活疫苗及生产方法 【技术领域】
本发明属于病毒学和疫苗发展的领域，涉及生产病毒疫苗的改良方法以及用该方法可获得的疫苗，所述病毒疫苗尤其是完整病毒疫苗，最好是减毒活疫苗。
【发明背景】
流感血凝素(HA)抗原是宿主对病毒的保护性免疫反应的主要靶。
回收新病毒分离物的一般实践涉及从鼻拭子和喉拭子或从类似来源回收，然后在含胚的鸡蛋中培养所述分离物。病毒适应其鸡蛋宿主，因此可以在鸡蛋中进行所述病毒的大规模生产。然而，所述涉及用含胚鸡蛋生产流感疫苗的常规方法异常麻烦，涉及每周处理成千上万个鸡蛋，以及大规模纯化从尿囊液获得的病毒悬浮液以保证不含鸡蛋蛋白。
使用鸡胚生产病毒的另一个缺点是该支持物非常倾向于选择在抗原特异性上与野生型病毒不同的病毒变异体，并且有时导致病毒由于表型改变而不适用于生产疫苗，所述表型改变包括，例如，免疫原性显著降低。
因此，本领域内已经进行许多努力，利用标准组织培养技术，用已经建立的哺乳动物细胞系如MDCK(Madin-Darby犬肾)细胞或Vero(非洲绿猴肾)细胞进行病毒生产，尤其是流感病毒的生产。
在组织细胞培养物中培养流感株的困难之一是需要对宿主中的流感血凝素进行蛋白水解切割。切割病毒HA前体成为HA1和HA2亚片段虽然对于装配病毒元件形成完整病毒粒不是必要的，但却是赋予病毒粒感染性(即使其能够感染新的细胞)所必需的。
已经报道(如Lazarowitz等，“通过蛋白水解切割血凝素多肽增强B型流感病毒的感染性”，Virology，68：440-454，1975)：通过在组织培养物的培养基中加入蛋白酶如胰蛋白酶，可以克服几个A型流感病毒株在标准细胞培养物中的有限复制。然而，在某些情况下，例如在使用Vero细胞的情况下，这仍然是有困难地。
Kaverian和Webster(J Virol 69/4：2700-2703，1995)报道：在Vero细胞培养物中，培养基中的胰蛋白酶活性从温育开始就快速降低，导致由于无法产生足够数量的感染性病毒粒而不能在培养基中积累病毒，但这种现象在MDCK、猪肾或恒河猴肾细胞培养物中不是那么明显。他们得出结论：从Vero细胞中释放出一种胰蛋白酶抑制因子。他们进一步显示：通过重复加入胰蛋白酶，可以恢复病毒复制，并使病毒复制维持多个循环，导致更好的病毒产量。
US5,753,489报道另一种有效生产疫苗的方法，其中使用无血清培养基在多种不同哺乳动物细胞系(包括MDCK细胞和Vero细胞)中增殖病毒。该文献公开的方法包括在无血清培养基中培养脊椎动物细胞、用病毒感染所述细胞培养物、温育病毒感染的所述细胞培养物、取出包含病毒的一部分培养基并用蛋白酶接触该部分、然后在该部分中加入蛋白酶抑制剂并将该部分返回所述细胞培养物。在该文献中，优选在第一个容器中提供培养、感染和温育的步骤，在第二个容器中进行接触胰蛋白酶和加入抑制剂的步骤，所述第二个容器与第一个容器形成循环，以便所述步骤能够在闭合循环内进行。该系统允许在培养物中以比细胞所能够通常承受的浓度更高的浓度使用胰蛋白酶或其它蛋白水解酶。
EP 0870508报道一种生产病毒抗原疫苗的方法，该方法包括用病毒感染一种动物细胞系(可选地是Vero细胞系)，在所述细胞培养物中繁殖病毒，在病毒增殖结束前不久在所述细胞培养物中加入核酸酶消化从裂解的宿主细胞释放到培养基中的核酸物质，然后收获所述病毒，通过提取获取病毒抗原，以制造病毒抗原疫苗。该方法没有提到用于增殖病毒的营养培养基种类以及蛋白酶的加入，而加入蛋白酶是最后加工流感病毒血凝素获得感染性病毒所需的。该方法还要求各种纯化步骤以提供现成的疫苗制备物。
然而，已知宿主支持物的性质和用于增殖病毒的营养培养基的组成可以显著影响由此获得的病毒后代的免疫原性和抗原性。尤其是含血清的培养基不仅降低病毒后代的抗原性，而且还降低培养基中的蛋白酶活性，由此抑制病毒成熟，并随后要求昂贵的纯化步骤。
发明简述
本发明克服在先技术的缺点。本发明涉及一种简单有效的方法，该方法用于从各种来源分离病毒，并在最小化或完全防止由于适应性选择而引起的所述病毒表面抗原改变的条件下，生产用作疫苗(尤其是活减毒流感疫苗)的病毒后代。
本发明的另一个目标是提供生产病毒(尤其是流感病毒)的方法，所述方法产生选择性凝集人红细胞而不凝集鸡红细胞的病毒后代，并且所述病毒后代的抗原特性最好与最初接种的病毒株(如初级临床野生型分离物)的抗原特性相同。
在一个优选实施方案中，所繁殖病毒的HA基因以及可选NA基因的核酸序列与最初接种的株(如流行株、感染患者的初级临床分离物)的所述核酸序列相同。
本发明的再一个目标是提供在单一步骤方法中有效生产完整病毒疫苗(尤其是活减毒疫苗)的方法，所述方法在通过离心从细胞培养物上清液收获病毒悬浮液后不需要任何层析或其它纯化步骤，尤其不需要蛋白分离或纯化步骤。
本发明的又一个目标是提供减毒、冷适应和温度敏感的A型和B型流感病毒株以及用它们制备的疫苗。
附图简述
图1是一个示意图，说明在Vero细胞培养物上清液中胰蛋白酶失活随时间变化的过程。
图2是一个示意图，说明在MDCK细胞培养物上清液中胰蛋白酶失活随时间变化的过程。
发明详述
使用含胚卵、MDCK和Vero细胞的比较试验清楚地证明：最初接种的病毒有可能在这些支持物中任何一种上生长时发生抗原性改变。
我们的试验证实：在无血清培养基内Vero细胞上培养的流感病毒株发生最少改变或不发生改变。此外，证实当A型流感病毒(至少H3N2亚型株)在无血清和无蛋白培养基内的Vero细胞上生长时表现选择性凝集人红细胞，但不凝集鸡红细胞。此外，它们不能在蛋内生长。首次表明与在MDCK细胞或蛋上培养的病毒相比，这些Vero培养的病毒可能与对应临床分离物的野生型病毒更相似。
事实上，从鼻拭子获取的野生型分离物的HA基因和NA基因与分别在Vero细胞和MDCK细胞上培养的同样病毒相比，揭示MDCK培养的病毒的HA或NA相对于拭子分离物的HA或NA发生改变，或相对于Vero培养的病毒发生改变，或相对于拭子分离物和Vero培养的病毒都发生改变。
此外，用Vero或MDCK培养的野生型病毒免疫雪貂的试验数据指出：Vero培养的病毒比MDCK培养的病毒的毒力强得多。另外，对猕猴进行动物试验测试的Vero培养的病毒的免疫原性显著优于在MDCK细胞或卵上培养的病毒的免疫原性。
这些发现综合在一起为下面的假说提供强有力的证据：根据下文更详细描述的本发明用于扩增和增殖病毒的方法产生与最初接种(如野生型)病毒相比未改变或仅发生很小程度修饰的病毒。
本发明的病毒扩增方法不仅避免抗原改变，而且异常简单，使其非常适于大规模工业化疫苗生产。
其它试验已经显示：胰蛋白酶(或胰蛋白酶原)的来源可能是影响感染性病毒粒总产量的另外一个因素。事实上，虽然本领域内已知的方法(如Kaverin和Webster，J Virol 69/4：2700-2703，1995；或US5,753,489)或者重复加入胰蛋白酶(Kaverin和Webster)，或者使用高胰蛋白酶浓度(US5,753,489)，但依照本发明的方法仅使用在先技术中报道的一半或更低胰蛋白酶浓度。此外，在温育感染宿主细胞开始前或开始时，在细胞培养物培养基中一次性加入低至0.5-10μg/ml、优选2-5μg/ml的胰蛋白酶足以达到最佳感染性病毒滴度。失活试验揭示：猪或人重组胰蛋白酶与牛胰蛋白酶相比，对Vero细胞或MDCK细胞失活的敏感性低得多。由于在本领域内最广泛使用牛胰蛋白酶，因此很有可能在先技术文献暗示牛胰蛋白酶，明确指出其他胰蛋白酶来源的除外。这也能帮助解释在例如Kaverin等和US5,753,489中陈述的应用胰蛋白酶的模式和浓度。
因此，根据本发明，在开始就使用猪或人重组胰蛋白酶或胰蛋白酶原补充用于Vero细胞培养物的无血清培养基，使得可以使用极低的胰蛋白酶或胰蛋白酶原浓度，并因此避免在收获含病毒的上清液后需要劳动密集型和昂贵的纯化步骤。
另一有利于使本方法简单并因此对于疫苗生产商有吸引力的步骤是：在温育感染Vero细胞用于病毒增殖开始前或开始时，在细胞培养物培养基中一次性加入高活性内切核酸酶。在培养基中一次性加入该内切核酸酶(最好是BenzonaseTM)的起始浓度非常低，为2-30单位/ml、优选5-15单位/ml，该内切核酸酶从细胞培养物培养基中有效清除主要由正在裂解或已裂解的宿主细胞产生的游离DNA和RNA。由离心后上清液获得的现成疫苗制备物的残余Benzonase酶浓度保持在每剂5ng或更低。
BenzonaseTM是Nycomed Pharma A/S Denmark的商标，指从粘质沙雷氏菌(Serratia marcescens)获得的一种细胞外非特异性内切核酸酶。Benzonase是一种遗传工程化的内切核酸酶，分解多种形式的DNA和RNA成为小寡核苷酸。它促进快速降低细胞裂解液的粘度，便利超速离心。它降低蛋白水解，并增加靶蛋白的产量，从例如重组蛋白中完全清除核酸。它具有400,000U/mg的异常高的活性。
本方法第三个重要优点是时间因子，这因此降低方法的费用。由于使用不含动物来源蛋白并且最好不含抗生素的无血清培养基，因此可以减少昂贵和耗时的纯化程序到最少或甚至完全避免。此外，因为外源酶例如蛋白酶(如胰蛋白酶或胰蛋白酶原)和核酸酶(如Benzonase)的添加一次性发生在病毒增殖期开始时，因此这节省了本领域目前使用的方法花费在含病毒的培养物上清液的温育后处理(如HA激活、RNA/DNA消化、蛋白纯化等)上的大量时间。
惊人的是，证实在病毒感染的Vero细胞培养物中早期加入蛋白酶(如胰蛋白酶或胰蛋白酶原)和核酸酶(如Benzonase)中的一种或两种对于病毒产量没有负面影响，这可能是由于本发明方法中应用的酶浓度非常低。
本发明的病毒增殖方法可用于扩增各种病毒，尤其是H3N2亚型A型流感病毒，但也适于分离和繁殖任何流行性或实验室流感病毒株，不论病毒接种物的种类如何(如血清样品、鼻洗液、鼻拭子、咽拭子、唾液等)。使用本发明方法的原理，已经产生多种A型流感和B型流感疫苗，这些疫苗构成本发明的一部分，并且在下面的实施例中更详细表征。
此外，已经证实根据本发明制造的疫苗的保护性功效和疫苗安全性，这将在实施例中得到证实。
本文使用的与按照本发明的病毒扩增或增殖方法有关的术语“不含蛋白”或“不含非血清蛋白”是指不含任何有功能活性的蛋白。然而，该术语应当不排除可能来自蛋白水解物例如酵母提取物或大豆提取物的非功能性肽。除另有陈述外，术语“不含蛋白”也不排除以本文公开和要求保护的浓度存在的蛋白酶和核酸酶。
在一个优选实施方案中，本发明涉及一种生产完整病毒疫苗的简单、可靠和高度经济的方法，所述完整病毒疫苗最好是一种减毒活疫苗，该方法包括下面步骤：
a)用所需病毒感染非洲绿猴肾(Vero)细胞，其中所述Vero细胞在无血清培养基中培养并从中分离，所述无血清培养基也不含非血清蛋白；
b)将所述感染细胞与合适的不含非血清蛋白(除蛋白酶和核酸酶外)的无血清细胞培养基混合；然后
c)在所述蛋白酶和所述核酸酶存在下温育所述细胞，产生感染性病毒，同时消化释放到细胞培养基中的核酸物质；
d)离心所述细胞培养物，收集包含病毒的上清液，从而收获感染性病毒；然后
e)如下制备其疫苗：用选自以下的至少一种加工步骤处理所述含病毒的上清液：过滤、浓缩、冷冻、冻干以及通过加入稳定剂稳定。
优选用于增殖的所述病毒未曾接触过除Vero细胞系外的宿主支持物。这将保证获得与最初病毒(如鼻拭子分离物)的最佳的免疫原性和抗原性同一性，以及增殖后获得最佳的病毒后代。
还优选用于增殖的病毒是选自以下的流感病毒：A/Sing/1/57ca株、A/Sing/1/57ca/ΔNS 87株、A/Sing/1/57ca/ΔNSPR8株、A/Sing/1/57ca/NS124PR8株、B/Vienna/1/99ca株、B/Vienna/99/ca37株以及来自这些病毒株中任一种的减毒变异体和重排列体，所述病毒尤其用于生产完整病毒疫苗，最好是一种流感减毒活疫苗。下面实施例详细地公开优选病毒株如原株(master strain)的遗传特征。
在另一实施方案中，本发明涉及完整病毒疫苗本身，最好涉及减毒活疫苗，所述疫苗的现成形式包含用于生产所述病毒的无血清和无蛋白Vero细胞培养物的含病毒上清液，所述上清液基本未调整处理、可选地经过过滤和/或浓缩。尤其优选所述疫苗根据本文公开和要求保护的本发明方法生产。
所述“一步”疫苗不需要进一步的加工，如除离心和/或常规过滤(即非凝胶过滤)外的纯化步骤，符合FDA批准的要求。
本文在按照本发明的疫苗制备中关于含病毒的上清液所使用的术语“基本未改变”应当指收获增殖病毒时上清液的组成，即在所述上清的液相中存在的可溶性组分和成分的组成。由于对所述含病毒的上清液进行例如过滤、无菌过滤、离心、浓缩、干燥或冻干等步骤而发生的成分组成的微小变化应当视为在“基本未改变”的范围内。此外，该术语应当不排除本领域内一般用于疫苗制备物的防腐剂和/或稳定剂的存在。
本发明的完整病毒疫苗可用于预防性或治疗性治疗病毒感染，尤其是治疗流感病毒感染。可以使用本领域内已知的途径给予，例如静脉内途径、皮下途径、肌内途径或最优选鼻内途径。然而，本文公开的病毒株以及用其制造的疫苗也可用作呈递异源抗原到免疫系统的载体或穿梭系统，例如呈递病毒被膜蛋白的抗原如HIV-1或肝炎抗原。
在从属权利要求中定义其它优选实施方案。
为更充分理解本文描述的本发明，陈述下面的实施例。它们仅用于举例说明的目的，不应当认为是在任何方面限制本发明。
实施例1：生产病毒
培养Vero/SF(＝无血清)细胞：
SF-培养基：DMEM(Biochrom F0435)，Ham’s F12(BiochromF0815)，5mM L-Gln，0.1％SF添加剂(a)或(b)；抗生素(仅用于病毒分离物的第一次传代)。
SF-添加剂：非动物来源的蛋白水解物，不含功能蛋白如胰岛素、转铁蛋白或生长因子：
a)62.5g hy-soy/UF，Quest 5X59100，加入500g HQ水，用PES
0.2μm滤器过滤；
b)12.5g hy-pep 1510，Quest，加入100g HQ水，用PES0.2μm
滤器过滤。
融化一支消毒过的(70％乙醇)深度冰冻(液氮)的盛装WCB Vero细胞的安瓿的内容物，将其加入10ml管内的9ml冷的无血清(SF)培养基，在1000rpm(170g)离心10分钟。将沉淀重悬浮于SF培养基中到总共30ml，然后转移到一个80cm2的Roux瓶，在37℃和7％CO2下温育至少15分钟。然后，移走所述培养基，用约0.1ml/cm2 PBS def.(＝不含Ca+和Mg2+的PBS)洗涤细胞。加入胰蛋白酶/EDTA溶液(8-10μl/cm2；0.1％胰蛋白酶/0.02％EDTA溶液)，在室温下温育约3分钟。用手掌温和推动所述Roux瓶，使细胞解吸附，然后加入约1/5胰蛋白酶/EDTA溶液体积的SF培养基和胰蛋白酶抑制剂(Sigma，T6522)。将所述细胞悬浮液重新分配到Roux瓶或滚瓶，在37℃和9％CO2下温育。
在DMEM/Ham’s F12+2％FCS(热失活)中培养MDCK细胞；含胚的鸡蛋11-12天大，SPF(不含特定病原体)来源。
繁殖病毒株：
从含Vero细胞的滚瓶中取出旧培养基，在每个滚瓶的SF培养基中加入5ml病毒悬浮液，使病毒感染细胞，MOI(感染复数)约0.01。在33℃温育45分钟后，用吸液管取出病毒接种物。在每个滚瓶中加入90ml补充0.5-10、优选2-5、最优选2μg/ml猪胰蛋白酶(供应商：AvP)或人重组胰蛋白酶或胰蛋白酶原(自产)以及0.5g/l碳酸氢钠的SF培养基，所述滚瓶在33℃和5％CO2下温育。为产生用于动物试验以及人体临床试验的减毒活疫苗样品，在所述SF培养基中补充胰蛋白酶以及BenzonaseTM，BenzonaseTM使用浓度为每ml培养基2-30、优选5-15、最优选10单位。感染后64小时，培养物上清液在10℃下50ml管中以4000rpm(3000g)离心5分钟，收获病毒。汇集每个病毒株的上清液，贮存在+4℃。其等分试样用于疫苗测试。
为贮存疫苗，可以冻干所述病毒制备物，或者可以加入稳定剂如海藻糖和HEPES缓冲液中的乳清蛋白酶促水解物。可以用无菌水进行重建。
实施例2：比较各细胞培养物中的胰蛋白酶失活
表1：Vero细胞与MDCK细胞培养物中胰蛋白酶失活的比较
Vero/MDCK0h 24h 48h 72h牛胰蛋白酶0.314/0.314 0.199/0.239 0.110/0.201 0.026/0.203猪胰蛋白酶(高)0.230/0.230 0.201/0.206 0.171/0.209 0.133/0.201猪胰蛋白酶(低)0.129/0.129 0.108/0.118 0.081/0.099 0.054/0.116人重组胰蛋白酶0.097/0.097 0.054/0.088 0.026/0.080 0.008/0.076
测试从未感染的Vero细胞培养物(如实施例1所述在SF培养基中培养)和MDCK细胞培养物(在如实施例所述在补充FCS的培养基中培养)中它们失活不同来源胰蛋白酶的能力，所述胰蛋白酶在时间0h以同等浓度加入上清液中。应用两种不同质量(从不同生产商获得)的猪胰蛋白酶，即高活性或低活性猪胰蛋白酶。结果显示于表1和图1和图2。
数据明白地显示：牛胰蛋白酶在Vero细胞培养物上清液中快速失活，在MDCK细胞培养物上清液中失活较慢。猪和人重组胰蛋白酶(本实验室生产)在MDCK上清液中保持完全活性，但它们在Vero上清液中逐渐失活，其速率是牛胰蛋白酶失活速率的约一半或更低。所测试的两种猪胰蛋白酶的差别仅是在247nm处的开始OD水平，而两批猪胰蛋白酶的失活特性基本相同。
实施例3：在不同宿主细胞支持物上培养后各种病毒特性的比较
如实施例1所述进行在不同宿主细胞支持物上的病毒增殖。从Vero细胞回收的七个分离物的每一个都与人红细胞反应，但不与鸡红细胞反应，并且没有一种在含胚卵中积累。另一方面，所述从MDCK细胞回收的分离物都与鸡和人红细胞反应，并且能够在蛋中生长。虽然这些差异没有在H1N1亚型A型流感病毒或B型流感病毒分离物中出现(见下面的表3和表4)，但仍然可以假设在Vero细胞上培养流感病毒将比在其它支持物上培养更好地维持抗原性特性。
表2：从临床材料分离的H3N2型病毒在Vero/SF细胞上的特征
分离物 抗原性相关于 分离自 HA滴度 在蛋中的 生长 鸡胚 人胚A/47/96 A/Johannesburg Vero MIDCK - + + + - +A/7729/98 A/Sydney/5/97 Vero MIDCK - + + + - +A/1143/99 A/Sydney/5/97 Vero MDCK - + + + - +A/1144/99 A/Sydney/5/97 Vero MDCK - + + + - +A/1179/99 A/Sydney/5/97 Vero MDCK - + + + - +A/1180/99 A/Sydney/5/97 Vero MDCK - + + + - +A/1182/99 A/Sydney/5/97 Vero MDCK - + + + - +
由表3的数据，看起来H1N1流感病毒对于适应性选择较不易感，因为Vero和MDCK培养的分离物在它们的血细胞凝集特性以及它们的HA序列上不显示显著差异。对于表4中列出的B分离物可以得出相似结论。
用于分离和复制病毒的临床起始材料(如血清样品、拭子)主要来自：
1.Institute of Virology，Vienna，Austria(Prof.F.Heinz)1995/96，1996/97
2.Unite de Genetique Moleculaire des Virus Respiratoires，Institute Pasteur，Paris，France(Prof.S.van der Werf)1996/97
3.Public Health Laboratory Service，London，UK(Dr.M.Zambon)1996/97
4.Laboratorie Central de Virologie，Hopitaux Universitaires deGeneve，Geneva，Switzerland(Dr.W.Wunderli)1996/97，1997/98
5.Virus Unit，Queen Mary Hospital，Hong Kong(Dr.W.L.Lim)1997/98
表3：分离自临床材料的H1N1病毒在Vero/SF细胞上的特性
分离物编 号 抗原性相 关于 分离自 HA滴度 在蛋中 的生长 HA1中下 面位置的变 化 鸡胚 人胚 225 A/5389/95 A/Bayern/7 /95 Vero MDCK + + + + + + D D A/1035/98 A/Beijing/2 62/95 Vero MDCK 蛋 拭子 + + + + + + + + + D D G D A/1131/98 A/Beijing/2 62/95 Vero MDCK 拭子 + + + + + + D D D A/1134/98 A/Beijing/2 62/95 Vero MDCK 蛋 拭子 + + + + + + + + + D D n.t. D
表4：分离自临床材料的B型病毒在Vero/SF细胞上的特征
分离物编号 抗原性相 关于分离自 HA滴度 在蛋中 的生长 HA1中下 面位置的变 化 鸡胚 人胚 198B/4291/97 B/Beijing/1 84/93VeroMDCK + + + + + + 相同B/1/99 B/Beijing/1 84/93VeroMDCK蛋拭子 + + + + + + + + + T(g.s.) T(g.s.) A T(g.s.)B/110/99 n.t.VeroMDCK + + + + + + 相同B/147/99 n.t.VeroMDCK + + + + + + 相同B/156/99 B/Beijing/1 84/93VeroMDCK + + + + + + 相同B/157/99 B/Beijing/1 84/93VeroMDCK + + + + + + 相同
表5：分离自不同宿主系统的H3N2流感病毒的HA、NA和M蛋白的氨基酸改变
分离物编号 下面位置的变化 HA NA M 128 129 229 133 218 220 136 151 A/47/96 Vero A/47/96 MDCK T(g.s) A A/7729/98 Vero A/7729/98 MDCK E G R K A/1143/99 拭子 A/1143/99 Vero A/1143/99 MDCK N(g.s) N(g.s) D G G E n.t n.t D G n.t 相同 A/1144/99 拭子 A/1144/99 Vero A/1144/99 MDCK R R G n.t 相同 n.t 相同 A/1179/99 拭子 A/1179/99 Vero A/1179/99 MDCK 相同 n.t 相同 n.t 相同 A/1180/99 拭子 A/1180/99 Vero A/1180/99 MDCK 相同 n.t Q R n.t n.t 相同 A/1182/99 拭子 A/1182/99 Vero A/1182/99 MDCK 相同 n.t n.t n.t n.t n.t n.t
结果显示：对于一些分离物，Vero或MDCK增殖的病毒的HA序列与通过PCR扩增直接得自棉拭材料的HA序列相比没有改变。在一些在MDCK细胞上生长的其它分离物中，HA和/或NA序列与从Vero细胞获得的对应序列不同。然而，从Vero得到的病毒在HA序列上并不显示与已经测定的拭子分离物的HA序列有任何差异。
表6：从Vero分离的病毒、从MDCK分离的病毒和从蛋分离的病毒对猕猴的免疫原性
动物编号 用于免疫的病毒 剂量，PFU/ml 血清HI滴度 96 A/Vienna/47/96V 5×104 256 88 A/Vienna/47/96V 5×104 128 15 A/Vienna/47/96V 1.0×106 128 95 A/Vienna/47/96V 1.0×106 256 93 A/Vienna/47/96M 1.0×106 16 128 A/Johannesburg/33/94E 5×106 32 110 A/Vienna/157/97V 5×104 128 78 A/Wuhan/359/95E 5×106 32
在不使用麻醉的情况下用1ml病毒悬浮液鼻内免疫猕猴
V-从Vero分离的病毒
M-从MDCK分离的病毒
E-从蛋分离的病毒
表7：从Vero和MDCK分离的A/Vienna/47/96野生型病毒变异体对雪貂的毒力
病毒 病毒剂量 在下面天数内发热的动物数量 PFU/ml 1 2 3 A/Vienna/47/96 Vero 2×102 1×103 FF FFF FFF FFF A/Vero/47/96 MDCK 5×102 5×103 5×104 FF FF F F
动物在用乙醚麻醉的情况下用1ml病毒悬浮液鼻内接种。
N-正常温度，从38.1℃到39.9℃
F-发热，超过40.0℃.
