CN112189017A - 水痘带状疱疹病毒的抗原变体及其用途 - Google Patents

水痘带状疱疹病毒的抗原变体及其用途 Download PDF

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CN112189017A
CN112189017A CN201980034772.8A CN201980034772A CN112189017A CN 112189017 A CN112189017 A CN 112189017A CN 201980034772 A CN201980034772 A CN 201980034772A CN 112189017 A CN112189017 A CN 112189017A
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南效廷
池可英
金恩美
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Abstract

本发明涉及一种抗原变体及其用途，该抗原变体是水痘带状疱疹病毒的表面蛋白(gE)中的一种蛋白质，该蛋白质表现出高表达水平和高免疫原性。因此，当使用该抗原变体作为疫苗组合物时，与活病毒疫苗相比，疫苗组合物具有更优异的安全性，并且抗原变体在宿主细胞中的表达水平高于其他抗原，因此可用作疫苗，所述疫苗用于预防或治疗水痘或由水痘带状疱疹病毒引起的带状疱疹。

Description

水痘带状疱疹病毒的抗原变体及其用途
技术领域
本发明涉及水痘带状疱疹病毒抗原变体及其用途，更具体地，涉及具有高表达水平和高免疫原性的水痘带状疱疹病毒抗原变体，其选自水痘带状疱疹病毒表面蛋白(gE)抗原变体，和一种用于预防或治疗水痘或带状疱疹的疫苗组合物，其包含水痘带状疱疹病毒抗原变体作为活性成分。
背景技术
水痘带状疱疹病毒(VZV)是一种主要在儿童和青少年中引起水痘的病毒。一旦发生感染，VZV会在感觉根和颅神经神经节细胞中保持休眠状态数年，并在免疫力降低时重新激活并在成年后引起带状疱疹。水痘具有高度传染性。一旦发生感染，就会在全身引起水疱样皮疹，并伴有发烧和全身不适。在大多数正常儿童中，水痘很少会发展到严重的状况，最终会发展为自限性疾病。然而，已知在接受器官移植或化学疗法的患者中，有很多水痘发展为严重症状的病例发生(Adriana Weinberg等，传染病杂志，200(7)：1068，2009；JudithBreuer等，疫苗专家评论，2017年，DOI：10.1080/14760584.2017.1394843)。
带状疱疹的最初症状是全身酸痛，如全身疼痛，或严重的瘙痒、刺痛和灼烧感，并伴有如同被刀刺伤一样剧烈的疼痛感。带状疱疹是一种疾病，几天后会出现水疱，随着皮肤损伤的增加疼痛会增加，老年患者往往抱怨疼痛更严重。即使在带状疱疹被治愈的情况下，也可能留下神经痛作为后遗症。众所周知，在60岁或60岁以上的人群中，神经痛可能使他们无法入睡，使他们抱怨慢性疲劳，即使在轻微接触或摩擦时也会感到剧烈疼痛，甚至导致抑郁，尽管这种神经痛在40岁或以下的成年人中相对罕见。
针对水痘的代表性预防疫苗包括例如VARIVAX(默克公司)和VARILRIX(葛兰素史克生物制品公司)等产品，它们是使用Oka菌株开发的，该菌株是1970年开发的减毒菌株。在韩国，产品如Suduvax(绿十字公司)，是使用1980年开发的MAV/06菌株生产的，可商购。所讨论的市售活疫苗显示出平均80％的保护效力，这意味着即使在接种疫苗后，仍有20％的疫苗接种者发生了感染。稳定性问题不断被指出，例如由疫苗中所含活病毒引起的水痘和带状疱疹的发生。
ZOSTAVAX(默克公司)是使用Oka毒株生产的减毒活疫苗，被开发为针对带状疱疹的预防性疫苗。由于该疫苗中含有大量病毒，因此该疫苗已在美国和韩国获得批准并销售的条件是应用于50岁或50岁以上的成年人，而不是儿童或青少年。最近，葛兰素史克生物制品公司开发了一种由病毒表面蛋白(gE)和佐剂组成的疫苗，该疫苗旨在用于50岁或以上的成年人，并已在临床试验中证明具有预防作用(美国专利号7,939,084，于2011年1月7日授权)。
在疫苗开发的早期，主要用减毒活细胞或死细胞作抗原。然而，由于安全性问题和病原体中存在的免疫抑制物质，此类抗原的开发正转向蛋白质抗原的开发，所述蛋白质抗原具有清楚的结构和组成，并可以为疾病防御建立所必需的免疫力。
因此，本发明人经过深入的努力，开发了一种在宿主细胞中具有高表达水平的蛋白抗原，所述蛋白抗原在不影响免疫原性的前提下，提高水痘带状疱疹病毒表面蛋白(gE)抗原的产量。结果，发明人生产了不同长度的水痘带状疱疹病毒表面蛋白抗原变体并测量了其表达水平，从而在抗原变体中鉴定了高表达水平的特定氨基酸序列。从而完成了本发明。
发明公开内容
技术问题
本发明的目的是提供一种具有高表达水平和高免疫原性的特异性抗原变体，其选自水痘带状疱疹病毒表面蛋白(gE)抗原变体。
本发明的另一个目的是提供编码抗原变体的基因，包含该基因的重组载体和用该重组载体转化的宿主细胞。
本发明的又一个目的是提供一种用于预防或治疗水痘或带状疱疹的疫苗组合物，其包含抗原变体作为活性成分。
本发明的又一个目的是提供一种预防或治疗水痘或带状疱疹的方法，其使用包含抗原变体作为活性成分的疫苗组合物。
本发明的又一个目的是提供包含抗原变体作为活性成分的疫苗组合物的用途，用于预防或治疗水痘或带状疱疹。
本发明的又一个目的是提供包含抗原变体作为活性成分的疫苗组合物的用途，用于制备预防或治疗水痘或带状疱疹的药物。
技术问题的解决方案
为了实现上述目的，在本发明中，提供了一种水痘带状疱疹病毒表面蛋白抗原变体，其特征在于该抗原变体包括选自以下组的任一氨基酸残基的羧基端截断的变体：氨基酸序列由SEQ ID NO：1所示的水痘带状疱疹病毒表面蛋白(gE)抗原中的第525位至第543位氨基酸残基。
另外，在本发明中，提供了编码抗原变体的基因，包含该基因的重组载体和用该重组载体转化的宿主细胞。
另外，在本发明中，提供了一种用于预防或治疗水痘或带状疱疹的疫苗组合物，其包含抗原变体作为活性成分。
另外，在本发明中，提供了一种预防或治疗水痘或带状疱疹的方法，使用包含抗原变体作为活性成分的疫苗组合物。
另外，在本发明中，提供了包含抗原变体作为活性成分的疫苗组合物的用途，用于预防或治疗水痘或带状疱疹。
另外，在本发明中，提供了包含抗原变体作为活性成分的疫苗组合物的用途，用于制备预防或治疗水痘或带状疱疹的药物。
附图说明
图1示意性地示出了本发明的实验过程。
图2显示了各种长度的水痘带状疱疹病毒表面蛋白(gE)抗原的氨基酸序列。
图3示出了比较水痘带状疱疹病毒抗原的gE片段表达水平的蛋白质印迹结果。
图4示出了通过蛋白质印迹比较包含在由葛兰素史克生物学公司生产的水痘带状疱疹病毒疫苗Shingrix中的表面蛋白抗原(GSK gE 546氨基酸)，和由本发明人生产的抗原变体(Mogam gE 537氨基酸)的表达水平的结果。
图5示出了通过抗gE特异性IgG ELISA比较包含在由葛兰素史克生物学公司生产的水痘带状疱疹病毒疫苗Shingrix中的表面蛋白抗原(GSK gE 546氨基酸)，和由本发明人生产的抗原变体(Mogam gE 537氨基酸)的体液免疫应答结果。
图6示出了通过小鼠IFN-γELISA比较包含在由葛兰素史克生物学公司生产的水痘带状疱疹病毒疫苗Shingrix中的表面蛋白抗原(GSK gE 546氨基酸)，和本发明人生产的抗原变体(Mogam gE 537氨基酸)的细胞介导的免疫应答(CMI)的结果。
图7说明了通过抗gE特异性IgG ELISA比较水痘带状疱疹病毒抗原的gE片段的体液免疫应答的结果。
图8示出了通过抗gE特异性IgG ELISA比较作为水痘带状疱疹病毒抗原的gE片段的体液免疫应答结果，随后总结显示出抗原特异性抗体反应的响应者的数量。
发明实施的最佳方式
除非另有定义，否则本文使用的所有技术和科学术语具有与本发明所属领域的技术人员通常理解的相同含义。通常，本文使用的术语是本领域公知的且通常使用的。
在本发明中，为了提高生产力而不影响免疫原性，从水痘带状疱疹病毒蛋白抗原中开发出在宿主细胞中具有高表达水平的抗原，进行通过产生不同长度的表面蛋白(gE)抗原变体，然后测量其表达水平的实验。在结果中，本发明人生产的水痘带状疱疹病毒表面蛋白抗原变体中，特定氨基酸序列为代表的抗原表现出比其他抗原更高的表达水平(图3)。
此外，通过抗体效价测量(图7)和抗原特异性响应者测量(图8)以选择具有高免疫原性的抗原变体。在结果中，确定了Mogam gE 534氨基酸至gE 540氨基酸具有更高的免疫原性。
因此，在本发明的一个方面，提供了一种水痘带状疱疹病毒表面蛋白抗原变体，包括由选自SEQ ID NO：1所示氨基酸序列表示的水痘带状疱疹病毒表面蛋白(gE)抗原的第525位至543位氨基酸的任一氨基酸残基的羧基末端的截断组成的变体。
具体地，在本发明中，提供了水痘带状疱疹病毒表面蛋白抗原变体，其特征在于该抗原变体包括选自下组的变体：
a)第525位氨基酸残基的羧基末端的截断；
b)第526位氨基酸残基的羧基末端的截断；
c)第527位氨基酸残基的羧基末端的截断；
d)第528位氨基酸残基的羧基末端的截断；
e)第529位氨基酸残基的羧基末端的截断；
f)第530位氨基酸残基的羧基末端的截断；
g)第531位氨基酸残基的羧基末端的截断；
h)第532位氨基酸残基的羧基末端的截断；
i)第533位氨基酸残基的羧基末端的截断；
j)第534位氨基酸残基的羧基末端的截断；
k)第535位氨基酸残基的羧基末端的截断；
l)第536位氨基酸残基的羧基末端的截断；
m)第537位氨基酸残基的羧基末端的截断；
n)第538位氨基酸残基的羧基末端的截断；
o)第539位氨基酸残基的羧基末端的截断；
p)第540位氨基酸残基的羧基末端的截断；
q)第541位氨基酸残基的羧基末端的截断；
r)第542位氨基酸残基的羧基末端的截断；和
s)第543位氨基酸残基的羧基末端的截断。
抗原变体的特征在于，其优选包括选自下组的变体：j)第534位氨基酸残基的羧基末端的截断；k)第535位氨基酸残基的羧基末端的截断；l)第536位氨基酸残基的羧基末端的截断；m)第537位氨基酸残基的羧基末端的截断；n)第538位氨基酸残基的羧基末端的截断；o)第539位氨基酸残基的羧基末端的截断；p)第540位氨基酸残基的羧基末端的截断；然而，变化不限于此。
