0.1毫升下面的抗原制剂对6-8周龄的四组12只雌性CD-1小鼠(Charles River,St-Constant,Quebec，加拿大)皮下注射三次：甲醛杀死的GBS菌株C388/90(6×107cfu),20μg来自不溶物(包函体)的硫氧还蛋白-His.Tag-GBS融合蛋白或者亲和纯化的(镍柱)来自大肠杆菌细胞质溶解级分的20μg融合蛋白，或者20μg亲和纯化的(镍柱)硫氧还蛋白-His.Tag对照多肽。向每一抗原制剂中加入20μgQuilATM(Cedarlane Laboratories Ltd.Hornby，加拿大)作为佐剂。在免疫之前(PB)和在免疫过程中第20天(TB1)，41天(TB2)和54天(TB3)从每一只小鼠获得血清样品。血清在-20℃下冷冻。
每一次注射融合蛋白之后记录ELISA效价的增加，表明好的初次应答和第二次和第三次施用后特异性体液免疫应答的加强。在免疫期最后，用从包函体获得的20μg融合蛋白免疫的组相应的ELISA效价的平均值是456145，相比之下，用得自大肠杆菌中可溶级分的蛋白质免疫的小鼠组是290133。后一结果提示从包函体获得的蛋白质比可溶蛋白质更具免疫原性。使用亲和纯化的硫氧还蛋白-His.Tag包被的平板在ELISA中的小鼠血清分析表明对于融合蛋白的硫氧还蛋白-His.Tag部分可忽略抗体效价。通过ELISA也试验了注射了重组融合蛋白的小鼠的血清对甲醛杀死的GBS菌株C388/90全细胞的反应性。重组融合蛋白免疫诱导的抗体也识别GBS细胞上它们的特异性表位，表明它们的构象足以接近天然的链球菌蛋白质来诱导交叉反应抗体。
为了证实免疫诱导的免疫应答能否保护抗GBS感染，用3.5×105cfuGBS菌株C388/90(Ⅰa/c)和1.2×105 cfu菌株NCS246(Ⅱ/R)攻击小鼠，其结果分别在表3和4中说明。用对照硫氧还蛋白-His.Tag肽免疫的小鼠都不能保护不受两种GBS菌株的攻击，而用甲醛杀死的GBS菌株C388/90全细胞免疫的那些小鼠只提供抗同种攻击的保护。本发明硫氧还蛋白-His.Tag-GBS融合蛋白保护小鼠不受两种GBS菌株的感染。这些小鼠的血和脾培养物表明不存在任何GBS。
表3在GBS菌株C388/90(Ⅰa/c)攻击下存活试验1
| 免疫试剂 | 攻击下存活的小鼠数目 | 存活百分率(％) |
| 硫氧还蛋白-His.Tag2 | 1/6 | 17 |
| 甲醛杀死的C388/90细胞3 | 6/6 | 100 |
| 硫氧还蛋白-His.Tag-GBS融合蛋白(包函体制剂)4 | 6/6 | 100 |
| 硫氧还蛋白-His.Tag-GBS融合蛋白(细胞质级分)4 | 6/6 | 100 |
1腹膜内施用调节到3.5×105cfu的1毫升Todd-Hewitt培养基；2施用20μg；存活小鼠后腿麻痹；对血液和脾检测GBS；3施用6×107cfu；4施用20μg。
表4在GBS菌株NCS246(Ⅱ/R)攻击下存活试验1
| 免疫试剂 | 攻击下存活的小鼠数目 | 存活百分率(％) |
| 硫氧还蛋白-His.Tag2 | 0/6 | 0 |
| 甲醛杀死的C388/90细胞3 | 2/6 | 34 |
| 硫氧还蛋白-His.Tag-GBS融合蛋白(包函体制剂)2 | 5/54 | 100 |
| 硫氧还蛋白-His.Tag-GBS融合蛋白(细胞质级分)2 | 6/6 | 100 |
1腹膜内施用调节到1.2×105cfu的1毫升含有GBSNC246(Ⅱ/R)悬浮液的Todd-Hewitt培养基；
2施用20μg；
3施用6×107cfu；
4一只小鼠在免疫期间死亡。
实施例8用重组GBS蛋白质免疫赋与抗试验性GBS感染的保护性
该实施例详细说明了通过用相应于SEQ ID NO:39的重组蛋白免疫保护小鼠抗致命GBS感染的保护作用。
以三星期间隔，使用从携带包含相应于SEQ ID NO:42的GBS基因的重组pURV22质粒载体的大肠杆菌菌株BLR(Novagen)纯化的20μg重组蛋白，在20μg QuilATM佐剂(Cedarlane Laboratories Ltd,Hornby，加拿大)存在下，对每组10只雌性CD-1小鼠(Charles River)皮下注射免疫三次，或者，作为对照，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)从眶下窦(orbital sinus)取血样。一星期之后，用大约104至106CFU的各种强毒株GBS攻击小鼠。在TSA/5％山羊血琼脂平板上平板培养GBS攻击种菌样品以确定CFU和核实攻击剂量。记录14天的死亡情况，并且在攻击后第14天，杀死存活的小鼠，并对血液和脾测定GBS微生物的存在。存活数据在表5中给出。
通过标准免疫测定法，对攻击前血清分析与GBS反应的抗体的存在。酶联免疫吸附测定和免疫印迹分析表明用大肠杆菌中产生的重组GBS蛋白免疫诱发了与重组和天然GBS蛋白质两者都反应的抗体。实施例9描述了对GBS应答的抗体。
表5相应于SEQ ID NO:39的重组GBS蛋白激发抗8种各异的GBS攻击菌株的保护作用的能力
| 攻击菌株 | |||
| 免疫原 | 编号 | 类型 | 存活数目：死亡数目1 |
| rGBS蛋白质没有 | C388/90 | Ia/c | 8∶2(P＜0.0001)0∶10 |
| rGBS蛋白质没有 | NCS246 | Ⅱ/R | 10∶0(P=0.0012)3∶7 |
| rGBS蛋白质没有 | ATCC12401 | Ib | 10∶0(P=0.001)3∶7 |
| rGBS蛋白质没有 | NCS535 | Ⅴ | 10∶0(P=0.01)5∶5 |
| rGBS蛋白质没有 | NC9842 | Ⅵ | 10∶0(P＜0.0001)0∶10 |
| rGBS蛋白质NCS 915-F3没有 | NCS915 | Ⅲ | 7∶3(P=0.0007)21∶94∶6 |
| rGBS蛋白质NCS 954-F没有 | NCS954 | Ⅲ/R | 7∶3(P=0.002)4∶61∶9 |
| rGBS蛋白质COH1-F没有 | COH1 | Ⅲ | 4∶6(P=0.0004)3∶70∶10 |
1使用每组10只小鼠的组，给出了感染存活的小鼠数目和死亡的小鼠的数目。应用对于非参数分析的logrank测定，将相应于重组GBS蛋白免疫的动物的成活率曲线与相应于模拟免疫的动物的成活率曲线相比较。
2对NCS915-F-免疫的动物的比较分析。
3在QuilATM佐剂存在下用甲醛杀死的GBS免疫动物。
在攻击后第14天，来自存活小鼠的所有血培养是阴性的。来自存活小鼠的所有脾培养是阴性的，除了实验MB-11的几只小鼠。
实施例9用重组GBS蛋白质接种激发对GBS的免疫应答
根据实施例8所述，用相应于SEQ ID NO:39的重组GBS蛋白质对10只雌性CD-1小鼠皮下免疫。为了评价对天然GBS蛋白的抗体反应，通过ELISA，使用选自Ⅲ型NCS954菌株，Ib菌株ATCC12401，Ⅴ型菌株NCS535或Ⅵ型菌株NCS9842的甲醛杀死的GBS细胞包被的平板，对在每一次免疫之前和在第三次免疫之后14天收集的血样血清测试抗体与GBS细胞的反应性。对GBS细胞提取物和纯化的重组抗原的蛋白质印迹测定证实产生的抗体对GBS蛋白质的特异性。图10中显示的结果清楚地证明动物对用作免疫原的重组GBS蛋白质强烈应答，相应的抗体效价中间值根据包被的抗原对于第三次免疫之后收集的血清来说在12000和128000之间变化。当在1∶100稀释度下测定时，所有免疫前血清是负值。在一次注射重组GBS蛋白之后每一只动物的血清中可检测到与GBS反应的抗体。
实施例10本发明GBS蛋白的抗原保守性
使用特异于本发明的GBS蛋白的单克隆抗体(MAbs)来证明所有的GBS产生该表面抗原，并且其抗原性高度保守。
收集68个GBS分离物来评价GBS-特异性MAbs的反应性。从加拿大Northern Alberta省立公共健康实验室，链球菌国家中心；魁北克大学中心医院，Pavillon CHUL，魁北克，加拿大；美国典型培养物保藏中心，USA；加拿大魁北克Laboratoire de Sante Publique，美国西雅图儿童医院和医药中心获得这些菌株。对下面菌株平行测试所有8个单克隆抗体：血清型Ⅰa或Ⅰa/c的6个分离物，血清型Ⅰb的3个分离物，血清型Ⅱ的4个分离物，血清型Ⅳ的2个分离物，血清型V的2个分离物，血清型Ⅵ的2个分离物，血清型Ⅶ的2个分离物，血清型Ⅷ的1个分离物，没有表征血清型的10个分离物和3个牛无菌链球菌菌株。Mab 3A2也与另外的GBS反应：血清型Ⅰa/c的9个分离物和血清型Ⅴ的10个分离物。37℃下5％CO2气氛下在血琼脂板上将菌株培养过夜。在-70℃下在含有20％(v/v)甘油的心浸液肉汤中保藏培养物。
为了获得GBS蛋白质特异性MAbs，以三星期间隔，用在20％QuilATM佐剂存在下的20μg纯化的重组GBS蛋白质(SEQ ID NO:44)三次免疫小鼠。从免疫小鼠回收的脾细胞与先前描述的非分泌SP2/0骨髓瘤细胞系(Hamel,J.等，1987,J.Med.Microbio1.23:163-170)融合产生杂交瘤细胞系。根据先前所述(Hamel,J.等，1987,J.Med.Microbiol.23:163-170)，使用甲醛灭活的GBS和纯化的重组GBS蛋白质(SEQ ID NO：39或44)作为包被抗原，通过ELISA对杂交克隆上清液测试特异抗体产生。通过限制稀释度克隆特异的杂化物，扩展，并且在液氮中冷冻。实施例4和5中存在重组GBS蛋白质的产生。通过使用不连续的Laemmli的缓冲体系根据生产商说明通过电泳解离纯化的重组GBS蛋白质或甲醛灭活的GBS，然后转移到硝基纤维素膜上用于蛋白质免疫印迹测定，如先前所述(Martin等，1992,Infect.Immun.60:2718-2725)。
蛋白质免疫印迹实验清楚地表明所有8个单克隆抗体识别相应于纯化的重组GBS蛋白质(SEQ ID NO:39)的蛋白质带。这些单克隆抗体还与迄今为止试验的每一种GBS分离物中存在的蛋白质带反应。表6中给出了这些GBS特异性单克隆抗体的反应性。每一种单克隆抗体与所有46种GBS很好地反应。另外，这些单克隆抗体也识别测试过的牛源的3个无菌链球菌菌株。MAb3A2还识别19种GBS；血清型Ⅰa/c的9个分离物和血清型Ⅴ的10个分离物。对其它MAbs没有测定抗这些另外的菌株的性能。
这些结果证明迄今为止测定的所有65种GBS和牛源的3个无菌链球菌菌株都产生GBS蛋白质(SEQ ID NO:39)。更重要的是，这些结果清楚地证明这8个GBS-特异性MAbs识别的表位分布广泛并且在GBS中是保守的。这些结果还表明这些表位不限制于血清学相关的分离物，因为测试了所有已知GBS血清型，包括主要疾病引发组的代表。
结论是，该实施例中存在的数据清楚地证明所有的GBS都产生本发明的GBS蛋白质并且其抗原性高度保守。
表6.根据蛋白质印迹评价的8个GBS蛋白质特异性单克隆抗体与不同的无菌链球菌菌株的反应性
| 图 | MAbs识别的无菌链球菌菌株各血清型的数目。 | |||||||||||
| Ia或Ia/C(6) | Ib(3) | Ⅱ(4) | Ⅲ(4) | Ⅳ(2) | Ⅴ(2) | Ⅵ(2) | Ⅶ(2) | Ⅷ(1) | NT(10)2 | 总计(26) | 牛(3) | |
| 3A21 | 6 | 3 | 4 | 4 | 2 | 2 | 2 | 2 | 1 | 10 | 46 | 3 |
| 5A12 | 6 | 3 | 4 | 4 | 2 | 2 | 2 | 2 | 1 | 10 | 46 | 3 |
| 6G11 | 6 | 3 | 4 | 4 | 2 | 2 | 2 | 2 | 1 | 10 | 46 | 2 |
| 889 | 6 | 3 | 4 | 4 | 2 | 2 | 2 | 2 | 1 | 10 | 46 | 3 |
| 8E11 | 6 | 3 | 4 | 4 | 2 | 2 | 2 | 2 | 1 | 10 | 46 | 3 |
| 12B12 | 6 | 3 | 4 | 4 | 2 | 2 | 2 | 2 | 1 | 10 | 46 | 3 |
| 18F11 | 6 | 3 | 4 | 4 | 2 | 2 | 2 | 2 | 1 | 10 | 46 | 3 |
| 20G2 | 6 | 3 | 4 | 4 | 2 | 2 | 2 | 2 | 1 | 10 | 46 | 3 |
1MAb3A2识别血清型Ⅰa/c的9个另外的菌株和血清型Ⅴ的10个菌株。
2这些菌株没有表征血清型。
序列表<110>BiOChem Vaccins
RIOUX,Clément
DENIS,Martin
BRODEUR,Bernard R.