表6中最惊人但重要的结果是从MDCK分离的A/Vienna/47/96病毒与从Vero分离的对应病毒相比免疫原性非常低。并不特别令人惊讶的是从蛋分离的病毒仅显示弱的免疫原性。
相似地，表7中的结果指出：从Vero分离的病毒与从MDCK分离的病毒相比，在它们的宿主支持物上受到的适应性选择改变(如果有的话)更少。这意味着与从MDCK分离的病毒相比，从Vero分离的病毒保持原始病毒更多或甚至全部的免疫相关特性，尤其是抗原性特性。
实施例4：用优选株生产疫苗
实施例1中描述的方法也用于生产用于动物测试和人类临床研究的疫苗样品。应当理解，其中描述的病毒增殖方法也包括本领域内一般技术人员不需创造性投入就可应用的变化，只要所述变化不改变本文和权利要求中所述的本发明的含义。
使用连续的Vero细胞系作为宿主细胞系统(除非为比较目的使用得自其它宿主支持物的样品)，在无血清培养基中专门生产包含一种或多种优选A型或B型野生型株、原株或重排列株(随后更详细描述)的疫苗样品，其中所述无血清培养基中另外补充实施例1中所述的营养成分和酶。
一些适于修饰野生型病毒的方法是本领域内已知的，包括减毒方法(如温度敏感性)、冷适应方法和重排列方法，这些方法的广泛综述见，例如，WO 99/64068。
下面表8-13给出用于生产减毒活疫苗的两个最优选的A型和B型流感原株候选物的其它特征，所述生产例如通过WHO推荐的对实际流行性流感病毒的HA和NA基因进行6/2重排列。
表8：活流感疫苗的原株候选物的特征
A型流感A/Singapore/1/57/caH2N2 B型流感 B/Vienna/1/99/ca传代史A/Singapore/1/57野生型从蛋分离的H2N237℃下在Vero/SF细胞上传代20次25℃下在Vero/SF细胞上传代25次 B/Vienna/1/99野生型 从Vero分离 33℃下在Vero/SF细胞上 另外传代1次 25℃下在Vero/SF细胞上 传代22次减毒方法在异源系统上在最佳温度和亚最佳温度下顺序传代 在异源系统上在最佳温度 和亚最佳温度下顺序传代表型标记温度敏感性(ts)冷适应(ca)在小鼠肺中非常少量地复制 温度敏感性(ts) 冷适应(ca) 在小鼠肺中非常少量地复 制基因型标记突变：13(8编码)PB2 3(2编码)PB1 2(1编码)PA 4(3编码)NP 1M 2(2编码)NS 1 突变：5(3编码) PB2 0 PB1 1 PA 0 NP 2(1编码) M 1 NS 1
表9：8个基因组区段和A/Singapore/1/57/ca株的10种对应蛋白的完整序列
A/Singapore/1/57/ca(H2N2)RNA区段 核苷酸序列(cDNA) 蛋白 氨基酸序列1 SEQ ID No.1 Pb2 SEQ ID No.92 SEQ ID No.2 Pb1 SEQ ID No.103 SEQ ID No.3 PA SEQ ID No.114 SEQ ID No.4 HA SEQ ID No.125 SEQ ID No.5 NP SEQ ID No.136 SEQ ID No.6 NA SEQ ID No.147 SEQ ID No.7 M1 SEQ ID No.15 M2 SEQ ID No.168 SEQ ID No.8 NS1 SEQ ID No.17 NS2 SEQ ID No.18
ca-冷适应
然而，应当注意到第4和第6基因组区段(即HA基因和NA基因)对于表征A型流感原株候选物不是必需的，因为这些基因可以与实际流行性流感病毒(如上文所述)的对应基因互换。对于疫苗安全性重要的特性(如温度敏感性)或允许鼻内给予疫苗的特性(既冷适应)(因为鼻内的平均温度低于实际体温)主要是由于其余6个基因组区段中的突变引起的。
下面表10列出A/Singapore/1/57/ca与对应野生型株A/Singapore/1/57wt相比在基因组区段中的突变。
表10：减毒、温度敏感性、冷适应流感株A/Singapore/1/57/ca与A/Singapore/1/57wt相比在基因组区段中的突变
RNA 长度 改变的核苷酸 蛋白 长度 改变的氨基酸 区段 (n′ds) 位置 wt ca (aa) 位置 wt ca 1 2341 252 581* 1046* at g gc t PB2 771 -185 340 -I R -T I 2 2341 1279* 1965 t a a c PB1 757 419 - L - I - 3 2233 707* 1425 1537* 1819* a t a g t a g c PA 716 228 - 505 598 I - V Q N - I E 5 1565 210 g a NP 506 - - - 7 1027 327* 499* g g a c M1 M2 252 97 101 158 - R Q - K R - 8 890 813 a g NS1 NS2 237 121 - - - - - -
突变总数-13(8编码)
*编码的突变
优选的A/Sing/1/57/ca的变异体包含下面表11列出的那些突变，其中“Δ”指“del”或“delta”，表示在其NS基因区段中包含至少一个“缺失”的突变异体。
表11：A/Sing/1/57/ca的优选变异体
*源自A/Singapore/1/57/ca的基因组区段
**源自流感A/PR8/34的基因组区段
ca-冷适应；ts-温度敏感性；
aa-氨基酸
IFN-induct.-在能够生产IFN的宿主支持物中以及在给予动物或人免疫系统时引起干扰素释放的株。
IFN-sensit.-对干扰素敏感的株；在生产干扰素的细胞中的复制减少或停止。
Sing ca/ΔNS 87-包含在NS1基因位置36-123缺失87个氨基酸的A/Singapore/1/57/ca株。
Sing ca/ΔNSPR8-包含来自A/PR8/34(本文也缩写为“PR8”)的NS基因区段的A/Singapore/1/57/ca株，所述区段缺失整个NS1基因。
Sing ca/NS124PR8-包含来自A/PR8/34的NS基因区段的A/Singapore/1/57/ca株，所述区段在NS1基因的氨基酸位置124包含一个终止密码子。
+/-指该表型需要进一步澄清，并且还不能明确地定义。
下表12、13和13A分别指出优选的B型流感原株候选物以及其变异和重排列体。
表12：B/Vienna/1/99/ca株的8个基因组区段和11个对应蛋白的完整序列
B/Vienna/1/99/caRNA 区段 核苷酸序列(cDNA) 蛋白 氨基酸序列1 SEQ ID No.19 PB2 SEQ ID No.272 SEQ ID No.20 PB1 SEQ ID No.283 SEQ ID No.21 PA SEQ ID No.294 SEQ ID No.22 HA0 SEQ ID No.305 SEQ ID No.23 NP SEQ ID No.316 SEQ ID No.24 NB NA SEQ ID No.32 SEQ ID No.337 SEQ ID No.25 M1 SEQ ID No.34 BM2 SEQ ID No.358 SEQ ID No.26 NS1 SEQ ID No.36 NS2 SEQ ID No.37
ca-冷适应
最初的B/Vienna/1/99株于1999年2月分离自用无血清培养基培养的Vero细胞培养物，来自奥地利维也纳一名患有急性流感的12岁女子。疾病控制中心(CDC)，Atlanta，USA将它评级为类似于B/Beijing/184/93。野生型株(名为B/Vienna/1/99wt)在33℃下再传一代后，通过在25℃下使用同样细胞培养系统进行22次连续传代而减毒。第一轮在25℃下进行噬菌斑纯化，随后在33℃下进行四轮纯化。扩增所获得的噬菌斑纯化克隆，贮存于-70℃，该克隆名为B/Vienna/1/99 ca或简称为BV22。通过用来自NIBSC的标准抗血清进行的HI测定确认B/Beijing/184/93样病毒的身份。
表13：B/Vienna/1/99/ca(＝BV22)与B/Vienna/1/99/wt(Bvie)相比的突变，在Vero/SF中第一代
区段(核苷酸长度) 改变的核苷酸蛋白(氨基酸长度) 改变的氨基酸 位置 Bvie BV2 2 位置 BVie BV221(2396) - - -PB2(770) - - -2(2369) 594 T CPB1(752) - - -3(2305) - - -PA(726) - - -4(1882) 457 1299 1595 G G G A T AHA0(584) 142 422 521 A K G T N E5(1844) 128 330 C T T CNP(560) 23 - S - F -6(1557) - 823 1135 - G T - A CNB(100)NA(466) - 257 361 - R I - Q T7(1190) - 831 - A - GM1(248)BM2(109) - 21 - M - V8(1097) 116 - G - A -NS1(281)NS2(122) 25 - A - T -
表26：根据Los Alamos National Library流感病毒库(db)(网址：www.flu.lanl.gov)，B/Vienna/1/99wt的表征
B/Vienna/1/99 wt编码基因 氨基酸序列 保藏号 核苷酸序列保 藏号备注 PB2，区段1 ISDACH017 ISDNCHB017数据库中列为区段2 PB1，区段2 ISDACH016 ISDNCHB016数据库中列为区段1 PA，区段3 ISDACH015 ISDNCHB015 HA，区段4 ISDACH018 ISDNCHB018 NP，区段5 ISDACH013 ISDNCHB013 NA，区段6 ISDACH012 ISDNCHB012 M，区段7 ISDACH011 ISDNCHB011 NS，区段8 ISDACH014 ISDNCHB014
此外，B/Vienna/1/99 ca继续传代另外15次后(即在无血清Vero细胞培养物上总共传代37次)导致突变异体B/Vienna/1/99 ca37(简称为BV37)，该株具有甚至由于某些BV22的特性。该突变异体包含与BV22相比数量增加的突变，并且目前看起来是生产基于非重组流感病毒突变异体的完整病毒疫苗(尤其是减毒流感活疫苗)最有希望的候选物。其它突变列于下面表13A：
表13A：BV22和BV37与B/Vienna/1/99wt相比的突变，在Vero/SF上第一次传代
区段(核苷酸 长度)改变的核苷酸 蛋白(氨基 酸长度) 改变的氨基酸 位置 BVie BV22 BV37 位置 BVie BV22 BV3 71(2396)----PB2(770)----2(2369)(BV37：2370)5942348T-CACAPB1(752)--------3(2305)----PA(726)----4(1882)457112212991595GCGGA*CTAA*AGAHA0(584)142363422521AFKGT+FNET+LKE5(1844)128212330CCTTCC#TTC#NP(560)2351-SP-FP-FL-6(1557)-8231135-GT-AC*-GC*NB(100)NA(466)-257361-RI-QT*-RT*7(1190)248318311029GAAAGGGAAGGGM1(248)BM2(109--2187--MI--VI--VV8(1097)116-G-A-A-NS1(281)NS2(122)25-A-T-T-
与流感序列数据库13.2.2001(www.flu-lanl.gov)的比较：
a)独特突变用粗体下划线表示；
b)共有突变：
*B/Lee/40、B/Osaka/70、B/Kadoma/1076/99(获得的氨基酸：
1)
+B/Lee/40、B/Osaka/70
#常常是：B/Lee/40、B/Ann Arbor/1/66 ca & wt、
B/Singapore/222/79、B/North Dakota/83、B/Norway/1/84、
B/Ibaraki/2/85、B/Ann Arbor/1/86、B/Victoria/2/87、B/Aichi/5/88
*B/Kanagawa/73
应当理解，根据本发明的A型和B型流感原株应当不限于在本文的表中明确列出的特点和遗传特征，而且应当还包括它的小变异，只要所述变异在本发明的意义内并且不显著改变该病毒的任何一种功能特征。所述变异可以例如由于病毒扩增或增殖的额外步骤而发生(如为获得进行序列分析的材料)。
此外，本文列出的基因序列包括与本发明一起使用的引物序列(位于每个基因组区段的开始或结束)，所述引物序列可以与野生型病毒株或减毒病毒株中一种或二者的病毒基因组区段的对应真序列不同。
实施例5：疫苗安全性和效能
下面数据证实按照本发明生产(如按照实施例1所述)的流感疫苗的温度敏感性和疫苗安全性。
表14：不麻醉的情况下进行一次鼻内免疫后小鼠的抗体反应
病毒 反应者数量 GMT3 攻击后的保护2PR8/Sing ca-2/6 0/6 ＜4 5/6PR8/Sing ca-ΔNS 4/6 6.7 5/6PR8-wt 5/6 16.0 5/6
1-阳性HI滴度＞1∶4的动物数量
2-在肺中未检测到病毒的动物数量
3-血清中抗体的几何平均滴度
PR8wt-流感株A/PR/8/34野生型(H1N1)，对小鼠致病
PR8/Sing ca-2/6-是减毒流感株A/Sing/1/57 ca和PR8wt之间重排列体，包含来自PR8wt的2个基因(HA和NA)以及来自A/Sing/1/57ca的所有其它基因。
PR8/Sing-ΔNS包含来自PR8wt的HA和NA基因、来自A/Sing/1/57ca的五个基因和来自PR8但缺乏NS1编码序列的NS基因(NS1缺失或剔除)。
表15：用A/Singapore/1/57病毒的不同变异体鼻内免疫(麻醉)后小鼠的抗体反应和保护
病毒 反应者1 两次免疫 后的GMT 攻击后的 保护4 第一次免疫 第二次免疫A/Sing/1/57/wt va2 9/9 9/9 103.9 9/9A/Sing/1/57/ca3 8/10 10/10 55.7 8/10A/Sing/57/ΔNS87 1/10 10/10 27.9 8/10
1-阳性HI滴度＞1∶4的动物数量
2-va-Vero适应的
3-ca-冷适应的
4-在肺中未检测到病毒的动物数量
表16：A/Singapore/1/57的wt、va和ca变异体在小鼠肺中的增殖a
病毒 感染后某天小鼠肺中的病毒滴度， PFU/mlb 2 4 6 A/Singapore/1/57/wt 1.6×106 2.2×105 1.4×103 A/Singapore/1/57/wt va 2.5×106 2.1×106 1.0×102 A/Singapore/1/57/ca ＜10 ＜10 ＜10
a小鼠鼻内感染50μl滴度为1.0×106PFU/ml的病毒液。
b每ml组织悬浮液的PFU，在MDCK细胞上滴定。
表17：A/Singapore/1/57病毒的wt和ca变异体对雪貂的毒力
病毒 感染后某天发热的动物数量 1 2 3A/Singapore/1/57wt FFF NNN NNNA/Singapore/1/57ca NNN NNN NNN
每日两次记录动物的直肠温度，特征如下：
N-正常温度，从38.1℃到39.9℃
F-发热，超过40℃。
每组包括3只动物，用乙醚麻醉动物，用1ml滴度为2×106PFU/ml的病毒液鼻内免疫动物。
表18：A/Hong Kong/1035/98wt和A/Singapore/1/57/ca的2/6重排列体在小鼠肺中的增殖a
病毒 感染后2-6天小鼠肺中的病毒滴度， PFU/mlb 2 4 6A/Hong Kong/1035/98wt H1N1 6.8×104 2.0×104 ＜10A/Singapore/1/57/caxA/Hong Kong/1035/98wt ＜10 ＜10 ＜10
a用乙醚麻醉小鼠，鼻内感染50μl病毒液。
b每ml 10％组织悬浮液的PFU/ml，在Vero/SF细胞上滴定，数据是6只小鼠的平均值(分别处理每只动物的肺)。
所述重排列体包含来自A/Hong Kong/1035/98wt野生型的HA和NA基因以及来自A/Singapore/1/57/ca的其它6个基因。
表19：A/Vienna/47/96wt和A/Singapore/1/57/ca的6/2重排列体对雪貂的毒力
病毒 病毒亚型 某天发热的动物数量 1 2 3 鼻炎b 原株 A/Singapore/1/57/ca H2N2 NNN NNN NNN ± 流行性病毒 A/Vienna/47/96wt H3N2 NNN FFF FFF +++ 重排列体 A/Singapore/1/57/cax Vienna/47/96wt H3N2 NNN NNN NNN ±
用乙醚麻醉动物，用1ml滴度为2×106PFU/ml的病毒进行鼻内免疫。
N-正常温度，从38.1℃到39.9℃
F-发热，超过40℃。
b+++-重度鼻炎
±无鼻炎
表16到19的结果清楚地证明包含所述减毒温度敏感性原株的疫苗的安全性，以及在重排列体的情况下，基于重排列病毒的疫苗的安全性，所述重排列病毒包含所述减毒温度敏感性原株的“骨架”(6个基因)以及来自如致病野生型株A/Hong Kong/1035/98wt的HA基因和NA基因。
表20：B/Vienna/1/99的Ts和ca表型
病毒 以下温度下在 Vero细胞上的 PFU/ml 以下温度下在MDCK细胞上 的PFU/ml 25℃ 33℃ 39℃B/Vienna/1/99wt ＜300 4×106 4×105B/Vienna/1/99ca(BV22) 1×106 2.4×106 ＜20
表21：B/Vienna/1/99 ca的ts表型的遗传稳定性
病毒 某温度下在MDCK细胞上的PFU/ml 33℃ 39℃B/Vienna/1/99wt 4×106 4×105B/Vienna/1/99ca(BV22) 2.4×106 ＜20B/Vienna/1/99ca(BV22)在33℃传代5次后 8×105 ＜20
BV22株以高MOI在Vero细胞上传代五次。然后再次控制ts表型。如表21所示，该株保持温度敏感性。
表22：B/Vienna/1/99ca和wt在小鼠肺中的毒力
感染后某天的PFU/ml* 病毒器官 2 3 4 B/Vienna/1/99ca(BV22)肺 ＜20 ＜20 ＜20鼻 1×102 1×102 20 B/Vienna/1/99wt肺 8×104 7×103 4.4×103鼻 3.8×104 3.4×104 1.4×104
*每株免疫9只OF1小鼠，用乙醚麻醉小鼠，以105PFU经鼻免疫。在感染后指定的天数，每组处死3只小鼠。匀浆肺和鼻甲，在PBS def.中制成10％(w/v)悬浮液。进行悬浮液的噬菌斑测定。
数据显示：ca原株候选物BV22在鼻粘膜中的温和复制是可能的，而该病毒的ts特性防止其在肺中增殖。
表23：B型流感株的重排列体的Ts和ca表型
病毒 某温度下在MDCK细胞上的PFU/ml 33℃ 39℃B/Vienna/1/99wt 4×106 4×105B/USSR/69wt 1.6×106 4×104B/Vienna/1/99ca(BV22) 1.4×106 ＜20BV22×B/USSR/69(6/2) 8×106 ＜20
建立一种6/2重排列体，该重排列体包含野生型流感株B/USSR/69wt的HA和NA以及来自B/Vienna/1/99ca(BV22)的其它6个基因组区段。通过HI测定测试血凝素的来源，使用本领域内已知方法通过RT-PCT和限制分析测试所有其它基因组区段。
表24：B型流感株重排列体在小鼠肺中的毒力
感染后某天的PFU/ml*病毒器官 2 3 4B/Vienna/1/99ca(BV22)肺 ＜20 ＜20 ＜20鼻 ＜20 1×102 40B/USSR/69wt肺 1.8×105 4×105 2.4×104鼻 1.6×105 2×105 1.6×105BV22×B/USSR/69wt(6/2)肺 ＜20 ＜20 ＜20鼻 2.8×103 2×103 4×102
*每株免疫9只OF1小鼠，用乙醚麻醉小鼠，以105PFU经鼻免疫。在感染后指定的天数，每组处死3只小鼠。匀浆肺和鼻甲，在PBS def.中制成10％(w/v)悬浮液。进行悬浮液的噬菌斑测定。
实施例6：临床研究
根据本发明(如按照实施例1所述)生产下面疫苗(采用鼻喷雾剂的形式)用于经鼻传递。每ml的组成(在重建冻干材料后)：
(1)安慰剂：2×SF培养基，40mM HEPES缓冲液，8％乳清蛋
白酶促水解物，4％海藻糖；
(2)Vero-Vac H1：A/Beijing/262/95(H1N1)样制备物，该制备物
包含4.3×107TCID50的A/Singapore/1/57/ca与A/Hong
Kong/1035/98的6/2重排列体；2x培养物上清液，40mM
HEPES缓冲液，8％乳清蛋白酶促水解物，4％海藻糖；
(3)Vero-Vac H3：A/Syndey/5/97(H3N2)样制备物，该制备物
包含2.1×107TCID50的A/Singapore/1/57/ca与A/SW/7729/98
的6/2重排列体；2x培养物上清液，40mM HEPES缓冲
液，8％乳清蛋白酶促水解物，4％海藻糖；
(4)俄罗斯三价疫苗(用于成人的活流感疫苗)：
A/17/Beijing/95/25(H1N1) 1.1×108EID50
A/17/Sidney/97/76(H3N2) 2.3×107EID50
B/60/Petersburg/95/20 1.1×107EID50
(5)单价Vero疫苗BV22：B/Beijing/184/93-样制备物，该制
备物包含2×106TCID50的原株候选物B/Vienna/1/99/ca
(＝BV22)；2x培养物上清液，40mM HEPES缓冲液，8％
乳清蛋白酶促水解物，4％海藻糖；
将所述疫苗给予每个接种组13名志愿者。将550μl重建的疫苗(或安慰剂，分别地)在第0天和在第22±1天第二次经鼻给予每名患者。结果在下面表25中综述。
安全性结果：
第一次和第二次接种后五天内不良反应(AE)的总数是14，其中包括9名轻度AE，4名中度AE。只有一名患者显示重度AE，包括体温在第一次接种后3小时内升到38.8℃，没有局部或系统性症状。随后四小时内他的体温再次回复正常。第一次接种后观察到7例AE。其中一例是局部的，六例是系统性的。第二次接种后观察到2例局部AE和5例系统性AE。
就安全性而言，揭示在研究的各组包括安慰剂组间没有显著差异。除上面提到的一例外，没有观察到与接种相关的重度AE。其中两例中度AE发生在H3N2组(一名志愿者在第三天体温升高到37.6℃并出现急性咽炎；另一名志愿者在第22-24天出现鼻塞、咽喉不适，并且体温升到37.5℃)，一例出现在H1N1组(从22-26天咽喉疼痛、鼻炎，体温在22-24天之间升到37-37.8℃)。
表25：血清反应阴性的志愿者对Vero Vac疫苗以及对三价俄罗斯冷适应的从蛋产生的疫苗的反应
编号 用于免疫的疫苗 病毒剂量， TCID50/ml 或EID50/ml 志愿 者数 目对抗原的血清HAI抗体滴度增加地至少4倍的志愿者％H1N1 H3N 2 B 1 安慰剂 13 (8) 2 Vero Vac H1(H1N1) 4.3×107 1338 3 Vero Vac H3(H3N2) 2.1×107 13 67 4 俄罗斯三价疫苗： A/17/Beijing/95/25 H1N1 A/17/Sidney/97/76 H3N2 B/60/Petersburg/95/20 1.1×108 2.3×107 1.1×107 1346 8 31 5 Vero疫苗BV22 2×106 13 33
(8)患者在研究期间发展出自发感染。
因此，从所述临床研究得出的结果证实：根据本发明生产的疫苗以及使用本发明优选的A型和B型流感原株候选物具有非常好的安全性。
【序列表】
<110>Polymun Scientific Immunbiologische Forschung GmbH
Katinger，Hermann
Katinger，Dietmar
Romanova，Julia
Egorov，Andrej
Ferko，Boris
<120>流感疫苗和生产方法
<130>BEP-4847-PC(流感疫苗)
<140>
<141>
<160>37
<170>PatentIn Ver.2.1
<210>1
<211>2341
<212>DNA
<213>流感病毒A/Singapore/1/57/ca
<400>1
agcaaaagca ggtcaattat attcaatatg gaaagaataa aagaactacg gaatctgatg 60
tcgcagtctc gcactcgcga gatactaaca aaaaccacag tggaccatat ggccataatt 120
aagaagtaca catcagggag acaggaaaag aacccgtcac ttaggatgaa atggatgatg 180
gcaatgaaat atccgattac agctgacaag aggataacag aaatgattcc tgagagaaat 240
gagcaagggc agactctatg gagtaaaatg aatgatgccg gatcggatcg agtgatggta 300
tcacctctgg ctgtgacatg gtggaataga aatggaccaa tgacaagtac ggttcattat 360
ccaaaaatct acaaaactta ttttgagaaa gtcgaaaggt taaaacatgg aacctttggc 420
cctgtccatt ttagaaacca agtcaaaata cgccgaagag ttgacataaa tcctggtcat 480
gcagacctca gtgccaagga ggcacaggat gtaatcatgg aagttgtttt ccctaacgaa 540
gtgggggcca ggatactaac gtcggaatcg caattaacaa caaccaaaga gaaaaaagaa 600
gaactccagg attgcaaaat ttctcctttg atggttgcgt acatgttaga gagagaactt 660
gtccgaaaaa cgagatttct cccagttgct ggtggaacaa gcagtgtgta cattgaagtg 720
ttgcacttaa ctcaaggaac atgctgggaa cagatgtaca ctccaggtgg agaagtgagg 780
aatgatgatg ttgatcaaag tctaattatt gcagccagga acatagtgag aagagcagca 840
gtatcagcag atccactagc atctttattg gagatgtgcc acagcacaca gattggcggg 900
acaaggatgg tggacattct taggcagaac ccaacggaag agcaagctgt ggatatatgc 960
aaggctgcaa tgggactgag aatcagctca tccttcagtt ttggcgggtt cacatttaag 1020
agaacaagcg gatcatcagt caagatagag gaagaagtgc ttacgggcaa tcttcaaaca 1080
ttgaaaataa gggtgcatga gggatacgag gagttcacaa tggttgggaa aagggcaaca 1140
gctatactca gaaaagcaac caggagattg attcagctga tagtgagtgg aagagacgaa 1200
cagtcgatag ccgaagcaat aattgtggcc atggtatttt cacaagaaga ttgtatgata 1260
aaagcagtta gaggtgatct gaatttcgtt aatagggcaa atcagcgatt gaatcccatg 1320
catcaacttt taagacattt tcagaaggat gcgaaagtgc tttttcaaaa ttggggaatt 1380
gaacatatcg acaatgtgat gggaatgatt ggggtattac cagacatgac tccaagcaca 1440
gagatgtcaa tgagaggggt aagagtcagc aaaatgggcg tagatgaata ctccagcgcg 1500
gagagagtag tggtgagcat tgaccggttt ttgagagttc gagaccaacg aggaaatgta 1560
ctactatctc ctgaggaggt cagtgaaaca cagggaacag agaaactgac aataacttac 1620
tcatcgtcaa tgatgtggga gattaatggc cctgagtcag tgttggtcaa tacctatcag 1680
tggatcatca gaaactggga aactgttaaa attcagtggt ctcagaatcc tacaatgcta 1740
tacaataaaa tggaatttga gccatttcag tctttagttc ctaaggccat tagaggccaa 1800
tacagtgggt ttgttaggac tctattccaa caaatgaggg atgtacttgg gacatttgat 1860
accacccaga taataaaact tcttcccttt gcagccgccc caccaaagca aagtagaatg 1920