SEQ ID NO:1:
mgtvnkpvvg vlmgfgiitg tlritnpvra svlryddfhX1 dedkldtnsv yepyyhsdhaesswvnrges srkaydhnsp yiwprndydg flenahehhg vynqgrgids gerlmqptqm saqedlgddtgihviptlng ddrhkivnvd qrqygdvfkg dlnpkpqgqr lievsveenh pftlrapiqr iygvrytetwsflpsltctg daapaiqhic lkhttcfqdv vvdvdcaent kedqlaeisy rfqgkkeadq pwivvntstlfdeleldppe iepgvlkvlr tekqylgvyi wnmrgsdgts tyatflvtwk gdektrnptp avtpqprgaefhmwnyhshv fsvgdtfsla mhlqykihea pfdlllewly vpidptcqpm rlystclyhp napqclshmnsgctftsphl aqrvastvyq ncehadnyta yclgishmep sfglilhdgg ttlkfvdtpe slsglyvfvvyfnghveava ytvvstvdhf vnaieergfp ptagqppatt kpkeitpvnp gtsplX2ryaa wtgglaavvllclviflict akrmrvkayr vdkspynqsm yyaglpvddf edsestdtee efgnaiggsh ggssytvyidktr(其中X1为T或I,以及X2为L或I)。
在本发明中，水痘带状疱疹病毒表面蛋白抗原变体的特征在于，它是由534至540个氨基酸组成的水痘带状疱疹病毒表面蛋白抗原变体，其衍生自SEQ ID NO：1所示氨基酸序列的水痘带状疱疹病毒表面蛋白抗原，其由623个氨基酸组成，或者是一些氨基酸残基中羧基末端被截断的变体。例如，如本文所用，术语“第534个氨基酸残基的羧基末端的截断的变体”是指从氨基末端(N末端)到羧基末端(C末端)的方向，第1个至第534个氨基酸残基保留，而从第535个氨基酸残基至羧基末端的连续氨基酸残基被截断。
根据本发明的一个具体实施方案，在由SEQ ID NO：1所示的氨基酸序列中的第40位氨基酸残基是苏氨酸。
根据本发明的一个具体实施方案，在由SEQ ID NO：1所示的氨基酸序列中的536位氨基酸残基是亮氨酸。
在本发明中，水痘带状疱疹病毒表面蛋白抗原变体的特征在于，由SEQ ID NO：2至8和SEQ ID NO：21至23中的任一种氨基酸序列表示，如下所示：
a)由SEQ ID NO：2所示的氨基酸序列表示的抗原变体，其通过截断第534位氨基酸残基的羧基末端获得；
b)由SEQ ID NO：3所示的氨基酸序列表示的抗原变体，其通过截断第535个氨基酸残基的羧基末端获得；
c)由SEQ ID NO：4所示的氨基酸序列表示的抗原变体，其通过截断第536个氨基酸残基的羧基末端获得；
d)由SEQ ID NO：5所示的氨基酸序列表示的抗原变体，其通过截断第537个氨基酸残基的羧基末端获得；
e)由SEQ ID NO：6所示的氨基酸序列表示的抗原变体，其通过截断第538个氨基酸残基的羧基末端获得；
f)由SEQ ID NO：7所示的氨基酸序列表示的抗原变体，其通过截断第539个氨基酸残基的羧基末端获得；
g)由SEQ ID NO：8所示的氨基酸序列表示的抗原变体，其通过截断第540位氨基酸残基的羧基末端获得；
h)由SEQ ID NO：21所示的氨基酸序列表示的抗原变体，其通过截断第525位氨基酸残基的羧基末端获得；
i)由SEQ ID NO：22所示的氨基酸序列表示的抗原变体，其通过截断第530位氨基酸残基的羧基末端获得；或
j)由SEQ ID NO：23所示的氨基酸序列表示的抗原变体，其通过截断543位氨基酸残基的羧基末端获得的。
本发明的表面蛋白源自的糖蛋白，为进化枝1的水痘带状疱疹病毒包膜的成分，是由534至540个氨基酸组成的肽片段(被截断的蛋白质)，其通过羧基末端的部分截断获得。
考虑到生物学上等价的氨基酸变化，将本发明使用的氨基酸序列理解为包括与SEQ ID NO：2至8和SEQ ID NO：21至23的序列具有基本同一性的序列。上述基本同一性是指在如上所述的本发明的序列和任何其他序列进行比对以获得最大对应性并且使用本领域常用的算法比对的序列进行分析的情况下，其他序列具有在具有相同功能的情况下，与本发明的序列具有至少70％的同源性，更特别地80％的同源性，甚至更特别地90％的同源性，最特别地95％的同源性。
在本发明的一个实施方案中，鉴定出以下抗原变体具有高免疫原性：a)由SEQ IDNO：2所示的氨基酸序列表示的抗原变体：其通过截断第534位氨基酸残基的羧基末端获得；b)由SEQ ID NO：3所示的氨基酸序列表示的抗原变体，其通过截断第535位氨基酸残基的羧基末端获得；c)由SEQ ID NO：4所示的氨基酸序列表示的抗原变体，其通过截断第536位氨基酸残基的羧基末端获得；d)由SEQ ID NO：5所示的氨基酸序列表示的抗原变体，其通过截断第537位氨基酸残基的羧基末端获得；e)由SEQ ID NO：6所示的氨基酸序列表示的抗原变体，其通过截断第538位氨基酸残基的羧基末端获得；f)由SEQ ID NO：7所示的氨基酸序列表示的抗原变体，其通过截断第539位氨基酸残基的羧基末端获得；或g)由SEQ ID NO：8所示的氨基酸序列表示的抗原变体，其通过截断第540位氨基酸残基的羧基末端获得。
在疫苗开发的早期，主要用减毒活细胞或死细胞作抗原。然而，由于安全问题，此类抗原的开发正转向开发具有清晰结构和组成的蛋白质抗原。然而，蛋白质抗原通常具有问题，因为与常规疫苗相比，蛋白质抗原具有低免疫原性的问题。从优异的稳定性和免疫原性以及在宿主细胞中高水平表达的观点出发，本发明的抗原变体可以有效地用作预防或治疗水痘带状疱疹病毒诱导的水痘或带状疱疹的疫苗。
在本发明的另一方面，提供了编码抗原变体的基因，包含该抗原变体的重组载体和用该重组载体转化的宿主细胞。
在本发明中，该基因的特征在于它可以由SEQ ID NO：9至15的任何一种核苷酸序列表示。
如本文所用，术语“载体”是指包含DNA序列的DNA构建体，该DNA序列可操作地连接至能够实现DNA序列在合适宿主中表达的合适控制序列。载体可以是质粒、噬菌体颗粒，或者仅仅是潜在的基因组插入物。一旦转化成合适的宿主，载体就可以独立于宿主基因组复制和发挥作用，或者在某些情况下可以整合到其基因组中。在本说明书中，“质粒”和“载体”有时可互换使用，因为质粒是目前最常用的载体形式。为了本发明的目的，优选使用质粒载体。可用于此目的的典型质粒载体具有以下结构，包括：(a)复制起点，允许有效复制，使每个宿主细胞可产生数百个质粒载体；(b)抗生素抗性基因，可允许选择用质粒载体转化的宿主细胞，和(c)允许插入外源DNA片段的限制性酶切位点。即使在载体中不存在适当的限制酶切割位点，根据常规方法使用合成的寡核苷酸衔接子或接头也可以使载体与外源DNA容易连接。
如本文所用，术语“重组载体”通常是指其中插入有异源DNA片段的重组载体，该重组载体通常为双链DNA片段的形式。在此，异源DNA是指在宿主细胞中非天然发现的外源DNA。重组载体一旦进入宿主细胞，即可独立于宿主染色体DNA复制，从而可产生该载体和插入其中的(异源)DNA的数个拷贝。
连接后，将基因或重组载体转化或转染到宿主细胞中。对于“转化”或“转染”，可以使用几种通常用于将外源核酸(DNA或RNA)引入原核或真核宿主细胞中的多种类型的技术，其实例包括电穿孔、磷酸钙沉淀、DEAE-葡聚糖转染和脂质转染。
如本领域众所周知的，为增加转染的基因在宿主细胞中的表达水平，所讨论的基因应可操作地连接至在选定的表达宿主中发挥其功能的转录和翻译表达控制序列。
如本文所用，术语“转化”是指将DNA引入宿主，使得DNA可以作为染色体外因子或通过染色体整合而复制。当然，应当理解，并不是所有的载体在表达本发明的基因序列时都具有相同的功能。同样，并非所有宿主细胞都相对于同一表达系统具有同等的功能。然而，本领域的技术人员可以在不脱离本发明的主旨的范围内没有过度实验负担地做出各种载体、表达控制序列和宿主之间的适当选择。例如，必须考虑宿主而选择载体，因为该载体必须在其中复制。在这方面，还必须考虑载体的拷贝数，其调节拷贝数的能力以及由载体编码的其他蛋白质的表达。
在本发明中，待转化的宿主细胞优选选自，但不限于下组：动物细胞、植物细胞、酵母、大肠杆菌和昆虫细胞。
具体地，在本发明中，当其应用于活体时，只要它是无毒或减毒的微生物，用作待转化宿主细胞的微生物可以使用任何微生物。其实例可包括革兰氏阴性细菌，例如大肠杆菌(E.coli)、伤寒沙门氏菌(Salmonella typhi)、鼠伤寒沙门氏菌(Salmonellatyphimurium)、霍乱弧菌(Vibrio cholerae)、牛分枝杆菌(Mycobacterium bovis)和志贺氏菌(Shigella)；革兰氏阳性细菌，如芽孢杆菌(Bacillus)、乳杆菌(Lactobacillus)、乳球菌(Lactococcus)、葡萄球菌(Staphylococcus)、李斯特菌(Listeria monocytogenes)和链球菌(Streptococcus)。其优选例可包括作为可食用微生物的乳酸菌。然而，微生物不限于此。
乳酸菌可包括乳杆菌属、链球菌属和双歧杆菌属。乳杆菌属的代表性例子可以包括嗜酸乳杆菌(Lactobacillus acidophilus)、植物乳杆菌(Lactobacillus plantarum)、发酵乳杆菌(Lactobacillus fermentum)、德氏乳杆菌(Lactobacillus delbrueckii)、强生乳杆菌(Lactobacillus johnsonii)、路透乳杆菌(Lactobacillus reuteri)、保加利亚乳杆菌(Lactobacillus bulgaricus)和干酪乳杆菌(Lactobacillus casei)；链球菌的代表性实子可能包括嗜热链球菌(Streptococcus thermophiles)；和双歧杆菌的代表性例子可包括婴儿双歧杆菌(Bifidobacterium infantis)、双歧双歧杆菌(Bifidobacteriumbifidum)、长双歧杆菌(Bifidobacterium longum)、短双歧杆菌(Bifidobacteriumbreve)、乳酸双歧杆菌(Bifidobacterium lactis)和青春双歧杆菌(Bifidobacteriumadolescentis)，更优选干酪乳杆菌(Lactobacillus casei)。然而，乳酸菌不限于此。