HAMEL,Josée
CHARLEBOIS,Isabelle
BOYER,Martine<120>新的B组链球菌抗原<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>链球菌<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 aaa 47Ser Gly Lys Glu Pro Ala Asn Arg Phe Ser Trp Ala Lys Asn Lys
1 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 Phe
20 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 Thr
35 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 Thr
50 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 Leu
65 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 Thr
100 105 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 Val
115 120 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 Ile
130 135 140ctt gct ctt ctt att tat cgt aaa caa gaa gat taatagaaag tatctagtga 484Leu Ala Leu Leu Ile Tyr Arg Lys Gln Glu Asp
145 150tagactaaca gtatgatatg gtatgtcaaa gtatttagga ggagaagat atg tct act 542
Met Ser Thr
155tta 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 Ile
160 165 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 Ile
175 180 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 Leu
210 215 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 Leu
225 230 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 Lys
240 245 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 Thr
255 260 265ttc tta ttt taatactact tagccgttcg atttagttga acggctttta 927Phe Leu Phe270gtaatcattt ttttctcata atacaggtag tttaagtaat ttgtctttaa aaatagtata 987atataactac gaattcaaag agaggtgact ttgatt atg act gag aac tgg tta 1041
Met Thr Glu ASn Trp Leu
275cat act aaa gat ggt tca gat att tat tat cgt gtc gtt ggt caa ggt 1089His Thr Lys Asp Gly Ser Asp Ile Tyr Tyr Arg Val Val Gly Gln Gly
280 285 290caa ccg att gtt ttt tta cat ggc aat agc tta agt agt cgc tat ttt 1137Gln 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 atg 1185Asp Lys Gln Ile Ala Tyr Phe Ser Lys Tyr Tyr Gln Val Ile Val Met
315 320 325gat agt aga ggg cat ggc aaa agt cat gca aag cta aat acc att agt 1233Asp Ser Arg Gly His Gly Lys Ser His Ala Lys Leu Asn Thr Ile Ser
330 335 340ttc agg caa ata gca gtt gac tta aag gat atc tta gtt cat tta gag 1281Phr Arg Gln Ile Ala Val Asp Leu Lys Asp Ile Leu Val His Leu Glu
345 350 355att gat aaa gtt ata ttg gta ggc cat agc gat ggt gcc aat tta gct 1329Ile Asp Lys Val Ile Leu Val Gly His Ser Asp Gly Ala Asn Leu Ala
360 365 370tta gtt ttt caa acg atg ttt cca ggt atg gtt aga ggg ctt ttg ctt 1377Leu 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 ctt 1425Asn Ser Gly ASn Leu Thr Ile His Gly Gln Arg Trp Trp Asp Ile Leu
395 400 405tta gta agg att gcc tat aaa ttc ctt cac tat tta ggg aaa ctc ttt 1473Leu Val Arg Ile Ala Tyr Lys Phe Leu His Tyr Leu Gly Lys Leu Phe
410 415 420ccg tat atg agg caa aaa gct caa gtt att tcg ctt atg ttg gag gat 1521Pro Tyr Met Arg Gln Lys Ala Gln Val Ile Ser Leu Met Leu Glu Asp
425 430 435ttg aag att agt cca gct gat tta cag cat gtg tca act cct gta atg 1569Leu Lys Ile Ser Pro Ala Asp Leu Gln His Val Ser Thr Pro Val Met
440 445 450gtt ttg gtt gga aat aag gac ata att aag tta aat cat tct aag aaa 1617Val 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 ttt 1665Leu Ala Ser Tyr Phe Pro Arg Gly Glu Phe Tyr Ser Leu Val Gly Phe
475 480 485ggg cat cac att att aag caa gat tcc cat gtt ttt aat att att gca 1713Gly His His Ile Ile Lys Gln Asp Ser His Val Phe Asn Ile Ile Ala
490 495 500aaa aag ttt atc aac gat acg ttg aaa gga gaa att gtt gaa aaa gct 1761Lys Lys Phe Ile ASn Asp Thr Leu Lys Gly Glu Ile Val Glu Lys Ala
505 510 515aat tga aaaagtcaaa tcactgactt ctgtgattaa aattgtattt tttatatctg 1817Asn *ttttagtgct tattattgtt gaa atg att cat ttg aaa cga act att tct gtt 1870
Met Ile His Leu Lys Arg Thr Ile Ser Val
520 525gag caa cta aag agt gtt ttt ggg caa tta tct cca atg aat ctt ttc 1918Glu 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 tat 1966Leu Ile Ile Leu Val Gly Val Ile Ala Val Leu Pro Thr Thr Gly Tyr
550 555 560gac ttt gta ctg aat gga ctt tta cgt aca gat aaa agc aaa agg tat 2014Asp Phe Val Leu Asn Gly Leu Leu Arg Thr Asp Lys Ser Lys Arg Tyr
565 570 575att tta cag act agt tgg tgt atc aac act ttt aat aac ttg tca gga 2062Ile Leu Gln Thr Ser Trp Cys Ile Asn Thr Phe ASn Asn Leu Ser Gly
580 585 590ttc ggt ggc tta atc gat att ggg ttg cgc atg gct ttt tat ggt aaa 2110Phe Gly Gly Leu Ile Asp Ile Gly Leu Arg Met Ala Phe Tyr Gly Lys
595 600 605aaa ggt caa gag aag agt gac cta aga gaa gtg act cgt ttt tta ccc 2158Lys 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 atg 2206Tyr Leu Ile Ser Gly Leu Ser Phe Ile Ser Val Ile Ala Leu Ile Met
630 635 640agc cat att ttt cat gcc aaa gct agt gtt gat tac tat tat ttg gta 2254Ser His Ile Phe His Ala Lys Ala Ser Val Asp Tyr Tyr Tyr Leu Val
645 650 655tta att ggt gct agt atg tat ttt cct gtt att tat tgg att tct ggt 2302Leu Ile Gly Ala Ser Met Tyr Phe Pro Val Ile Tyr Trp Ile Ser Gly
660 665 670cat aaa gga agc cat tat ttc gga gat atg cca tct agt act cgt ata 2350His Lys Gly Ser His Tyr Phe Gly Asp Met Pro Ser Ser Thr Arg Ile
675 680 685aaa tta ggt gtt gtt tct ttt ttt gaa tgg gga tgt gcg gcc gca gca 2398Lys 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 aaa 2446Phe Ile Ile Ile Gly Tyr Leu Met Gly Ile His Leu Pro Val Tyr Lys
710 715 720att tta cca cta ttt tgt att ggt tgt gcc gtc ggg att gta tcc ctt 2494Ile Leu Pro Leu Phe Cys Ile Gly Cys Ala Val Gly Ile Val Ser Leu
725 730 735att ccc ggt gga tta gga agt ttt gaa tta gtt cta ttt aca ggg ttt 2542Ile Pro Gly Gly Leu Gly Ser Phe Glu Leu Val Leu Phe Thr Gly Phe
740 745 750gct gcc gag gga cta cct aaa gaa act gtg gtt gca tgg tta tta ctt 2590Ala Ala Glu Gly Leu Pro Lys Glu Thr Val Val Ala Trp Leu Leu Leu
755 760 765tat cgt tta gcc tac tat att att cca ttc ttt gca ggt atc tat ttc 2638Tyr 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 gtc 2686Phe Ile His Tyr Leu Gly Ser Gln Ile Asn Gln Arg Tyr Glu Asn Val
790 795 800ccg aaa gag tta gta tca act gtt cta caa acc atg gtg agc cat ttg 2734Pro Lys Glu Leu Val Ser Thr Val Leu Gln Thr Met Val Ser His Leu
805 810 815atg cgt att tta ggt gca ttc tta ata ttt tca aca gca ttt ttt gaa 2782Met Arg Ile Leu Gly Ala Phe Leu Ile Phe Ser Thr Ala Phe Phe Glu
820 825 830aat att act tat att atg tgg ttg cag aag cta ggc ttg gac cca tta 2830Asn Ile Thr Tyr Ile Met Trp Leu Gln Lys Leu Gly Leu Asp Pro Leu
835 840 845caa gaa caa atg tta tgg cag ttt cca ggt tta ttg ctg ggg gtt tgt 2878Gln 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 ttt 2926Phe Ile Leu Leu Ala Arg Thr Ile Asp Gln Lys Val Lys Asn Ala Phe
870 875 880cca att gct att atc tgg att act ttg aca ttg ttt tat ctt aat tta 2974Pro Ile Ala Ile Ile Trp Ile Thr Leu Thr Leu Phe Tyr Leu Asn Leu
885 890 895ggt cat att agt tgg cga cra tct ttc tgg ttt att tta cta ttg tta 3022Gly His Ile Ser Trp Arg Leu Ser Phe Trp Phe Ile Leu Leu Leu Leu
900 905 910ggc tta tta gtc att aag cca act ctc tat aaa aaa caa ttt att tat 3070Gly Leu Leu Val Ile Lys Pro Thr Leu Tyr Lys Lys Gln Phe Ile Tyr
915 920 925agc tgg gaa gag cgt att aag gat gga atc att atc gtt agt tta atg 3118Ser 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 cat 3166Gly Val Leu Phe Tyr Ile Ala Gly Leu Leu Phe Pro Ile Arg Ala His
950 955 960att aca ggt ggt agt att gaa cgc ctg cat tat atc ata gca tgg gag 3214Ile Thr Gly Gly Set Ile Glu Arg Leu His Tyr Ile Ile Ala Trp Glu
965 970 975ccg ata gca ttg gct acg ttg att ctt act crc gtt tat tta tgt ttg 3262Pro Ile Ala Leu Ala Thr Leu Ile Leu Thr Leu Val Tyr Leu Cys Leu
980 985 990gtt aag att tta caa gga aaa tct tgt cag att ggt gat gtg ttc aat 3310Val Lys Ile Leu Gln Gly Lys Ser Cys Gln Ile Gly Asp Val Phe Asn
995 1000 1005gtg gat cgt tat aaa aaa cta ctt caa gct tac ggt ggt tct tcg gat 3358Val Asp Arg Tyr Lys Lys Leu Leu Gln Ala Tyr Gly Gly Ser Ser Asp1010 1015 1020 1025agc ggt tta gcc ttt tta aat gat aaa agg ctc tac tgg tac caa aaa 3406Ser Gly Leu Ala Phe Leu Asn Asp Lys Arg Leu Tyr Trp Tyr Gln Lys
1030 1035 1040aat gga gaa gat tgc gtt gcg ttc caa ttt gta att gtc aat aat aaa 3454Asn Gly Glu Asp Cys Val Ala Phe Gln Phe Val Ile Val Asn Asn Lys
1045 1050 1055tgt ctt att atg ggg gaa cca gcc ggt gat gac act tat att cgt gaa 3502Cys Leu Ile Met Gly Glu Pro Ala Gly Asp Asp Thr Tyr Ile Arg Glu
1060 1065 1070gct att gaa tcg ttt att gat gat gct gat aag cta gac tat gac ctt 3550Ala Ile Glu Set Phe Ile Asp Asp Ala Asp Lys Leu Asp Tyr Asp Leu1075 1080 1085gtt ttt tac agt att gga cag aag ttg aca cta ctt tta cat gag tat 3598Val Phe Tyr 5er Ile Gly Gln Lys Leu Thr Leu Leu Leu His Glu Tyr1090 1095 1100 1105ggt ttt gac ttt atg aaa gtt ggt gag gat gct tta gtt aat tta gaa 3646Gly Phe Asp Phe Met Lys Val Gly Glu Asp Ala Leu Val Asn Leu Glu
1110 1115 1120acg ttt att ctt aaa ggg aat aag tac aaa cct ttc aga aat gcc cta 3694Thr Phe Thr Ltu Lys Gly Asn Lys Tyr Lys Pro Phe Arg Asn Ala Leu
1125 1130 1135aat aga gtt gaa aag gat ggt ttc tat ttc gaa gtt gta caa tcg cca 3742Asn Arg Val Glu Lys Asp Gly Phe Tyr Phe Glu Val Val Gln Ser Pro
1140 1145 1150cat agt caa gag cta cta aat agt ttg gaa gag att tct aat act tgg 3790His Ser Gln Glu Leu Leu Asn Ser Leu Glu Glu Ile Ser Asn Thr Trp
1155 1160 1165tta gaa gga cgc cct gaa aaa ggt ttc tca cta gga tat ttt aat aaa 3838Leu Glu Gly Arg Pro Glu Lys Gly Phe Ser Leu Gly Tyr Phe Asn Lys1170 1175 1180 1185gat tat ttc caa caa gcc cca ata gct ttg gta aaa aat gct gaa cac 3886Asp Tyr Phe Gln Gln Ala Pro Ile Ala Leu Val Lys Asn Ala Glu His
1190 1195 1200gaa gtt gct gct ttt gct aat att atg cca aac tat gaa aag agt att 3934Glu Val Val Ala Phe Ala Asn Ile Met Pro Asn Tyr Glu Lys Ser Ile
1205 1210 1215atc tct att gat tta atg cgt cac gat aaa cag aaa att ccg aat ggc 3982Ile Ser Ile Asp Leu Met Arg His Asp Lys Gln Lys Ile Pro Asn Gly
1220 1225 1230gtt atg gat ttc ctc ttt tta tca tta ttc tct tat tat caa gag aag 4030Val Met Asp Phe Leu Phe Leu Ser Leu Phe Ser Tyr Tyr Gln Glu Lys
1235 1240 1245gga tac cac tat ttt gat ttg ggg atg gca cct tta tca gga gtt ggt 4078Gly Tyr His Tyr Phe Asp Leu Gly Met Ala Pro Leu Ser Gly Val Gly1250 1255 1260 1265cgc gtt gaa aca agt ttt gct aaa gag aga atg gcg tat ctt gtc tat 4126Arg Val Glu Thr Ser Phe Ala Lys Glu Arg Met Ala Tyr Leu Val Tyr
1270 1275 1280cat ttc ggt agt cat trc tac tca ttt aat ggt tta cac aag tat aag 4174His Phe Gly Ser His Phe Tyr Ser Phe Asn Gly Leu His Lys Tyr Lys
1285 1290 1295aag aag ttt aca cca ttg tgg tcg gaa cgt tat att tct tgt tct cgt 4222Lys Lys Phe Thr Pro Leu Trp Ser Glu Arg Tyr Ile Ser Cys Ser Arg
1300 1305 1310tcg tcc tgg tta att tgt gct att tgt gcc cta tta atg gaa gat agt 4270Ser Ser Trp Leu Ile Cys Ala Ile Cys Ala Leu Leu Met Glu Asp Ser
1315 1320 1325aaa att aag att gtt aaa taagctttat ttggcaatta aaaagagcat 4318Lys Ile Lys Ile Val Lys1330 1335gtcatgcgac 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>蛋白质<213>链球菌<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 Phe
20 25 30Ala Val Gln Phe Ile Gly Leu Lys Pro Asp Tyr Pro Gly Lys Thr Tyr
35 40 45Phe Ile Ile Leu Leu Thr Ala Trp Thr Leu Met Ala Leu Val Thr Ala
50 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 Leu
85 90 95Ser Pro Lys Phe Phe Gln Thr Ile Gln Pro Phe Leu Pro Met Thr Tyr
100 105 110Ser Val Ser Gly Leu Arg Glu Thr Ile Ser Leu Thr Gly Asp Val Asn