cagttctctt cattgactgt gaatgtgagg ggatcaggaa tgagaatact tgtaaggggc 1980
aattctcctg tattcaacta caacaagacc actaagagac taacaattct cggaaaggat 2040
gctggcactt taactgaaga cccagatgaa ggcacatctg gagtggagtc cgctgttctg 2100
agaggattcc tcattctggg caaagaagat aggagatatg gaccagcatt aagcatcaat 2160
gaactgagta accttgcgaa aggagaaaag gctaatgtac taattgggca aggagacgtg 2220
gtgttggtaa tgaaacgaaa acgggactct agcatactta ctgacagcca gacagcgacc 2280
aaaagaattc ggatggccat caattaatgt tgaatagttt aaaaacgacc ttgtttctac 2340
t 2341
<210>2
<211>2341
<212>DNA
<213>流感病毒A/Singapore/1/57/ca
<400>2
agcaaaagca ggcaaaccat ttgaatggat gtcaatccga ccttactttt cttgaaagtt 60
ccagcgcaaa atgccataag tactacattc ccttatactg gagatcctcc atacagccat 120
ggaacaggaa caggatacac catggacaca gtcaacagaa cacatcaata ttcagaaaag 180
gggaagtgga caacaaacac ggaaactgga gcgccccaac ttaacccaat tgatggacca 240
ctacctgagg acaatgaacc aagtggatat gcacaaacag actgcgtcct ggaagcaatg 300
gctttccttg aagaatccca cccgggaatc tttgaaaact cgtgtcttga aacgatggaa 360
gttattcaac aaacaagagt ggacaaactg acccaaggtc gtcagaccta tgattggaca 420
ttgaacagaa atcagccggc tgcaactgcg ctagccaaca ctatagaggt cttcagatcg 480
aatggtctga cagctaatga atcgggaagg ctaatagatt tcctcaagga tgtgatagaa 540
tcaatggata aagaggagat ggaaataaca acacacttcc aaagaaaaag aagagtaaga 600
gacaacatga ccaagaaaat ggtcacacaa cgaacaatag gaaaaaagaa gcaaagattg 660
aacaagagaa gctatctaat aagagcactg acattgaaca caatgactaa agatgcagag 720
agaggtaaat taaagagaag agcaattgca acacccggta tgcagatcag agggttcgtg 780
tactttgtcg aaacactagc gagaagtatt tgtgagaagc ttgaacagtc tgggcttccg 840
gttggaggta atgaaaagaa ggctaaactg gcaaatgttg tgagaaaaat gatgactaat 900
tcacaagaca cagagctctc tttcacaatt actggagaca ataccaaatg gaatgagaat 960
caaaatcctc ggatgttcct ggcgatgata acatacatca caagaaatca acctgaatgg 1020
tttagaaacg tcctgagcat cgcacctata atgttctcaa ataaaatggc aagactaggg 1080
aaaggataca tgttcgaaag caagagcatg aagctccgaa cacaaatacc agcagaaatg 1140
ctagcaagta ttgacctgaa atactttaat gaatcaacaa gaaagaaaat cgagaaaata 1200
aggcctctcc taatagatgg cacagtctca ttgagtcctg gaatgatgat gggcatgttc 1260
aacatgctaa gtacagtcat aggagtctca atcctgaatc ttggacaaaa gaagtacacc 1320
aaaacaacat actggtggga cggactccaa tcctctgatg acttcgccct catagtgaat 1380
gcaccaaatc atgagggaat acaagcagga gtggatagat tctacagaac ctgcaagcta 1440
gtcggaatca atatgagcaa aaagaagtcc tacataaata ggacagggac atttgaattc 1500
acaagctttt tctatcgcta tggatttgta gccaatttta gcatggagct gcccagtttt 1560
ggagtgtctg gaattaatga atcggctgat atgagcattg gggtaacagt gataaagaac 1620
aacatgataa acaatgacct tgggccagca acagcccaaa tggctcttca actattcatc 1680
aaagactaca gatatacgta ccggtgccac agaggagaca cacaaattca gacaaggaga 1740
tcattcgagc taaagaagct gtgggagcaa acccgctcaa aggcaggact tttggtttcg 1800
gatggaggac caaacttata caatatccgg aatctccaca ttccagaagt ctgcttgaag 1860
tgggagctaa tggatgaaga ctatcagggg aggctttgta atcccctgaa tccatttgtc 1920
agtcataagg agattgagtc tgtaaacaat gctgtggtaa tgcccgctca cggtccagcc 1980
aagagcatgg aatatgatgc tgttgctact acacactcct ggatccctaa gaggaaccgc 2040
tccattctca acacaagcca aaggggaatt cttgaggatg aacagatgta tcagaagtgt 2100
tgcaatctat tcgagaaatt cttccctagc agttcgtaca ggagaccagt tggaatttcc 2160
agcatggtgg aggccatggt gtctagggcc cggattgatg cacggattga cttcgagtct 2220
ggacggatta agaaagagga gttcgctgag atcatgaaga tctgttccac cattgaagag 2280
ctcagacggc aaaaatagtg aatttagctt gtccttcatg aaaaaatgcc ttgtttctac 2340
t 2341
<210>3
<211>2233
<212>DNA
<213>流感病毒A/Singapore/1/57/ca
<400>3
agcaaaagca ggtactgatc cgaaatggaa gattttgtgc gacaatgctt caatccgatg 60
attgtcgagc ttgcggaaag ggcaatgaaa gagtatggag aggatctgaa aatcgaaaca 120
aacaaatttg cagcaatatg cactcacttg gaagtatgct tcatgtattc agattttcat 180
ttcatcaatg agcaaggcga gtcaataata gtagagcttg atgatccaaa tgcacttttg 240
aagcacagat ttgaaataat agagggaaga gatcgcacaa tggcctggac agtagtaaac 300
agtatttgca acactacagg agctgagaaa ccgaagtttc tgccagattt gtatgattac 360
aaggagaata gattcatcga gattggagtg acaaggaggg aagtccacat atactatctt 420
gaaaaggcca ataaaattaa atctgagaag acacacatcc acattttctc attcactggg 480
gaagaaatgg ccacaaaggc cgactacact ctcgatgagg aaagcagggc taggatcaaa 540
accagactat tcaccataag acaagaaatg gctagcagag gcctctggga ttcctttcgt 600
cagtccgaaa gaggcgaaga aacaattgaa gaaagatttg aaatcacagg gacaatgcgc 660
aggctcgccg accaaagtct cccgccgaac ttctcctgcc ttgagatttt tagagcctat 720
gtggatggat tcgaaccgaa cggctacatt gagggcaagc tttctcaaat gtccaaagaa 780
gtaaatgcta aaattgaacc ttttctgaaa acaacaccaa gaccaattag acttccggat 840
gggcctcctt gttctcagcg gtccaaattc ctgctgatgg atgctttaaa attaagcatt 900
gaggacccaa gtcacgaagg agagggaata ccactatatg atgcgatcaa gtgtatgaga 960
acattctttg gatggaaaga accctatgtt gttaaaccac acgaaaaggg aataaatcca 1020
aattatctgc tgtcatggaa gcaagtactg gcagaactgc aggacattga gaatgaggag 1080
aagattccaa gaaccaaaaa catgaagaaa acgagtcagc taaagtgggc acttggtgag 1140
aacatggcac cagagaaggt agactttgac gactgtagag atataagcga tttgaagcaa 1200
tatgatagtg atgaacctga attaaggtca ctttcaagct ggatccagaa tgagttcaac 1260
aaggcatgcg agctgaccaa ttcaatctgg atagagctcg atgagattgg agaagatgtg 1320
gctccaattg aacacattgc aagcatgaga aggaattact tcacagcaga ggtgtctcat 1380
tgcagagcca cagaatatat aatgaagggg gtatacatta atacagcctt gcttaatgca 1440
tcctgtgcag caatggacga tttccaacta attcccatga taagcaaatg tagaactaaa 1500
gagggaaggc gaaagaccaa tttatatggt ttcatcgtaa aaggaagatc tcacttaagg 1560
aatgacaccg acgtggtaaa ctttgtgagc atggagtttt ctctcactga cccaagactt 1620
gagccacaca aatgggagaa gtactgtgtt cttgagatag gagatatgct actaagaagt 1680
gccataggcc aggtgtcaag gcccatgttc ttgtatgtga ggacaaatgg aacatcaaag 1740
attaaaatga aatggggaat ggagatgagg cgttgcctcc ttcagtcact ccaacaaatc 1800
gagagcatga ttgaagccca gtcctctgtc aaggagaaag acatgaccaa agagtttttc 1860
gagaataaat cagaaacatg gcccattgga gagtccccta aaggagtgga agaaggttcc 1920
attgggaagg tctgcaggac tttattagcc aagtcggtat tcaatagcct gtatgcatct 1980
ccacaattag aaggattttc agctgaatca agaaaactgc tccttgtcgt tcaggctctt 2040
agggacaatc ttgaacctgg gacctttgat cttggggggc tatatgaagc aattgaggag 2100
tgcctgatta atgatccctg ggttttgctt aatgcgtctt ggttcaactc cttcctaaca 2160
catgcattaa gatagttgtg gcaatgctac tatttgctat ccatactgtc caaaaaagta 2220
ccttgtttct act 2233
<210>4
<211>1773
<212>DNA
<213>流感病毒A/Singapore/1/57/ca
<400>4
agcaaaagca ggggttatac catagacaac cagaagcaaa acaatggcca tcatttatct 60
cattctcctg ttcacagcag tgagagggga ccagatatgc attggatacc atgccaataa 120
ttccacagag aaggtcgaca caattctaga gcagaacgtc actgtgactc atgccaagga 180
cattcttgag aagacccata acggaaagtt atgcaaacta aacggaatcc ctccacttga 240
actaggggac tgtagcattg ccggatggct ccttggaaat ccagaatgtg ataggcttct 300
aagtgtgcca gaatggtcct atataatgga gaaagaaaac ccgagagacg gtttgtgtta 360
tccaggcagc ttcaatgatt atgaagaatt gaaacatctc ctcagcagcg tgaaacattt 420
cgagaaagta aagattctgc ccaaagatag atggacacag catacaacaa ctggaggttc 480
acgggcctgc gcggtgtctg gtaatccatc attcttcagg aacatggtct ggctgacaaa 540
gaaagaatca aattatccgg ttgccaaagg atcgtacaac aatacaagcg gagaacaaat 600
gctaataat ttggggggtgc accatcccaa tgatgagaca gaacaaagaa cattgtacca 660
gaatgtggga acctatgttt ccgtaggcac atcaacattg aacaaaaggt caaccccaga 720
catagcaaca aggcctaaag tgaatggact aggaagtaga atggaattct cttggaccct 780
attggatatg tgggacacca taaattttga gagtactggt aatctaattg caccagagta 840
tggattcaaa atatcgaaaa gaggtaattc agggatcatg aaaacagaag gaacacttga 900
gaactgtgag accaaatgcc aaactccttt gggagcaata aatacaacat tgccttttca 960
caatgtccac ccactgacaa taggtgagtg ccccaaatat gtaaaatcgg agaagttggt 1020
cttagcaaca ggaccaagga atgttcccca gattgaatca agaggattgt ttggggcaat 1080
agctggtttt atagaaggag gatggcaagg aatggttgat ggttggtatg gataccatca 1140
cagcaatgac cagggatcag ggtatgcagc agacaaagaa tccactcaaa aggcatttga 1200
tggaatcacc aacaaggtaa attctgtgat tgaaaagatg aacacccaat ttgaagctgt 1260
tgggaaagaa ttcagtaact tagagagaag actggagaac ttgaacaaaa agatggaaga 1320
cgggtttcta gatgtgtgga catacaatgc tgagcttcta gttctgatgg aaaatgagag 1380
gacacttgac tttcatgatt ctaatgtcaa gaatctgtat gataaagtca gaatgcagct 1440
gagagacaac gtcaaagaac taggaaatgg atgttttgaa ttttatcaca aatgtgatga 1500
tgaatgcatg aatagtgtga aaaacgggac gtatgattat cccaagtatg aagaagagtc 1560
taaactaaat agaaatgaaa tcaaaggggt aaaattgagc agcatggggg tttatcaaat 1620
ccttgccatt tatgctacag tagcaggttc tctgtcactg gcaatcatga tggctgggat 1680
ctctttctgg atgtgctcca acgggtctct gcagtgcagg atctgcatat gattataagt 1740
cattttataa ttaaaaacac ccttgtttct act 1773
<210>5
<211>1565
<212>DNA
<213>流感病毒A/Singapore/1/57/ca
<400>5
agcaaaagca gggtagataa tcactcactg agtgacatca aaatcatggc gtcccaaggc 60
accaaacggt cttatgaaca gatggaaact gatggggaac gccagaatgc aactgaaatc 120
agagcatccg tcgggaagat gattgatgga attggacgat tctacatcca aatgtgcacc 180
gaacttaaac tcagtgatta tgaggggcga ctgatccaga acagcttaac aatagagaga 240
atggtgctct ctgcttttga cgagaggagg aataaatatc tggaagaaca tcccagcgcg 300
gggaaggatc ctaagaaaac tggaggaccc atatacaaga gagtaaatgg aaagtggatg 360
agggaactcg tcctttatga caaagaagaa ataaggcgaa tctggcgcca agctaataat 420
ggtgatgatg caacagctgg tctgactcac atgatgatct ggcattccaa tttgaatgat 480
acaacatacc agaggacaag agctcttgtt cgcaccggaa tggatcccag gatgtgctct 540
ttgatgcagg gttcgactct ccctaggagg tctggagccg caggcgctgc agtcaaagga 600
gttgggacaa tggtgatgga gttgatcagg atgatcaaac gtgggatcaa tgatcggaac 660
ttctggagag gtgagaatgg gcggaaaaca aggattgctt atgagagaat gtgcaacatt 720
ctcaaaggaa aatttcaaac agctgcacaa agagcaatga tggatcaagt gagagaaagc 780
cggaacccag gaaatgctga gatcgaagat ctcatctttc tggcacggtc tgcactcata 840
ttgagagggt cagttgctca caaatcttgt ctgcctgcct gtgtgtatgg aactgccgta 900
gccagtgggt acgacttcga aaaagaggga tactctttag tagggataga ccctttcaaa 960
ctgcttcaaa acagccaagt atacagccta atcagaccga acgagaatcc agcacacaag 1020
agtcagctgg tgtggatggc atgcaattct gctgcatttg aagatctaag agtatcaagc 1080
ttcatcagag ggaccaaagtaatcccaagg gggaaacttt ccactagagg agtacaaatt 1140
gcttcaaatg aaaacatgga tactatggaa tcaagtactc ttgaactgag aagcaggtac 1200
tgggccataa ggaccagaag tggaggaaac actaatcaac agagggcctc tgcaggtcaa 1260
atcagtgtac aacctacgtt ttctgtgcaa agaaacctcc catttgacaa aacaaccatc 1320
atggcagcat tcactgggaa tgcagaggga agaacatcag acatgagggc agaaatcata 1380
aggatgatgg aaggtgcaaa accagaagaa gtgtccttcc aggggcgggg agtcttcgag 1440
ctctcggacg aaaaggcaac gaacccgatc gtgccctctt ttgacatgag taatgaagga 1500
tcttatttct tcggagacaa tgcagaggag tacgacaatt aaggaaaaat acccttgttt 1560
ctact 1565
<210>6
<211>1466
<212>DNA
<213>流感病毒A/Singapore/1/57/ca
<400>6
agcaaaagca ggagtgaaga tgaatccaaa tcaaaagata ataacaattg gctctgtctc 60
tctcaccatt gcaacagtat gcttcctcat gcagattgcc atcctggcaa ctactgtgac 120
attgcatttt aaacaacatg agtgcgactc ccccgcgagc aaccaagtaa tgccatgtga 180
accaataata atagaaagga acataacaga gatagtgtat ttgaataaca ccaccataga 240
gaaagagatt tgccccgaag tagtggaata cagaaattgg tcaaagccgc aatgtcaaat 300
tacaggattt gcaccttttt ctaaggacaa ttcaatccgg ctttctgctg gtggggacat 360
ttgggtgacg agagaacctt atgtgtcatg cgatcctggc aagtgttatc aatttgcact 420
cgggcagggg accacactat acaacaaaca ttcaaatggc acaatacatg atagaatccc 480
tcatcgaacc ctattaatga atgagttggg tgttccattt catttaggaa ccaaacaagt 540
gtgtgtagca tggtccagct caagttgtca cgatggaaaa gcatggttgc atgtttgtgt 600
cactggggat gatagaaatg cgactgctag cttcatttat gacgggaggc ttgtggacag 660
tattggttca tggtctcaaa atatcctcag gacccaggag tcggaatgcg tttgtatcaa 720
tgggacttgc acagtagtaa tgactgatgg aagtgcatca ggaagagccg atactagaat 780
actattcatt aaagagggga aaattgtccg tattagccca ttgtcaggaa gtgctcagca 840
tatagaggag tgttcctgtt accctcgata tcctgacgtc agatgtatct gcagagacaa 900
ctggaaaggc tctaataggc ccgttataga cataaatatg gaagattata gcattgattc 960
cagttatgtg tgctcagggc ttgttggcga cacacccagg aacgacgaca gctctagcaa 1020
tagcaattgc agggatccta acaatgagag agggaatcca ggagtgaaag gctgggcctt 1080
tgacaatgga gatgatgtat ggatgggaag aacaatcaac aaagattcac gctcaggtta 1140
tgaaactttc aaagtcattg gtggttggtc cacacctaat tccaaatcgc aggtcaatag 1200
acaggtcata gttgacaaca ataattggtc tggttactct ggtattttct ctgttgaggg 1260
caaaagctgc atcaataggt gcttttatgt ggagttgata aggggaaggc cacaggagac 1320
tagagtatgg tggacctcaa acagtattgt tgtgttttgt ggcacttcag gtacttatgg 1380
aacaggctca tggcctgatg gggcgaacat caatttcatg cctatataag ctttcgcaat 1440
tttagaaaaa actccttgtt tctact 1466
<210>7
<211>1027
<212>DNA
<213>流感病毒A/Singapore/1/57/ca
<400>7
agcaaaagca ggtagatatt gaaagatgag tcttctaacc gaggtcgaaa cgtatgttct 60
ctctatcgtc ccgtcaggcc ccctcaaagc cgagatcgca cagagacttg aagatgtctt 120
tgctgggaag aacaccgatc ttgaggctct catggaatgg ctaaagacaa gaccaatcct 180
gtcacctctg actaagggga ttttgggatt tgtattcacg ctcaccgtgc ccagtgagcg 240
aggactgcag cgtagacgct ttgtccaaaa tgccctcaat gggaatgggg atccaaataa 300
catggacaga gcagttaaac tgtataaaaa gcttaagagg gagataacat tccatggggc 360
caaagaaata gcgctcagtt attctgctgg tgcacttgcc agttgtatgg gcctcatata 420
caacaggatg ggggctgtga ccactgaagt ggcctttggc ctggtatgtg caacctgtga 480
acagattgct gactcccacc ataggtctca taggcaaatg gtgacaacaa ccaatccact 540
aataagacat gagaacagaa tggttctggc cagcactaca gctaaggcta tggagcaaat 600
ggctggatcg agtgagcaag cagcagaggc catggaggtt gctagtcagg ccaggcaaat 660
ggtgcaggca atgagagcca ttgggactca tcctagctcc agtgctggtc taaaagatga 720
tcttcttgaa aatttgcagg cctatcagaa acgaatgggg gtgcagatgc aacgattcaa 780