微生物也可以是真核细胞，包括真菌，例如曲霉属(Aspergillus sp.)；酵母，例如巴斯德毕赤酵母(Pichia pastoris)，酿酒酵母(Saccharomyces cerevisiae)，裂殖酵母属(Schizosaccharomyces sp.)和克雷索氏菌(Neurospora crassa)，其他较低级的真核细胞，以及较高级的真核细胞，例如来自昆虫的细胞。
微生物可以源自植物或哺乳动物。其优选实例可包括猴肾细胞(COS-7细胞)、NS0细胞、SP2/0、中国仓鼠卵巢(CHO)细胞、W138、仓鼠肾(BHK)细胞、MDCK、骨髓瘤细胞系、HuT78细胞和HEK293细胞，其中CHO细胞是优选的。然而，微生物不限于此。
本发明的另一方面，提供了一种生产水痘带状疱疹病毒抗原的方法，该方法包括培养宿主细胞的步骤。
在将能够表达水痘带状疱疹病毒抗原的重组载体引入宿主细胞的情况下，可以通过将宿主细胞培养足以使抗原在其中表达的时间来产生抗原，或更优选地，足以使抗原分泌到其中培养宿主细胞的培养基中的时间。
在某些情况下，可以从宿主细胞中分离表达的抗原并纯化至同质。抗原的分离或纯化可以通过用于蛋白质的常规分离和纯化方法，例如色谱法来进行。色谱的实例可以包括亲和色谱，包括蛋白A柱或蛋白G柱、离子交换色谱和疏水色谱。除了上述色谱法之外，还可以通过进一步组合过滤、超滤、盐析、透析等来分离和纯化抗原。
在本发明的又一方面，提供了用于预防或治疗水痘或带状疱疹的疫苗组合物，其包含水痘带状疱疹病毒抗原变体作为活性成分。
在本发明的又一方面，提供了一种预防或治疗水痘或带状疱疹的方法，所述疫苗组合物包含水痘带状疱疹病毒抗原变体作为活性成分。
在本发明的又一方面，提供了一种疫苗组合物的用途，该疫苗组合物包含水痘带状疱疹病毒抗原变体作为活性成分，用于预防或治疗水痘或带状疱疹。
在本发明的又一方面，提供了包含作为活性成分的抗原变体的疫苗组合物在制备用于预防或治疗水痘或带状疱疹的药物中的用途。
如本文所用，术语“预防”是指在尚未被诊断为患有该疾病或疾病并且很可能患有这种疾病或疾病的受试者中抑制该病症或疾病的发生。
如本文所用，术语“治疗”是指(a)抑制病症或疾病或其症状的进展；(b)减轻病症或疾病或其症状；(c)消除病症或疾病或其症状。本发明的组合物在患有由水痘带状疱疹病毒感染引起的水痘或带状疱疹的个体中激活针对水痘带状疱疹病毒的免疫应答，从而起到抑制疾病进展、消除或减轻其症状的作用。因此，本发明的组合物本身可以是用于水痘或带状疱疹的治疗组合物，或者可以用作与其他药理成分组合施用的疾病的治疗辅助剂。
因此，在本说明书中，术语“治疗”或“治疗剂”也包括“辅助治疗”或“治疗辅助”的含义。
如本文所用，术语“活性成分”是指足以产生所需作用的疫苗组合物，其包括但不限于在患者中诱导或增加针对水痘带状疱疹病毒的免疫应答、预防、减轻或消除感染相同病毒或接种活水痘带状疱疹病毒疫苗的患者中的水痘带状疱疹病毒的再激活，可预防带状疱疹(HZ)和/或疱疹后神经痛(PHN)，并降低HZ和/或PHN的严重性或持续时间。本领域技术人员可知晓，这种期望效果的水平可以变化。
如本文所用，术语“免疫应答”是指细胞介导的(T细胞)免疫应答和/或抗体(B细胞)应答。
本发明的疫苗组合物可用于在免疫活性和免疫功能低下的患者群体中预防水痘和/或HZ和/或PHN，或降低水痘和/或HZ和/或PHN的严重程度或持续时间，包括但不限于接受造血细胞移植(HCT)或实体器官移植(SOT)的健康个体和免疫功能低下的患者，感染了HIV的患者，患有自身免疫性疾病的患者以及患有血液癌的患者；接受过各种实体恶性肿瘤化疗的个人；以及针对各种疾病(包括类风湿关节炎(RA)、系统性红斑狼疮(SLE)、克罗恩病、牛皮癣和多发性硬化症)进行慢性免疫抑制治疗的患者。
在本发明中，疫苗组合物的特征在于它进一步包含药学上可接受的载体，赋形剂或稀释剂。
根据本发明所属领域的技术人员可以容易地实施的方法，通过使用药学上可接受的载体和/或赋形剂进行配制，可以将本发明的疫苗组合物制备成单位剂型，也可以制成多剂量容器的形式制备。在此，剂型可以按照常规方法配制成口服制剂的形式，例如散剂、颗粒剂、片剂、胶囊剂、混悬剂、乳剂、糖浆剂和气雾剂，制备外用制剂，栓剂和按常规方法制备和使用无菌注射溶液。本领域已知的合适制剂可以是在马克出版公司出版的《雷明顿的医药科学》，宾夕法尼亚州伊斯顿，中公开的那些。
用于口服的固体制剂包括片剂、丸剂、散剂、颗粒剂、胶囊剂等，并且这些固体制剂通过与至少一种赋形剂如淀粉、碳酸钙、蔗糖、乳糖和明胶混合而制备。除简单的赋形剂外，固体制剂还使用润滑剂，例如硬脂酸镁和滑石粉。
用于口服的液体制剂包括混悬剂、口服液体、乳剂、糖浆等，并且这些液体制剂除了通常使用的简单的稀释剂如水和液体石蜡以外，还可以包含各种赋形剂，例如润湿剂、甜味剂、香料和防腐剂。
肠胃外给药的制剂包括无菌水溶液，非水溶剂、悬浮液、乳剂、冻干制剂和栓剂。作为栓剂的基质，可以使用维特索尔、聚乙二醇、吐温61、可可脂、月桂酸、甘油明胶等。
在本发明中，疫苗组合物的特征在于它还包含佐剂。通常，单独的蛋白质抗原不能强烈地诱导免疫应答，因此通过与佐剂混合可以提高疫苗组合物的效果。
如本文所用，术语“佐剂”是指在免疫细胞的初始活化过程中非特异性地促进对抗原的免疫应答的物质，包括试剂、分子等，类似的，每一种都不是对宿主的免疫原而是通过增加免疫系统中细胞的活性来增强免疫(Warren等，Annu.Rev.Immunol，4：369，1986)。可以增强免疫应答的本发明中使用的佐剂可以与疫苗组合物同时给药，或者可以一定时间间隔依次给药。
本发明的佐剂的特征在于，其选自但不限于下组：氢氧化钙磷酸钙、矿物油、角鲨烯、toll样受体(TLR)拮抗剂、去污剂、脂质体、皂角苷、细胞因子，及其组合。
在本发明的又一方面，提供了一种用于治疗或预防患者的疾病或障碍的方法，该方法包括向患者施用治疗有效量的疫苗组合物的步骤。
可以通过标准研究确定本发明疫苗组合物的最佳剂量，所述标准研究包括在受试者中观察合适的免疫应答。初次接种疫苗后，可以适当的间隔对受试者进行一次或多次加强免疫。
本发明的疫苗组合物的合适剂量取决于诸如制剂方法，给药方式，患者的年龄、体重、性别、病理状况、饮食、给药时间、给药途径、排泄率和反应灵敏度等因素，并且可以由本领域技术人员考虑上述因素来适当确定。
本发明的疫苗组合物可以通过医学领域中常用的途径来施用。肠胃外给药是优选的，并且可以例如通过静脉内、腹膜内、肌内、动脉内、口服、心内、髓内、硬膜内、透皮、肠、皮下、舌下或局部途径进行给药。通常，本发明的疫苗组合物的特征在于其包含治疗有效量的根据本发明的水痘带状疱疹病毒表面蛋白抗原变体作为活性成分。
发明方式
在下文中，将通过实施例更详细地描述本发明。这些实施例仅出于说明目的，并且对于本领域普通技术人员将显而易见的是，本发明的范围不解释为受这些实施例的限制。
实施例1：表面蛋白(gE)构建体的产生
为了产生表面蛋白(gE)构建体，通过PCR以获得所需的gE片段。然后，将每个gE片段用限制酶切割，并插入pcDNA3.1载体中。通过测序鉴定插入到pcDNA3.1载体中的gE片段的序列。通过midiprep获得了包含序列鉴定的gE片段的pcDNA3.1载体的DNA。表面蛋白(gE)片段的氨基酸序列示于下表1中。
[表1]
Figure BDA0002793525870000141
实施例2：瞬时转染
为了鉴定gE片段的表达水平，使用脂质体3000将其瞬时转染到293细胞中。将5×105个细胞添加到6孔板的每个孔中，并进行培养。然后，第二天，准备用于转染的样品。将0.2μgDNA和0.4μgP3000添加到试管中，然后用125μLoptiMEM稀释；然后将3.75μL的脂质体3000添加到另一个试管中，然后用125μL的optiMEM稀释。将稀释的DNA与等量的稀释的脂质体3000转移到试管中。然后，将试管在室温下混合孵育10分钟，以制备DNA-脂质体混合物。温育完成后，将DNA-脂质体混合物添加至含293细胞的6孔板中，然后在CO2温育器中进行培养2天。培养完成后，获得上清液，向其中加入4×含有β-巯基乙醇的样品缓冲液，然后在100℃下加热5分钟。加热后，将所得物保持冷冻直至进行蛋白质印迹。
实施例3：免疫印迹
进行蛋白质印迹以比较gE片段的表达水平。每个样品在NuPAGE 4-12％Bis-Tris凝胶上电泳，然后转移到PVDF膜上。将膜在5％脱脂奶中封闭1小时，与单克隆gE抗体(1μg/mL)孵育2小时，用TBST(吐温0.05％)洗涤，然后与稀释5000×的山羊抗小鼠IgG-HRP孵育1小时。温育的膜用TBST洗涤，然后用ECL底物显影。用Chemidoc机器进行检测。作为进行蛋白质印迹的结果，如图3所示，发现抗原、gE 534氨基酸、gE 537氨基酸和gE 540氨基酸表现出更高的表达水平。
实施例4：与葛兰素史克生物学公司的水痘带状疱疹病毒表面蛋白抗原就表达水平方面的比较
就表达水平而言，比较作为当前商业化的葛兰素史克生物学公司的水痘带状疱疹疫苗的Shingrix中包含的表面蛋白抗原(GSK gE 546氨基酸)和本发明人产生的抗原(Mogam gE 537氨基酸)。以与实施例3中所述相同的方式进行蛋白质印迹。葛兰素史克生物学公司SA的Shingrix中包含的表面蛋白抗原(GSK gE 546氨基酸)与本发明人生产的抗原(Mogam gE 537氨基酸)之间的差异如下表2所示。作为进行蛋白质印迹的结果，如图4所示，发现本发明人产生的抗原比葛兰素史克生物学公司的表面蛋白抗原具有更高的表达水平。
[表2]
Mogam gE 537氨基酸 GSK gE 546氨基酸