115 120 125His Gln Trp Arg Met Leu Val Ile Phe Leu Val Ser Ser Met Ile Leu
130 135 140Ala Leu Leu Ile Tyr Arg Lys Gln Glu Asp145 150<210>3<211>118<212>蛋白质<213>链球菌<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 Thr
20 25 30Gly Lys Ile Phe Ser Met Ser Lys Glu Glu Leu Ser Tyr Leu Pro Val
35 40 45Ile Lys Leu Phe Lys Asn Gln Gly Val Tyr Asn Gly Leu Ile Gly Leu
50 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 Thr
85 90 95Val Asp Lys Lys Ile Leu Leu Lys Gln Gly Gly Leu Pro Ile Leu Ala
100 105 110Leu Leu Thr Phe Leu Phe
115<210>4<211>247<212>蛋白质<213>链球菌<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 Ser
20 25 30Leu Ser Ser Arg Tyr Phe Asp Lys Gln Ile Ala Tyr Phe Ser Lys Tyr
35 40 45Tyr Gln Val Ile Val Met Asp Ser Arg Gly His Gly Lys Ser His Ala
50 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 Ser
85 90 95Asp Gly Ala Asn Leu Ala Leu Val Phe Gln Thr Met Phe Pro Gly Met
100 105 110Val Arg Gly Leu Leu Leu Asn Ser Gly Asn Leu Thr Ile His Gly Gln
115 120 125Arg Trp Trp Asp Ile Leu Leu Val Arg Ile Ala Tyr Lys Phe Leu His
130 135 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 His
165 170 175Val Ser Thr Pro Val Met Val Leu Val Gly Asn Lys Asp Ile Ile Lys
180 185 190Leu Asn His Ser Lys Lys Leu Ala Ser Tyr Phe Pro Arg Gly Glu Phe
195 200 205Tyr Ser Leu Val Gly Phe Gly His His Ile Ile Lys Gln Asp Ser His
210 215 220Val Phe Asn Ile Ile Ala Lys Lys Phe Ile Asn Asp Thr Leu Lys Gly225 230 235 240Glu Ile Val Glu Lys Ala Asn
245<210>5<211>816<212>蛋白质<213>链球菌<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 Gly
20 25 30Val Ile Ala Val Leu Pro Thr Thr Gly Tyr Asp Phe Val Leu Asn Gly
35 40 45Leu Leu Arg Thr Asp Lys Ser Lys Arg Tyr Ile Leu Gln Thr Ser Trp
50 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 Ser
85 90 95Asp Leu Arg Glu Val Thr Arg Phe Leu Pro Tyr Leu Ile Ser Gly Leu
100 105 110Ser Phe Ile Ser Val Ile Ala Leu Ile Met Ser His Ile Phe His Ala
115 120 125Lys Ala Ser Val Asp Tyr Tyr Tyr Leu Val Leu Ile Gly Ala Ser Met
130 135 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 Ser
165 170 175Phe Phe Glu Trp Gly Cys Ala Ala Ala Ala Phe Ile Ile Ile Gly Tyr
180 185 190Leu Met Gly Ile His Leu Pro Val Tyr Lys Ile Leu Pro Leu Phe Cys
195 200 205Ile Gly Cys Ala Val Gly Ile Val Ser Leu Ile Pro Gly Gly Leu Gly
210 215 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 Tyr
245 250 255Ile Ile Pro Phe Phe Ala Gly Ile Tyr Phe Phe Ile His Tyr Leu Gly
260 265 270Ser Gln Ile Asn Gln Arg Tyr Glu Asn Val Pro Lys Glu Leu Val Ser
275 280 285Thr Val Leu Gln Thr Met Val Ser His Leu Met Arg Ile Leu Gly Ala
290 295 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 Trp
325 330 335Gln Phe Pro Gly Leu Leu Leu Gly Val Cys Phe Ile Leu Leu Ala Arg
340 345 350Thr Ile Asp Gln Lys Val Lys Asn Ala Phe Pro Ile Ala Ile Ile Trp
355 360 365Ile Thr Leu Thr Leu Phe Tyr Leu Asn Leu Gly His Ile Ser Trp Arg
370 375 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 Ile
405 410 415Lys Asp Gly Ile Ile Ile Val Ser Leu Met Gly Val Leu Phe Tyr Ile
420 425 430Ala Gly Leu Leu Phe Pro Ile Arg Ala His Ile Thr Gly Gly Ser Ile
435 440 445Glu Arg Leu His Tyr Ile Ile Ala Trp Glu Pro Ile Ala Leu Ala Thr
450 455 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 Lys
485 490 495Leu Leu Gln Ala Tyr Gly Gly Ser Ser Asp Ser Gly Leu Ala Phe Leu
500 505 510Asn Asp Lys Arg Leu Tyr Trp Tyr Gln Lya Asn Gly Glu Asp Cys Val
515 520 525Ala Phe Gln Phe Val Ile Val Asn Asn Lys Cys Leu Ile Met Gly Glu
530 535 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 Gly
565 570 575Gln Lys Leu Thr Leu Leu Leu His Glu Tyr Gly Phe Asp Phe Met Lys
580 585 590Val Gly Glu Asp Ala Leu Val Asn Leu Glu Thr Phe Thr Leu Lys Gly
595 600 605Asn Lys Tyr Lys Pro Phe Arg Asn Ala Leu Asn Arg Val Glu Lys Asp
610 615 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 Glu
645 650 655Lys Gly Phe Ser Leu Gly Tyr Phe Asn Lys Asp Tyr Phe Gln Gln Ala
660 665 670Pro Ile Ala Leu Val Lys Asn Ala Glu His Glu Val Val Ala Phe Ala
675 680 685Asn Ile Met Pro Asn Tyr Glu Lys Ser Ile Ile Ser Ile Asp Leu Met
690 695 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 Asp
725 730 735Leu Gly Met Ala Pro Leu Ser Gly Val Gly Arg Val Glu Thr Ser Phe
740 745 750Ala Lys Glu Arg Met Ala Tyr Leu Val Tyr His Phe Gly Ser His Phe
755 760 765Tyr Ser Phe Asn Gly Leu His Lys Tyr Lys Lys Lys Phe Thr Pro Leu
770 775 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 Lys
805 810 815<210>6<211>518<212>蛋白质<213>链球菌<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 Leu
20 25 30Gln Glu Gln Met Leu Trp Gln Phe Pro Gly Leu Leu Leu Gly Val Cys
35 40 45Phe Ile Leu Leu Ala Arg Thr Ile Asp Gln Lys Val Lys Asn Ala Phe
50 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 Leu
85 90 95Gly Leu Leu Val Ile Lys Pro Thr Leu Tyr Lys Lys Gln Phe Ile Tyr
100 105 110Ser Trp Glu Glu Arg Ile Lys Asp Gly Ile Ile Ile Val Ser Leu Met
115 120 125Gly Val Leu Phe Tyr Ile Ala Gly Leu Leu Phe Pro Ile Arg Ala His
130 135 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 Leu
165 170 175Val Lys Ile Leu Gln Gly Lys Ser Cys Gln Ile Gly Asp Val Phe Asn
180 185 190Val Asp Arg Tyr Lys Lys Leu Leu Gln Ala Tyr Gly Gly Ser Ser Asp
195 200 205Ser Gly Leu Ala Phe Leu Asn Asp Lys Arg Leu Tyr Trp Tyr Gln Lys
210 215 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 Glu
245 250 255Ala Ile Glu Ser Phe Ile Asp Asp Ala Asp Lys Leu Asp Tyr Asp Leu
260 265 270Val Phe Tyr Ser Ile Gly Gln Lys Leu Thr Leu Leu Leu His Glu Tyr
275 280 285Gly Phe Asp Phe Met Lys Val Gly Glu Asp Ala Leu Val Asn Leu Glu
290 295 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 Pro
325 330 335His Ser Gln Glu Leu Leu Asn Ser Leu Glu Glu Ile Ser Asn Thr Trp
340 345 350Leu Glu Gly Arg Pro Glu Lys Gly Phe Ser Leu Gly Tyr Phe Asn Lys
355 360 365Asp Tyr Phe Gln Gln Ala Pro Ile Ala Leu Val Lys Asn Ala Glu His
370 375 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 Gly
405 410 415Val Met Asp Phe Leu Phe Leu Ser Leu Phe Ser Tyr Tyr Gln Glu Lys
420 425 430Gly Tyr His Tyr Phe Asp Leu Gly Met Ala Pro Leu Ser Gly Val Gly
435 440 445Arg Val Glu Thr Ser Phe Ala Lys Glu Arg Met Ala Tyr Leu Val Tyr
450 455 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 Arg
485 490 495Ser Ser Trp Leu Ile Cys Ala Ile Cys Ala Leu Leu Met Glu Asp Ser
500 505 510Lys Ile Lys Ile Val Lys
515<210>7<211>5126<212>DNA<213>链球菌<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 Asp
20 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 Ile
35 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 Leu
50 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 Glu
85 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 Gln
l00 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 Met
115 120 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 Lys
130 135 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 Glu
165 170 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 Pro
180 185 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 Val
195 200 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 Lys
210 215 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 Ser
240 245 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 Gly
255 260 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 Phe
270 275 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 Asp
305 310 315acc gta ctt tct tct gac tta aga gag atg gct tta att aaa gaa tat 1009Thr Val Leu Ser Ser Asp Leu Arg Glu Net Ala Leu Ile Lys Glu Tyr
320 325 330gaa tta ttg ctt aat cac tac gaa gaa agt aag caa tca cgt cta gag 1057Glu Leu Leu Leu Asn His Tyr Glu Glu Ser Lys Gln Ser Arg Leu Glu
335 340 345aaa gta atg gca gaa atg gat tct tta gat gct tgg tct att gag agc 1105Lys Val Met Ala Glu Met Asp Ser Leu Asp Ala Trp Ser Ile Glu Ser
350 355 360gaa gtc aaa aca gta tta tcc aaa tta ggt att act gat ttg cag ttg 1153Glu 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 gcg 1201Set Val Gly Glu Leu Set Gly Gly Leu Arg Arg Arg Val Gln Leu Ala
385 390 395caa gta tta tta aat gat gca gat tta ttg ctc tta gac gaa cct act 1249Gln Val Leu Leu Asn Asp Ala Asp Leu Leu Leu Leu Asp Glu Pro Thr
400 405 410aac cac tta gat att gac act att gca tgg tta acg aat ttt ttg aaa 1297Asn His Leu Asp Ile Asp Thr Ile Ala Trp Leu Thr Asn Phe Leu Lys
415 420 425aat agt aaa aag aca gtg ctt ttt ata act cat gat cgt tat ttt cta 1345Asn Set Lys Lys Thr Val Leu Phe Ile Thr His Asp Arg Tyr Phe Leu
430 435 440gac aat gtt gca aca cgt att ttt gaa tta gat aag gca cag att aca 1393Asp 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 caa 1441Glu Tyr Gln Gly Asn Tyr Gln Asp Tyr Val Arg Leu Arg Ala Glu Gln
465 470 475gac gag cgt gat gct gct agt tta cat aaa aag aaa cag ctt tat aaa 1489Asp Glu Arg Asp Ala Ala Ser Leu His Lys Lys Lys Gln Leu Tyr Lys
480 485 490cag gaa cta gct tgg atg cgt act cag cca caa gct cgt gca acg aaa 1537Gln Glu Leu Ala Trp Met Arg Thr Gln Pro Gln Ala Arg Ala Thr Lys
495 500 505caa cag gct cgt att aat cgt ttt caa aat cta aaa aac gat tta cac 1585Gln Gln Ala Arg Ile Asn Arg Phe Gln Asn Leu Lys Asn Asp Leu His
510 515 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 Pro
545 550 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 Asp
560 565 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 Leu
575 580 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 Ile
590 595 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 Glu
625 630 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 Gln
640 645 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 Ser
655 660 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 Lys
670 675 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 Val
705 710 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 Lys
720 725 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 Tyr
735 740 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 Val
750 755 760atc agt aaa aaa gag agt acc aag aca agt cgt gaa aag caa agt cgt 2353Ile 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 gac 2401Lys Arg Met Ser Tyr Phe Glu Lys Gln Glu Trp Ala Thr Ile Glu Asp
785 790 795gat att atg ata ttg gaa aat act atc act cgt ata gaa aat gat atg 2449Asp Ile Met Ile Leu Glu Asn Thr Ile Thr Arg Ile Glu Asn Asp Met
800 805 810caa aca tgt ggt agt gat ttt aca agg tta tct gat tta caa aag gaa 2497Gln Thr Cys Gly Ser Asp Phe Thr Arg Leu Ser Asp Leu Gln Lys Glu
815 820 825tta gat gca aaa aat gaa gca ctt cta gaa aag tat gac cgt tat gag 2545Leu Asp Ala Lys Asn Glu Ala Leu Leu Glu Lys Tyr Asp Arg Tyr Glu
830 835 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 Asp845 850 855 860gac caa gca gtt gca caa tta att cga caa agt tta cgc gcc tat gat 2643Asp Gln Ala Val Ala Gln Leu Ile Arg Gln Sar Leu Arg Ala Tyr Asp
865 870 875tta gat aaa cct gat aca gca tat tca gac cct cac tta gat cat ttg 2691Leu Asp Lys Pro Asp Thr Ala Tyr Ser Asp Pro His Leu Asp His Leu
880 885 890acc tca tac tac gaa aaa ata gag aag tca gga ttc ttt gtc att gag 2739Thr Ser Tyr Tyr Glu Lys Ile Glu Lys Ser Gly Phe Phe Val Ile Glu