gtgaccctct tgttgttgcc gcgagtatca ttgggatctt gcacttgata ttgtggattc 840
ttgatcgtct ttttttcaaa tgcatttatc gcttctttaa acacggtctg aaaagagggc 900
cttctacgga aggagtacca gagtctatga gggaagaata tcgaaaggaa cagcagagtg 960
ctgtggatgc tgacgatagt cattttgtca gcatagagct ggagtaaaaa actaccttgt 1020
ttctact 1027
<210>8
<211>890
<212>DNA
<213>流感病毒A/Singapore/1/57/ca
<400>8
agcaaaagca gggtgacaaa gacataatgg atcctaacac tgtgtcaagc tttcaggtag 60
attgcttcct ttggcatgtc cgcaaacaag ttgcagacca agaactaggt gatgccccat 120
tccttgatcg gcttcgccga gatcagaagt ccctaagggg aagaggcagc actctcggtc 180
tgaacatcga aacagccacc cgtgttggaa agcagatagt ggagaggatt ctgaaggaag 240
aatccgatga ggcacttaaa atgaccatgg cctccgcacc tgcttcgcga tacctaactg 300
acatgactat tgaggaaatg tcaagggact ggttcatgct aatgcccaag cagaaagtgt 360
caggccctct ttgtatcaga atggaccagg caatcatgga taagaacatc atattgaaag 420
cgaatttcag tgtgattttt gaccggctag agaccctaat attactaagg gctttcaccg 480
aagagggagc aattgttggc gaaatttcac cattgccttc tcttccagga catactaatg 540
aggatgtcaa aaatgcaatt ggggtcctca tcggaggact tgaatggaat gataacacag 600
ttcgagtctc taaaactcta cagagattcg cttggagaaa cagtaatgag aatgggagac 660
ctccactcac tccaaaacag aaacggaaaa tggcgagaac aattaggtca aaagttcgaa 720
gaaataagat ggctgattga agaagtgaga cacaaattga agataacaga gaatagtttt 780
gagcaaataa catttatgca agccttacag ctgctatttg aagtggaaca agagataaga 840
actttctcgt ttcagcttat ttaatgataa aaaacaccct tgtttctact 890
<210>9
<211>771
<212>PRT
<213>流感病毒A/Singapore/1/57/ca
<400>9
Met Glu Arg Ile Lys Glu Leu Arg Asn Leu Met Ser Gln Ser Arg Thr
1 5 10 15
Arg Glu Ile Leu Thr Lys Thr Thr Val Asp His Met Ala Ile Ile Lys
20 25 30
Lys Tyr Thr Ser Gly Arg Gln Glu Lys Asn Pro Ser Leu Arg Met Lys
35 40 45
Trp Met Met Ala Met Lys Tyr Pro Ile Thr Ala Asp Lys Arg Ile Thr
50 55 60
Glu Met Ile Pro Glu Arg Asn Glu Gln Gly Gln Thr Leu Trp Ser Lys
65 70 75 80
Met Asn Asp Ala Gly Ser Asp Arg Val Met Val Ser Pro Leu Ala Val
85 90 95
Thr Trp Trp Asn Arg Asn Gly Pro Met Thr Ser Thr Val His Tyr Pro
100 105 110
Lys Ile Tyr Lys Thr Tyr Phe Glu Lys Val Glu Arg Leu Lys His Gly
115 120 125
Thr Phe Gly Pro Val His Phe Arg Asn Gln Val Lys Ile Arg Arg Arg
130 135 140
Val Asp Ile Asn Pro Gly His Ala Asp Leu Ser Ala Lys Glu Ala Gln
145 150 155 160
Asp Val Ile Met Glu Val Val Phe Pro Asn Glu Val Gly Ala Arg Ile
165 170 175
Leu Thr Ser Glu Ser Gln Leu Thr Thr Thr Lys Glu Lys Lys Glu Glu
180 185 190
Leu Gln Asp Cys Lys Ile Ser Pro Leu Met Val Ala Tyr Met Leu Glu
195 200 205
Arg Glu Leu Val Arg Lys Thr Arg Phe Leu Pro Val Ala Gly Gly Thr
210 215 220
Ser Ser Val Tyr Ile Glu Val Leu His Leu Thr Gln Gly Thr Cys Trp
225 230 235 240
Glu Gln Met Tyr Thr Pro Gly Gly Glu Val Arg Asn Asp Asp Val Asp
245 250 255
Gln Ser Leu Ile Ile Ala Ala Arg Asn Ile Val Arg Arg Ala Ala Val
260 265 270
Ser Ala Asp Pro Leu Ala Ser Leu Leu Glu Met Cys His Ser Thr Gln
275 280 285
Ile Gly Gly Thr Arg Met Val Asp Ile Leu Arg Gln Asn Pro Thr Glu
290 295 300
Glu Gln Ala Val Asp Ile Cys Lys Ala Ala Met Gly Leu Arg Ile Ser
305 310 315 320
Ser Ser Phe Ser Phe Gly Gly Phe Thr Phe Lys Arg Thr Ser Gly Ser
325 330 335
Ser Val Lys Ile Glu Glu Glu Val Leu Thr Gly Asn Leu Gln Thr Leu
340 345 350
Lys Ile Arg Val His Glu Gly Tyr Glu Glu Phe Thr Met Val Gly Lys
355 360 365
Arg Ala Thr Ala Ile Leu Arg Lys Ala Thr Arg Arg Leu Ile Gln Leu
370 375 380
Ile Val Ser Gly Arg Asp Glu Gln Ser Ile Ala Glu Ala Ile Ile Val
385 390 395 400
Ala Met Val Phe Ser Gln Glu Asp Cys Met Ile Lys Ala Val Arg Gly
405 410 415
Asp Leu Asn Phe Val Asn Arg Ala Asn Gln Arg Leu Asn Pro Met His
420 425 430
Gln Leu Leu Arg His Phe Gln Lys Asp Ala Lys Val Leu Phe Gln Asn
435 440 445
Trp Gly Ile Glu His Ile Asp Asn Val Met Gly Met Ile Gly Val Leu
450 455 460
Pro Asp Met Thr Pro Ser Thr Glu Met Ser Met Arg Gly Val Arg Val
465 470 475 480
Ser Lys Met Gly Val Asp Glu Tyr Ser Ser Ala Glu Arg Val Val Val
485 490 495
Ser Ile Asp Arg Phe Leu Arg Val Arg Asp Gln Arg Gly Asn Val Leu
500 505 510
Leu Ser Pro Glu Glu Val Ser Glu Thr Gln Gly Thr Glu Lys Leu Thr
515 520 525
Ile Thr Tyr Ser Ser Ser Met Met Trp Glu Ile Asn Gly Pro Glu Ser
530 535 540
Val Leu Val Asn Thr Tyr Gln Trp Ile Ile Arg Asn Trp Glu Thr Val
545 550 555 560
Lys Ile Gln Trp Ser Gln Asn Pro Thr Met Leu Tyr Asn Lys Met Glu
565 570 575
Phe Glu Pro Phe Gln Ser Leu Val Pro Lys Ala Ile Arg Gly Gln Tyr
580 585 590
Ser Gly Phe Val Arg Thr Leu Phe Gln Gln Met Arg Asp Val Leu Gly
595 600 605
Thr Phe Asp Thr Thr Gln Ile Ile Lys Leu Leu Pro Phe Ala Ala Ala
610 615 620
Pro Pro Lys Gln Ser Arg Met Gln Phe Ser Ser Leu Thr Val Asn Val
625 630 635 640
Arg Gly Ser Gly Met Arg Ile Leu Val Arg Gly Asn Ser Pro Val Phe
645 650 655
Asn Tyr Asn Lys Thr Thr Lys Arg Leu Thr Ile Leu Gly Lys Asp Ala
660 665 670
Gly Thr Leu Thr Glu Asp Pro Asp Glu Gly Thr Ser Gly Val Glu Ser
675 680 685
Ala Val Leu Arg Gly Phe Leu Ile Leu Gly Lys Glu Asp Arg Arg Tyr
690 695 700
Gly Pro Ala Leu Ser Ile Asn Glu Leu Ser Asn Leu Ala Lys Gly Glu
705 710 715 720
Lys Ala Asn Val Leu Ile Gly Gln Gly Asp Val Val Leu Val Met Lys
725 730 735
Arg Lys Arg Asp Ser Ser Ile Leu Thr Asp Ser Gln Thr Ala Thr Lys
740 745 750
Arg Ile Arg Met Ala Ile Asn Xaa Cys Xaa Ile Val Xaa Lys Arg Pro
755 760 765
Cys Phe Tyr
770
<210>10
<211>757
<212>PRT
<213>流感病毒A/Singapore/1/57/ca
<400>10
Met Asp Val Asn Pro Thr Leu Leu Phe Leu Lys Val Pro Ala Gln Asn
1 5 10 15
Ala Ile Ser Thr Thr Phe Pro Tyr Thr Gly Asp Pro Pro Tyr Ser His
20 25 30
Gly Thr Gly Thr Gly Tyr Thr Met Asp Thr Val Asn Arg Thr His Gln
35 40 45
Tyr Ser Glu Lys Gly Lys Trp Thr Thr Asn Thr Glu Thr Gly Ala Pro
50 55 60
Gln Leu Asn Pro Ile Asp Gly Pro Leu Pro Glu Asp Asn Glu Pro Ser
65 70 75 80
Gly Tyr Ala Gln Thr Asp Cys Val Leu Glu Ala Met Ala Phe Leu Glu
85 90 95
Glu Ser His Pro Gly Ile Phe Glu Asn Ser Cys Leu Glu Thr Met Glu
100 105 110
Val Ile Gln Gln Thr Arg Val Asp Lys Leu Thr Gln Gly Arg Gln Thr
115 120 125
Tyr Asp Trp Thr Leu Asn Arg Asn Gln Pro Ala Ala Thr Ala Leu Ala
130 135 140
Asn Thr Ile Glu Val Phe Arg Ser Asn Gly Leu Thr Ala Asn Glu Ser
145 150 155 160
Gly Arg Leu Ile Asp Phe Leu Lys Asp Val Ile Glu Ser Met Asp Lys
165 170 175
Glu Glu Met Glu Ile Thr Thr His Phe Gln Arg Lys Arg Arg Val Arg
180 185 190
Asp Asn Met Thr Lys Lys Met Val Thr Gln Arg Thr Ile Gly Lys Lys
195 200 205
Lys Gln Arg Leu Asn Lys Arg Ser Tyr Leu Ile Arg Ala Leu Thr Leu
210 215 220
Asn Thr Met Thr Lys Asp Ala Glu Arg Gly Lys Leu Lys Arg Arg Ala
225 230 235 240
Ile Ala Thr Pro Gly Met Gln Ile Arg Gly Phe Val Tyr Phe Val Glu
245 250 255
Thr Leu Ala Arg Ser Ile Cys Glu Lys Leu Glu Gln Ser Gly Leu Pro
260 265 270
Val Gly Gly Asn Glu Lys Lys Ala Lys Leu Ala Asn Val Val Arg Lys
275 280 285
Met Met Thr Asn Ser Gln Asp Thr Glu Leu Ser Phe Thr Ile Thr Gly
290 295 300
Asp Asn Thr Lys Trp Asn Glu Asn Gln Asn Pro Arg Met Phe Leu Ala
305 310 315 320
Met Ile Thr Tyr Ile Thr Arg Asn Gln Pro Glu Trp Phe Arg Asn Val
325 330 335
Leu Ser Ile Ala Pro Ile Met Phe Ser Asn Lys Met Ala Arg Leu Gly
340 345 350
Lys Gly Tyr Met Phe Glu Ser Lys Ser Met Lys Leu Arg Thr Gln Ile
355 360 365
Pro Ala Glu Met Leu Ala Ser Ile Asp Leu Lys Tyr Phe Asn Glu Ser
370 375 380
Thr Arg Lys Lys Ile Glu Lys Ile Arg Pro Leu Leu Ile Asp Gly Thr
385 390 395 400
Val Ser Leu Ser Pro Gly Met Met Met Gly Met Phe Asn Met Leu Ser
405 4l0 415
Thr Val Ile Gly Val Ser Ile Leu Asn Leu Gly Gln Lys Lys Tyr Thr
420 425 430
Lys Thr Thr Tyr Trp Trp Asp Gly Leu Gln Ser Ser Asp Asp Phe Ala
435 440 445
Leu Ile Val Asn Ala Pro Asn His Glu Gly Ile Gln Ala Gly Val Asp
450 455 460
Arg Phe Tyr Arg Thr Cys Lys Leu Val Gly Ile Asn Met Ser Lys Lys
465 470 475 480
Lys Ser Tyr Ile Asn Arg Thr Gly Thr Phe Glu Phe Thr Ser Phe Phe
485 490 495
Tyr Arg Tyr Gly Phe Val Ala Asn Phe Ser Met Glu Leu Pro Ser Phe
500 505 510
Gly Val Ser Gly Ile Asn Glu Ser Ala Asp Met Ser Ile Gly Val Thr
515 520 525
Val Ile Lys Asn Asn Met Ile Asn Asn Asp Leu Gly Pro Ala Thr Ala
530 535 540
Gln Met Ala Leu Gln Leu Phe Ile Lys Asp Tyr Arg Tyr Thr Tyr Arg
545 550 555 560
Cys His Arg Gly Asp Thr Gln Ile Gln Thr Arg Arg Ser Phe Glu Leu
565 570 575
Lys Lys Leu Trp Glu Gln Thr Arg Ser Lys Ala Gly Leu Leu Val Ser
580 585 590
Asp Gly Gly Pro Asn Leu Tyr Asn Ile Arg Asn Leu His Ile Pro Glu
595 600 605
Val Cys Leu Lys Trp Glu Leu Met Asp Glu Asp Tyr Gln Gly Arg Leu
6l0 615 620
Cys Asn Pro Leu Asn Pro Phe Val Ser His Lys Glu Ile Glu Ser Val
625 630 635 640
Asn Asn Ala Val Val Met Pro Ala His Gly Pro Ala Lys Ser Met Glu
645 650 655
Tyr Asp Ala Val Ala Thr Thr His Ser Trp Ile Pro Lys Arg Asn Arg
660 665 670
Ser Ile Leu Asn Thr Ser Gln Arg Gly Ile Leu Glu Asp Glu Gln Met
675 680 685
Tyr Gln Lys Cys Cys Asn Leu Phe Glu Lys Phe Phe Pro Ser Ser Ser
690 695 700
Tyr Arg Arg Pro Val Gly Ile Ser Ser Met Val Glu Ala Met Val Ser
705 710 715 720
Arg Ala Arg Ile Asp Ala Arg Ile Asp Phe Glu Ser Gly Arg Ile Lys
725 730 735
Lys Glu Glu Phe Ala Glu Ile Met Lys Ile Cys Ser Thr Ile Glu Glu
740 745 750
Leu Arg Arg Gln Lys
755
<210>11
<211>716
<212>PRT
<213>流感病毒A/Singapore/1/57/ca
<400>11
Met Glu Asp Phe Val Arg Gln Cys Phe Asn Pro Met Ile Val Glu Leu
1 5 10 15
Ala Glu Arg Ala Met Lys Glu Tyr Gly Glu Asp Leu Lys Ile Glu Thr
20 25 30
Asn Lys Phe Ala Ala Ile Cys Thr His Leu Glu Val Cys Phe Met Tyr
35 40 45
Ser Asp Phe His Phe Ile Asn Glu Gln Gly Glu Ser Ile Ile Val Glu
50 55 60
Leu Asp Asp Pro Asn Ala Leu Leu Lys His Arg Phe Glu Ile Ile Glu
65 70 75 80
Gly Arg Asp Arg Thr Met Ala Trp Thr Val Val Asn Ser Ile Cys Asn
85 90 95
Thr Thr Gly Ala Glu Lys Pro Lys Phe Leu Pro Asp Leu Tyr Asp Tyr
100 105 110
Lys Glu Asn Arg Phe Ile Glu Ile Gly Val Thr Arg Arg Glu Val His
115 120 125
Ile Tyr Tyr Leu Glu Lys Ala Asn Lys Ile Lys Ser Glu Lys Thr His
130 135 140
Ile His Ile Phe Ser Phe Thr Gly Glu Glu Met Ala Thr Lys Ala Asp
145 150 155 160
Tyr Thr Leu Asp Glu Glu Ser Arg Ala Arg Ile Lys Thr Arg Leu Phe
165 170 175
Thr Ile Arg Gln Glu Met Ala Ser Arg Gly Leu Trp Asp Ser Phe Arg
180 185 190
Gln Ser Glu Arg GIy Glu Glu Thr Ile Glu Glu Arg Phe Glu Ile Thr
195 200 205
Gly Thr Met Arg Arg Leu Ala Asp Gln Ser Leu Pro Pro Asn Phe Ser
210 215 220
Cys Leu Glu Ile Phe Arg Ala Tyr Val Asp Gly Phe Glu Pro Asn Gly
225 230 235 240
Tyr Ile Glu Gly Lys Leu Ser Gln Met Ser Lys Glu Val Asn Ala Lys
245 250 255
Ile Glu Pro Phe Leu Lys Thr Thr Pro Arg Pro Ile Arg Leu Pro Asp
260 265 270
Gly Pro Pro Cys Ser Gln Arg Ser Lys Phe Leu Leu Met Asp Ala Leu
275 280 285
Lys Leu Ser Ile Glu Asp Pro Ser His Glu Gly Glu Gly Ile Pro Leu
290 295 300
Tyr Asp Ala Ile Lys Cys Met Arg Thr Phe Phe Gly Trp Lys Glu Pro
305 310 315 320
Tyr Val Val Lys Pro His Glu Lys Gly Ile Asn Pro Asn Tyr Leu Leu
325 330 335
Ser Trp Lys Gln Val Leu Ala Glu Leu Gln Asp Ile Glu Asn Glu Glu
340 345 350
Lys Ile Pro Arg Thr Lys Asn Met Lys Lys Thr Ser Gln Leu Lys Trp
355 360 365
Ala Leu Gly Glu Asn Met Ala Pro Glu Lys Val Asp Phe Asp Asp Cys
370 375 380
Arg Asp Ile Ser Asp Leu Lys Gln Tyr Asp Ser Asp Glu Pro Glu Leu
385 390 395 400
Arg Ser Leu Ser Ser Trp Ile Gln Asn Glu Phe Asn Lys Ala Cys Glu
405 410 415
Leu Thr Asn Ser Ile Trp Ile Glu Leu Asp Glu Ile Gly Glu Asp Val
420 425 430
Ala Pro Ile Glu His Ile Ala Ser Met Arg Arg Asn Tyr Phe Thr Ala
435 440 445
Glu Val Ser His Cys Arg Ala Thr Glu Tyr Ile Met Lys Gly Val Tyr
450 455 460
Ile Asn Thr Ala Leu Leu Asn Ala Ser Cys Ala Ala Met Asp Asp Phe
465 470 475 480
Gln Leu Ile Pro Met Ile Ser Lys Cys Arg Thr Lys Glu Gly Arg Arg
485 490 495
Lys Thr Asn Leu Tyr Gly Phe Ile Val Lys Gly Arg Ser His Leu Arg
500 505 510
Asn Asp Thr Asp Val Val Asn Phe Val Ser Met Glu Phe Ser Leu Thr
515 520 525
Asp Pro Arg Leu Glu Pro His Lys Trp Glu Lys Tyr Cys Val Leu Glu
530 535 540
Ile Gly Asp Met Leu Leu Arg Ser Ala Ile Gly Gln Val Ser Arg Pro
545 550 555 560
Met Phe Leu Tyr Val Arg Thr Asn Gly Thr Ser Lys Ile Lys Met Lys
565 570 575
Trp Gly Met Glu Met Arg Arg Cys Leu Leu Gln Ser Leu Gln Gln Ile
580 585 590
Glu Ser Met Ile Glu Ala Gln Ser Ser Val Lys Glu Lys Asp Met Thr
595 600 605
Lys Glu Phe Phe Glu Asn Lys Ser Glu Thr Trp Pro Ile Gly Glu Ser
610 615 620
Pro Lys Gly Val Glu Glu Gly Ser Ile Gly Lys Val Cys Arg Thr Leu
625 630 635 640
Leu Ala Lys Ser Val Phe Asn Ser Leu Tyr Ala Ser Pro Gln Leu Glu
645 650 655
Gly Phe Ser Ala Glu Ser Arg Lys Leu Leu Leu Val Val Gln Ala Leu
660 665 670
Arg Asp Asn Leu Glu Pro Gly Thr Phe Asp Leu Gly Gly Leu Tyr Glu
675 680 685
Ala Ile Glu Glu Cys Leu Ile Asn Asp Pro Trp Val Leu Leu Asn Ala
690 695 700
Ser Trp Phe Asn Ser Phe Leu Thr His Ala Leu Arg
705 710 715
<210>12
<211>562
<212>PRT
<213>流感病毒A/Singapore/1/57/ca
<400>12
Met Ala Ile Ile Tyr Leu Ile Leu Leu Phe Thr Ala Val Arg Gly Asp
1 5 l0 15
Gln Ile Cys Ile Gly Tyr His Ala Asn Asn Ser Thr Glu Lys Val Asp
20 25 30
Thr Ile Leu Glu Gln Asn Val Thr Val Thr His Ala Lys Asp Ile Leu
35 40 45
Glu Lys Thr His Asn Gly Lys Leu Cys Lys Leu Asn Gly Ile Pro Pro
50 55 60
Leu Glu Leu Gly Asp Cys Ser Ile Ala Gly Trp Leu Leu Gly Asn Pro
65 70 75 80
Glu Cys Asp Arg Leu Leu Ser Val Pro Glu Trp Ser Tyr Ile Met Glu
85 90 95
Lys Glu Asn Pro Arg Asp Gly Leu Cys Tyr Pro Gly Ser Phe Asn Asp