来源 进化枝1(野生型，杜马斯株) 进化枝3(野生型)
第40位氨基酸 T I
第536位氨基酸 L I
C端 w/o YAAWTGGLA YAAWTGGLA
实施例5：与葛兰素史克生物学公司的水痘带状疱疹病毒表面蛋白抗原就免疫原性的比较
在免疫原性方面，比较作为当前商业化的葛兰素史克生物学公司的水痘带状疱疹疫苗的Shingrix中包含的表面蛋白抗原(GSK gE 546氨基酸)和本发明人生产的抗原(Mogam gE 537氨基酸)，进行了动物实验。从人类具有水痘感染史的观点出发，为了模仿小鼠中的水痘感染，通过对皮下注射减毒活疫苗(LAV.3,000pfu)一次，对雌性C57BL/6小鼠进行初次免疫(LAV初免)。从LAV初次免疫(第0天)的28天后，通过肌肉内注射含或不含佐剂的Mogam gE或GSK gE抗原组合物对小鼠进行二次免疫。LAV初免后42天(第42天)收集血液样本，以测量对gE的体液免疫反应；LAV初免后42天(第42天)从脾脏样本中收集白细胞，以测量针对gE或VZV的细胞介导的免疫应答(CMI)。从LAV初免日(即第0天)开始计算免疫日，采血日和脾脏样本日。总体动物实验方法如下表3所述。
[表3]
Figure BDA0002793525870000161
Figure BDA0002793525870000171
实施例5-1：体液免疫反应的比较
进行初次和二次免疫后，进行酶联免疫吸附测定(ELISA)以测量gE抗原特异性IgG效能。将重组gE蛋白(1μg/mL)分配到ELISA板上，并在4℃下进行过夜温育以使蛋白抗原被包被在其上。将每个抗原包被的ELISA板洗涤3次，然后用含有2％牛血清白蛋白(BSA)的磷酸盐缓冲盐水(PBS)溶液封闭1小时。用BSA完成封闭反应后，洗涤ELISA板。然后，向其中加入稀释的血清样品，并进行孵育2小时。向其中添加辣根过氧化物酶(HRP)共轭的山羊抗小鼠IgG、IgG1或IgG2c抗体，并孵育1小时。在最终温育后，洗涤ELISA板，并通过添加作为HRP底物的3,3'，5,5'-四甲基联苯胺(TMB，由KPL制造)来诱导HRP反应。然后，添加TMB终止溶液以终止HRP反应，并使用ELISA酶标仪(Spectramax 250，酶标仪)在450nm下测量光密度(OD)，以检查产生的抗体量。结果，如图5所示，含有本发明人生产的抗原(Mogam gE 537氨基酸)和佐剂的G6的发光强度最高。从这些结果，发现在G6中诱导了最高的体液免疫应答。
实施例5-2：细胞介导的免疫反应的比较
进行初次和二次免疫后，进行IFN-γELISA以检查分泌的IFN-γ量，它是抗原刺激后T细胞分泌的代表性细胞因子。用VZV裂解物刺激从小鼠收集的白细胞3天。然后，进行离心以获得上清液，并且用小鼠IFN-γELISA试剂盒分析上清液。将IFN-γ捕获抗体(4μg/mL)分配到ELISA板上，并且在室温下进行过夜温育以允许将IFN-γ捕获抗体包被在其上。将每个抗体包被的ELISA板洗涤3次，然后用含有1％牛血清白蛋白(BSA)的PBS封闭1小时。用BSA完成封闭反应后，洗涤ELISA板。然后，向其中加入刺激白细胞后获得的上清液，并在室温下孵育2小时。培养结束后，洗涤ELISA板，在室温下与生物素化的小鼠IFN-γ检测抗体(400ng/mL)一起培养2小时。进行洗涤，然后与抗生蛋白链菌素-HRP一起温育另外20分钟。洗涤最终孵育的ELISA板，然后在室温下与底物溶液反应20分钟。向其中加入终止溶液以终止反应，然后使用ELISA酶标仪(Spectramax 250，酶标仪)在450nm处测量光密度(OD)，以检查产生的IFN-γ细胞因子的量。结果，如图6所示，含有本发明人生产的抗原(Mogam gE 537氨基酸)和佐剂的G6显示出最大量的IFN-γ细胞因子。从这些结果，发现在G6中诱导了最高的细胞介导的免疫应答。
实施例6：鉴定gE抗原片段的抗原特异性免疫原性
实施例6-1：瞬时转染用于抗原生产
为了表达gE片段，使用Expifectamine TM 293转染试剂盒将其瞬时转染到293细胞中。将细胞以2 106×细胞/mL的浓度置于125mL烧瓶中进行培养。然后，在第二天进行转染。将293细胞以2.9×106细胞/mL稀释至总计25.5mL，并制备用于转染的复合物。将μ30g DNA放入15mL试管中，并使用Opti-MEM调节至1.5mL。这样，制备了复合物1。将81μLExpiFectamine TM 293试剂放入另一个15mL试管中，并使用Opti-MEM调节至1.5mL。这样，制备了复合物2。在室温下孵育5分钟。5分钟后，将复合物1转移至含复合物2的管中并进行混合。然后，将试管在室温下孵育20分钟以制备DNA-脂质复合物。温育完成后，将DNA-脂质复合物全部置于含293细胞的125mL烧瓶中，并在温育器中进行培养。20小时后，用增强剂进行处理。将150μL ExpiFectamine TM 293转染增强剂1放入1.5mL试管中，并向其中加入ExpiFectamine TM 293转染增强剂2至1.5mL。然后，将所得物添加至293细胞，并在培养箱中进行5天的培养。5天后，获得培养上清液，通过0.45μm过滤器过滤，并冷冻保存直至纯化。
实施例6-2：纯化以获得抗原
将已经冷冻保存的培养液解冻，并向培养液中加入等量的PBS。使用0.22μm的过滤器进行过滤，然后进行阴离子交换色谱。向洗脱液中加入5M NaCl，然后进行疏水作用色谱。进行色谱分离的洗脱液通过0.22μm的过滤器过滤，冷冻保存直至进行动物实验。
实施例6-3：免疫
进行动物实验以鉴定gE抗原片段的免疫原性。以2周的间隔向雌性C57BL/6小鼠肌肉内注射gE抗原片段，并在二次免疫后2周从小鼠中采集血液样本。从收集的血样中分离出血清，然后冷冻保存直至测定抗体效价。
实施例6-4：抗原特异性IgG效能和响应者的测量
进行酶联免疫吸附测定(ELISA)以测量抗原特异性IgG效能。将VZV表面蛋白(1μg/mL)包被在ELISA板上，在4℃下进行过夜温育，并将每个ELISA板洗涤3次。然后，用含有2％牛血清白蛋白(BSA)的磷酸盐缓冲盐水(PBS)溶液封闭ELISA板1小时。洗涤ELISA板。然后，向其中加入稀释的血清样品，并进行孵育2小时。向其中添加辣根过氧化物酶(HRP)偶联的山羊抗小鼠IgG抗体，孵育1小时。最后的温育后，洗涤ELISA板，并通过加入作为底物的3,3'，5,5'-四甲基联苯胺(TMB)来诱导HRP反应。然后，向其中加入ELISA终止溶液以终止HRP反应，并使用分光计在450nm下测量光密度(OD)。
为了鉴定gE抗原片段之间的序列差异是否引起抗原特异性免疫原性的差异，通过实施例6-3中所述的方法进行动物免疫。使用根据上述抗体效价测量方法在动物实验中获得的血清来测量抗原特异性抗体效价。结果如图7所示，通过gE 534氨基酸或gE 543氨基酸免疫后的OD值高于用gE 500氨基酸，gE 510氨基酸，gE 525氨基酸或gE 546氨基酸免疫后的OD值。
测定抗体滴度后，将OD值为0.6或更高的个体作为响应者，对结果进行总结。结果，没有针对gE 500氨基酸的抗原特异性响应，并且用gE 510氨基酸免疫后的响应者数目与用gE 546氨基酸免疫后的响应者数目相同。用gE 525氨基酸、gE 534氨基酸、gE 537氨基酸或gE 540氨基酸免疫后的响应者数量高于用gE 510氨基酸或gE 546氨基酸免疫后的响应者数量。即，已确定gE 525氨基酸至gE 540氨基酸表现出更高数量的响应者(图8)。
结果，在抗体效价测定中，gE 534氨基酸至gE 543氨基酸显示出较高的值，而在抗原特异性响应者中，gE 525氨基酸至gE 540氨基酸显示出较高的值。因此，在将以上两个结果放在一起的情况下，可以确定gE 534氨基酸至540氨基酸具有更高的免疫原性。
工业适用性
本发明的水痘带状疱疹病毒表面蛋白(gE)抗原变体是蛋白抗原。在用作疫苗组合物的情况下，与活病毒疫苗相比，抗原变体在宿主细胞中表现出优异的安全性和高表达水平。因此，这种抗原变体可用作预防或治疗水痘带状疱疹病毒诱导的水痘或带状疱疹的疫苗。
如上所述，已经详细描述了本发明的特定部分。然而，对于本领域技术人员显而易见的是，这样的具体描述仅用于示出优选实施例，并且本发明的范围不限于此。因此，本发明的实质范围将由所附权利要求及其等同物来限定。
序列表自由文本
附电子文件。
<110> 财团法人牧岩生命科学研究所
<120> 水痘带状疱疹病毒的抗原变体及其用途
<130> PP-B2204
<150> KR 2018-0058219
<151> 2018-05-23
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Arg Leu Met Gln Pro Thr Gln Met Ser Ala Gln Glu Asp Leu Gly Asp
115 120 125
Asp Thr Gly Ile His Val Ile Pro Thr Leu Asn Gly Asp Asp Arg His
130 135 140
Lys Ile Val Asn Val Asp Gln Arg Gln Tyr Gly Asp Val Phe Lys Gly
145 150 155 160
Asp Leu Asn Pro Lys Pro Gln Gly Gln Arg Leu Ile Glu Val Ser Val
165 170 175
Glu Glu Asn His Pro Phe Thr Leu Arg Ala Pro Ile Gln Arg Ile Tyr
180 185 190
Gly Val Arg Tyr Thr Glu Thr Trp Ser Phe Leu Pro Ser Leu Thr Cys
195 200 205
Thr Gly Asp Ala Ala Pro Ala Ile Gln His Ile Cys Leu Lys His Thr
210 215 220
Thr Cys Phe Gln Asp Val Val Val Asp Val Asp Cys Ala Glu Asn Thr
225 230 235 240
Lys Glu Asp Gln Leu Ala Glu Ile Ser Tyr Arg Phe Gln Gly Lys Lys