895 900 905gag aga gat gag att att ggc tgt ggc ggc ttt ggt ccg ctg aaa aat 2787Glu 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 ggt 2835Leu Ile Ala Glu Met Gln Lys Val Tyr Ile Ala Glu Arg Phe Arg Gly
930 935 940aag ggg ctt gct act gat tta gtg aaa atg att gaa gta gaa gct cga 2883Lys Gly Leu Ala Thr Asg Leu Val Lys Met Ile Glu Val Glu Ala Arg
945 950 955aaa att ggg tat aga caa ctt tat tta gag aca gcc agt act ttg agt 2931Lys Ile Gly Tyr Arg Gln Leu Tyr Leu Glu Thr Ala Ser Thr Leu Ser
960 965 970agg gca act gcg gtt tat aag cat atg gga tat tgt gcc tta tcg caa 2979Arg Ala Thr Ala Val Tyr Lys His Met Gly Tyr Cys Ala Leu Ser Gln
975 980 985 cca ata gca aat gat caa ggt cat aca gct atg gat att tgg atg att 3027Pro Ile Ala Asn Asp Gln Gly His Thr Ala Met Asp Ile Trp Met Ile990 995 1000 1005aaa gat tta taagttgaaa gtggattagt gaacatggat taattatttt 3076Lys Asp Leugagataagag gaaagaaaag gagacatat atg gca tat att tgg tct tat ttg 3129
Met Ala Tyr Ile Trp Ser Tyr Leu
1010 1015aaa agg tac ccc aat tgg tta tgg ctt gat tta cta gga gct atg ctt 3177Lys Arg Tyr Pro Asn Trp Leu Trp Leu Asp Leu Leu Gly Ala Met Leu
1020 1025 1030ttt gtg acg gtt atc cta gga atg ccc aca gcc tta gcg ggt atg att 3225Phe Val Thr Val Ile Leu Gly Met Pro Thr Ala Leu Ala Gly Met Ile
1035 1040 1045gat aat ggc gtt aca aaa ggt gat cgg act gga gtt tat ctg tgg acg 3273Asp Asn Gly Val Thr Lys Gly Asp Arg Thr Gly Val Tyr Leu Trp Thr
1050 1055 1060
ttc atc atg ttt ata ttt gtt gta cta ggt att att ggg cgt att acg 3321Phe Ile Met Phe Ile Phe Val Val Leu Gly Ile Ile Gly Arg Ile Thr1065 1070 1075 1080atg gct tac gca tct agt cgc tta acg aca aca atg att aga gat atg 3369Met Ala Tyr Ala Ser Ser Arg Leu Thr Thr Thr Met Ile Arg Asp Met
1085 1090 1095cgt aat gat atg tat gct aag ctt caa gaa tac tcc cat cat gaa tat 3417Arg Asn Asp Met Tyr Ala Lys Leu Gln Glu Tyr Ser His His Glu Tyr
1100 1105 1110gaa cag ata ggt gta tct tca cta gtg aca cgt atg aca agc gat act 3465Glu Gln Ile Gly Val Ser Ser Leu Val Thr Arg Met Thr Ser Asp Thr
1115 1120 1125ttt gtt ttg atg caa ttt gct gaa atg tct tta cgt tta ggc cta gta 3513Phe Val Leu Met Gln Phe Ala Glu Met Ser Leu Arg Leu Gly Leu Val
1130 1135 1140act cct atg gta atg att ttt agc gtg gtt atg ata cta att acg agt 3561Thr Pro Met Val Met Ile Phe Ser Val Val Met Ile Leu Ile Thr Ser1145 1150 1155 1160cca tct ttg gct tgg ctt gta gcg gtt gcg atg cct ctt ttg gta gga 3609Pro Ser Leu Ala Trp Leu Val Ala Val Ala Met Pro Leu Leu Val Gly
1165 1170 1175gtc gtt tta tat gta gct ata aaa aca aaa cct tta tct gaa aga caa 3657Val Val Leu Tyr Val Ala Ile Lys Thr Lys Pro Leu Ser Glu Arg Gln
1180 1185 1190 cag 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 Thr
1195 1200 1205ggg 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 Gln
1210 1215 1220aaa 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 Leu1225 1230 1235 1240ttt 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 Ile
1245 1250 1255gca 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 Arg
1260 1265 1270ggt 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 Phe
1275 1280 1285cat 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 Tyr
1290 1295 1300cct 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 Met1305 1310 1315 1320cca 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 Leu
1325 1330 1335aaa 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 Glu
1340 1345 1350aca 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 Gly
1355 1360 1365gaa 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 Leu
1370 1375 1380gtt 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 Leu1385 1390 1395 1400gta 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 Gln
1405 1410 1415 aag att gga ttt atc ccc caa aaa gct ctt tta ttt aca ggg aca ata 4377Lys Ile Gly Phe Ile Pro Gln Lys Ala Leu Leu Phe Thr Gly Thr Ile
1420 1425 1430gga gag aat tta aaa tat gga aaa gct gat gct act att gat gat ctt 4425Gly Glu Asn Leu Lys Tyr Gly Lys Ala Asp Ala Thr Ile Asp Asp Leu
1435 1440 1445aga caa gcg gtt gat att tct caa gct aaa gag ttt att gag agt cac 4473Arg Gln Ala Val Asp Ile Ser Gln Ala Lys Glu Phe Ile Glu Ser His
1450 1455 1460caa gaa gcc ttt gaa acg cat tta gct gaa ggt ggg agc aat ctt tct 4521Gln Glu Ala Phe Glu Thr His Leu Ala Glu Gly Gly Ser ASn Leu Ser1465 1470 1475 1480ggg ggt caa aaa caa cgg tta tct att gct agg gct gtt gtt aaa gat 4569Gly Gly Gln Lys Gln Arg Leu Ser Ile Ala Arg Ala Val Val Lys Asp
1485 1490 1495cca gat tta tat att ttt gat gat tca ttt tct gct ctc gat tat aag 4617Pro Asp Leu Tyr Ile Phe Asp Asp Ser Phe Ser Ala Leu Asp Tyr Lys
1500 1505 1510aca gac gct act tta aga gcg cgt cta aaa gaa gta acc ggt gat tct 4665Thr Asp Ala Thr Leu Arg Ala Arg Leu Lys Glu Val Thr Gly Asp Ser
1515 1520 1525aca gtt ttg ata gtt gct caa agg gtg ggt acg att atg gat gct gat 4713Thr Val Leu Ile Val Ala Gln Arg Val Gly Thr Ile Met Asp Ala Asp
1530 1535 1540cag att att gtc ctt gat gaa ggc gaa att gtc ggt cgt ggt acc cac 4761Gln Ile Ile Val Leu Asp Glu Gly Glu Ile Val Gly Arg Gly Thr His1545 1550 1555 1560gct caa tta ata gaa aat aat gct att tat cgt gaa atc gct gag tca 4809Ala Gln Leu Ile Glu Asn Asn Ala Ile Tyr Arg Glu Ile Ala Glu Ser
1565 1570 1575caa ctg aag aac caa aac tta tca gaa gga gag tgattgt atg aga aaa 4858Gln Leu Lys Asn Gln Asn Leu Ser Glu Gly Glu Met Arg Lys
1580 1585 1590aaa tct gtt ttt ttg aga tta tgg tct tac cta act cgc tac aaa gct 4906Lys Ser Val Phe Leu Arg Leu Trp Ser Tyr Leu Thr Arg Tyr Lys Ala
1595 1600 1605act ctt ttc tta gcg att ttt ttg aaa gtt tta tct agt ttt atg agt 4954Thr Leu Phe Leu Ala Ile Phe Leu Lys Val Leu Ser Ser Phe Met Ser
1610 1615 1620gtt ctg gag cct ttt att tta ggg tta gcg ata aca gag ttg act gct 5002Val Leu Glu Pro Phe Ile Leu Gly Leu Ala Ile Thr Glu Leu Thr Ala
1625 1630 1635 aac ctt gtt gat atg gct aag gga gtt tct ggg gca gaa ttg aac gtt 5050Asn Leu Val Asp Met Ala Lys Gly Val Ser Gly Ala Glu Leu Asn Val
1640 1645 1650cct tat att gct ggt att ttg att att tat ttt ttc aga ggt gtt ttc 5098Pro Tyr Ile Ala Gly Ile Leu Ile Ile Tyr Phe Phe Arg Gly Val Phe1655 1660 1665 1670tat gaa tta ggt tct tat ggc tca aat t 5126Tyr Glu Leu Gly Ser Tyr Gly Ser Asn
1675<220>8<211>229<212>蛋白质<213>链球菌<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 Asp
20 25 30Lys Leu Leu Leu Ala Pro Tyr Trp Arg Lys Gly Met Leu Ala Leu Ile
35 40 45Asp Ser His Ala Phe Asn Tyr Leu Pro Cys Leu Lys Asn Arg Glu Leu
50 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 Glu
85 90 95His Thr Lys Thr Phe Leu Lys Lys Trp Lys Thr Ser Thr His Phe Gln
100 105 110Lys Asp Val Glu His Ile Val Asp Val Tyr Arg Ile Arg Glu Gln Met
115 120 125Gly Leu Ala Lys Glu His Leu Tyr Arg Tyr Gly Lys Thr Ile Ile Lys
130 135 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 Glu
165 170 175Ile Val Val Asn Gly Gly Thr Leu Ile Lys Lys Leu Gly Ile Lys Pro
180 185 190Gly Pro Gln Met Gly Asp Ile Ile Ser Gln Ile Glu Leu Ala Ile Val
195 200 205Leu Gly Gln Leu Ile Asn Glu Glu Glu Ala Ile Leu His Phe Val Lys
210 215 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 Ile
20 25 30Gly Ile Ile Gly Val Asn Gly Thr Gly Lys Thr Thr Leu Leu Asp Val
35 40 45Ile Ser Gly Glu Leu Gly Phe Asp Gly Asp Arg Ser Pro Phe Ser Ser
50 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 Glu
85 90 95Met Ala Leu Ile Lys Glu Tyr Glu Leu Leu Leu Asn His Tyr Glu Glu
100 105 110Ser Lys Gln Ser Arg Leu Glu Lys Val Met Ala Glu Met Asp Ser Leu
115 120 125Asp Ala Trp Ser Ile Glu Ser Glu Val Lys Thr Val Leu Ser Lys Leu
130 135 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 Leu
165 170 175Leu Leu Leu Asp Glu Pro Thr Asn His Leu Asp Ile Asp Thr Ile Ala
180 185 190Trp Leu Thr Asn Phe Leu Lys Asn Ser Lys Lys Thr Val Leu Phe Ile
195 200 205Thr His Asp Arg Tyr Phe Leu Asp Asn Val Ala Thr Arg Ile Phe Glu
210 215 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 His
245 250 255Lys Lys Lys Gln Leu Tyr Lys Gln Glu Leu Ala Trp Met Arg Thr Gln
260 265 270Pro Gln Ala Arg Ala Thr Lys Gln Gln Ala Arg Ile Asn Arg Phe Gln
275 280 285Asn Leu Lys Asn Asp Leu His Gln Thr Ser Asp Thr Ser Asp Leu Glu
290 295 300
Met 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 Asn
325 330 335Leu Leu Ile Gln Asn Lys Asp Arg Ile Gly Ile Val Gly Asp Asn Gly
340 345 350Val Gly Lys Ser Thr Leu Leu Asn Leu Ile Val Gln Asp Leu Gln Pro
355 360 365Asp Ser Gly Asn Val Ser Ile Gly Glu Thr Ile Arg Val Gly Tyr Phe
370 375 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 Ser
405 410 415Val Thr Glu Leu Leu Glu Gln Phe Leu Phe Pro Arg Ser Thr His Gly
420 425 430Thr Gln Ile Ala Lys Leu Ser Gly Gly Glu Lys Lys Arg Leu Tyr Leu
435 440 445Leu Lys Ile Leu Ile Glu Lys Pro Asn Val Leu Leu Leu Asp Glu Pro
450 455 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 Phe
485 490 495Leu Asp Lys Val Ala Asn Lys Ile Ile Ala Phe Glu Asp Asn Asp Ile
500 505 510Arg Glu Phe Phe Gly Asn Tyr Thr Asp Tyr Leu Asp Glu Lys Ala Phe
515 520 525Asn Glu Gln Asn Asn Glu Val Ile Ser Lys Lys Glu Ser Thr Lys Thr
530 535 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 Ile
565 570 575Thr Arg Ile Glu Asn Asp Met Gln Thr Cys Gly Ser Asp Phe Thr Arg
580 585 590Leu Ser Asp Leu Gln Lys Glu Leu Asp Ala Lys Asn Glu Ala Leu Leu
595 600 605Glu Lys Tyr Asp Arg Tyr Glu Tyr Leu Ser Glu Leu Asp Thr
610 615 620<210>10<211>157<212>蛋白质<213>链球菌<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 Thr
20 25 30Ala Tyr Ser Asp Pro His Leu Asp His Leu Thr Ser Tyr Tyr Glu Lys
35 40 45Ile Glu Lys Ser Gly Phe Phe Val Ile Glu Glu Arg Asp Glu Ile Ile
50 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 Asp
85 90 95Leu Val Lys Met Ile Glu Val Glu Ala Arg Lys Ile Gly Tyr Arg Gln
100 105 110Leu Tyr Leu Glu Thr Ala Ser Thr Leu Ser Arg Ala Thr Ala Val Tyr
115 120 125Lys His Met Gly Tyr Cys Ala Leu Ser Gln Pro Ile Ala Asn Asp Gln
130 135 140Gly His Thr Ala Met Asp Ile Trp Met Ile Lys Asp Leu145 150 155<210>11<211>579<212>蛋白质<213>链球菌<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 Met
20 25 30Pro Thr Ala Leu Ala Gly Met Ile Asp Asn Gly Val Thr Lys Gly Asp
35 40 45Arg Thr Gly Val Tyr Leu Trp Thr Phe Ile Met Phe Ile Phe Val Val
50 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 Leu
85 90 95Gln Glu Tyr Ser His His Glu Tyr Glu Gln Ile Gly Val Ser Ser Leu
100 105 110Val Thr Arg Met Thr Ser Asp Thr Phe Val Leu Met Gln Phe Ala Glu
115 120 125Met Ser Leu Arg Leu Gly Leu Val Thr Pro Met Val Met Ile Phe Ser
130 135 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 Lys
165 170 175Thr Lys Pro Leu Ser Glu Arg Gln Gln Thr Met Leu Asp Lys Ile Asn
180 185 190Gln Tyr Val Arg Glu Asn Leu Thr Gly Leu Arg Val Val Arg Ala Phe
195 200 205Ala Arg Glu Asn Phe Gln Ser Gln Lys Phe Gln Val Ala Asn Gln Arg
210 215 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 Trp
245 250 255Phe Ala Leu Asp Pro Leu Gln Arg Gly Ala Ile Lys Ile Gly Asp Leu
260 265 270Val Ala Phe Ile Glu Tyr Ser Phe His Ala Leu Phe Ser Phe Leu Leu
275 280 285Phe Ala Asn Leu Phe Thr Met Tyr Pro Arg Met Val Val Ser Ser His
290 295 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 Asn
325 330 335Val Thr Phe Ala Tyr Pro Gly Glu Thr Glu Ser Pro Val Leu His Asp
340 345 350Ile Ser Phe Lys Ala Lys Pro Gly Glu Thr Ile Ala Phe Ile Gly Ser