100 105 110
Tyr Glu Glu Leu Lys His Leu Leu Ser Ser Val Lys His Phe Glu Lys
115 120 125
Val Lys Ile Leu Pro Lys Asp Arg Trp Thr Gln His Thr Thr Thr Gly
130 135 140
Gly Ser Arg Ala Cys Ala Val Ser Gly Asn Pro Ser Phe Phe Arg Asn
145 150 155 160
Met Val Trp Leu Thr Lys Lys Glu Ser Asn Tyr Pro Val Ala Lys Gly
165 170 175
Ser Tyr Asn Asn Thr Ser Gly Glu Gln Met Leu Ile Ile Trp Gly Val
180 185 190
His His Pro Asn Asp Glu Thr Glu Gln Arg Thr Leu Tyr Gln Asn Val
195 200 205
Gly Thr Tyr Val Ser Val Gly Thr Ser Thr Leu Asn Lys Arg Ser Thr
210 215 220
Pro Asp Ile Ala Thr Arg Pro Lys Val Asn Gly Leu Gly Ser Arg Met
225 230 235 240
Glu Phe Ser Trp Thr Leu Leu Asp Met Trp Asp Thr Ile Asn Phe Glu
245 250 255
Ser Thr Gly Asn Leu Ile Ala Pro Glu Tyr Gly Phe Lys Ile Ser Lys
260 265 270
Arg Gly Asn Ser Gly Ile Met Lys Thr Glu Gly Thr Leu Glu Asn Cys
275 280 285
Glu Thr Lys Cys Gln Thr Pro Leu Gly Ala Ile Asn Thr Thr Leu Pro
290 295 300
Phe His Asn Val His Pro Leu Thr Ile Gly Glu Cys Pro Lys Tyr Val
305 310 315 320
Lys Ser Glu Lys Leu Val Leu Ala Thr Gly Pro Arg Asn Val Pro Gln
325 330 335
Ile Glu Ser Arg Gly Leu Phe Gly Ala Ile Ala Gly Phe Ile Glu Gly
340 345 350
Gly Trp Gln Gly Met Val Asp Gly Trp Tyr Gly Tyr His His Ser Asn
355 360 365
Asp Gln Gly Ser Gly Tyr Ala Ala Asp Lys Glu Ser Thr Gln Lys Ala
370 375 380
Phe Asp Gly Ile Thr Asn Lys Val Asn Ser Val Ile Glu Lys Met Asn
385 390 395 400
Thr Gln Phe Glu Ala Val Gly Lys Glu Phe Ser Asn Leu Glu Arg Arg
405 410 415
Leu Glu Asn Leu Asn Lys Lys Met Glu Asp Gly Phe Leu Asp Val Trp
420 425 430
Thr Tyr Asn Ala Glu Leu Leu Val Leu Met Glu Asn Glu Arg Thr Leu
435 440 445
Asp Phe His Asp Ser Asn Val Lys Asn Leu Tyr Asp Lys Val Arg Met
450 455 460
Gln Leu Arg Asp Asn Val Lys Glu Leu Gly Asn Gly Cys Phe Glu Phe
465 470 475 480
Tyr His Lys Cys Asp Asp Glu Cys Met Asn Ser Val Lys Asn Gly Thr
485 490 495
Tyr Asp Tyr Pro Lys Tyr Glu Glu Glu Ser Lys Leu Asn Arg Asn Glu
500 505 510
Ile Lys Gly Val Lys Leu Ser Ser Met Gly Val Tyr Gln Ile Leu Ala
515 520 525
Ile Tyr Ala Thr Val Ala Gly Ser Leu Ser Leu Ala Ile Met Met Ala
530 535 540
Gly Ile Ser Phe Trp Met Cys Ser Asn Gly Ser Leu Gln Cys Arg Ile
545 550 555 560
Cys Ile
<210>13
<21l>506
<212>PRT
<213>流感病毒A/Singapore/1/57/ca
<400>13
Met Ala Ser Gln Gly Thr Lys Arg Ser Tyr Glu Gln Met Glu Thr Asp
1 5 10 15
Gly Glu Arg Gln Asn Ala Thr Glu Ile Arg Ala Ser Val Gly Lys Met
20 25 30
Ile Asp Gly Ile Gly Arg Phe Tyr Ile Gln Met Cys Thr Glu Leu Lys
35 40 45
Leu Ser Asp Tyr Glu Gly Arg Leu Ile Gln Asn Ser Leu Thr Ile Glu
50 55 60
Arg Met Val Leu Ser Ala Phe Asp Glu Arg Arg Asn Lys Tyr Leu Glu
65 70 75 80
Glu His Pro Ser Ala Gly Lys Asp Pro Lys Lys Thr Gly Gly Pro Ile
85 90 95
Tyr Lys Arg Val Asn Gly Lys Trp Met Arg Glu Leu Val Leu Tyr Asp
100 105 110
Lys Glu Glu Ile Arg Arg Ile Trp Arg Gln Ala Asn Asn Gly Asp Asp
115 120 125
Ala Thr Ala Gly Leu Thr His Met Met Ile Trp His Ser Asn Leu Asn
130 135 140
Asp Thr Thr Tyr Gln Arg Thr Arg Ala Leu Val Arg Thr Gly Met Asp
145 150 155 160
Pro Arg Met Cys Ser Leu Met Gln Gly Ser Thr Leu Pro Arg Arg Ser
165 170 175
Gly Ala Ala Gly Ala Ala Val Lys Gly Val Gly Thr Met Val Met Glu
180 185 190
Leu Ile Arg Met Ile Lys Arg Gly Ile Asn Asp Arg Asn Phe Trp Arg
195 200 205
Gly Glu Asn Gly Arg Lys Thr Arg Ile Ala Tyr Glu Arg Met Cys Asn
210 215 220
Ile Leu Lys Gly Lys Phe Gln Thr Ala Ala Gln Arg Ala Met Met Asp
225 230 235 240
Gln Val Arg Glu Ser Arg Asn Pro Gly Asn Ala Glu Ile Glu Asp Leu
245 250 255
Ile Phe Leu Ala Arg Ser Ala Leu Ile Leu Arg Gly Ser Val Ala His
260 265 270
Lys Ser Cys Leu Pro Ala Cys Val Tyr Gly Thr Ala Val Ala Ser Gly
275 280 285
Tyr Asp Phe Glu Lys Glu Gly Tyr Ser Leu Val Gly Ile Asp Pro Phe
290 295 300
Lys Leu Leu Gln Asn Ser Gln Val Tyr Ser Leu Ile Arg Pro Asn Glu
305 310 315 320
Asn Pro Ala His Lys Ser Gln Leu Val Trp Met Ala Cys Asn Ser Ala
325 330 335
Ala Phe Glu Asp Leu Arg Val Ser Ser Phe Ile Arg Gly Thr Lys Val
340 345 350
Ile Pro Arg Gly Lys Leu Ser Thr Arg Gly Val Gln Ile Ala Ser Asn
355 360 365
Glu Asn Met Asp Thr Met Glu Ser Ser Thr Leu Glu Leu Arg Ser Arg
370 375 380
Tyr Trp Ala Ile Arg Thr Arg Ser Gly Gly Asn Thr Asn Gln Gln Arg
385 390 395 400
Ala Ser Ala Gly Gln Ile Ser Val Gln Pro Thr Phe Ser Val Gln Arg
405 410 415
Asn Leu Pro Phe Asp Lys Thr Thr Ile Met Ala Ala Phe Thr Gly Asn
420 425 430
Ala Glu Gly Arg Thr Ser Asp Met Arg Ala Glu Ile Ile Arg Met Met
435 440 445
Glu Gly Ala Lys Pro Glu Glu Val Ser Phe Gln Gly Arg Gly Val Phe
450 455 460
Glu Leu Ser Asp Glu Lys Ala Thr Asn Pro Ile Val Pro Ser Phe Asp
465 470 475 480
Met Ser Asn Glu Gly Ser Tyr Phe Phe Gly Asp Asn Ala Glu Glu Tyr
485 490 495
Asp Asn Xaa Gly Lys Ile Pro Leu Phe Leu
500 505 <210>14 <211>469 <212>PRT
<213>流感病毒A/Singapore/1/57/ca
<400>14
Met Asn Pro Asn Gln Lys Ile Ile Thr Ile Gly Ser Val Ser Leu Thr
1 5 10 15
Ile Ala Thr Val Cys Phe Leu Met Gln Ile Ala Ile Leu Ala Thr Thr
20 25 30
Val Thr Leu His Phe Lys Gln His Glu Cys Asp Ser Pro Ala Ser Asn
35 40 45
Gln Val Met Pro Cys Glu Pro Ile Ile Ile Glu Arg Asn Ile Thr Glu
50 55 60
Ile Val Tyr Leu Asn Asn Thr Thr Ile Glu Lys Glu Ile Cys Pro Glu
65 70 75 80
Val Val Glu Tyr Arg Asn Trp Ser Lys Pro Gln Cys Gln Ile Thr Gly
85 90 95
Phe Ala Pro Phe Ser Lys Asp Asn Ser Ile Arg Leu Ser Ala Gly Gly
100 105 110
Asp Ile Trp Val Thr Arg Glu Pro Tyr Val Ser Cys Asp Pro Gly Lys
115 120 125
Cys Tyr Gln Phe Ala Leu Gly Gln Gly Thr Thr Leu Tyr Asn Lys His
130 135 140
Ser Asn Gly Thr Ile His Asp Arg Ile Pro His Arg Thr Leu Leu Met
145 150 155 160
Asn Glu Leu Gly Val Pro Phe His Leu Gly Thr Lys Gln Val Cys Val
165 170 175
Ala Trp Ser Ser Ser Ser Cys His Asp Gly Lys Ala Trp Leu His Val
180 185 190
Cys Val Thr Gly Asp Asp Arg Asn Ala Thr Ala Ser Phe Ile Tyr Asp
195 200 205
Gly Arg Leu Val Asp Ser Ile Gly Ser Trp Ser Gln Asn Ile Leu Arg
210 215 220
Thr Gln Glu Ser Glu Cys Val Cys Ile Asn Gly Thr Cys Thr Val Val
225 230 235 240
Met Thr Asp Gly Ser Ala Ser Gly Arg Ala Asp Thr Arg Ile Leu Phe
245 250 255
Ile Lys Glu Gly Lys Ile Val Arg Ile Ser Pro Leu Ser Gly Ser Ala
260 265 270
Gln His Ile Glu Glu Cys Ser Gys Tyr Pro Arg Tyr Pro Asp Val Arg
275 280 285
Cys Ile Cys Arg Asp Asn Trp Lys Gly Ser Asn Arg Pro Val Ile Asp
290 295 300
Ile Asn Met Glu Asp Tyr Ser Ile Asp Ser Ser Tyr Val Cys Ser Gly
305 310 315 320
Leu Val Gly Asp Thr Pro Arg Asn Asp Asp Ser Ser Ser Asn Ser Asn
325 330 335
Cys Arg Asp Pro Asn Asn Glu Arg Gly Asn Pro Gly Val Lys Gly Trp
340 345 350
Ala Phe Asp Asn Gly Asp Asp Val Trp Met Gly Arg Thr Ile Asn Lys
355 360 365
Asp Ser Arg Ser Gly Tyr Glu Thr Phe Lys Val Ile Gly Gly Trp Ser
370 375 380
Thr Pro Asn Ser Lys Ser Gln Val Asn Arg Gln Val Ile Val Asp Asn
385 390 395 400
Asn Asn Trp Ser Gly Tyr Ser Gly Ile Phe Ser Val Glu Gly Lys Ser
405 410 415
Cys Ile Asn Arg Cys Phe Tyr Val Glu Leu Ile Arg Gly Arg Pro Gln
420 425 430
Glu Thr Arg Val Trp Trp Thr Ser Asn Ser Ile Val Val Phe Cys Gly
435 440 445
Thr Ser Gly Thr Tyr Gly Thr Gly Ser Trp Pro Asp Gly Ala Asn Ile
450 455 460
Asn Phe Met Pro Ile
465
<210>15
<211>252
<212>PRT
<213>流感病毒A/Singapore/1/57/ca
<400>15
Met Ser Leu Leu Thr Glu Val Glu Thr Tyr Val Leu Ser Ile Val Pro
1 5 10 15
Ser Gly Pro Leu Lys Ala Glu Ile Ala Gln Arg Leu Glu Asp Val Phe
20 25 30
Ala Gly Lys Asn Thr Asp Leu Glu Ala Leu Met Glu Trp Leu Lys Thr
35 40 45
Arg Pro Ile Leu Ser Pro Leu Thr Lys Gly Ile Leu Gly Phe Val Phe
50 55 60
Thr Leu Thr Val Pro Ser Glu Arg Gly Leu Gln Arg Arg Arg Phe Val
65 70 75 80
Gln Asn Ala Leu Asn Gly Asn Gly Asp Pro Asn Asn Met Asp Arg Ala
85 90 95
Val Lys Leu Tyr Lys Lys Leu Lys Arg Glu Ile Thr Phe His Gly Ala
100 105 110
Lys Glu Ile Ala Leu Ser Tyr Ser Ala Gly Ala Leu Ala Ser Cys Met
115 120 125
Gly Leu Ile Tyr Asn Arg Met Gly Ala Val Thr Thr Glu Val Ala Phe
130 135 140
Gly Leu Val Cys Ala Thr Cys Glu Gln Ile Ala Asp Ser His His Arg
145 150 155 160
Ser His Arg Gln Met Val Thr Thr Thr Asn Pro Leu Ile Arg His Glu
165 170 175
Asn Arg Met Val Leu Ala Ser Thr Thr Ala Lys Ala Met Glu Gln Met
180 185 190
Ala Gly Ser Ser Glu Gln Ala Ala Glu Ala Met Glu Val Ala Ser Gln
195 200 205
Ala Arg Gln Met Val Gln Ala Met Arg Ala Ile Gly Thr His Pro Ser
210 215 220
Ser Ser Ala Gly Leu Lys Asp Asp Leu Leu Glu Asn Leu Gln Ala Tyr
225 230 235 240
Gln Lys Arg Met Gly Val Gln Met Gln Arg Phe Lys
245 250
<210>16
<211>97
<212>PRT
<213>流感病毒A/Singapore/1/57/ca
<400>16
Met Ser Leu Leu Thr Glu Val Glu Thr Pro Ile Arg Asn Glu Trp Gly
1 5 10 15
Cys Arg Cys Asn Asp Ser Ser Asp Pro Leu Val Val Ala Ala Ser Ile
20 25 30
Ile Gly Ile Leu His Leu Ile Leu Trp Ile Leu Asp Arg Leu Phe Phe
35 40 45
Lys Cys Ile Tyr Arg Phe Phe Lys His Gly Leu Lys Arg Gly Pro Ser
50 55 60
Thr Glu Gly Val Pro Glu Ser Met Arg Glu Glu Tyr Arg Lys Glu Gln
65 70 75 80
Gln Ser Ala Val Asp Ala Asp Asp Ser His Phe Val Ser Ile Glu Leu
85 90 95
Glu
<210>17
<211>237
<212>PRT
<213>流感病毒A/Singapore/1/57/ca
<400>17
Met Asp Pro Asn Thr Val Ser Ser Phe Gln Val Asp Cys Phe Leu Trp
1 5 10 15
His Val Arg Lys Gln Val Ala Asp Gln Glu Leu Gly Asp Ala Pro Phe
20 25 30
Leu Asp Arg Leu Arg Arg Asp Gln Lys Ser Leu Arg Gly Arg Gly Ser
35 40 45
Thr Leu Gly Leu Asn Ile Glu Thr Ala Thr Arg Val Gly Lys Gln Ile
50 55 60
Val Glu Arg Ile Leu Lys Glu Glu Ser Asp Glu Ala Leu Lys Met Thr
65 70 75 80
Met Ala Ser Ala Pro Ala Ser Arg Tyr Leu Thr Asp Met Thr Ile Glu
85 90 95
Glu Met Ser Arg Asp Trp Phe Met Leu Met Pro Lys Gln Lys Val Ser
100 105 110
Gly Pro Leu Cys Ile Arg Met Asp Gln Ala Ile Met Asp Lys Asn Ile
115 120 125
Ile Leu Lys Ala Asn Phe Ser Val Ile Phe Asp Arg Leu Glu Thr Leu
130 135 140
Ile Leu Leu Arg Ala Phe Thr Glu Glu Gly Ala Ile Val Gly Glu Ile
145 150 155 160
Ser Pro Leu Pro Ser Leu Pro Gly His Thr Asn Glu Asp Val Lys Asn
165 170 175
Ala Ile Gly Val Leu Ile Gly Gly Leu Glu Trp Asn Asp Asn Thr Val
180 185 190
Arg Val Ser Lys Thr Leu Gln Arg Phe Ala Trp Arg Asn Ser Asn Glu
195 200 205
Asn Gly Arg Pro Pro Leu Thr Pro Lys Gln Lys Arg Lys Met Ala Arg
210 215 220
Thr Ile Arg Ser Lys Val Arg Arg Asn Lys Met Ala Asp
225 230 235
<210>18
<211>121
<212>PRT
<213>流感病毒A/Singapore/1/57/ca
<400>18
Met Asp Pro Asn Thr Val Ser Ser Phe Gln Asp Ile Leu Met Arg Met
1 5 10 15
Ser Lys Met Gln Leu Gly Ser Ser Ser Glu Asp Leu Asn Gly Met Ile
20 25 30
Thr Gln Phe Glu Ser Leu Lys Leu Tyr Arg Asp Ser Leu Gly Glu Thr
35 40 45
Val Met Arg Met Gly Asp Leu His Ser Leu Gln Asn Arg Asn Gly Lys
50 55 60
Trp Arg Glu Gln Leu Gly Gln Lys Phe Glu Glu Ile Arg Trp Leu Ile
65 70 75 80
Glu Glu Val Arg His Lys Leu Lys Ile Thr Glu Asn Ser Phe Glu Gln
85 90 95
Ile Thr Phe Met Gln Ala Leu Gln Leu Leu Phe Glu Val Glu Gln Glu
100 105 110
Ile Arg Thr Phe Ser Phe Gln Leu Ile
115 120
<210>19
<211>2396
<212>DNA
<213>流感B/Vienna/1/99/ca
<400>19
agcagaagcg gagcgttttc aagatgacat tggctaaaat tgaattgtta aaacaactgt 60
taagggacaa tgaagccaaa acagtattga aacaaacaac agtagatcaa tataacataa 120
taagaaaatt caatacatca agaattgaaa agaacccttc attaaggatg aagtgggcaa 180
tgtgttctaa ttttcccttg gctttgacca agggtgacat ggcaaacaga atccccttgg 240
aatacaaggg aatacaactt aaaacaaatg ctgaagacat aggaaccaaa ggccaaatgt 300
gctcaatagc agcagttacc tggtggaata catatggacc aataggagat actgaaggtt 360
tcgaaaaagt ctacgaaagc ttttttctca gaaagatgag acttgacaat gccacttggg 420
gccgaataac ttttggccca gttgaaagag taagaaaaag ggtactgcta aaccctctca 480
ccaaggaaat gcctccagat gaagcaagta atgtgataat ggaaatattg ttccctaagg 540
aagcaggaat accaagagaa tctacttgga tacataggga actgataaaa gaaaaaagag 600
aaaaattgaa aggaacgatg ataactccca ttgtactggc atacatgctt gagagggaat 660
tggttgccag gagaaggttc ctgccggtag caggagcaac atcagctgag ttcatagaaa 720
tgctacactg cttacaaggt gaaaattgga gacaaatata tcacccggga gggaataaac 780
taactgaatc taggtctcaa tcgatgattg tggcttgtag aaagataatc agaagatcaa 840
tagtcgcatc aaacccattg gagctagctg tagaaattgc aaacaagact gtaatagata 900
ctgaaccttt aaaatcatgt ctgacagcca tagacggagg tgatgtcgcc tgtgacataa 960
taagggctgc attaggacta aagatcagac aaagacaaag atttggacga cttgaactaa 1020
agagaatatc aggaagagga ttcaaaaatg atgaagaaat attaatcggg aacggaacaa 1080
tacagaagat tggaatatgg gacggagaag aggagttcca tgtaagatgt ggtgaatgca 1140
ggggaatatt aaaaaagagc aaaatgagaa tggaaaaact actaataaat tcagctaaaa 1200
aggaagacat gaaagattta ataatcttgt gcatggtatt ttctcaagac actaggatgt 1260
tccaaggagt gaggggagaa ataaattttc ttaatagagc aggccaactt ttatctccaa 1320
tgtatcaact ccaaagatat tttttgaata gaagtaatga tctctttgat caatgggggt 1380
atgaggaatc acccaaagca agtgagctac atgggataaa tgaattaatg aatgcatctg 1440
actacacttt gaaaggggtt gtagtaacaa aaaatgtgat tgatgatttt agttctactg 1500
aaacagaaaa agtatctata acaaaaaatc ttagtttaat aaaaaggact ggggaagtca 1560
taatgggagc caatgacgta agtgaattag aatcacaagc tcagctaatg ataacatatg 1620
atacacctaa gatgtgggag atgggaacaa ccaaagaact ggtgcaaaac acctatcaat 1680
gggtgctgaa aaatttggta acactgaagg ctcagtttct tctaggaaaa gaagacatgt 1740
tccaatggga tgcatttgaa gcatttgaaa gcataatccc ccagaagatg gctggccaat 1800
acagtggatt tgcaagagca gtgctcaaac aaatgagaga ccaagaggtc atgaaaactg 1860
accagttcat aaagttgttg cccttttgtt tctcaccacc aaagttaagg agcaatgggg 1920