245 250 255
Glu Ala Asp Gln Pro Trp Ile Val Val Asn Thr Ser Thr Leu Phe Asp
260 265 270
Glu Leu Glu Leu Asp Pro Pro Glu Ile Glu Pro Gly Val Leu Lys Val
275 280 285
Leu Arg Thr Glu Lys Gln Tyr Leu Gly Val Tyr Ile Trp Asn Met Arg
290 295 300
Gly Ser Asp Gly Thr Ser Thr Tyr Ala Thr Phe Leu Val Thr Trp Lys
305 310 315 320
Gly Asp Glu Lys Thr Arg Asn Pro Thr Pro Ala Val Thr Pro Gln Pro
325 330 335
Arg Gly Ala Glu Phe His Met Trp Asn Tyr His Ser His Val Phe Ser
340 345 350
Val Gly Asp Thr Phe Ser Leu Ala Met His Leu Gln Tyr Lys Ile His
355 360 365
Glu Ala Pro Phe Asp Leu Leu Leu Glu Trp Leu Tyr Val Pro Ile Asp
370 375 380
Pro Thr Cys Gln Pro Met Arg Leu Tyr Ser Thr Cys Leu Tyr His Pro
385 390 395 400
Asn Ala Pro Gln Cys Leu Ser His Met Asn Ser Gly Cys Thr Phe Thr
405 410 415
Ser Pro His Leu Ala Gln Arg Val Ala Ser Thr Val Tyr Gln Asn Cys
420 425 430
Glu His Ala Asp Asn Tyr Thr Ala Tyr Cys Leu Gly Ile Ser His Met
435 440 445
Glu Pro Ser Phe Gly Leu Ile Leu His Asp Gly Gly Thr Thr Leu Lys
450 455 460
Phe Val Asp Thr Pro Glu Ser Leu Ser Gly Leu Tyr Val Phe Val Val
465 470 475 480
Tyr Phe Asn Gly His Val Glu Ala Val Ala Tyr Thr Val Val Ser Thr
485 490 495
Val Asp His Phe Val Asn Ala Ile Glu Glu Arg Gly Phe Pro Pro Thr
500 505 510
Ala Gly Gln Pro Pro Ala Thr Thr Lys Pro Lys Glu Ile Thr Pro Val
515 520 525
Asn Pro Gly Thr Ser Pro
530
<210> 3
<211> 535
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> 人工序列（Artificial Sequence）
<400> 3
Met Gly Thr Val Asn Lys Pro Val Val Gly Val Leu Met Gly Phe Gly
1 5 10 15
Ile Ile Thr Gly Thr Leu Arg Ile Thr Asn Pro Val Arg Ala Ser Val
20 25 30
Leu Arg Tyr Asp Asp Phe His Thr Asp Glu Asp Lys Leu Asp Thr Asn
35 40 45
Ser Val Tyr Glu Pro Tyr Tyr His Ser Asp His Ala Glu Ser Ser Trp
50 55 60
Val Asn Arg Gly Glu Ser Ser Arg Lys Ala Tyr Asp His Asn Ser Pro
65 70 75 80
Tyr Ile Trp Pro Arg Asn Asp Tyr Asp Gly Phe Leu Glu Asn Ala His
85 90 95
Glu His His Gly Val Tyr Asn Gln Gly Arg Gly Ile Asp Ser Gly Glu
100 105 110
Arg Leu Met Gln Pro Thr Gln Met Ser Ala Gln Glu Asp Leu Gly Asp
115 120 125
Asp Thr Gly Ile His Val Ile Pro Thr Leu Asn Gly Asp Asp Arg His
130 135 140
Lys Ile Val Asn Val Asp Gln Arg Gln Tyr Gly Asp Val Phe Lys Gly
145 150 155 160
Asp Leu Asn Pro Lys Pro Gln Gly Gln Arg Leu Ile Glu Val Ser Val
165 170 175
Glu Glu Asn His Pro Phe Thr Leu Arg Ala Pro Ile Gln Arg Ile Tyr
180 185 190
Gly Val Arg Tyr Thr Glu Thr Trp Ser Phe Leu Pro Ser Leu Thr Cys
195 200 205
Thr Gly Asp Ala Ala Pro Ala Ile Gln His Ile Cys Leu Lys His Thr
210 215 220
Thr Cys Phe Gln Asp Val Val Val Asp Val Asp Cys Ala Glu Asn Thr
225 230 235 240
Lys Glu Asp Gln Leu Ala Glu Ile Ser Tyr Arg Phe Gln Gly Lys Lys
245 250 255
Glu Ala Asp Gln Pro Trp Ile Val Val Asn Thr Ser Thr Leu Phe Asp
260 265 270
Glu Leu Glu Leu Asp Pro Pro Glu Ile Glu Pro Gly Val Leu Lys Val
275 280 285
Leu Arg Thr Glu Lys Gln Tyr Leu Gly Val Tyr Ile Trp Asn Met Arg
290 295 300
Gly Ser Asp Gly Thr Ser Thr Tyr Ala Thr Phe Leu Val Thr Trp Lys
305 310 315 320
Gly Asp Glu Lys Thr Arg Asn Pro Thr Pro Ala Val Thr Pro Gln Pro
325 330 335
Arg Gly Ala Glu Phe His Met Trp Asn Tyr His Ser His Val Phe Ser
340 345 350
Val Gly Asp Thr Phe Ser Leu Ala Met His Leu Gln Tyr Lys Ile His
355 360 365
Glu Ala Pro Phe Asp Leu Leu Leu Glu Trp Leu Tyr Val Pro Ile Asp
370 375 380
Pro Thr Cys Gln Pro Met Arg Leu Tyr Ser Thr Cys Leu Tyr His Pro
385 390 395 400
Asn Ala Pro Gln Cys Leu Ser His Met Asn Ser Gly Cys Thr Phe Thr
405 410 415
Ser Pro His Leu Ala Gln Arg Val Ala Ser Thr Val Tyr Gln Asn Cys
420 425 430
Glu His Ala Asp Asn Tyr Thr Ala Tyr Cys Leu Gly Ile Ser His Met
435 440 445
Glu Pro Ser Phe Gly Leu Ile Leu His Asp Gly Gly Thr Thr Leu Lys
450 455 460
Phe Val Asp Thr Pro Glu Ser Leu Ser Gly Leu Tyr Val Phe Val Val
465 470 475 480
Tyr Phe Asn Gly His Val Glu Ala Val Ala Tyr Thr Val Val Ser Thr
485 490 495
Val Asp His Phe Val Asn Ala Ile Glu Glu Arg Gly Phe Pro Pro Thr
500 505 510
Ala Gly Gln Pro Pro Ala Thr Thr Lys Pro Lys Glu Ile Thr Pro Val
515 520 525
Asn Pro Gly Thr Ser Pro Leu
530 535
<210> 4
<211> 536
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> 人工序列（Artificial Sequence）
<400> 4
Met Gly Thr Val Asn Lys Pro Val Val Gly Val Leu Met Gly Phe Gly
1 5 10 15
Ile Ile Thr Gly Thr Leu Arg Ile Thr Asn Pro Val Arg Ala Ser Val
20 25 30
Leu Arg Tyr Asp Asp Phe His Thr Asp Glu Asp Lys Leu Asp Thr Asn
35 40 45
Ser Val Tyr Glu Pro Tyr Tyr His Ser Asp His Ala Glu Ser Ser Trp