355 360 365Thr Gly Ser Gly Lys Ser Ser Leu Val Asn Leu Ile Pro Arg Phe Tyr
370 375 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 Lys
405 410 415Ala Leu Leu Phe Thr Gly Thr Ile Gly Glu Asn Leu Lys Tyr Gly Lys
420 425 430Ala Asp Ala Thr Ile Asp Asp Leu Arg Gln Ala Val Asp Ile Ser Gln
435 440 445Ala Lys Glu Phe Ile Glu Ser His Gln Glu Ala Phe Glu Thr His Leu
450 455 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 Asp
485 490 495Ser Phe Ser Ala Leu Asp Tyr Lys Thr Asp Ala Thr Leu Arg Ala Arg
500 505 510Leu Lys Glu Val Thr Gly Asp Ser Thr Val Leu Ile Val Ala Gln Arg
515 520 525Val Gly Thr Ile Met Asp Ala Asp Gln Ile Ile Val Leu Asp Glu Gly
530 535 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 Ser
565 570 575Glu Gly Glu<210>12<211>92<212>蛋白质<213>链球菌<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 Ser
20 25 30Phe Met Ser Val Leu Glu Pro Phe Ile Leu Gly Leu Ala Ile Thr Glu
35 40 45Leu Thr Ala Asn Leu Val Asp Met Ala Lys Gly Val Ser Gly Ala Glu
50 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 Asn
85 90<210>13<211>5215<212>DNA<213>链球菌<220><221>CDS<222>(3)…(122)<221>CDS<222>(133)…(2511)<221>CDS<222>(367)…(2511)<221>CDS<222>(2946)(2716)<223>互补链<221>CDS<222>(3252)…(2995)<223>互补链<221>CDS<222>(3676)…(3299)<223>互补链<221>CDS<222>(4124)…(3837)<223>互补链<221>CDS<222>(5214)…(4351)<223>互补链<400>13aa ttt gga agt gct cta tca aca gtt gaa gta aag gag att att agt 47Phe Gly Ser Ala Leu Ser Thr Val Glu Val Lys Glu Ile Ile Ser
1 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 Ser
20 25 30tac tca tat tgg aag tta cca act tgg taagcatcat atg ggt cta gca 144Tyr Ser Tyr Trp Lys Leu Pro Thr Trp Met Gly Leu Ala
35 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 Glu
65 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 Tyr
80 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 Pro
95 100 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 Tyr
110 115 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 Lys
145 150 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 Lys
160 165 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 Leu
175 180 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 Arg
190 195 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 Tyr
225 230 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 Ala
240 245 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 Arg
255 260 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 Thr
270 275 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 Asp
305 310 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 Arg
320 325 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 Glu
335 340 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 His
350 355 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 Asp
385 390 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 Thr
400 405 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 Lys
415 420 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 His
430 435 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 Ala
465 470 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 Leu
480 485 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 Val
495 500 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 Lys
510 515 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 Tyr
545 550 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 Asp
560 565 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 Gly
575 580 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 Tyr
590 595 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 Glu
625 630 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 Gly
640 645 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 Ala
655 660 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 Pro
670 675 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 Ser
705 710 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 Lys
720 725 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 Gln
735 740 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 Lys
750 755 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 Lys
785 790 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 Gln
800 805 810ttt tat aat aaa aat ggt gaa ttg gta act tat gat atc aag aca ctt 2496Phe Tyr Asn Lys Asn Gly Glu Leu Val Thr Tyr Asp Ile Lys Thr Leu
815 820 825caa caa ata aac cct taaccaaaag aagatctcat tgttaaagca ctgctttgtc 2551Gln Gln Ile Asn Pro
830aaagcaagtt acggtgattt tgaagtcatt ctatgtaacg agtagtgata aaagttggat 2611aatagcggtt ttcttttgca aagaaatggt atccatgtta gaatagtaaa aaaagaggag 2671gattcttgga ctaatgtcaa ataagtagac agaaaactgt gttattttattgcgt 2726taaaataatt 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 tacaggcgac 3025tggtcgttta 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>蛋白质<213>链球菌<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 Tyr
20 25 30Ser Tyr Trp Lys Leu Pro Thr Trp
35 40<210>15<211>793<212>蛋白质<213>链球菌<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 Ile
20 25 30Ser Ala Glu Glu Gly Ile Ser Ala Glu Gln Ile Val Val Lys Ile Thr
35 40 45Asp Gln Gly Tyr Val Thr Ser His Gly Asp His Tyr His Phe Tyr Asn
50 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 Leu
85 90 95Asp Gly Tyr Val Ile Lys Val Asn Gly Asn Tyr Tyr Val Tyr Leu Lys
100 105 110Pro Gly Ser Lys Arg Lys Asn Ile Arg Thr Lys Gln Gln Ile Ala Glu
115 120 125Gln Val Ala Lys Gly Thr Lys Glu Ala Lys Glu Lys Gly Leu Ala Gln
130 135 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 Thr
165 170 175Asp Ile Ile Asp Asp Leu Gly Asp Ala Tyr Leu Val Pro His Gly Asn
180 185 190His Tyr His Tyr Ile Pro Lys Lys Asp Leu Ser Pro Ser Glu Leu Ala
195 200 205Ala Ala Gln Ala Tyr Trp Ser Gln Lys Gln Gly Arg Gly Ala Arg Pro
210 215 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 Gly
245 250 255Gly Tyr His Pro Ala Pro Pro Arg Pro Asn Asp Ala Ser Gln Asn Lys
260 265 270His Gln Arg Asp Glu Phe Lys Gly Lys Thr Phe Lys Glu Leu Leu Asp
275 280 285Gln Leu His Arg Leu Asp Leu Lys Tyr Arg His Val Glu Glu Asp Gly
290 295 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 Leu
325 330 335Ser Pro Leu Glu Met Glu Leu Ala Asp Arg Tyr Leu Ala Gly Gln Thr
340 345 350Glu Asp Asn Asp Ser Gly Ser Glu His Ser Lys Pro Ser Asp Lys Glu
355 360 365Val Thr His Thr Phe Leu Gly His Arg Ile Lys Ala Tyr Gly Lys Gly
370 375 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 Gly
405 410 415Asp His Phe His Tyr Ile Gly Phe Gly Glu Leu Glu Gln Tyr Glu Leu
420 425 430Asp Glu Val Ala Asn Trp Val Lys Ala Lys Gly Gln Ala Asp Glu Leu
435 440 445Ala Ala Ala Leu Asp Gln Glu Gln Gly Lys Glu Lys Pro Leu Phe Asp
450 455 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 Leu
485 490 495Asp Leu Thr Gln Ile Ala Phe Ala Glu Gln Glu Leu Met Leu Lys Asp
500 505 510Lys Lys His Tyr Arg Tyr Asp Ile Val Asp Thr Gly Ile Glu Pro Arg
515 520 525Leu Ala Val Asp Val Ser Ser Leu Pro Met His Ala Gly Asn Ala Thr
530 535 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 Lys
565 570 575Tyr Val Met Gln His Pro Glu Val Arg Pro Asp Val Trp Ser Lys Pro
580 585 590Gly His Glu Glu Ser Gly Ser Val Ile Pro Asn Val Thr Pro Leu Asp
595 600 605Lys Arg Ala Gly Met Pro Asn Trp Gln Ile Ile His Ser Ala Glu Glu
610 615 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 Lys
645 650 655Asp Gly Ser Phe Ser Ile Pro Arg Ala Asp Gly Ser Ser Leu Arg Thr
660 665 670Ile Asn Lys Ser Asp Leu Ser Gln Ala Glu Trp Gln Gln Ala Gln Glu
675 680 685Leu Leu Ala Lys Lys Asn Thr Gly Asp Ala Thr Asp Thr Asp Lys Pro
690 695 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 Leu
725 730 735Pro Asp Tyr Gly Leu Asp Arg Ala Thr Leu Glu Asp His Ile Asn Gln
740 745 750Leu Ala Gln Lys Ala Asn Ile Asp Pro Lys Tyr Leu Ile Phe Gln Pro
755 760 765Glu Gly Val Gln Phe Tyr Asn Lys Asn Gly Glu Leu Val Thr Tyr Asp
770 775 780Ile Lys Thr Leu Gln Gln Ile Asn Pro785 790<210>16<211>715<212>蛋白质<213>链球菌<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 Tyr
20 25 30Leu Lys Pro Gly Ser Lys Arg Lys Asn Ile Arg Thr Lys Gln Gln Ile
35 40 45Ala Glu Gln Val Ala Lys Gly Thr Lys Glu Ala Lys Glu Lys Gly Leu
50 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 Ser
85 90 95Pro Thr Asp Ile Ile Asp Asp Leu Gly Asp Ala Tyr Leu Val Pro His
100 105 110Gly Asn His Tyr His Tyr Ile Pro Lys Lys Asp Leu Ser Pro Ser Glu
115 120 125Leu Ala Ala Ala Gln Ala Tyr Trp Ser Gln Lys Gln Gly Arg Gly Ala
130 135 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 Asp
165 170 175Asn Gly Gly Tyr His Pro Ala Pro Pro Arg Pro Asn Asp Ala Ser Gln
180 185 190Asn Lys His Gln Arg Asp Glu Phe Lys Gly Lys Thr Phe Lys Glu Leu
195 200 205Leu Asp Gln Leu His Arg Leu Asp Leu Lys Tyr Arg His Val Glu Glu
210 215 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 Ser
245 250 255Gln Leu Ser Pro Leu Glu Met Glu Leu Ala Asp Arg Tyr Leu Ala Gly
260 265 270Gln Thr Glu Asp Asn Asp Ser Gly Ser Glu His Ser Lys Pro Ser Asp
275 280 285Lys Glu Val Thr His Thr Phe Leu Gly His Arg Ile Lys Ala Tyr Gly
290 295 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 Lys
325 330 335His Gly Asp His Phe His Tyr Ile Gly Phe Gly Glu Leu Glu Gln Tyr
340 345 350Glu Leu Asp Glu Val Ala Asn Trp Val Lys Ala Lys Gly Gln Ala Asp
355 360 365Glu Leu Ala Ala Ala Leu Asp Gln Glu Gln Gly Lys Glu Lys Pro Leu
370 375 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 Asp
405 410 415Gln Leu Asp Leu Thr Gln Ile Ala Phe Ala Glu Gln Glu Leu Met Leu
420 425 430Lys Asp Lys Lys His Tyr Arg Tyr Asp Ile Val Asp Thr Gly Ile Glu
435 440 445Pro Arg Leu Ala Val Asp Val Ser Ser Leu Pro Met His Ala Gly Asn
450 455 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 Thr
485 490 495Val Lys Tyr Val Met Gln His Pro Glu Val Arg Pro Asp Val Trp Ser
500 505 510Lys Pro Gly His Glu Glu Ser Gly Ser Val Ile Pro Asn Val Thr Pro
515 520 525Leu Asp Lys Arg Ala Gly Met Pro Asn Trp Gln Ile Ile His Ser Ala
530 535 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 Val
565 570 575Trp Lys Asp Gly Ser Phe Ser Ile Pro Arg Ala Asp Gly Ser Ser Leu
580 585 590Arg Thr Ile Asn Lys Ser Asp Leu Ser Gln Ala Glu Trp Gln Gln Ala
595 600 605Gln Glu Leu Leu Ala Lys Lys Asn Thr Gly Asp Ala Thr Asp Thr Asp
610 615 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 Asp
645 650 655Ser Leu Pro Asp Tyr Gly Leu Asp Arg Ala Thr Leu Glu Asp His Ile
660 665 670Asn Gln Leu Ala Gln Lys Ala Asn Ile Asp Pro Lys Tyr Leu Ile Phe
675 680 685Gln Pro Glu Gly Val Gln Phe Tyr Asn Lys Asn Gly Glu Leu Val Thr
690 695 700Tyr Asp Ile Lys Thr Leu Gln Gln Ile Asn Pro705 710 715<210>17<211>77<212>蛋白质<213>链球菌<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 His
20 25 30Tyr Gln Asn Ile Lys Gln Val Gln Leu Asp Cys Phe Glu Tyr Ile Glu
35 40 45Asn Phe Tyr Asn Asn Tyr Asn Pro His Thr Ala Asn Leu Gly Leu Thr
50 55 60Pro Asn Gln Lys Glu Glu Asn Tyr Phe Asn Ala Ile Lys65 70 75<210>18<211>86<212>蛋白质<213>链球菌<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 His
20 25 30Thr Val Leu Lys Thr Glu Thr Phe Tyr Phe His Asn Arg Arg Lys Tyr
35 40 45Asn Lys Asp Ser Ile Thr Asn Ile Val Lys Asn Tyr Ile Thr Phe Tyr
50 55 60Asn Glu Thr Arg Ile Gln Gln Arg Leu Asn Asp Gln Ser Pro Val Gln65 70 75 80Tyr Arg Lys Leu Ile Ala