agccttatca gttcttgagg cttgtattga agggaggagg agaaaatttc atcgaagtaa 1980
ggaaagggtc tcctctattc tcttacaatc cacaaacaga agtcctaact atatgcggca 2040
gaatgatgtc attaaaaggg aaaattgaag atgaagaaag gaatagatca atggggaatg 2100
cagtgttggc gggttttctt gttagtggca agtatgaccc agatcttgga gatttcaaaa 2160
ctattgaaga gcttgaaaag ctaaaaccgg gggagaaagc aaacatctta ctttatcaag 2220
gaaagcccgt taaagtagtt aaaaggaaaa gatatagtgc tttatccaat gacatttcac 2280
aaggaattaa gagacaaaga atgacagttg agtccatggg gtgggccttg agctaatata 2340
aatttatcca ttaattcaat gaatgcaatt gagtgaaaaa tgctcgtgtt tctcat 2396
<210>20
<211>2369
<212>DNA
<213>流感B/Vienna/1/99/ca
<400>20
agcagaagcg gagcctttaa gatgaatata aatccttatt ttctcttcat agatgtaccc 60
atacaggcag caatttcaac aacattccca tacaccggtg ttccccctta ttcccatgga 120
acgggaacag gccacacaat agacaccgtg atcagaacac atgagtactc gaacaaagga 180
aaacagtatg tttctgacat cacaggatgt acaatggtag atccaacaaa tggaccatta 240
cccgaagaca atgagccaag tgcctatgca caattagatt gcgttctgga ggctttggat 300
agaatggatg aggaacatcc aggtctgttt caagcagcct cacagaatgc catggaggca 360
ctaatggtca caactgtaga caaattaacc caggggagac agactttcga ttggacagta 420
tgcagaaatc agcctgctgc aacggcacta aacacaacaa taacctcctt taggttgaat 480
gatttgaatg gagctgacaa gggtggattg gtaccctttt gccaagatat cattgattca 540
ttagacaagc ctgaaatgac tttcttctca gtaaagaata taaagaaaaa attccctgct 600
aaaaacagaa agggtttcct cataaagaga ataccaatga aagtaaaaga caggatatcc 660
agagtggaat acatcaaaag agcattgtca ttaaacacaa tgacaaaaga tgctgaaagg 720
ggcaaactaa aaagaagagc gattgcaacc gctggaatac aaatcagagg gtttgtatta 780
gtagttgaaa acttggctaa aaatatctgt gaaaatctag aacaaagtgg tttgcccgta 840
ggtggaaatg aaaagaaggc caaactgtca aatgcagtgg ccaaaatgct cagtaactgc 900
ccaccaggag ggatcagcat gacagtaaca ggagacaata ctaaatggaa tgaatgctta 960
aatccaagag tctttttggc tatgactgaa agaataacca gagacagccc aatttggttc 1020
cgggattttt gtagtatagc accggtcttg ttctccaata aaatagccag attgggaaaa 1080
ggatttatga taacaagtaa aacaaaaaga ctgaaggctc aaataccttg tcctgatctg 1140
tttagcatac cattagaaag atataatgaa gaaacaaggg caaaattaaa aaagctgaaa 1200
ccattcttca atgaagaagg aacggcatct ttgtcgcctg gaatgatgat gggaatgttt 1260
aatatgctat ctaccgtgtt gggagtagca gcactaggta tcaaaaacat tggaaacaag 1320
gaatacttat gggatggact gcaatcttct gatgattttg ctctgtttgt taatgcaaaa 1380
gatgaagaga catgtatgga aggaataaac gatttttacc gaacatgtaa attattggga 1440
ataaacatga gcaaaaagaa aagttactgt aacgaaactg gaatgtttga atttacaagc 1500
atgttctata gagatggatt tgtatctaac tttgcaatgg aaattccttc atttggagtt 1560
gctggagtaa atgaatcagc agatatggca ataggaatga caataataaa gaacaatatg 1620
attaacaatg ggatgggtcc agcaacagca caaacagcca tacaattgtt catagctgat 1680
tataggtaca cctacaaatg ccacagagga gattccaaag tggaaggaaa aagaatgaaa 1740
attataaagg agctatggga aaacactaaa ggaagagatg gtctgttagt ggcagatggt 1800
gggcccaaca tttacaattt gagaaactta catatcccag aaatagtatt gaagtacaat 1860
ctaatggacc ctgaatacaa agggcggtta cttcaccctc aaaatccctt tgtaggacat 1920
ttgtctattg aaggcatcaa agaagcagat ataaccccag cacatggtcc tgtgaagaaa 1980
atggattatg atgcagtgtc tggaactcat agttggagaa ccaaaaggaa cagatctata 2040
ctaaatactg atcagaggaa catgattctt gaggaacaat gctacgctaa atgttgcaac 2100
ctttttgagg cctgttttaa cagtgcatca tacaggaaac cagtagggca gcacagcatg 2160
cttgaggcta tggcccatag attaagaatg gatgcacgac tggattatga atcaggaaga 2220
atgtcaaagg atgattttga gaaagcaatg gctcaccttg gtgagattgg gtacacataa 2280
gctccgaaga tgtccatggg gttattggtc atcattggat acatgtgata aacaaatgat 2340
taaaatgaaa aaaggctcgt gtttctact 2369
<210>21
<211>2305
<212>DNA
<213>流感B/Vienna/1/99/ca
<400>21
agcagaagcg gtgcgtttga tttgtcataa tggatacttt tattacaaga aacttccaga 60
ctacaataat acaaaaggcc aaaaacacaa tggcagaatt tagtgaagat cctgaattac 120
aaccagcaat gctattcaac atctgcgtcc atctagaggt ttgctatgta ataagtgaca 180
tgaattttct tgacgaagaa ggaaaagcat atacagcatt agaaggacaa gggaaagaac 240
aaaatttgag accacaatat gaagtaattg agggaatgcc aagaaccata gcatggatgg 300
tccaaagatc cttagctcaa gagcatggaa tagagactcc caagtatctg gctgatttgt 360
ttgattataa aaccaagaga tttatagaag ttggaataac aaaaggattg gctgatgatt 420
acttttggaa aaagaaagaa aagctgggaa atagcatgga actgatgata ttcagctaca 480
atcaagacta ttcgttaagt aatgaatcct cattggatga ggaagggaaa gggagagtgc 540
taagcagact cacagaactt caggctgaat taagtctgaa aaacctatgg caagttctca 600
taggagaaga agatgttgaa aagggaattg actttaaact tggacaaaca atatctagac 660
taagggatat atctgttcca gctggtttct ccaattttga aggaatgagg agctacatag 720
acaatataga cccgaaagga gcaatagaga gaaatctagc aaggatgtct cccttagtat 780
cagtcacacc taaaaagttg aaatgggagg acctaagacc aatagggcct cacatttaca 840
accatgagct accagaagtt ccatataatg cctttcttct aatgtctgat gaactggggc 900
tggccaatat gactgaggga aagtccaaaa aaccgaagac attagccaaa gaatgtctag 960
aaaagtactc aacactacgg gatcaaactg acccaatatt aataatgaaa agcgaaaaag 1020
ctaacgaaaa tttcctatgg aagctttgga gagactgtgt aaatacaata agtaatgagg 1080
aaatgagtaa cgagttacag aaaaccaatt atgccaagtg ggccacaggg gatggattaa 1140
cataccagaa aataatgaaa gaagtagcaa tagatgacga aacaatgtgc caagaagagc 1200
ctaaaatccc taacaaatgt agagtggctg cttgggttca aacagagatg aatctattga 1260
gcactctgac aagtaaaaaa gctctggacc taccagaaat agggccagac gtagcacccg 1320
tggagcatgt agggagtgaa agaaggaaat actttgttaa tgaaatcaac tactgtaagg 1380
cctctacagt tatgatgaag tatgtgcttt ttcacacttc attgttgaat gaaagcaatg 1440
ccagcatggg aaaatacaaa gtaataccaa taaccaatag agtagtaaat gaaaaaggag 1500
aaagtttcga catgctctat ggtctggcgg ttaaaggaca atctcatctg aggggagata 1560
ctgatgttgt aacagttgta actttcgaat ttagtagtac agaccccaga gtggactcag 1620
gaaagtggcc aaaatatact gtgtttagga ttggctccct atttgtgagt gggagggaaa 1680
agtctgtgta cctatattgc cgagtgaatg gcacaaataa gatccaaatg aaatggggaa 1740
tggaagctag aagatgtctg cttcaatcaa tgcaacaaat ggaagcaatt gttgaacaag 1800
aatcatcgat acaaggatat gacatgacca aagcttgttt caagggagac agagtaaata 1860
gccccaaaac tttcagtatt ggaactcaag aaggaaaact agtaaaagga tcctttggaa 1920
aagcactaag agtaatattt actaaatgtt tgatgcacta tgtatttgga aatgcccaat 1980
tggaggggtt tagtgccgag tctaggagac ttctactgtt gattcaagca ttaaaggaca 2040
gaaagggccc ttgggtgttc gacttagagg gaatgtattc tggaatagaa gaatgtatta 2100
gtaacaaccc ttgggtaata cagagtgcat actggttcaa tgaatggttg ggctttgaaa 2160
aggaggggag taaagtatta gaatcagtag atgaaataat ggatgaataa aaggacatag 2220
tactcaattt agtactattt tgttcattat gtatctaaac atccaataaa aaggacaaag 2280
aattaaaaat gcacgtgttt ctact 2305
<210>22
<211>1882
<212>DNA
<213>流感B/Vienna/1/99/ca
<400>22
agcagaagca gagcattttc taatatccac aaaatgaagg caataattgt actactcatg 60
gtagtaacat ccaatgcaga tcgaatctgc actgggataa catcgtcaaa ctcacctcat 120
gtggtcaaaa cagctactca aggggaggtc aatgtgactg gtgcgatacc actgacaaca 180
acaccaacaa aatctcattt tgcaaatctc aaaggaacaa agaccagagg gaaactatgc 240
ccaacctgtc tcaactgcac agatctggat gtggccttgg gcagaccaat gtgtgtgggg 300
atcacacctt cggcaaaagc ttcaatactc cacgaagtca gacctgttac atccggatgc 360
tttcctataa tgcacgacag aacaaaaatc agacagctac ccaatcttct cagaggatat 420
gaaaaaatca gattatcaac ccaaaacgtt atcaacacag aaaaggcacc aggaggaccc 480
tacagacttg gaacttcagg atcttgccct aacgctacca gtaaaagcgg atttttcgca 540
acaatggctt gggctgtccc aagggacaac aacaaaacag caacgaatcc actaacagta 600
gaagtaccac acatctgtac aaaagaagaa gaccaaatta ctgtttgggg gttccattct 660
gataacaaaa cccaaatgaa aaacctctat ggagactcaa atcctcaaaa gttcacctca 720
tctgctaatg ggataaccac acattatgtt tctcagattg gcggcttccc ggaccaaaca 780
gaagacggag ggctaccaca aagcggcaga attgttgttg attacatggt gcaaaaacct 840
gggaaaacag gaacaattgt ctatcaaaga gggattttgt tgcctcaaaa ggtgtggtgc 900
gcgagtggca ggagcaaagt aataaaaggg tccttgcctt taattggtga agcagattgc 960
cttcacgaaa aatacggtgg attaaacaaa agcaagcctt actacacagg agaacatgca 1020
aaagccatag gaaattgccc aatatgggtg aaaacacctt tgaagcttgc caatggaacc 1080
aaatatagac ctcctgcaaa actattaaag gaaaggggtt tcttcggagc tattgctggt 1140
ttcttagaag gaggatggga aggaatgatt gcaggttggc acggatacac atctcacgga 1200
gcacatggag tggcagtggc agcagacctt aagagtacgc aagaagccat aaacaagata 1260
acaaaaaatc tcaattcttt gagtgagcta gaagtaaata accttcaaag actaagtggt 1320
gccatggatg aactccataa cgaaatactc gagctggatg agaaagtgga tgatctcaga 1380
gctgacacaa taagctcaca aatagaactt gcagtcttgc tttccaacga aggaataata 1440
aacagtgaag atgagcatct attggcactt gagagaaaac taaagaaaat gctgggtccc 1500
tctgctgtag acatagggaa tggatgcttc gaaaccaaac acaagtgcaa ccagacctgc 1560
ttagacagga tagctgctgg cacctttaat gcagaagaat tttctcttcc cacttttgat 1620
tcactgaaca ttactgctgc atctttaaat gatgatggat tggataacca tactatactg 1680
ctctactact caactgctgc ttctagtttg gctgtaacat tgatgatagc tatttttatt 1740
gtttatatga tctccagaga caatgtttct tgctccatct gtctataagg aaaattaagc 1800
cctgtatttt cctttattgt ggtgcttgtt tgcttgttat cattacaaag aaacgttatt 1860
gaaaaatgct cttgttacta ct 1882
<210>23
<211>1844
<212>DNA
<213>流感B/Vienna/1/99/ca
<400>23
agcagaagca cagcattttc ttgtgaactt caagtaccag taaaagaact gaaaatcaaa 60
atgtccaaca tggatattga cggtatcaac actgggacaa ttgacaaaac accggaagaa 120
ataacttttg gaaccagtgg gacaaccaga ccaatcatca gaccagcaac ccttgcccca 180
ccaagcaaca aacgaacccg taacccatcc ccggaaagag caaccacaag cagtgaagct 240
gatgtcggga ggaaaaccca aaagaaacag accccgacag agataaagaa gagcgtctac 300
aacatggtag tgaaactggg cgaattctac aaccagatga tggtcaaagc tggactcaac 360
gatgacatgg agagaaacct aatccaaaat gcgcatgctg tggaaagaat tctattggct 420
gccactgatg acaagaaaac tgaattccag aagaaaaaga ataccagaga tgtcaaagaa 480
gggaaagaag aaatagatca caacaaaaca ggaggcacct tttacaagat ggtaagagat 540
gataaaacca tctacttcag ccctataaga attacctttt taaaagaaga ggtgaaaaca 600
atgtacaaaa ccaccatggg gagtgatggc ttcagtggac taaatcacat aatgattggg 660
cattcacaga tgaatgatgt ctgtttccaa agatcaaagg cactaaaaag agttggactt 720
gacccttcat taatcagtac ctttgcggga agcacaatcc ccagaagatc aggtgcaact 780
ggtgttgcaa tcaaaggagg tggaacttta gtggctgaag ccattcgatt tataggaaga 840
gcaatggcag acagagggct attgagagac atcaaagcca agactgccta tgaaaagatt 900
cttctgaatc taaaaaacaa atgctctgcg ccccaacaaa aggctctagt tgatcaagtg 960
atcggaagta gaaatccagg gattgcagac attgaagatc taaccctgct tgctcgtagt 1020
atggtcgttg ttaggccctc tgtggcgagc aaagtggtgc ttcccataag catttacgcc 1080
aaaatacctc aactagggtt caatgttgaa gagtactcta tggttgggta cgaagccatg 1140
gctctttaca atatggcaac acctgtttcc atattaagaa tgggagatga tgcaaaagat 1200
aagtcgcaat tattcttcat gtcttgcttc ggagctgcct atgaagacct gagagttttg 1260
tctgcattaa caggcacaga attcaagcct agatcagcat taaaatgcaa gggtttccat 1320
gttccagcaa aggaacaggt ggaaggaatg ggggcagctc tgatgtccat caagctccag 1380
ttttgggctc caatgaccag atctgggggg aacgaagtag gtggagacgg agggtctggc 1440
caaataagct gcagcccagt gtttgcagtg gaaagaccta ttgctctaag caagcaagct 1500
gtaagaagaa tgctgtcaat gaatattgag ggacgtgatg cagatgtcaa aggaaatcta 1560
ctcaagatga tgaatgactc aatggctaag aaaaccagtg gaaatgcttt cattgggaag 1620
aaaatgtttc aaatatcaga caaaaacaaa accaatcccg ttgaaattcc aattaagcag 1680
accatcccca atttcttctt tgggagggac acagcagagg attatgatga cctcgattat 1740
taaagcaaca aaatagacac tatgactgtg attgtttcaa tacgtttgga atgtgggtgt 1800
ttattcttat taaaataaat ataaaaaatg ctgttgtttc tact 1844
<210>24
<211>1557
<212>DNA
<213>流感B/Vienna/1/99/ca
<400>24
agcagaagca gagcatcttc tcaaaactga ggcaaatagg ccaaaaatga acaatgctac 60
cttcaactat acaaacgtta accctattcc tcacatcagg gggagtgtta ttatcactat 120
atgtgtcagc ttcactgtca tacttattat attcggatat attgctaaaa ttttcaccaa 180
cagaaataac tgcaccaaca atgccattgg attgtgcaaa cgcatcaaat gttcaggctg 240
tgaaccgttc tgcaacaaaa ggggtgacac ttcttctccc agaaccggag tggacatacc 300
cgcgtttatc ttgcccgggc tcaacctttc agaaagcact cctaattagc cctcatagat 360
tcggagaaac caaaggaaac tcagctccct tgataataag ggaacctttt attgcttgtg 420
gaccaaagga atgcaaacac tttgctctaa cccattatgc agcccaacca gggggatact 480
acaatggaac aagagaagac agaaacaagc tgaggcatct aatttcagtc aaattgggca 540
aaatcccaac agtagaaaac tccattttcc acatggcagc atggagcggg tccgcatgcc 600
atgatggtaa agaatggaca tatatcggag ttgatggccc tgacagtaat gcattgctca 660
aaataaaata tggagaagca tatactgaca cataccattc ctatgcaaac aacatcctaa 720
gaacacaaga aagtgcctgc aattgcatcg ggggaaattg ttatcttatg ataactgatg 780
gctcagcttc aggtattagt gaatgcagat ttcttaagat tcaagagggc cgaataataa 840
aagaaatatt tccaacagga agagtagaac atactgaaga atgcacatgc ggatttgcca 900
gcaataaaac catagaatgt gcctgtagag ataacagtta cacagcaaaa agaccctttg 960
tcaaattaaa tgtggagact gatacagcag aaataagatt gatgtgcaca gagacttact 1020
tggacacccc cagaccagat gatggaagca taacagggcc ttgtgaatct aatggggata 1080
aagggagtgg aggcatcaag ggaggatttg ttcatcaaag aatggcatcc aagactggaa 1140
ggtggtactc tcgaacaatg tctaaaacta aaaggatggg gatgggactg tatgtcaagt 1200
atgatggaga cccatggact gacagtgatg cccttgctct tagtggagta atggtttcaa 1260
tggaagaacc tggttggtac tcctttggct tcgaaataaa agataagaaa tgtgatgtcc 1320
cctgtattgg gatagagatg gtacatgatg gtggaaagga gacttggcac tcagcagcaa 1380
cagccattta ctgtttaatg ggctcaggac aactgctatg ggacactgtc acaggtgtta 1440
atatggctct gtaatggagg aatggttgag tctgttctaa accctttgtt cctattttgt 1500
ttgaacaatt gtccttactg aacttaattg tttctgaaaa atgctcttgt tactact 1557
<210>25
<211>1190
<212>DNA
<213>流感B/Vienna/1/99/ca
<400>25
agcagaagca cgcactttct taagatgtcg ctgtttggag acacaattgc ctacctgctt 60
tcattgacag aagatggaga aggcaaagca gaactagcag aaaaattaca ctgttggttc 120