50 55 60
Val Asn Arg Gly Glu Ser Ser Arg Lys Ala Tyr Asp His Asn Ser Pro
65 70 75 80
Tyr Ile Trp Pro Arg Asn Asp Tyr Asp Gly Phe Leu Glu Asn Ala His
85 90 95
Glu His His Gly Val Tyr Asn Gln Gly Arg Gly Ile Asp Ser Gly Glu
100 105 110
Arg Leu Met Gln Pro Thr Gln Met Ser Ala Gln Glu Asp Leu Gly Asp
115 120 125
Asp Thr Gly Ile His Val Ile Pro Thr Leu Asn Gly Asp Asp Arg His
130 135 140
Lys Ile Val Asn Val Asp Gln Arg Gln Tyr Gly Asp Val Phe Lys Gly
145 150 155 160
Asp Leu Asn Pro Lys Pro Gln Gly Gln Arg Leu Ile Glu Val Ser Val
165 170 175
Glu Glu Asn His Pro Phe Thr Leu Arg Ala Pro Ile Gln Arg Ile Tyr
180 185 190
Gly Val Arg Tyr Thr Glu Thr Trp Ser Phe Leu Pro Ser Leu Thr Cys
195 200 205
Thr Gly Asp Ala Ala Pro Ala Ile Gln His Ile Cys Leu Lys His Thr
210 215 220
Thr Cys Phe Gln Asp Val Val Val Asp Val Asp Cys Ala Glu Asn Thr
225 230 235 240
Lys Glu Asp Gln Leu Ala Glu Ile Ser Tyr Arg Phe Gln Gly Lys Lys
245 250 255
Glu Ala Asp Gln Pro Trp Ile Val Val Asn Thr Ser Thr Leu Phe Asp
260 265 270
Glu Leu Glu Leu Asp Pro Pro Glu Ile Glu Pro Gly Val Leu Lys Val
275 280 285
Leu Arg Thr Glu Lys Gln Tyr Leu Gly Val Tyr Ile Trp Asn Met Arg
290 295 300
Gly Ser Asp Gly Thr Ser Thr Tyr Ala Thr Phe Leu Val Thr Trp Lys
305 310 315 320
Gly Asp Glu Lys Thr Arg Asn Pro Thr Pro Ala Val Thr Pro Gln Pro
325 330 335
Arg Gly Ala Glu Phe His Met Trp Asn Tyr His Ser His Val Phe Ser
340 345 350
Val Gly Asp Thr Phe Ser Leu Ala Met His Leu Gln Tyr Lys Ile His
355 360 365
Glu Ala Pro Phe Asp Leu Leu Leu Glu Trp Leu Tyr Val Pro Ile Asp
370 375 380
Pro Thr Cys Gln Pro Met Arg Leu Tyr Ser Thr Cys Leu Tyr His Pro
385 390 395 400
Asn Ala Pro Gln Cys Leu Ser His Met Asn Ser Gly Cys Thr Phe Thr
405 410 415
Ser Pro His Leu Ala Gln Arg Val Ala Ser Thr Val Tyr Gln Asn Cys
420 425 430
Glu His Ala Asp Asn Tyr Thr Ala Tyr Cys Leu Gly Ile Ser His Met
435 440 445
Glu Pro Ser Phe Gly Leu Ile Leu His Asp Gly Gly Thr Thr Leu Lys
450 455 460
Phe Val Asp Thr Pro Glu Ser Leu Ser Gly Leu Tyr Val Phe Val Val
465 470 475 480
Tyr Phe Asn Gly His Val Glu Ala Val Ala Tyr Thr Val Val Ser Thr
485 490 495
Val Asp His Phe Val Asn Ala Ile Glu Glu Arg Gly Phe Pro Pro Thr
500 505 510
Ala Gly Gln Pro Pro Ala Thr Thr Lys Pro Lys Glu Ile Thr Pro Val
515 520 525
Asn Pro Gly Thr Ser Pro Leu Leu
530 535
<210> 5
<211> 537
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> 人工序列（Artificial Sequence）
<400> 5
Met Gly Thr Val Asn Lys Pro Val Val Gly Val Leu Met Gly Phe Gly
1 5 10 15
Ile Ile Thr Gly Thr Leu Arg Ile Thr Asn Pro Val Arg Ala Ser Val
20 25 30
Leu Arg Tyr Asp Asp Phe His Thr Asp Glu Asp Lys Leu Asp Thr Asn
35 40 45
Ser Val Tyr Glu Pro Tyr Tyr His Ser Asp His Ala Glu Ser Ser Trp
50 55 60
Val Asn Arg Gly Glu Ser Ser Arg Lys Ala Tyr Asp His Asn Ser Pro
65 70 75 80
Tyr Ile Trp Pro Arg Asn Asp Tyr Asp Gly Phe Leu Glu Asn Ala His
85 90 95
Glu His His Gly Val Tyr Asn Gln Gly Arg Gly Ile Asp Ser Gly Glu
100 105 110
Arg Leu Met Gln Pro Thr Gln Met Ser Ala Gln Glu Asp Leu Gly Asp
115 120 125
Asp Thr Gly Ile His Val Ile Pro Thr Leu Asn Gly Asp Asp Arg His
130 135 140
Lys Ile Val Asn Val Asp Gln Arg Gln Tyr Gly Asp Val Phe Lys Gly
145 150 155 160
Asp Leu Asn Pro Lys Pro Gln Gly Gln Arg Leu Ile Glu Val Ser Val
165 170 175
Glu Glu Asn His Pro Phe Thr Leu Arg Ala Pro Ile Gln Arg Ile Tyr
180 185 190
Gly Val Arg Tyr Thr Glu Thr Trp Ser Phe Leu Pro Ser Leu Thr Cys
195 200 205
Thr Gly Asp Ala Ala Pro Ala Ile Gln His Ile Cys Leu Lys His Thr
210 215 220
Thr Cys Phe Gln Asp Val Val Val Asp Val Asp Cys Ala Glu Asn Thr
225 230 235 240
Lys Glu Asp Gln Leu Ala Glu Ile Ser Tyr Arg Phe Gln Gly Lys Lys
245 250 255
Glu Ala Asp Gln Pro Trp Ile Val Val Asn Thr Ser Thr Leu Phe Asp
260 265 270
Glu Leu Glu Leu Asp Pro Pro Glu Ile Glu Pro Gly Val Leu Lys Val
275 280 285
Leu Arg Thr Glu Lys Gln Tyr Leu Gly Val Tyr Ile Trp Asn Met Arg
290 295 300
Gly Ser Asp Gly Thr Ser Thr Tyr Ala Thr Phe Leu Val Thr Trp Lys
305 310 315 320
Gly Asp Glu Lys Thr Arg Asn Pro Thr Pro Ala Val Thr Pro Gln Pro
325 330 335
Arg Gly Ala Glu Phe His Met Trp Asn Tyr His Ser His Val Phe Ser
340 345 350
Val Gly Asp Thr Phe Ser Leu Ala Met His Leu Gln Tyr Lys Ile His
355 360 365
Glu Ala Pro Phe Asp Leu Leu Leu Glu Trp Leu Tyr Val Pro Ile Asp
370 375 380
Pro Thr Cys Gln Pro Met Arg Leu Tyr Ser Thr Cys Leu Tyr His Pro
385 390 395 400
Asn Ala Pro Gln Cys Leu Ser His Met Asn Ser Gly Cys Thr Phe Thr
405 410 415