85<210>19<211>126<212>蛋白质<213>链球菌<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 Ile
20 25 30Met Lys Asn Asn Gly Trp Leu Cys Arg Thr Arg Thr Lys Lys Val Pro
35 40 45Asn Leu Gly Lys Ala Tyr Tyr Leu Thr Asp Asn Lys Leu Ser Arg Asp
50 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 Leu
85 90 95Tyr Asn Arg Glu Ile Ile Ala Tyr Thr Ile Ser Asp Cys Gln Asp Thr
100 105 110Asp Phe Val Leu Asp Thr Leu Asn Gln Leu Lys Leu Pro Lys
115 120 125<210>20<211>96<212>蛋白质<213>链球菌<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 Gly
20 25 30Ile Lys Asn Asn Ser Gln Ile Tyr Thr Trp Met Lys Trp Tyr Glu Asn
35 40 45Glu Glu Leu Tyr Arg Phe His Gln Gly Val Gly Lys Gln Tyr Thr Tyr
50 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 Lys
85 90 95<210>21<211>288<212>蛋白质<213>链球菌<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 Arg
20 25 30Trp Leu Asn Met Pro His Ser Ser Tyr Tyr Tyr Gln Ala Val Glu Ser
35 40 45Val Ser Glu Thr Glu Phe Glu Glu Thr Ile Lys Arg Ile Phe Leu Asp
50 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 Arg
85 90 95Leu Asn Leu Val Ser Val Tyr Gln Lys Ala Thr Phe Lys Pro His Ser
100 105 110Arg Gly Lys Asn Glu Ala Pro Ile Pro Asn His Leu Asp Arg Gln Phe
115 120 125Lys Gln Glu Arg Pro Leu Gln Ala Leu Val Thr Asp Leu Thr Tyr Val
130 135 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 Glu
165 170 175Leu Val Lys Gln Ala Ile Gln Ser Ile Pro Tyr Ala Leu Thr Lys Val
180 185 190Lys Met Phe His Ser Asp Arg Gly Lys Glu Phe Asp Asn Gln Leu Ile
195 200 205Asp Glu Ile Leu Glu Ala Phe Gly Ile Thr Arg Ser Leu Ser Gln Ala
210 215 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 Leu
245 250 255Ala Leu Lys Thr Lys Asp Tyr Val His Trp Trp Asn Tyr His Arg Ile
260 265 270His Gly Ser Leu Asn Tyr Gln Thr Pro Met Thr Lys Arg Leu Ile Ala
275 280 285<210>22<211>5058<212>DNA<213>链球菌<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 Asn
20 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 Gly
35 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 Asp
50 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 Asp
85 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 Ala
100 105 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 Val
115 120 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 Asp
130 135 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 Leu
165 170 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 Thr
180 185 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 Ser
195 200 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 Lys
210 215 220cttaaactaa atatgtgatc aatgataaag ggtggtgaag acatgaaagt gtctttgcct 733cttttcataa ggttagattt ggagacttt atg act gac ttg gaa aaa att att 786
Met Thr Asp Leu Glu Lys Ile Ile
225aaa 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 Val
250 255 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 Lys
265 270 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 Glu
280 285 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 Tyr
295 300 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 Gln
330 335 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 Ser
345 350 355tcc gca cat aaa aat cta aca gaa aca gtt aaa gct tac aaa gac tat 1218Ser Ala His Lys Asn Leu Thr Glu Thr Val Lys Ala Tyr Lys Asp Tyr
360 365 370cta ccc gat tat tta ccc ctg gtt cac cca tca ccg cga aat caa att 1266Leu Pro Asp Tyr Leu Pro Leu Val His Pro Ser Pro Arg Asn Gln Ile
375 380 385tgg cta aag aag aat cca tgg ttt gaa aaa gat cta atc gtt gat tta 1314Trp 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 Met
410 415aga gat aat cat cta cac acg tat ttt tcc tat gat tgt caa acg gca 1412Arg Asp Asn His Leu His Thr Tyr Phe Ser Tyr Asp Cys Gln Thr Ala
420 425 430ttt gag gac tat att aat ggt ttt aca ggt gaa ttt atc acg aca gaa 1460Phe Glu Asp Tyr Ile Asn Gly Phe Thr Gly Glu Phe Ile Thr Thr Glu
435 440 445cat ttt gat tta tca aat cct tac acc ggt caa gac gat gtt cct gat 1508His Phe Asp Leu Ser Asn Pro Tyr Thr Gly Gln Asp Asp Val Pro Asp
450 455 460tat agt gct tat tgt caa aaa ata gat tat ctt aat cag aaa tat gga 1556Tyr 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 gaa 1604Asn Arg Phe Lys Lys Gly Ile Glu Ile Gly Tyr Phe Lys Asp Arg Glu
485 490 495tca gat att tta gat tat tta aaa aat aaa gaa ttt gat tta aaa cta 1652Ser Asp Ile Leu Asp Tyr Leu Lys Asn Lys Glu Phe Asp Leu Lys Leu
500 505 510ttg tca atc cat cat aat ggt agg tat gat tat ctg caa gaa gaa gct 1700Leu Ser Ile His His Asn Gly Arg Tyr Asp Tyr Leu Gln Glu Glu Ala
515 520 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 Leu
530 535 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 tta 2283
Met Lys Arg Lys Asp Leu
545ttt ggt gat aaa caa act caa tac acg att aga aag tta agt gtt gga 2331Phe Gly Asp Lys Gln Thr Gln Tyr Thr Ile Arg Lys Leu Ser Val Gly
550 555 560gta gct tca gtt aca aca ggg gta tgt att ttt ctt cat agt cca cag 2379Val Ala Ser Val Thr Thr Gly Val Cys Ile Phe Leu His Ser Pro Gln
565 570 575gta ttt gct gaa gaa gta agt gtt tct cct gca act aca gcg att gca 2427Val 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 aaa 2475Glu Ser Asn Ile Asn Gln Val Asp Asn Gln Gln Ser Thr Asn Leu Lys
600 605 610gat gac ara aac tca aac tct gag acg gtt gtg aca ccc tca gat atg 2523Asp Asp Ile Asn Ser Asn Ser Glu Thr Val Val Thr Pro Ser Asp Met
615 620 625ccg gat acc aag caa tta gta tca gat gaa act gac act caa aag gga 2571Pro Asp Thr Lys Gln Leu Val Ser Asp Glu Thr Asp Thr Gln Lys Gly
630 635 640gtg aca gag ccg gat aag gcg aca agc ctg ctt gaa gaa aat aaa ggt 2619Val Thr Glu Pro Asp Lys Ala Thr Ser Leu Leu Glu Glu Asn Lys Gly
645 650 655cct gtt tca gat aaa aat acc tta gat tta aaa gta gca cca tct aca 2667Pro 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 agc 2715Leu Gln Asn Thr Pro Asp Lys Thr Ser Gln Ala Ile Gly Ala Pro Ser
680 685 690cct acc ttg aaa gta gct aat caa gct cca cgg att gaa aat ggt tac 2763Pro Thr Leu Lys Val Ala Asn Gln Ala Pro Arg Ile Glu Asn Gly Tyr
695 700 705ttt agg cta cat ctt aaa gaa ttg cct caa ggt cat cct gta gaa agc 2811Phe Arg Leu His Leu Lys Glu Leu Pro Gln Gly His Pro Val Glu Ser
710 715 720act gga ctt tgg ata tgg gga gat gtt gat caa ccg tct agt aat tgg 2859Thr Gly Leu Trp Ile Trp Gly Asp Val Asp Gln Pro Ser Ser Asn Trp
725 730 735cca aat ggt gct att cct atg act gat gct aag aaa gat gat tac ggt 2907Pro 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 tct 2955Tyr Tyr Val Asp Phe Lys Leu Ser Glu Lys Gln Arg Lys Gln Ile Ser
760 765 770ttt tta att aat aac aaa gca ggg aca aat tta agc ggc gat cat cat 3003Phe Leu Ile Asn Asn Lys Ala Gly Thr Asn Leu Ser Gly Asp His His
775 780 785att cca tta tta cga cct gag atg aac caa gtt tgg att gat gaa aag 3051Ile Pro Leu Leu Arg Pro Glu Met Asn Gln Val Trp Ile Asp Glu Lys
790 795 800tac ggt ata cat act tat caa ccc ctc aaa gaa ggg tat gtc cgt att 3099Tyr Gly Ile His Thr Tyr Gln Pro Leu Lys Glu Gly Tyr Val Arg Ile
805 810 815aac tat ttg agt tcc tcu agt aac tat gac cac tta tca gca tgg ctc 3147Asn 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 aat 3195Phe Lys Asp Val Ala Thr Xaa Ser Thr Thr Trp Pro Asp Gly Ser Asn
840 845 850ttt gtg aat caa gga cta tat gga agg tat att gat gta tca cta aaa 3243Phe Val Asn Gln Gly Leu Tyr Gly Arg Tyr Ile Asp Val Ser Leu Lys
855 860 865act aac gcc aaa gag att ggt ttt cta atc tta gat gaa agt aag aca 3291Thr Asn Ala Lys Glu Ile Gly Phe Leu Ile Leu Asp Glu Ser Lys Thr
870 875 880gga gat gca gtg aaa gtt caa ccc aac gac tat gtt ttt aga gat tta 3339Gly Asp Ala Val Lys Val Gln Pro Asn Asp Tyr Val Phe Arg Asp Leu
885 890 895gct aac cat aac caa att ttt gta aaa gat aag gat cca aag gtt tat 3387Ala 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 caa 3435Asn Asn Pro Tyr Tyr Ile Asp Gln Val Gln Leu Lys Asp Ala Gln Gln
920 925 930att gat tta aca agt att caa gca agt ttt aca act cta gat ggg gta 3483Ile Asp Leu Thr Ser Ile Gln Ala Ser Phe Thr Thr Leu Asp Gly Val
935 940 945gat aaa act gaa att tta aaa gaa ttg aaa gtg act gat aaa aat caa 3531Asp Lys Thr Glu Ile Leu Lys Glu Leu Lys Val Thr Asp Lys Asn Gln
950 955 960aat gct ata caa att tct gat atc act ctc gat act agt aaa tct ctt 3579Asn Ala Ile Gln Ile Ser Asp Ile Thr Leu Asp Thr Ser Lya Ser Leu
965 970 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 Lys
1000 1005 1010gac 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 Gln
1015 1020 1025gat 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 Ser
1030 1035 1040gtc 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 Ala
1045 1050 1055act 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 Leu1060 1065 1070 1075gat 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 Tyr
1080 1085 1090gaa 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 Ala
1095 1100 1105aag 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 Lys
1110 1115 1120acg 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 Asn
1125 1130 1135tta 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 Val1140 1145 1150 1155ata 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 Asp
1160 1165 1170gga 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 Lys
1175 1180 1185cta 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 Pro
1190 1195 1200gta 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 Thr
1205 1210 1215gct 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 Gln1220 1225 1230 1235agc 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 Pro
1240 1245 1250tca 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 Lys
1255 1260 1265cgt 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 Lys
1270 1275 1280act 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 Asn
1285 1290 1295gaa 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 Thr1300 1305 1310 1315act 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 Leu
1320 1325 1330aca 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 Asp
1335 1340 1345cat 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 Ile
1350 1355 1360aat 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 Gly
1365 1370 1375gat 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 Ser1380 1385 1390 1395acc 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 Lys
1400 1405 1410tct 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 Pro
1415 1420 1425caa 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 Asn
1430 1435 1440ttt 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 His
1445 1450 1455gat aac ctt acc ttg cat gat gtg att gca aaa tca att 5058Asp Asn Leu Thr Leu His Asp Val Ile Ala Lys Ser Ile1460 1465 1470<210>23<211>221<212>蛋白质<213>链球菌<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 Asn
20 25 30Gly Ala Thr Tyr Leu Ala Leu Tyr Leu Ser Gln Phe Asp Phe Glu Gly
35 40 45Pro Ala Arg Ala Phe Leu Asp Val Thr Ala Asn Ile Ile His Glu Asp
50 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 Asp
85 90 95Asp Asn Thr Ile Ala Leu Asn Phe Arg Tyr Pro Gln Gly Thr Asp Ala
100 105 110Lys Thr Ile Gln Thr Lys Leu Glu Lys Leu Asn Gly Val Glu Lys Val
115 120 125Thr Leu Ser Asp His Glu His Thr Pro His Tyr Val Pro Met Asp Asp