ggtgggaaag aatttgacct agactctgcc ttggaatgga taaaaaacaa aagatgctta 180
actgatatac aaaaagcact aattggtgcc tctatctgct ttttaaaacc caaagaccag 240
gaaagaaaaa gaagattcat cacagagccc ctatcaggaa tgggaacaac agcaacaaaa 300
aagaaaggcc tgattctagc tgagagaaaa atgagaagat gtgtgagctt tcatgaagca 360
tttgaaatag cagaaggcca tgaaagctca gcgctactat attgtctcat ggtcatgtac 420
ctgaatcctg gaaattattc aatgcaagta aaactaggaa cgctctgtgc tttgtgcgag 480
aaacaagcat cacattcaca cagggctcat agcagagcag cgagatcttc agtgcccgga 540
gtgagacgag aaatgcagat ggtctcagct atgaacacag caaaaacaat gaatggaatg 600
ggaaaaggag aagacgtcca aaaactggca gaagagctgc aaagcaacat tggagtactg 660
agatctcttg gggcaagtca aaagaatggg gaaggaattg caaaggatgt aatggaagtg 720
ctaaagcaga gctctatggg aaattcagct cttgtgaaga aatatctata atgctcgaac 780
catttcagat tctttcaatt tgttctttta tcttatcagc tctccatttc gtggcttgga 840
caatagggca tttgaatcaa ataaaaagag gagtaaacat gaaaatacga ataaaaagtc 900
caaacaaaga gacaataaac agagaggtat caattttgag acacagttac caaaaagaaa 960
tccaggccaa agaaacaatg aaggaagtac tctctgacaa catggaggta ttgggtgacc 1020
acatagtaat tgaggggctt tctgccgaag agataataaa aatgggtgaa acagttttgg 1080
agatagaaga attgcattaa attcaatttt tactgtattt cttactatgc atttaagcaa 1140
attgtaatca atgtcagcaa ataaactgga aaaagtgcgt tgtttctact 1190
<210>26
<211>1097
<212>DNA
<213>流感B/Vienna/1/99/ca
<400>26
agcagaagca gagcatttgt ttagtcactg gcaaacagga aaaatggcga acaacataac 60
cacaacacaa attgaggtgg gtccgggagc aaccaatgcc accataaact ttgaaacagg 120
aattctggag tgctatgaaa ggctttcatg gcaaagagcc cttgactacc ctggtcaaga 180
ccgcctaaac agactaaaga gaaaattaga gtcaagaata aagactcaca acaaaagtga 240
gcctgaaagt aaaaggatgt ctcttgaaga gaggaaagca attggagtaa aaatgatgaa 300
agtactccta tttatgaatc catctgctgg aattgaaggg tttgagccat actatatgaa 360
aagttcctca aatagcaact gtccgaaata caattggacc gattaccctt caacaccagg 420
gaggtgcctt gatgacatag aagaagaacc agaggatgtt gatggcccaa ctgaaatagt 480
attaagggac atgaacaaca aagatgcaag gcaaaagata aaagaggaag taaacactca 540
gaaagaaggg aagttccgtt tgacaataaa aagggatata cgtaatgtat tgtccttgag 600
agtgttggta aacggaacat tcctcaaaca ccccaatgga tacaagtcct tatcaactct 660
gcatagattg aatgcatatg accagagtgg aaggcttgtt gctaaacttg ttgctactga 720
tgatcttaca gtggaggatg aagaagatgg ccatcggatc ctcaactcac tcttcgagcg 780
tcttaatgaa ggacattcaa agccaattcg agcagctgaa actgcggtgg gagtcttatc 840
ccaatttggt caagagcacc gattatcacc agaagaggga gacaattaaa ctggtcacag 900
aagaacttta tcttttaagt aaaagaattg atgataacat attgttccac aaaacagtaa 960
tagctaacag ctccataata gctgacatgg ttgtatcatt atcattatta gaaacattgt 1020
atgaaatgaa ggatgtggtt gaagtgtaca gcaggcagtg cttgtgaatt taaaataaaa 1080
atcctcttgt tactact 1097
<210>27
<211>770
<212>PRT
<213>流感B/Vienna/1/99/ca
<400>27
Met Thr Leu Ala Lys Ile Glu Leu Leu Lys Gln Leu Leu Arg Asp Asn
1 5 10 15
Glu Ala Lys Thr Val Leu Lys Gln Thr Thr Val Asp Gln Tyr Asn Ile
20 25 30
Ile Arg Lys Phe Asn Thr Ser Arg Ile Glu Lys Asn Pro Ser Leu Arg
35 40 45
Met Lys Trp Ala Met Cys Ser Asn Phe Pro Leu Ala Leu Thr Lys Gly
50 55 60
Asp Met Ala Asn Arg Ile Pro Leu Glu Tyr Lys Gly Ile Gln Leu Lys
65 70 75 80
Thr Asn Ala Glu Asp Ile Gly Thr Lys Gly Gln Met Cys Ser Ile Ala
85 90 95
Ala Val Thr Trp Trp Asn Thr Tyr Gly Pro Ile Gly Asp Thr Glu Gly
100 105 110
Phe Glu Lys Val Tyr Glu Ser Phe Phe Leu Arg Lys Met Arg Leu Asp
115 120 125
Asn Ala Thr Trp Gly Arg Ile Thr Phe Gly Pro Val Glu Arg Val Arg
130 135 140
Lys Arg Val Leu Leu Asn Pro Leu Thr Lys Glu Met Pro Pro Asp Glu
145 150 155 160
Ala Ser Asn Val Ile Met Glu Ile Leu Phe Pro Lys Glu Ala Gly Ile
165 170 175
Pro Arg Glu Ser Thr Trp Ile His Arg Glu Leu Ile Lys Glu Lys Arg
180 185 190
Glu Lys Leu Lys Gly Thr Met Ile Thr Pro Ile Val Leu Ala Tyr Met
195 200 205
Leu Glu Arg Glu Leu Val Ala Arg Arg Arg Phe Leu Pro Val Ala Gly
210 215 220
Ala Thr Ser Ala Glu Phe Ile Glu Met Leu His Cys Leu Gln Gly Glu
225 230 235 240
Asn Trp Arg Gln Ile Tyr His Pro Gly Gly Asn Lys Leu Thr Glu Ser
245 250 255
Arg Ser Gln Ser Met Ile Val Ala Cys Arg Lys Ile Ile Arg Arg Ser
260 265 270
Ile Val Ala Ser Asn Pro Leu Glu Leu Ala Val Glu Ile Ala Asn Lys
275 280 285
Thr Val Ile Asp Thr Glu Pro Leu Lys Ser Cys Leu Thr Ala Ile Asp
290 295 300
Gly Gly Asp Val Ala Cys Asp Ile Ile Arg Ala Ala Leu Gly Leu Lys
305 310 315 320
Ile Arg Gln Arg Gln Arg Phe Gly Arg Leu Glu Leu Lys Arg Ile Ser
325 330 335
Gly Arg Gly Phe Lys Asn Asp Glu Glu Ile Leu Ile Gly Asn Gly Thr
340 345 350
Ile Gln Lys Ile Gly Ile Trp Asp Gly Glu Glu Glu Phe His Val Arg
355 360 365
Cys Gly Glu Cys Arg Gly Ile Leu Lys Lys Ser Lys Met Arg Met Glu
370 375 380
Lys Leu Leu Ile Asn Ser Ala Lys Lys Glu Asp Met Lys Asp Leu Ile
385 390 395 400
Ile Leu Cys Met Val Phe Ser Gln Asp Thr Arg Met Phe Gln Gly Val
405 410 415
Arg Gly Glu Ile Asn Phe Leu Asn Arg Ala Gly Gln Leu Leu Ser Pro
420 425 430
Met Tyr Gln Leu Gln Arg Tyr Phe Leu Asn Arg Ser Asn Asp Leu Phe
435 440 445
Asp Gln Trp Gly Tyr Glu Glu Ser Pro Lys Ala Ser Glu Leu His Gly
450 455 460
Ile Asn Glu Leu Met Asn Ala Ser Asp Tyr Thr Leu Lys Gly Val Val
465 470 475 480
Val Thr Lys Asn Val Ile Asp Asp Phe Ser Ser Thr Glu Thr Glu Lys
485 490 495
Val Ser Ile Thr Lys Asn Leu Ser Leu Ile Lys Arg Thr Gly Glu Val
500 505 510
Ile Met Gly Ala Asn Asp Val Ser Glu Leu Glu Ser Gln Ala Gln Leu
515 520 525
Met Ile Thr Tyr Asp Thr Pro Lys Met Trp Glu Met Gly Thr Thr Lys
530 535 540
Glu Leu Val Gln Asn Thr Tyr Gln Trp Val Leu Lys Asn Leu Val Thr
545 550 555 560
Leu Lys Ala Gln Phe Leu Leu Gly Lys Glu Asp Met Phe Gln Trp Asp
565 570 575
Ala Phe Glu Ala Phe Glu Ser Ile Ile Pro Gln Lys Met Ala Gly Gln
580 585 590
Tyr Ser Gly Phe Ala Arg Ala Val Leu Lys Gln Met Arg Asp Gln Glu
595 600 605
Val Met Lys Thr Asp Gln Phe Ile Lys Leu Leu Pro Phe Cys Phe Ser
610 615 620
Pro Pro Lys Leu Arg Ser Asn Gly Glu Pro Tyr Gln Phe Leu Arg Leu
625 630 635 640
Val Leu Lys Gly Gly Gly Glu Asn Phe Ile Glu Val Arg Lys Gly Ser
645 650 655
Pro Leu Phe Ser Tyr Asn Pro Gln Thr Glu Val Leu Thr Ile Cys Gly
660 665 670
Arg Met Met Ser Leu Lys Gly Lys Ile Glu Asp Glu Glu Arg Asn Arg
675 680 685
Ser Met Gly Asn Ala Val Leu Ala Gly Phe Leu Val Ser Gly Lys Tyr
690 695 700
Asp Pro Asp Leu Gly Asp Phe Lys Thr Ile Glu Glu Leu Glu Lys Leu
705 710 715 720
Lys Pro Gly Glu Lys Ala Asn Ile Leu Leu Tyr Gln Gly Lys Pro Val
725 730 735
Lys Val Val Lys Arg Lys Arg Tyr Ser Ala Leu Ser Asn Asp Ila Ser
740 745 750
Gln Gly Ile Lys Arg Gln Arg Met Thr Val Glu Ser Met Gly Trp Ala
755 760 765
Leu Ser
770
<210>28
<211>752
<212>PRT
<213>流感B/Vienna/1/99/ca
<400>28
Met Asn Ile Asn Pro Tyr Phe Leu Phe Ile Asp Val Pro Ile Gln Ala
1 5 10 15
Ala Ile Ser Thr Thr Phe Pro Tyr Thr Gly Val Pro Pro Tyr Ser His
20 25 30
Gly Thr Gly Thr Gly His Thr Ile Asp Thr Val Ile Arg Thr His Glu
35 40 45
Tyr Ser Asn Lys Gly Lys Gln Tyr Val Ser Asp Ile Thr Gly Cys Thr
50 55 60
Met Val Asp Pro Thr Asn Gly Pro Leu Pro Glu Asp Asn Glu Pro Ser
65 70 75 80
Ala Tyr Ala Gln Leu Asp Cys Val Leu Glu Ala Leu Asp Arg Met Asp
85 90 95
Glu Glu His Pro Gly Leu Phe Gln Ala Ala Ser Gln Asn Ala Met Glu
100 105 110
Ala Leu Met Val Thr Thr Val Asp Lys Leu Thr Gln Gly Arg Gln Thr
115 120 125
Phe Asp Trp Thr Val Cys Arg Asn Gln Pro Ala Ala Thr Ala Leu Asn
130 135 140
Thr Thr Ile Thr Ser Phe Arg Leu Asn Asp Leu Asn Gly Ala Asp Lys
145 150 155 160
Gly Gly Leu Val Pro Phe Cys Gln Asp Ile Ile Asp Ser Leu Asp Lys
165 170 175
Pro Glu Met Thr Phe Phe Ser Val Lys Asn Ile Lys Lys Lys Phe Pro
180 185 190
Ala Lys Asn Arg Lys Gly Phe Leu Ile Lys Arg Ile Pro Met Lys Val
195 200 205
Lys Asp Arg Ile Ser Arg Val Glu Tyr Ile Lys Arg Ala Leu Ser Leu
210 215 220
Asn Thr Met Thr Lys Asp Ala Glu Arg Gly Lys Leu Lys Arg Arg Ala
225 230 235 240
Ile Ala Thr Ala Gly Ile Gln Ile Arg Gly Phe Val Leu Val Val Glu
245 250 255
Asn Leu Ala Lys Asn Ile Cys Glu Asn Leu Glu Gln Ser Gly Leu Pro
260 265 270
Val Gly Gly Asn Glu Lys Lys Ala Lys Leu Ser Asn Ala Val Ala Lys
275 280 285
Met Leu Ser Asn Cys Pro Pro Gly Gly Ile Ser Met Thr Val Thr Gly
290 295 300
Asp Asn Thr Lys Trp Asn Glu Cys Leu Asn Pro Arg Val Phe Leu Ala
305 310 315 320
Met Thr Glu Arg Ile Thr Arg Asp Ser Pro Ile Trp Phe Arg Asp Phe
325 330 335
Cys Ser Ile Ala Pro Val Leu Phe Ser Asn Lys Ile Ala Arg Leu Gly
340 345 350
Lys Gly Phe Met Ile Thr Ser Lys Thr Lys Arg Leu Lys Ala Gln Ile
355 360 365
Pro Cys Pro Asp Leu Phe Ser Ile Pro Leu Glu Arg Tyr Asn Glu Glu
370 375 380
Thr Arg Ala Lys Leu Lys Lys Leu Lys Pro Phe Phe Asn Glu Glu Gly
385 390 395 400
Thr Ala Ser Leu Ser Pro Gly Met Met Met Gly Met Phe Asn Met Leu
405 410 415
Ser Thr Val Leu Gly Val Ala Ala Leu Gly Ile Lys Asn Ile Gly Asn
420 425 430
Lys Glu Tyr Leu Trp Asp Gly Leu Gln Ser Ser Asp Asp Phe Ala Leu
435 440 445
Phe Val Asn Ala Lys Asp Glu Glu Thr Cys Met Glu Gly Ile Asn Asp
450 455 460
Phe Tyr Arg Thr Cys Lys Leu Leu Gly Ile Asn Met Ser Lys Lys Lys
465 470 475 480
Ser Tyr Cys Asn Glu Thr Gly Met Phe Glu Phe Thr Ser Met Phe Tyr
485 490 495
Arg Asp Gly Phe Val Ser Asn Phe Ala Met Glu Ile Pro Ser Phe Gly
500 505 510
Val Ala Gly Val Asn Glu Ser Ala Asp Met Ala Ile Gly Met Thr Ile
515 520 525
Ile Lys Asn Asn Met Ile Asn Asn Gly Met Gly Pro Ala Thr Ala Gln
530 535 540
Thr Ala Ile Gln Leu Phe Ile Ala Asp Tyr Arg Tyr Thr Tyr Lys Cys
545 550 555 560
His Arg Gly Asp Ser Lys Val Glu Gly Lys Arg Met Lys Ile Ile Lys
565 570 575
Glu Leu Trp Glu Asn Thr Lys Gly Arg Asp Gly Leu Leu Val Ala Asp
580 585 590
Gly Gly Pro Asn Ile Tyr Asn Leu Arg Asn Leu His Ile Pro Glu Ile
595 600 605
Val Leu Lys Tyr Asn Leu Met Asp Pro Glu Tyr Lys Gly Arg Leu Leu
610 615 620
His Pro Gln Asn Pro Phe Val Gly His Leu Ser Ile Glu Gly Ile Lys
625 630 635 640
Glu Ala Asp Ile Thr Pro Ala His Gly Pro Val Lys Lys Met Asp Tyr
645 650 655
Asp Ala Val Ser Gly Thr His Ser Trp Arg Thr Lys Arg Asn Arg Ser
660 665 670
Ile Leu Asn Thr Asp Gln Arg Asn Met Ile Leu Glu Glu Gln Cys Tyr
675 680 685
Ala Lys Cys Cys Asn Leu Phe Glu Ala Cys Phe Asn Ser Ala Ser Tyr
690 695 700
Arg Lys Pro Val Gly Gln His Ser Met Leu Glu Ala Met Ala His Arg
705 710 715 720
Leu Arg Met Asp Ala Arg Leu Asp Tyr Glu Ser Gly Arg Met Ser Lys
725 730 735
Asp Asp Phe Glu Lys Ala Met Ala His Leu Gly Glu Ile Gly Tyr Thr
740 745 750
<210>29
<211>726
<212>PRT
<213>流感B/Vienna/1/99/ca
<400>29
Met Asp Thr Phe Ile Thr Arg Asn Phe Gln Thr Thr Ile Ile Gln Lys
1 5 10 15
Ala Lys Asn Thr Met Ala Glu Phe Ser Glu Asp Pro Glu Leu Gln Pro
20 25 30
Ala Met Leu Phe Asn Ile Cys Val His Leu Glu Val Cys Tyr Val Ile
35 40 45
Ser Asp Met Asn Phe Leu Asp Glu Glu Gly Lys Ala Tyr Thr Ala Leu
50 55 60
Glu Gly Gln Gly Lys Glu Gln Asn Leu Arg Pro Gln Tyr Glu Val Ile
65 70 75 80
Glu Gly Met Pro Arg Thr Ile Ala Trp Met Val Gln Arg Ser Leu Ala
85 90 95
Gln Glu His Gly Ile Glu Thr Pro Lys Tyr Leu Ala Asp Leu Phe Asp
100 105 110
Tyr Lys Thr Lys Arg Phe Ile Glu Val Gly Ile Thr Lys Gly Leu Ala
115 120 125
Asp Asp Tyr Phe Trp Lys Lys Lys Glu Lys Leu Gly Asn Ser Met Glu
130 135 140
Leu Met Ile Phe Ser Tyr Asn Gln Asp Tyr Ser Leu Ser Asn Glu Ser
145 150 155 160
Ser Leu Asp Glu Glu Gly Lys Gly Arg Val Leu Ser Arg Leu Thr Glu
165 170 175
Leu Gln Ala Glu Leu Ser Leu Lys Asn Leu Trp Gln Val Leu Ile Gly
180 185 190
Glu Glu Asp Val Glu Lys Gly Ile Asp Phe Lys Leu Gly Gln Thr Ile
195 200 205
Ser Arg Leu Arg Asp Ile Ser Val Pro Ala Gly Phe Ser Asn Phe Glu
210 215 220
Gly Met Arg Ser Tyr Ile Asp Asn Ile Asp Pro Lys Gly Ala Ile Glu
225 230 235 240
Arg Asn Leu Ala Arg Met Ser Pro Leu Val Ser Val Thr Pro Lys Lys
245 250 255
Leu Lys Trp Glu Asp Leu Arg Pro Ile Gly Pro His Ile Tyr Asn His
260 265 270
Glu Leu Pro Glu Val Pro Tyr Asn Ala Phe Leu Leu Met Ser Asp Glu
275 280 285
Leu Gly Leu Ala Asn Met Thr Glu Gly Lys Ser Lys Lys Pro Lys Thr
290 295 300
Leu Ala Lys Glu Cys Leu Glu Lys Tyr Ser Thr Leu Arg Asp Gln Thr
305 310 315 320
Asp Pro Ile Leu Ile Met Lys Ser Glu Lys Ala Asn Glu Asn Phe Leu
325 330 335
Trp Lys Leu Trp Arg Asp Cys Val Asn Thr Ile Ser Asn Glu Glu Met
340 345 350
Ser Asn Glu Leu Gln Lys Thr Asn Tyr Ala Lys Trp Ala Thr Gly Asp
355 360 365
Gly Leu Thr Tyr Gln Lys Ile Met Lys Glu Val Ala Ile Asp Asp Glu
370 375 380
Thr Met Cys Gln Glu Glu Pro Lys Ile Pro Asn Lys Cys Arg Val Ala
385 390 395 400
Ala Trp Val Gln Thr Glu Met Asn Leu Leu Ser Thr Leu Thr Ser Lys
405 410 415
Lys Ala Leu Asp Leu Pro Glu Ile Gly Pro Asp Val Ala Pro Val Glu
420 425 430
His Val Gly Ser Glu Arg Arg Lys Tyr Phe Val Asn Glu Ile Asn Tyr
435 440 445
Cys Lys Ala Ser Thr Val Met Met Lys Tyr Val Leu Phe His Thr Ser
450 455 460
Leu Leu Asn Glu Ser Asn Ala Ser Met Gly Lys Tyr Lys Val Ile Pro
465 470 475 480
Ile Thr Asn Arg Val Val Asn Glu Lys Gly Glu Ser Phe Asp Met Leu
485 490 495
Tyr Gly Leu Ala Val Lys Gly Gln Ser His Leu Arg Gly Asp Thr Asp
500 505 510
Val Val Thr Val Val Thr Phe Glu Phe Ser Ser Thr Asp Pro Arg Val
515 520 525
Asp Ser Gly Lys Trp Pro Lys Tyr Thr Val Phe Arg Ile Gly Ser Leu
530 535 540
Phe Val Ser Gly Arg Glu Lys Ser Val Tyr Leu Tyr Cys Arg Val Asn
545 550 555 560
Gly Thr Asn Lys Ile Gln Met Lys Trp Gly Met Glu Ala Arg Arg Cys
565 570 575
Leu Leu Gln Ser Met Gln Gln Met Glu Ala Ile Val Glu Gln Glu Ser