Ser Pro His Leu Ala Gln Arg Val Ala Ser Thr Val Tyr Gln Asn Cys
420 425 430
Glu His Ala Asp Asn Tyr Thr Ala Tyr Cys Leu Gly Ile Ser His Met
435 440 445
Glu Pro Ser Phe Gly Leu Ile Leu His Asp Gly Gly Thr Thr Leu Lys
450 455 460
Phe Val Asp Thr Pro Glu Ser Leu Ser Gly Leu Tyr Val Phe Val Val
465 470 475 480
Tyr Phe Asn Gly His Val Glu Ala Val Ala Tyr Thr Val Val Ser Thr
485 490 495
Val Asp His Phe Val Asn Ala Ile Glu Glu Arg Gly Phe Pro Pro Thr
500 505 510
Ala Gly Gln Pro Pro Ala Thr Thr Lys Pro Lys Glu Ile Thr Pro Val
515 520 525
Asn Pro Gly Thr Ser Pro Leu Leu Arg
530 535
<210> 6
<211> 538
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> 人工序列（Artificial Sequence）
<400> 6
Met Gly Thr Val Asn Lys Pro Val Val Gly Val Leu Met Gly Phe Gly
1 5 10 15
Ile Ile Thr Gly Thr Leu Arg Ile Thr Asn Pro Val Arg Ala Ser Val
20 25 30
Leu Arg Tyr Asp Asp Phe His Thr Asp Glu Asp Lys Leu Asp Thr Asn
35 40 45
Ser Val Tyr Glu Pro Tyr Tyr His Ser Asp His Ala Glu Ser Ser Trp
50 55 60
Val Asn Arg Gly Glu Ser Ser Arg Lys Ala Tyr Asp His Asn Ser Pro
65 70 75 80
Tyr Ile Trp Pro Arg Asn Asp Tyr Asp Gly Phe Leu Glu Asn Ala His
85 90 95
Glu His His Gly Val Tyr Asn Gln Gly Arg Gly Ile Asp Ser Gly Glu
100 105 110
Arg Leu Met Gln Pro Thr Gln Met Ser Ala Gln Glu Asp Leu Gly Asp
115 120 125
Asp Thr Gly Ile His Val Ile Pro Thr Leu Asn Gly Asp Asp Arg His
130 135 140
Lys Ile Val Asn Val Asp Gln Arg Gln Tyr Gly Asp Val Phe Lys Gly
145 150 155 160
Asp Leu Asn Pro Lys Pro Gln Gly Gln Arg Leu Ile Glu Val Ser Val
165 170 175
Glu Glu Asn His Pro Phe Thr Leu Arg Ala Pro Ile Gln Arg Ile Tyr
180 185 190
Gly Val Arg Tyr Thr Glu Thr Trp Ser Phe Leu Pro Ser Leu Thr Cys
195 200 205
Thr Gly Asp Ala Ala Pro Ala Ile Gln His Ile Cys Leu Lys His Thr
210 215 220
Thr Cys Phe Gln Asp Val Val Val Asp Val Asp Cys Ala Glu Asn Thr
225 230 235 240
Lys Glu Asp Gln Leu Ala Glu Ile Ser Tyr Arg Phe Gln Gly Lys Lys
245 250 255
Glu Ala Asp Gln Pro Trp Ile Val Val Asn Thr Ser Thr Leu Phe Asp
260 265 270
Glu Leu Glu Leu Asp Pro Pro Glu Ile Glu Pro Gly Val Leu Lys Val
275 280 285
Leu Arg Thr Glu Lys Gln Tyr Leu Gly Val Tyr Ile Trp Asn Met Arg
290 295 300
Gly Ser Asp Gly Thr Ser Thr Tyr Ala Thr Phe Leu Val Thr Trp Lys
305 310 315 320
Gly Asp Glu Lys Thr Arg Asn Pro Thr Pro Ala Val Thr Pro Gln Pro
325 330 335
Arg Gly Ala Glu Phe His Met Trp Asn Tyr His Ser His Val Phe Ser
340 345 350
Val Gly Asp Thr Phe Ser Leu Ala Met His Leu Gln Tyr Lys Ile His
355 360 365
Glu Ala Pro Phe Asp Leu Leu Leu Glu Trp Leu Tyr Val Pro Ile Asp
370 375 380
Pro Thr Cys Gln Pro Met Arg Leu Tyr Ser Thr Cys Leu Tyr His Pro
385 390 395 400
Asn Ala Pro Gln Cys Leu Ser His Met Asn Ser Gly Cys Thr Phe Thr
405 410 415
Ser Pro His Leu Ala Gln Arg Val Ala Ser Thr Val Tyr Gln Asn Cys
420 425 430
Glu His Ala Asp Asn Tyr Thr Ala Tyr Cys Leu Gly Ile Ser His Met
435 440 445
Glu Pro Ser Phe Gly Leu Ile Leu His Asp Gly Gly Thr Thr Leu Lys
450 455 460
Phe Val Asp Thr Pro Glu Ser Leu Ser Gly Leu Tyr Val Phe Val Val
465 470 475 480
Tyr Phe Asn Gly His Val Glu Ala Val Ala Tyr Thr Val Val Ser Thr
485 490 495
Val Asp His Phe Val Asn Ala Ile Glu Glu Arg Gly Phe Pro Pro Thr
500 505 510
Ala Gly Gln Pro Pro Ala Thr Thr Lys Pro Lys Glu Ile Thr Pro Val
515 520 525
Asn Pro Gly Thr Ser Pro Leu Leu Arg Tyr
530 535
<210> 7
<211> 539
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> 人工序列（Artificial Sequence）
<400> 7
Met Gly Thr Val Asn Lys Pro Val Val Gly Val Leu Met Gly Phe Gly
1 5 10 15
Ile Ile Thr Gly Thr Leu Arg Ile Thr Asn Pro Val Arg Ala Ser Val
20 25 30
Leu Arg Tyr Asp Asp Phe His Thr Asp Glu Asp Lys Leu Asp Thr Asn
35 40 45
Ser Val Tyr Glu Pro Tyr Tyr His Ser Asp His Ala Glu Ser Ser Trp
50 55 60
Val Asn Arg Gly Glu Ser Ser Arg Lys Ala Tyr Asp His Asn Ser Pro
65 70 75 80
Tyr Ile Trp Pro Arg Asn Asp Tyr Asp Gly Phe Leu Glu Asn Ala His
85 90 95
Glu His His Gly Val Tyr Asn Gln Gly Arg Gly Ile Asp Ser Gly Glu
100 105 110
Arg Leu Met Gln Pro Thr Gln Met Ser Ala Gln Glu Asp Leu Gly Asp
115 120 125
Asp Thr Gly Ile His Val Ile Pro Thr Leu Asn Gly Asp Asp Arg His
130 135 140
Lys Ile Val Asn Val Asp Gln Arg Gln Tyr Gly Asp Val Phe Lys Gly
145 150 155 160
Asp Leu Asn Pro Lys Pro Gln Gly Gln Arg Leu Ile Glu Val Ser Val
165 170 175
Glu Glu Asn His Pro Phe Thr Leu Arg Ala Pro Ile Gln Arg Ile Tyr
180 185 190
Gly Val Arg Tyr Thr Glu Thr Trp Ser Phe Leu Pro Ser Leu Thr Cys
195 200 205
Thr Gly Asp Ala Ala Pro Ala Ile Gln His Ile Cys Leu Lys His Thr