130 135 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 Leu
165 170 175Glu Arg Gly Val Ala Tyr Gly Ala Met Phe Pro Gly Asp Glu Asn Thr
180 185 190Met His Gln Ala Asn Glu Tyr Met Pro Leu Glu Asn Ile Phe Arg Ser
195 200 205Ala Ala Ile Tyr Ala Glu Ala Ile Tyr Glu Leu Ile Lys
210 215 220<210>24<211>194<212>蛋白质<213>链球菌<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 Pro
20 25 30Lys Thr Ala Arg Ile Asn Ile Val Gly Gln Ala Pro Gly Leu Lys Thr
35 40 45Gln Glu Ala Arg Leu Tyr Trp Lys Asp Lys Ser Gly Asp Arg Leu Arg
50 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 Gly
85 90 95Asp Leu Pro Pro Arg Lys Gly Phe Ala Glu Lys Trp His Pro Leu Ile
100 105 110Leu Lys Glu Met Pro Asn Val Gln Leu Thr Leu Leu Val Gly Gln Tyr
115 120 125Ala Gln Lys Tyr Tyr Leu Gly Ser Ser Ala His Lys Asn Leu Thr Glu
130 135 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 Phe
165 170 175Glu Lys Asp Leu Ile Val Asp Leu Gln Lys Ile Val Ala Asp Ile Leu
180 185 190Lys Asp<210>25<211>126<212>蛋白质<213>链球菌<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 Thr
20 25 30Glu His Phe Asp Leu Ser Asn Pro Tyr Thr Gly Gln Asp Asp Val Pro
35 40 45Asp Tyr Ser Ala Tyr Cys Gln Lys Ile Asp Tyr Leu Asn Gln Lys Tyr
50 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 Lys
85 90 95Leu Leu Ser Ile His His Asn Gly Arg Tyr Asp Tyr Leu Gln Glu Glu
100 105 110Ala Leu Lys Val Pro Thr Lys Gly Ala Phe Ser Arg Leu Leu
115 120 125<210>26<211>931<212>蛋白质<213>链球菌<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 Ile
20 25 30Phe Leu His Ser Pro Gln Val Phe Ala Glu Glu Val Ser Val Ser Pro
35 40 45Ala Thr Thr Ala Ile Ala Glu Ser Asn Ile Asn Gln Val Asp Asn Gln
50 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 Glu
85 90 95Thr Asp Thr Gln Lys Gly Val Thr Glu Pro Asp Lys Ala Thr Ser Leu
100 105 110Leu Glu Glu Asn Lys Gly Pro Val Ser Asp Lys Asn Thr Leu Asp Leu
115 120 125Lys Val Ala Pro Ser Thr Leu Gln Asn Thr Pro Asp Lys Thr Ser Gln
130 135 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 Gln
165 170 175Gly His Pro Val Glu Ser Thr Gly Leu Trp Ile Trp Gly Asp Val Asp
180 185 190Gln Pro Ser Ser Asn Trp Pro Asn Gly Ala Ile Pro Met Thr Asp Ala
195 200 205Lys Lys Asp Asp Tyr Gly Tyr Tyr Val Asp Phe Lys Leu Ser Glu Lys
210 215 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 Gln
245 250 255Val Trp Ile Asp Glu Lys Tyr Gly Ile His Thr Tyr Gln Pro Leu Lys
260 265 270Glu Gly Tyr Val Arg Ile Asn Tyr Leu Ser Ser Ser Ser Asn Tyr Asp
275 280 285His Leu Ser Ala Trp Leu Phe Lys Asp Val Ala Thr Xaa Ser Thr Thr
290 295 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 Ile
325 330 335Leu Asp Glu Ser Lys Thr Gly Asp Ala Val Lys Val Gln Pro Asn Asp
340 345 350Tyr Val Phe Arg Asp Leu Ala Asn His Asn Gln Ile Phe Val Lys Asp
355 360 365Lys Asp Pro Lys Val Tyr Asn Asn Pro Tyr Tyr Ile Asp Gln Val Gln
370 375 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 Lys
405 410 415Val Thr Asp Lys Asn Gln Asn Ala Ile Gln Ile Ser Asp Ile Thr Leu
420 425 430Asp Thr Ser Lys Ser Leu Leu Ile Ile Lys Gly Asp Phe Asn Pro Lys
435 440 445Gln Gly His Phe Asn Ile Ser Tyr Asn Gly Asn Asn Val Met Thr Arg
450 455 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 Trp
485 490 495Ser Pro Ser Ala Asp Ser Val Thr Met Ile Ile Tyr Asp Lys Asp Asn
500 505 510Gln Asn Arg Val Val Ala Thr Thr Pro Leu Val Lys Asn Asn Lys Gly
515 520 525Val Trp Gln Thr Ile Leu Asp Thr Lys Leu Gly Ile Lys Asn Tyr Thr
530 535 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 Thr
565 570 575Val Asn Asp Asp Ile Lys Thr Ala Lys Ala Ala Phe Val Asn Pro Ser
580 585 590Gln Leu Gly Pro Gln Asn Leu Ser Phe Ala Lys Ile Ala Asn Phe Lys
595 600 605Gly Arg Gln Asp Ala Val Ile Tyr Glu Ala His Val Arg Asp Phe Thr
610 615 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 Thr
645 650 655His Ile Gln Leu Leu Pro Val Leu Ser Tyr Phe Tyr Val Asn Glu Met
660 665 670Asp Lys Ser Arg Ser Thr Ala Tyr Thr Ser Ser Asp Asn Asn Tyr Asn
675 680 685Trp Gly Tyr Asp Pro Gln Ser Tyr Phe Ala Leu Ser Gly Met Tyr Ser
690 695 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 Val
725 730 735Tyr Asn His Thr Ala Lys Thr Tyr Leu Phe Glu Asp Ile Glu Pro Asn
740 745 750Tyr Tyr His Phe Met Asn Glu Asp Gly Ser Pro Arg Glu Ser Phe Gly
755 760 765Gly Gly Arg Leu Gly Thr Thr His Ala Met Ser Arg Arg Val Leu Val
770 775 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 Tyr
805 810 815Lys Glu Ala Lys Ala Ile Asn Pro Asn Met Ile Met Ile Gly Glu Gly
820 825 830Trp Arg Thr Phe Gln Gly Asp Gln Gly Gln Pro Val Lys Pro Ala Asp
835 840 845Gln Asp Trp Met Lys Ser Thr Asp Thr Val Gly Val Phe Ser Asp Asp
850 855 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 Ile
885 890 895Lys Ala Gln Pro Gly Asn Phe Glu Ala Asp Ser Pro Gly Asp Val Val
900 905 910Gln Tyr Ile Ala Ala His Asp Asn Leu Thr Leu His Asp Val Ile Ala
915 920 925Lys Ser Ile
930<210>27<211>5607<212>DNA<213>链球菌<220><221>CDS<222>(2)…(301)<400>27a att caa agt ttg aca gaa ggt caa ctt cgt tct gat atc cct gag ttc 49Ile 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 Thr
20 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 Gly
35 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 Ile
50 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 Leu
85 90 95cgc gca ttg caa ggtaaagctg cacgtattaa agaaatccgt cgttaatttt 341Arg Ala Leu Gln
100gatgatcaga 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>蛋白质<213>链球菌<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 Thr
20 25 30Arg Glu Arg Ile Gln Ile Phe Glu Gly Val Val Ile Ser Arg Lys Gly
35 40 45Gln Gly Ile Ser Glu Met Tyr Thr Val Arg Lys Ile Ser Gly Gly Ile
50 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 Leu
85 90 95Arg Ala Leu Gln Gly Lys Ala Ala Arg Ile Lys Glu Ile Arg Arg
100 105 110<210>29<211>173<212>蛋白质<213>链球菌<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 Asn
20 25 30Gln Arg Tyr Leu Pro Thr Lys Asn Lys Ser Ser Ile Arg Asn Ile Pro
35 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 His
85 90 95Thr Phe Ala Ser Tyr Leu Ile Ser Ile Ser Gln Val Leu Asp His Glu
100 105 110Asn Leu Asn Ile Thr Leu Glu Val Tyr Ala His Gln Leu Gln Glu Gln
115 120 125Lys Asp Arg Asn Asp Lys Leu Asn Gln Arg Asn Leu Gly Gln Asn Ser
130 135 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 His
165 170<210>30<211>389<212>蛋白质<213>链球菌<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 Asn
20 25 30Gln Phe Lys Asn Ile Glu Lys Ile Lys Glu Val Glu Glu Lys Ile Phe
35 40 45Gln Tyr Asp Gly Leu Ala Lys Leu Lys Asp Leu Lys Val Val Ser Gly
50 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 Ser
85 90 95Ser Gly Val Phe Ser Phe Lys Glu Gly Lys Asn Ile Glu Glu Asn Asp
100 105 110Lys Asn Ser Ile Leu Val His Glu Glu Phe Ala Lys Gln Asn Lys Leu
115 120 125Lys Leu Gly Asp Glu Ile Asp Leu Glu Leu Leu Asp Thr Glu Lys Ser
130 135 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 Asn
165 170 175Met Val Phe Val Asp Tyr Ser Thr Ser Gln Glu Ile Leu Asn Lys Ser
180 185 190Glu Asn Asn Arg Ile Ala Asn Lys Ile Leu Met Tyr Ser Gly Ser Leu
195 200 205Glu Ser Thr Glu Leu Ala Leu Asn Lys Leu Lys Asp Phe Lys Ile Asp
210 215 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 Tyr
245 250 255Ser Ile Met Leu Gly Gly Ile Val Val Leu Ser Leu Ile Leu Ile Leu
260 265 270Trp Leu Arg Glu Arg Ile Tyr Glu Ile Gly Ile Phe Leu Ser Ile Gly
275 280 285Thr Thr Lys Ile Gln Ile Ile Arg Gln Phe Ile Phe Glu Leu Ile Phe
290 295 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 Ile
325 330 335Phe Gly Gly Ser Leu Ile Asn Lys Ser Ser Phe Met Leu Asn Ile Thr
340 345 350Thr Leu Ala Glu Ser Tyr Leu Ile Leu Ile Ser Ile Ile Val Leu Ser
355 360 365Val Val Met Ala Ser Ser Leu Ile Leu Phe Lys Lys Pro Gln Glu Ile
370 375 380Leu Ser Lys Ile Ser385<210>31<211>169<212>蛋白质<213>链球菌<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 Lys
20 25 30Phe Tyr Ala Ile Val Gly Lys Ser Gly Thr Gly Lys Ser Thr Leu Leu
35 40 45Ser Leu Leu Ala Gly Leu Asp Lys Val Gln Thr Gly Lys Ile Leu Phe
50 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 Ser
85 90 95Pro Ile Glu Asn Ile Arg Leu Val Asn Lys Ser Val Asp Glu Ser Ile
100 105 110Leu Phe Glu Leu Gly Leu Asp Lys Lys Gln Ile Lys Arg Asn Val Met
115 120 125Lys Leu Ser Gly Gly Gln Gln Gln Arg Val Ala Ile Ala Arg Ala Leu
130 135 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 Ile
165<210>32<211>4171<212>DNA<213>链球菌<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>蛋白质<213>链球菌<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 Leu
20 25 30His Ala Asp Asn His Tyr His Phe Phe Asn Gly Lys Ser Leu Ala Thr
35 40 45Phe Asn Thr Asn Gln Leu Ile Arg Glu Val Val Tyr Val Glu Ile Ser
50 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 Ser
85 90 95Pro Tyr His Gln Gly Ile Asn Asp Pro Ala Ala Asp Gln Lys Thr Tyr
100 105 110Gln Met Glu Gly Ala Leu Ala Val Lys Gln Pro Lys His Ile Gln Val
115 120 125Asp Thr Lys Pro Phe Lys Glu Glu Val Lys His Pro Ser Lys Leu Pro
130 135 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 Gly
165 170 175Val Gly Thr Ala Gly Ser Thr Gly Phe Met Thr Ser Gly Asp Tyr Gln
180 185 190Gln Ile Gln Ser Phe Lys Ala Val Ile Asp Trp Leu Asn Gly Lys Val
195 200 205Thr Ala Phe Thr Ser His Lys Arg Asp Lys Gln Val Lys Ala Asp Trp
210 215 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 Ile
245 250 255Ala Glu Ala Ala Ile Ser Thr Trp Tyr Asp Tyr Tyr Arg Glu Asn Gly
260 265 270Leu Val Cys Ser Pro Gly Gly Tyr Pro Gly Glu Asp Leu Asp Val Leu
275 280 285Thr Glu Leu Thr Tyr Ser Arg Asn Leu Leu Ala Gly Asp Tyr Ile Lys
290 295 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 Tyr
325 330 335Leu Thr His Val Asn Asn Val Lys Ser Arg Val Val Tyr Thr His Gly
340 345 350Leu Gln Asp Trp Asn Val Lys Pro Arg His Val Tyr Lys Val Phe Asn
355 360 365Ala Leu Pro Gln Thr Ile Lys Lys His Leu Phe Leu His Gln Gly Gln
370 375 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 Gln
405 410 415Leu Glu Glu Val Ile Trp Gln Asp Asn Thr Thr Glu Gln Thr Trp Gln
420 425 430Val Leu Asp Ala Phe Gly Gly Asn His Gln Glu Gln Ile Gly Leu Gly
435 440 445Asp Ser Lys Lys Leu Ile Asp Asn His Tyr Asp Lys Glu Ala Phe Asp
450 455 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 Asn
485 490 495Tyr Leu Leu Asn Gly Gln Cys Lys Leu His Leu Arg Val Lys Thr Ser
500 505 510Asp Lys Lys Ala Ile Leu Ser Ala Gln Ile Leu Asp Tyr Gly Pro Lys
515 520 525Lys Arg Phe Lys Asp Thr Pro Thr Ile Lys Phe Leu Asn Ser Leu Asp
530 535 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 Asn
565 570 575Arg Thr Asp Leu Leu Thr Ile Glu Ala Ile Glu Pro Glu Gln Trp Phe
580 585 590Asp Ile Glu Phe Ser Leu Gln Pro Ser Ile Tyr Gln Leu Ser Lys Gly
595 600 605Asp Asn Leu Arg Ile Ile Leu Tyr Thr Thr Asp Phe Glu His Thr Ile
610 615 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 Asn
645<210>34<211>119<212>蛋白质<213>链球菌<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 Asp
20 25 30His Asp Asn Lys Asn Ile Pro Gln Val Val Ala Thr Ile Val Asp Asp
35 40 45Leu Gln Gly Ser Gly Ser Ser Asn His Phe Trp Tyr Phe Gly Asn Thr
50 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 Ala
85 90 95Ile Asn Asn Trp Lys Ser Leu Leu Gln Thr Gln Leu Glu Ala Leu Asn