580 585 590
Ser Ile Gln Gly Tyr Asp Met Thr Lys Ala Cys Phe Lys Gly Asp Arg
595 600 605
Val Asn Ser Pro Lys Thr Phe Ser Ile Gly Thr Gln Glu Gly Lys Leu
610 615 620
Val Lys Gly Ser Phe Gly Lys Ala Leu Arg Val Ile Phe Thr Lys Cys
625 630 635 640
Leu Met His Tyr Val Phe Gly Asn Ala Gln Leu Glu Gly Phe Ser Ala
645 650 655
Glu Ser Arg Arg Leu Leu Leu Leu Ile Gln Ala Leu Lys Asp Arg Lys
660 665 670
Gly Pro Trp Val Phe Asp Leu Glu Gly Met Tyr Ser Gly Ile Glu Glu
675 680 685
Cys Ile Ser Asn Asn Pro Trp Val Ile Gln Ser Ala Tyr Trp Phe Asn
690 695 700
Glu Trp Leu Gly Phe Glu Lys Glu Gly Ser Lys Val Leu Glu Ser Val
705 710 715 720
Asp Glu Ile Met Asp Glu
725
<210>30
<211>584
<212>PRT
<213>流感B/Vienna/1/99/ca
<400>30
Met Lys Ala Ile Ile Val Leu Leu Met Val Val Thr Ser Asn Ala Asp
1 5 10 15
Arg Ile Cys Thr Gly Ile Thr Ser Ser Asn Ser Pro His Val Val Lys
20 25 30
Thr Ala Thr Gln Gly Glu Val Asn Val Thr Gly Ala Ile Pro Leu Thr
35 40 45
Thr Thr Pro Thr Lys Ser His Phe Ala Asn Leu Lys Gly Thr Lys Thr
50 55 60
Arg Gly Lys Leu Cys Pro Thr Cys Leu Asn Cys Thr Asp Leu Asp Val
65 70 75 80
Ala Leu Gly Arg Pro Met Cys Val Gly Ile Thr Pro Ser Ala Lys Ala
85 90 95
Ser Ile Leu His Glu Val Arg Pro Val Thr Ser Gly Cys Phe Pro Ile
100 105 110
Met His Asp Arg Thr Lys Ile Arg Gln Leu Pro Asn Leu Leu Arg Gly
115 120 125
Tyr Glu Lys Ile Arg Leu Ser Thr Gln Asn Val Ile Asn Thr Glu Lys
130 135 140
Ala Pro Gly Gly Pro Tyr Arg Leu Gly Thr Ser Gly Ser Cys Pro Asn
145 150 155 160
Ala Thr Ser Lys Ser Gly Phe Phe Ala Thr Met Ala Trp Ala Val Pro
165 170 175
Arg Asp Asn Asn Lys Thr Ala Thr Asn Pro Leu Thr Val Glu Val Pro
180 185 190
His Ile Cys Thr Lys Glu Glu Asp Gln Ile Thr Val Trp Gly Phe His
195 200 205
Ser Asp Asn Lys Thr Gln Met Lys Asn Leu Tyr Gly Asp Ser Asn Pro
210 215 220
Gln Lys Phe Thr Ser Ser Ala Asn Gly Ile Thr Thr His Tyr Val Ser
225 230 235 240
Gln Ile Gly Gly Phe Pro Asp Gln Thr Glu Asp Gly Gly Leu Pro Gln
245 250 255
Ser Gly Arg Ile Val Val Asp Tyr Met Val Gln Lys Pro Gly Lys Thr
260 265 270
Gly Thr Ile Val Tyr Gln Arg Gly Ile Leu Leu Pro Gln Lys Val Trp
275 280 285
Cys Ala Ser Gly Arg Ser Lys Val Ile Lys Gly Ser Leu Pro Leu Ile
290 295 300
Gly Glu Ala Asp Cys Leu His Glu Lys Tyr Gly Gly Leu Asn Lys Ser
305 310 315 320
Lys Pro Tyr Tyr Thr Gly Glu His Ala Lys Ala Ile Gly Asn Cys Pro
325 330 335
Ile Trp Val Lys Thr Pro Leu Lys Leu Ala Asn Gly Thr Lys Tyr Arg
340 345 350
Pro Pro Ala Lys Leu Leu Lys Glu Arg Gly Phe Phe Gly Ala Ile Ala
355 360 365
Gly Phe Leu Glu Gly Gly Trp Glu Gly Met Ile Ala Gly Trp His Gly
370 375 380
Tyr Thr Ser His Gly Ala His Gly Val Ala Val Ala Ala Asp Leu Lys
385 390 395 400
Ser Thr Gln Glu Ala Ile Asn Lys Ile Thr Lys Asn Leu Asn Ser Leu
405 410 415
Ser Glu Leu Glu Val Asn Asn Leu Gln Arg Leu Ser Gly Ala Met Asp
420 425 430
Glu Leu His Asn Glu Ile Leu Glu Leu Asp Glu Lys Val Asp Asp Leu
435 440 445
Arg Ala Asp Thr Ile Ser Ser Gln Ile Glu Leu Ala Val Leu Leu Ser
450 455 460
Asn Glu Gly Ile Ile Asn Ser Glu Asp Glu His Leu Leu Ala Leu Glu
465 470 475 480
Arg Lys Leu Lys Lys Met Leu Gly Pro Ser Ala Val Asp Ile Gly Asn
485 490 495
Gly Cys Phe Glu Thr Lys His Lys Cys Asn Gln Thr Cys Leu Asp Arg
500 505 510
Ile Ala Ala Gly Thr Phe Asn Ala Glu Glu Phe Ser Leu Pro Thr Phe
515 520 525
Asp Ser Leu Asn Ile Thr Ala Ala Ser Leu Asn Asp Asp Gly Leu Asp
530 535 540
Asn His Thr Ile Leu Leu Tyr Tyr Ser Thr Ala Ala Ser Ser Leu Ala
545 550 555 560
Val Thr Leu Met Ile Ala Ile Phe Ile Val Tyr Met Ile Ser Arg Asp
565 570 575
Asn Val Ser Cys Ser Ile Cys Leu
580
<210>31
<211>560
<212>PRT
<213>流感B/Vienna/1/99/ca
<400>31
Met Ser Asn Met Asp Ile Asp Gly Ile Asn Thr Gly Thr Ile Asp Lys
1 5 10 15
Thr Pro Glu Glu Ile Thr Phe Gly Thr Ser Gly Thr Thr Arg Pro Ile
20 25 30
Ile Arg Pro Ala Thr Leu Ala Pro Pro Ser Asn Lys Arg Thr Arg Asn
35 40 45
Pro Ser Pro Glu Arg Ala Thr Thr Ser Ser Glu Ala Asp Val Gly Arg
50 55 60
Lys Thr Gln Lys Lys Gln Thr Pro Thr Glu Ile Lys Lys Ser Val Tyr
65 70 75 80
Asn Met Val Val Lys Leu Gly Glu Phe Tyr Asn Gln Met Met Val Lys
85 90 95
Ala Gly Leu Asn Asp Asp Met Glu Arg Asn Leu Ile Gln Asn Ala His
100 105 110
Ala Val Glu Arg Ile Leu Leu Ala Ala Thr Asp Asp Lys Lys Thr Glu
115 120 125
Phe Gln Lys Lys Lys Asn Thr Arg Asp Val Lys Glu Gly Lys Glu Glu
130 135 140
Ile Asp His Asn Lys Thr Gly Gly Thr Phe Tyr Lys Met Val Arg Asp
145 150 155 160
Asp Lys Thr Ile Tyr Phe Ser Pro Ile Arg Ile Thr Phe Leu Lys Glu
165 170 175
Glu Val Lys Thr Met Tyr Lys Thr Thr Met Gly Ser Asp Gly Phe Ser
180 185 190
Gly Leu Asn His Ile Met Ile Gly His Ser Gln Met Asn Asp Val Cys
195 200 205
Phe Gln Arg Ser Lys Ala Leu Lys Arg Val Gly Leu Asp Pro Ser Leu
210 215 220
Ile Ser Thr Phe Ala Gly Ser Thr Ile Pro Arg Arg Ser Gly Ala Thr
225 230 235 240
Gly Val Ala Ile Lys Gly Gly Gly Thr Leu Val Ala Glu Ala Ile Arg
245 250 255
Phe Ile Gly Arg Ala Met Ala Asp Arg Gly Leu Leu Arg Asp Ile Lys
260 265 270
Ala Lys Thr Ala Tyr Glu Lys Ile Leu Leu Asn Leu Lys Asn Lys Cys
275 280 285
Ser Ala Pro Gln Gln Lys Ala Leu Val Asp Gln Val Ile Gly Ser Arg
290 295 300
Asn Pro Gly Ile Ala Asp Ile Glu Asp Leu Thr Leu Leu Ala Arg Ser
305 310 315 320
Met Val Val Val Arg Pro Ser Val Ala Ser Lys Val Val Leu Pro Ile
325 330 335
Ser Ile Tyr Ala Lys Ile Pro Gln Leu Gly Phe Asn Val Glu Glu Tyr
340 345 350
Ser Met Val Gly Tyr Glu Ala Met Ala Leu Tyr Asn Met Ala Thr Pro
355 360 365
Val Ser Ile Leu Arg Met Gly Asp Asp Ala Lys Asp Lys Ser Gln Leu
370 375 380
Phe Phe Met Ser Cys Phe Gly Ala Ala Tyr Glu Asp Leu Arg Val Leu
385 390 395 400
Ser Ala Leu Thr Gly Thr Glu Phe Lys Pro Arg Ser Ala Leu Lys Cys
405 410 415
Lys Gly Phe His Val Pro Ala Lys Glu Gln Val Glu Gly Met Gly Ala
420 425 430
Ala Leu Met Ser Ile Lys Leu Gln Phe Trp Ala Pro Met Thr Arg Ser
435 440 445
Gly Gly Asn Glu Val Gly Gly Asp Gly Gly Ser Gly Gln Ile Ser Cys
450 455 460
Ser Pro Val Phe Ala Val Glu Arg Pro Ile Ala Leu Ser Lys Gln Ala
465 470 475 480
Val Arg Arg Met Leu Ser Met Asn Ile Glu Gly Arg Asp Ala Asp Val
485 490 495
Lys Gly Asn Leu Leu Lys Met Met Asn Asp Ser Met Ala Lys Lys Thr
500 505 510
Ser Gly Asn Ala Phe Ile Gly Lys Lys Met Phe Gln Ile Ser Asp Lys
515 520 525
Asn Lys Thr Asn Pro Val Glu Ile Pro Ile Lys Gln Thr IIe Pro Asn
530 535 540
Phe Phe Phe Gly Arg Asp Thr Ala Glu Asp Tyr Asp Asp Leu Asp Tyr
545 550 555 560
<210>32
<211>100
<212>PRT
<213>流感B/Vienna/1/99/ca
<400>32
Met Asn Asn Ala Thr Phe Asn Tyr Thr Asn Val Asn Pro Ile Pro His
1 5 10 15
Ile Arg Gly Ser Val Ile Ile Thr Ile Cys Val Ser Phe Thr Val Ile
20 25 30
Leu Ile Ile Phe Gly Tyr Ile Ala Lys Ile Phe Thr Asn Arg Asn Asn
35 40 45
Cys Thr Asn Asn Ala Ile Gly Leu Cys Lys Arg Ile Lys Cys Ser Gly
50 55 60
Cys Glu Pro Phe Cys Asn Lys Arg Gly Asp Thr Ser Ser Pro Arg Thr
65 70 75 80
Gly Val Asp Ile Pro Ala Phe Ile Leu Pro Gly Leu Asn Leu Ser Glu
85 90 95
Ser Thr Pro Asn
100
<210>33
<211>466
<212>PRT
<213>流感B/Vienna/1/99/ca
<400>33
Met Leu Pro Ser Thr Ile Gln Thr Leu Thr Leu Phe Leu Thr Ser Gly
1 5 10 15
Gly Val Leu Leu Ser Leu Tyr Val Ser Ala Ser Leu Ser Tyr Leu Leu
20 25 30
Tyr Ser Asp Ile Leu Leu Lys Phe Ser Pro Thr Glu Ile Thr Ala Pro
35 40 45
Thr Met Pro Leu Asp Cys Ala Asn Ala Ser Asn Val Gln Ala Val Asn
50 55 60
Arg Ser Ala Thr Lys Gly Val Thr Leu Leu Leu Pro Glu Pro Glu Trp
65 70 75 80
Thr Tyr Pro Arg Leu Ser Cys Pro Gly Ser Thr Phe Gln Lys Ala Leu
85 90 95
Leu Ile Ser Pro His Arg Phe Gly Glu Thr Lys Gly Asn Ser Ala Pro
100 105 110
Leu Ile Ile Arg Glu Pro Phe Ile Ala Cys Gly Pro Lys Glu Cys Lys
115 120 125
His Phe Ala Leu Thr His Tyr Ala Ala Gln Pro Gly Gly Tyr Tyr Asn
130 135 140
Gly Thr Arg Glu Asp Arg Asn Lys Leu Arg His Leu Ile Ser Val Lys
145 150 155 160
Leu Gly Lys Ile Pro Thr Val Glu Asn Ser Ile Phe His Met Ala Ala
165 170 175
Trp Ser Gly Ser Ala Cys His Asp Gly Lys Glu Trp Thr Tyr Ile Gly
180 185 190
Val Asp Gly Pro Asp Ser Asn Ala Leu Leu Lys Ile Lys Tyr Gly Glu
195 200 205
Ala Tyr Thr Asp Thr Tyr His Ser Tyr Ala Asn Asn Ile Leu Arg Thr
210 215 220
Gln Glu Ser Ala Cys Asn Cys Ile Gly Gly Asn Cys Tyr Leu Met Ile
225 230 235 240
Thr Asp Gly Ser Ala Ser Gly Ile Ser Glu Cys Arg Phe Leu Lys Ile
245 250 255
Gln Glu Gly Arg Ile Ile Lys Glu Ile Phe Pro Thr Gly Arg Val Glu
260 265 270
His Thr Glu Glu Cys Thr Cys Gly Phe Ala Ser Asn Lys Thr Ile Glu
275 280 285
Cys Ala Cys Arg Asp Asn Ser Tyr Thr Ala Lys Arg Pro Phe Val Lys
290 295 300
Leu Asn Val Glu Thr Asp Thr Ala Glu Ile Arg Leu Met Cys Thr Glu
305 310 315 320
Thr Tyr Leu Asp Thr Pro Arg Pro Asp Asp Gly Ser Ile Thr Gly Pro
325 330 335
Cys Glu Ser Asn Gly Asp Lys Gly Ser Gly Gly Ile Lys Gly Gly Phe
340 345 350
Val His Gln Arg Met Ala Ser Lys Thr Gly Arg Trp Tyr Ser Arg Thr
355 360 365
Met Ser Lys Thr Lys Arg Met Gly Met Gly Leu Tyr Val Lys Tyr Asp
370 375 380
Gly Asp Pro Trp Thr Asp Ser Asp Ala Leu Ala Leu Ser Gly Val Met
385 390 395 400
Val Ser Met Glu Glu Pro Gly Trp Tyr Ser Phe Gly Phe Glu Ile Lys
405 410 415
Asp Lys Lys Cys Asp Val Pro Cys Ile Gly Ile Glu Met Val His Asp
420 425 430
Gly Gly Lys Glu Thr Trp His Ser Ala Ala Thr Ala Ile Tyr Cys Leu
435 440 445
Met Gly Ser Gly Gln Leu Leu Trp Asp Thr Val Thr Gly Val Asn Met
450 455 460
Ala Leu
465
<210>34
<211>248
<212>PRT
<213>流感B/Vienna/1/99/ca
<400>34
Met Ser Leu Phe Gly Asp Thr Ile Ala Tyr Leu Leu Ser Leu Thr Glu
1 5 10 15
Asp Gly Glu Gly Lys Ala Glu Leu Ala Glu Lys Leu His Cys Trp Phe
20 25 30
Gly Gly Lys Glu Phe Asp Leu Asp Ser Ala Leu Glu Trp Ile Lys Asn
35 40 45
Lys Arg Cys Leu Thr Asp Ile Gln Lys Ala Leu Ile Gly Ala Ser Ile
50 55 60
Cys Phe Leu Lys Pro Lys Asp Gln Glu Arg Lys Arg Arg Phe Ile Thr
65 70 75 80
Glu Pro Leu Ser Gly Met Gly Thr Thr Ala Thr Lys Lys Lys Gly Leu
85 90 95
Ile Leu Ala Glu Arg Lys Met Arg Arg Cys Val Ser Phe His Glu Ala
100 105 110
Phe Glu Ile Ala Glu Gly His Glu Ser Ser Ala Leu Leu Tyr Cys Leu
115 120 125
Met Val Met Tyr Leu Asn Pro Gly Asn Tyr Ser Met Gln Val Lys Leu
130 135 140
Gly Thr Leu Cys Ala Leu Cys Glu Lys Gln Ala Ser His Ser His Arg
145 150 155 160
Ala His Ser Arg Ala Ala Arg Ser Ser Val Pro Gly Val Arg Arg Glu
165 170 175
Met Gln Met Val Ser Ala Met Asn Thr Ala Lys Thr Met Asn Gly Met
180 185 190
Gly Lys Gly Glu Asp Val Gln Lys Leu Ala Glu Glu Leu Gln Ser Asn
195 200 205
Ile Gly Val Leu Arg Ser Leu Gly Ala Ser Gln Lys Asn Gly Glu Gly
210 215 220
Ile Ala Lys Asp Val Met Glu Val Leu Lys Gln Ser Ser Met Gly Asn
225 230 235 240
Ser Ala Leu Val Lys Lys Tyr Leu
245
<210>35
<211>109
<212>PRT
<213>流感B/Vienna/1/99/ca
<400>35
Met Leu Glu Pro Phe Gln Ile Leu Ser Ile Cys Ser Phe Ile Leu Ser
1 5 10 15
Ala Leu His Phe Val Ala Trp Thr Ile Gly His Leu Asn Gln Ile Lys
20 25 30
Arg Gly Val Asn Met Lys Ile Arg Ile Lys Ser Pro Asn Lys Glu Thr
35 40 45
Ile Asn Arg Glu Val Ser Ile Leu Arg His Ser Tyr Gln Lys Glu Ile
50 55 60
Gln Ala Lys Glu Thr Met Lys Glu Val Leu Ser Asp Asn Met Glu Val
65 70 75 80
Leu Gly Asp His Ile Val Ile Glu Gly Leu Ser Ala Glu Glu Ile Ile
85 90 95
Lys Met Gly Glu Thr Val Leu Glu Ile Glu Glu Leu His
100 105
<210>36
<211>281
<212>PRT
<213>流感B/Vienna/1/99/ca
<400>36
Met Ala Asn Asn Ile Thr Thr Thr Gln Ile Glu Val Gly Pro Gly Ala
1 5 10 15
Thr Asn Ala Thr Ile Asn Phe Glu Thr Gly Ile Leu Glu Cys Tyr Glu
20 25 30
Arg Leu Ser Trp Gln Arg Ala Leu Asp Tyr Pro Gly Gln Asp Arg Leu
35 40 45
Asn Arg Leu Lys Arg Lys Leu Glu Ser Arg Ile Lys Thr His Asn Lys
50 55 60
Ser Glu Pro Glu Ser Lys Arg Met Ser Leu Glu Glu Arg Lys Ala Ile
65 70 75 80
Gly Val Lys Met Met Lys Val Leu Leu Phe Met Asn Pro Ser Ala Gly
85 90 95
Ile Glu Gly Phe Glu Pro Tyr Tyr Met Lys Ser Ser Ser Asn Ser Asn
l00 105 110
Cys Pro Lys Tyr Asn Trp Thr Asp Tyr Pro Ser Thr Pro Gly Arg Cys
115 120 125
Leu Asp Asp Ile Glu Glu Glu Pro Glu Asp Val Asp Gly Pro Thr Glu
130 135 140
Ile Val Leu Arg Asp Met Asn Asn Lys Asp Ala Arg Gln Lys Ile Lys
145 150 155 160
Glu Glu Val Asn Thr Gln Lys Glu Gly Lys Phe Arg Leu Thr Ile Lys
165 170 175
Arg Asp Ile Arg Asn Val Leu Ser Leu Arg Val Leu Val Asn Gly Thr
180 185 190
Phe Leu Lys His Pro Asn Gly Tyr Lys Ser Leu Ser Thr Leu His Arg
195 200 205
Leu Asn Ala Tyr Asp Gln Ser Gly Arg Leu Val Ala Lys Leu Val Ala
210 215 220
Thr Asp Asp Leu Thr Val Glu Asp Glu Glu Asp Gly His Arg Ile Leu
225 230 235 240
Asn Ser Leu Phe Glu Arg Leu Asn Glu Gly His Ser Lys Pro Ile Arg
245 250 255
Ala Ala Glu Thr Ala Val Gly Val Leu Ser Gln Phe Gly Gln Glu His
260 265 270
Arg Leu Ser Pro Glu Glu Gly Asp Asn
275 280
<210>37
<211>122
<212>PRT
<213>流感B/Vienna/1/99/ca
<400>37
Met Ala Asn Asn Ile Thr Thr Thr Gln Ile Glu Trp Arg Met Lys Lys
1 5 10 15
Met Ala Ile Gly Ser Ser Thr His Ser Ser Ser Val Leu Met Lys Asp
20 25 30
Ile Gln Ser Gln Phe Glu Gln Leu Lys Leu Arg Trp Glu Ser Tyr Pro
35 40 45
Asn Leu Val Lys Ser Thr Asp Tyr His Gln Lys Arg Glu Thr Ile Lys
50 55 60
Leu Val Thr Glu Glu Leu Tyr Leu Leu Ser Lys Arg Ile Asp Asp Asn
65 70 75 80
Ile Leu Phe His Lys Thr Val Ile Ala Asn Ser Ser Ile Ile Ala Asp
85 90 95
Met Val Val Ser Leu Ser Leu Leu Glu Thr Leu Tyr Glu Met Lys Asp
100 105 110
Val Val Glu Val Tyr Ser Arg Gln Cys Leu
115 120