100 105 110Asp Thr Leu Ala Ile Phe Gln
115<210>35<211>326<212>蛋白质<213>链球菌<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 Asp
20 25 30lle Glu Ala Ala Leu Ser Gln Leu Thr Ala Ala Gly Gly Lys Gln Leu
35 40 45Arg Pro Ala Phe Phe Tyr Leu Phe Ser Gln Leu Gly Asn Lys Glu Asn
50 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 Leu
85 90 95Arg Arg Gly Asn Met Thr Ile Gln Ser Lys Phe Gly Lys Asp Ile Ala
100 105 110Val Tyr Thr Gly Asp Leu Leu Phe Thr Val Phe Phe Asp Leu Ile Leu
115 120 125Glu Ser Met Thr Asp Thr Pro Phe Met Arg Ile Asn Ala Lys Ser Met
130 135 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 Thr
165 170 175Ala Glu Leu Phe Lys Leu Ala Ser Lys Glu Gly Ala Tyr Phe Gly Gly
180 185 190Ala Glu Lys Glu Val Val Arg Leu Ala Gly His Ile Gly Phe Asn Ile
195 200 205Gly Met Thr Phe Gln Ile Leu Asp Asp Ile Leu Asp Tyr Thr Ala Asp
210 215 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 Phe
245 250 255Lys Pro Ile Leu Asp Lys Lys Thr Asp Met Ala Thr Glu Asp Met Glu
260 265 270Lys Ile Ala Tyr Leu Val Val Ser His Arg Gly Val Asp Lys Ala Arg
275 280 285His Leu Ala Arg Lys Phe Thr Glu Lys Ala Ile Ser Asp Ile Asn Lys
290 295 300Leu Pro Gln Asn Ser Ala Lys Lys Gln Leu Leu Gln Leu Thr Asn Tyr305 310 315 320Leu Leu Lys Arg Lys Ile
325<210>36<211>247<212>蛋白质<213>链球菌<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 Ile
20 25 30Lys Glu Gly Glu Lys Ile Ala Ile Leu Gly Arg Ser Gly Ser Gly Lys
35 40 45Ser Thr Leu Ala Ser Leu Leu Arg Gly Asp Leu Lys Ala Ser Gln Gly
50 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 Thr
85 90 95Thr Leu Leu Asn Asn Ile Arg Ile Gly Asn Gln Asp Ala Ser Glu Glu
100 105 110Asp Val Trp Lys Val Leu Glu Arg Val Gly Leu Lys Glu Met Val Thr
115 120 125Asp Leu Ser Asp Gly Leu Tyr Thr Met Val Asp Glu Ala Gly Leu Arg
130 135 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 Asp
165 170 175Pro Ile Thr Glu Gln Ala Leu Leu Arg Val Phe Met Lys Glu Leu Glu
180 185 190Gly Lys Thr Leu Val Trp Ile Thr His His Leu Lys Gly Ile Glu His
195 200 205Ala Asp Arg Ile Leu Phe Ile Glu Asn Gly Gln Leu Glu Leu Glu Gly
210 215 220Ser Pro Gln Glu Leu Ser Gln Ser Ser Gln Arg Tyr Arg Gln Leu Lys225 230 235 240Ala Ala Asp Asp Gly Asp Leu
245<210>37<211>3480<212>DNA<213>链球菌<400>37aattctattt ggaggttttt cttgaataaa tggttagtta aggcaagttc cttagttgtt 50ttaggtggta 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>蛋白质<213>链球菌<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 Val
20 25 30Leu Ala Asp Thr Tyr Val Arg Pro Ile Asp Asn Gly Arg Ile Thr Thr
35 40 45Gly Phe Asn Gly Tyr Pro Gly His Cys Gly Val Asp Tyr Ala Val Pro
50 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 Cys
85 90 95Val Met Ile Gln His Ala Asp Gly Met His Ser Gly Tyr Ala His Met
100 105 110Ser Arg Val Val Ala Arg Thr Gly Glu Lys Val Lys Gln Gly Asp Ile
115 120 125Ile Gly Tyr Val Gly Ala Thr Gly Met Ala Thr Gly Pro His Leu His
130 135 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 Gly
165 170 175Lys Thr Gln Ala Ser Ala Pro Ser Ile Lys Pro Leu Gln Ser Ala Pro
180 185 190Val Gln Asn Gln Ser Ser Lys Leu Lys Val Tyr Arg Val Asp Glu Leu
195 200 205Gln Lys Val Asn Gly Val Trp Leu Val Lys Asn Asn Thr Leu Thr Pro
210 215 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 Lys
245 250 255Gly Gly Tyr Phe Ile Phe Asn Pro Lys Thr Leu Lys Thr Val Glu Lys
260 265 270Pro Ile Gln Gly Thr Ala Gly Leu Thr Trp Ala Lys Thr Arg Phe Ala
275 280 285Asn Gly Ser Ser Val Trp Leu Arg Val Asp Asn Ser Gln Glu Leu Leu
290 295 300Tyr Lys305<210>39<211>434<212>蛋白质<213>链球菌<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 Trp
20 25 30Thr Ala Arg Thr Val Ser Glu Val Lys Ala Asp Leu Val Lys Gln Asp
35 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 Thr
85 90 95Tyr Asp Gln Lys Ser His Thr Ala Thr Ser Met Lys Ile Glu Thr Pro
100 105 110Ala Thr Asn Ala Ala Gly Gln Thr Thr Ala Thr Val Asp Leu Lys Thr
115 120 125Asn Gln Val Ser Val Ala Asp Gln Lys Val Ser Leu Asn Thr Ile Ser
130 135 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 Gln
165 170 175Glu Gln Ala Val Ser Gln Ala Ala Ala Asn Glu Gln Val Ser Thr Ala
180 185 190Pro Val Lys Ser Ile Thr Ser Glu Val Pro Ala Ala Lys Glu Glu Val
195 200 205Lys Pro Thr Gln Thr Ser Val Ser Gln Ser Thr Thr Val Ser Pro Ala
210 215 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 Pro
245 250 255Lys Val Glu Thr Gly Ala Ser Pro Glu His Val Ser Ala Pro Ala Val
260 265 270Pro Val Thr Thr Thr Ser Thr Ala Thr Asp Ser Lys Leu Gln Ala Thr
275 280 285Glu Val Lys Ser Val Pro Val Ala Gln Lys Ala Pro Thr Ala Thr Pro
290 295 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 Ala
325 330 335Ser Thr Tyr Gly Val Asn Glu Phe Ser Thr Tyr Arg Ala Gly Asp Pro
340 345 350Gly Asp His Gly Lys Gly Leu Ala Val Asp Phe Ile Val Gly Lys Asn
355 360 365Gln Ala Leu Gly Asn Glu Val Ala Gln Tyr Ser Thr Gln Asn Met Ala
370 375 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 Asp
405 410 415Arg Gly Gly Val Thr Ala Asn His Tyr Asp His Val His Val Ser Phe
420 425 430Asn Lys<210>40<211>232<212>蛋白质<213>链球菌<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 Glu
20 25 30Ser Gly Gly Val Met Pro Leu Leu Ala Leu Ala Ala Gln Glu Ala Gly
35 40 45Ala Val Gly Ile Arg Ala Asn Ser Val Arg Asp Ile Lys Glu Ile Gln
50 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 Leu
85 90 95Ala Ser Leu Asp Ile Ala Val Ile Ala Leu Asp Cys Thr Leu Arg Glu
100 105 110Arg His Asp Gly Leu Ser Val Ala Glu Phe Ile Gln Lys Ile Lys Gly
115 120 125Lys Tyr Pro Glu Gln Leu Leu Met Ala Asp Ile Ser Thr Phe Glu Glu
130 135 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 Glu
165 170 175Leu Leu Asn Lys Leu Cys Gln Ala Gly Ile Asp Val Ile Ala Glu Gly
180 185 190Lys Ile His Thr Pro Lys Gln Ala Asn Glu Ile Asn His Ile Gly Val
195 200 205Ala Gly Ile Val Val Gly Gly Ala Ile Thr Arg Pro Lys Glu Ile Ala
210 215 220Glu Arg Phe Ile Ser Gly Leu Ser225 230<210>41<211>39<212>蛋白质<213>链球菌<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 Ala
20 25 30Glu Lys Asp Lys Val Glu Ile
35<210>42<211>1305<212>DNA<213>链球菌<400>42atgaaaatga ataaaaaggt actattgaca tcgacaatgg cagcttcgct attatcagtc 60gcaagtgttc 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>链球菌<400>43caagaaacag atacgacgtg gacagcacgt actgtttcag aggtaaaggc tgatttggta 60aagcaagaca 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>蛋白质<213>链球菌<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 Tyr
20 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 Pro
50 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 Thr
85 90 95Ala Thr Val Asp Leu Lys Thr Asn Gln Val Ser Val Ala Asp Gln Lys
100 105 110Val Ser Leu Asn Thr Ile Ser Glu Gly Met Thr Pro Glu Ala Ala Thr
115 120 125Thr Ile Val Ser Pro Met Lys Thr Tyr Ser Ser Ala Pro Ala Leu Lys
130 135 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 Val
165 170 175Pro Ala Ala Lys Glu Glu Val Lys Pro Thr Gln Thr Ser Val Ser Gln
180 185 190Ser Thr Thr Val Ser Pro Ala Ser Val Ala Ala Glu Thr Pro Ala Pro
195 200 205Val Ala Lys Val Ala Pro Val Arg Thr Val Ala Ala Pro Arg Val Ala
210 215 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 Thr
245 250 255Asp Ser Lys Leu Gln Ala Thr Glu Val Lys Ser Val Pro Val Ala Gln
260 265 270Lys Ala Pro Thr Ala Thr Pro Val Ala Gln Pro Ala Ser Thr Thr Asn
275 280 285Ala Val Ala Ala His Pro Glu Asn Ala Gly Leu Gln Pro His Val Ala
290 295 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 Val
325 330 335Asp Phe Ile Val Gly Lys Asn Gln Ala Leu Gly Asn Glu Val Ala Gln
340 345 350Tyr Ser Thr Gln Asn Met Ala Ala Asn Asn Ile Ser Tyr Val Ile Trp
355 360 365Gln Gln Lys Phe Tyr Ser Asn Thr Asn Ser Ile Tyr Gly Pro Ala Asn
370 375 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 Lys
405
Claims (48)
1．分离的多核苷酸，其编码的多肽与选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，类似物或衍生物。
2．根据权利要求1的多核苷酸，其中所述多核苷酸编码的多肽与第二多肽有至少95％的一致性。
3．分离的多核苷酸，其所编码的多肽能产生对含有选自如下的序列的多肽有特异性结合性的抗体：
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，类似物或衍生物。
4．与权利要求1的多核苷酸互补的分离的多核苷酸。
5．与权利要求3的多核苷酸互补的分离的多核苷酸。
6．权利要求1的多核苷酸，其中所述多核苷酸是DNA。
7．权利要求3的多核苷酸，其中所述多核苷酸是DNA。
8．权利要求1的多核苷酸，其中所述多核苷酸是RNA。
9．权利要求3的多核苷酸，其中所述多核苷酸是RNA。
10．在严格条件下与含有选自下组之序列的第二多核苷酸杂交的多核苷酸：
SEQ ID NO:1,SEQ ID NO:7,SEQ ID NO:13,SEQ ID
NO: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，类似物或衍生物。
11．在严格条件下与具有选自下面的一个序列的第二多核苷酸杂交的多核苷酸：SEQ ID NO:37,SEQ ID NO:42和SEQ ID NO:43。
12．根据权利要求11的多核苷酸，其在严格条件下与具有SEQ IDNO:37序列的第二多核苷酸杂交。
13．根据权利要求11的多核苷酸，其在严格条件下与具有SEQ IDNO:42序列的第二多核苷酸杂交。
14．根据权利要求11的多核苷酸，其在严格条件下与具有SEQ IDNO:43序列的第二多核苷酸杂交。
15．根据权利要求10的多核苷酸，其中所述多核苷酸具有与所述第二多核苷酸至少95％的互补性。
16．根据权利要求11的多核苷酸，其中所述多核苷酸具有与所述第二多核苷酸至少95％的互补性。
17．包含权利要求1的多核苷酸的载体，其中所述多核苷酸与表达调控区操作连接。
18．包含权利要求3的多核苷酸的载体，其中所述多核苷酸与表达调控区操作连接。
19．用权利要求17的载体转染的宿主细胞。
20．用权利要求18的载体转染的宿主细胞。
21．制备多肽的方法，包括在适合表达所述多肽的条件下培养权利要求19的宿主细胞。
22．制备多肽的方法，包括在适合表达所述多肽的条件下培养权利要求20的宿主细胞。
23．与含有选自下组之序列的第二多肽有至少70％一致性的分离的多肽：
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，类似物或衍生物。
24．具有SEQ ID NO：39序列的权利要求23的分离的多肽。
25．具有SEQ ID NO：44序列的权利要求23的分离的多肽。
26．能产生对于含有选自下组之序列的第二多肽有结合特异性的抗体的分离的多肽：
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，类似物或衍生物。
27．具有SEQ ID NO:39序列的权利要求26的分离的多肽。
28．具有SEQ ID NO:44序列的权利要求26的分离的多肽。
29．具有选自下组之氨基酸序列的分离的多肽：
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，类似物或衍生物。
30．具有SEQ ID NO:39的氨基酸序列的权利要求29的分离的多肽。
31．具有SEQ ID NO:44的氨基酸序列的分离的多肽。
32．权利要求29-31任一项的分离的多肽，其中N-末端Met残基缺失。
33．权利要求29-30任一项的分离的多肽，其中分泌氨基酸序列缺失。
34．疫苗组合物，包括根据权利要求23-31任一项的一种多肽和药学可接受载体，稀释剂或佐剂。
35．疫苗组合物，包括根据权利要求32的一种多肽和药学可接受载体，稀释剂或佐剂。
36．疫苗组合物，包括根据权利要求33的一种多肽和药学可接受载体，稀释剂或佐剂。
37．一种治疗或预防怀疑感染链球菌的动物链球菌感染的方法，包括对所述动物施用治疗或预防量的权利要求34的组合物。
38．一种治疗或预防怀疑感染链球菌的动物链球菌感染的方法，包括对所述动物施用治疗或预防量的权利要求35的组合物。
39．一种治疗或预防怀疑感染链球菌的动物链球菌感染的方法，包括对所述动物施用治疗或预防量的权利要求36的组合物。
40．根据权利要求37-39任一项的方法，其中所述动物是牛。
41．根据权利要求37-39任一项的方法，其中所述动物是人。
42．根据权利要求37-39任一项的方法，其中所述细菌感染选自A组链球菌和B组链球菌。
43．根据权利要求42的方法，其中所述细菌感染是B组链球菌。
44．根据权利要求34的一种疫苗组合物治疗或预防怀疑或者感染了链球菌的动物链球菌感染的用途，包括对所述动物施用治疗或预防量的该组合物。
45．根据权利要求35-36任一项的一种疫苗组合物治疗或预防怀疑或者感染了链球菌的动物链球菌感染的用途，包括对所述动物施用治疗或预防量的该组合物。
46．根据权利要求23-31任一项的一种疫苗组合物制备用于治疗或预防怀疑感染了链球菌的动物链球菌感染的疫苗的用途，所述治疗或预防包括对所述动物施用治疗或预防量的该组合物。
47．根据权利要求32的一种疫苗组合物在制备用于治疗或预防怀疑感染了链球菌的动物链球菌感染的疫苗中的用途，所述治疗或预防包括对所述动物施用治疗或预防量的该组合物。
48．根据权利要求33的一种疫苗组合物在制备用于治疗或预防怀疑感染了链球菌的动物链球菌感染的疫苗中的用途，所述治疗或预防包括对所述动物施用治疗或预防量的该组合物。
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| GB0105922D0 (en) * | 2001-03-09 | 2001-04-25 | Microscience Ltd | Genes and proteins, and their use |
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| DE60124132T2 (de) * | 2000-06-12 | 2007-09-06 | The University Of Saskatchewan | Immunisation von milchvieh mit gapc protein gegen streptococcus infektion |
| US6833134B2 (en) | 2000-06-12 | 2004-12-21 | University Of Saskacthewan | Immunization of dairy cattle with GapC protein against Streptococcus infection |
| US6866855B2 (en) | 2000-06-12 | 2005-03-15 | University Of Saskatchewan | Immunization of dairy cattle with GapC protein against Streptococcus infection |
| EP1294771B1 (en) | 2000-06-12 | 2008-10-29 | University Of Saskatchewan | Chimeric GapC protein from Streptococcus and its use in vaccination and diagnosis |
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