210 215 220
Thr Cys Phe Gln Asp Val Val Val Asp Val Asp Cys Ala Glu Asn Thr
225 230 235 240
Lys Glu Asp Gln Leu Ala Glu Ile Ser Tyr Arg Phe Gln Gly Lys Lys
245 250 255
Glu Ala Asp Gln Pro Trp Ile Val Val Asn Thr Ser Thr Leu Phe Asp
260 265 270
Glu Leu Glu Leu Asp Pro Pro Glu Ile Glu Pro Gly Val Leu Lys Val
275 280 285
Leu Arg Thr Glu Lys Gln Tyr Leu Gly Val Tyr Ile Trp Asn Met Arg
290 295 300
Gly Ser Asp Gly Thr Ser Thr Tyr Ala Thr Phe Leu Val Thr Trp Lys
305 310 315 320
Gly Asp Glu Lys Thr Arg Asn Pro Thr Pro Ala Val Thr Pro Gln Pro
325 330 335
Arg Gly Ala Glu Phe His Met Trp Asn Tyr His Ser His Val Phe Ser
340 345 350
Val Gly Asp Thr Phe Ser Leu Ala Met His Leu Gln Tyr Lys Ile His
355 360 365
Glu Ala Pro Phe Asp Leu Leu Leu Glu Trp Leu Tyr Val Pro Ile Asp
370 375 380
Pro Thr Cys Gln Pro Met Arg Leu Tyr Ser Thr Cys Leu Tyr His Pro
385 390 395 400
Asn Ala Pro Gln Cys Leu Ser His Met Asn Ser Gly Cys Thr Phe Thr
405 410 415
Ser Pro His Leu Ala Gln Arg Val Ala Ser Thr Val Tyr Gln Asn Cys
420 425 430
Glu His Ala Asp Asn Tyr Thr Ala Tyr Cys Leu Gly Ile Ser His Met
435 440 445
Glu Pro Ser Phe Gly Leu Ile Leu His Asp Gly Gly Thr Thr Leu Lys
450 455 460
Phe Val Asp Thr Pro Glu Ser Leu Ser Gly Leu Tyr Val Phe Val Val
465 470 475 480
Tyr Phe Asn Gly His Val Glu Ala Val Ala Tyr Thr Val Val Ser Thr
485 490 495
Val Asp His Phe Val Asn Ala Ile Glu Glu Arg Gly Phe Pro Pro Thr
500 505 510
Ala Gly Gln Pro Pro Ala Thr Thr Lys Pro Lys Glu Ile Thr Pro Val
515 520 525
Asn Pro Gly Thr Ser Pro Leu Leu Arg Tyr Ala
530 535
<210> 8
<211> 540
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> 人工序列（Artificial Sequence）
<400> 8
Met Gly Thr Val Asn Lys Pro Val Val Gly Val Leu Met Gly Phe Gly
1 5 10 15
Ile Ile Thr Gly Thr Leu Arg Ile Thr Asn Pro Val Arg Ala Ser Val
20 25 30
Leu Arg Tyr Asp Asp Phe His Thr Asp Glu Asp Lys Leu Asp Thr Asn
35 40 45
Ser Val Tyr Glu Pro Tyr Tyr His Ser Asp His Ala Glu Ser Ser Trp
50 55 60
Val Asn Arg Gly Glu Ser Ser Arg Lys Ala Tyr Asp His Asn Ser Pro
65 70 75 80
Tyr Ile Trp Pro Arg Asn Asp Tyr Asp Gly Phe Leu Glu Asn Ala His
85 90 95
Glu His His Gly Val Tyr Asn Gln Gly Arg Gly Ile Asp Ser Gly Glu
100 105 110
Arg Leu Met Gln Pro Thr Gln Met Ser Ala Gln Glu Asp Leu Gly Asp
115 120 125
Asp Thr Gly Ile His Val Ile Pro Thr Leu Asn Gly Asp Asp Arg His
130 135 140
Lys Ile Val Asn Val Asp Gln Arg Gln Tyr Gly Asp Val Phe Lys Gly
145 150 155 160
Asp Leu Asn Pro Lys Pro Gln Gly Gln Arg Leu Ile Glu Val Ser Val
165 170 175
Glu Glu Asn His Pro Phe Thr Leu Arg Ala Pro Ile Gln Arg Ile Tyr
180 185 190
Gly Val Arg Tyr Thr Glu Thr Trp Ser Phe Leu Pro Ser Leu Thr Cys
195 200 205
Thr Gly Asp Ala Ala Pro Ala Ile Gln His Ile Cys Leu Lys His Thr
210 215 220
Thr Cys Phe Gln Asp Val Val Val Asp Val Asp Cys Ala Glu Asn Thr
225 230 235 240
Lys Glu Asp Gln Leu Ala Glu Ile Ser Tyr Arg Phe Gln Gly Lys Lys
245 250 255
Glu Ala Asp Gln Pro Trp Ile Val Val Asn Thr Ser Thr Leu Phe Asp
260 265 270
Glu Leu Glu Leu Asp Pro Pro Glu Ile Glu Pro Gly Val Leu Lys Val
275 280 285
Leu Arg Thr Glu Lys Gln Tyr Leu Gly Val Tyr Ile Trp Asn Met Arg
290 295 300
Gly Ser Asp Gly Thr Ser Thr Tyr Ala Thr Phe Leu Val Thr Trp Lys
305 310 315 320
Gly Asp Glu Lys Thr Arg Asn Pro Thr Pro Ala Val Thr Pro Gln Pro
325 330 335
Arg Gly Ala Glu Phe His Met Trp Asn Tyr His Ser His Val Phe Ser
340 345 350
Val Gly Asp Thr Phe Ser Leu Ala Met His Leu Gln Tyr Lys Ile His
355 360 365
Glu Ala Pro Phe Asp Leu Leu Leu Glu Trp Leu Tyr Val Pro Ile Asp
370 375 380
Pro Thr Cys Gln Pro Met Arg Leu Tyr Ser Thr Cys Leu Tyr His Pro
385 390 395 400
Asn Ala Pro Gln Cys Leu Ser His Met Asn Ser Gly Cys Thr Phe Thr
405 410 415
Ser Pro His Leu Ala Gln Arg Val Ala Ser Thr Val Tyr Gln Asn Cys
420 425 430
Glu His Ala Asp Asn Tyr Thr Ala Tyr Cys Leu Gly Ile Ser His Met
435 440 445
Glu Pro Ser Phe Gly Leu Ile Leu His Asp Gly Gly Thr Thr Leu Lys
450 455 460
Phe Val Asp Thr Pro Glu Ser Leu Ser Gly Leu Tyr Val Phe Val Val
465 470 475 480
Tyr Phe Asn Gly His Val Glu Ala Val Ala Tyr Thr Val Val Ser Thr
485 490 495
Val Asp His Phe Val Asn Ala Ile Glu Glu Arg Gly Phe Pro Pro Thr
500 505 510
Ala Gly Gln Pro Pro Ala Thr Thr Lys Pro Lys Glu Ile Thr Pro Val
515 520 525
Asn Pro Gly Thr Ser Pro Leu Leu Arg Tyr Ala Ala
530 535 540
<210> 9
<211> 1602
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> 人工序列（Artificial Sequence）
<400> 9
atgggaacag tcaacaaacc agtcgtcggc gtgctgatgg gcttcggtat tattacagga 60
actctgagga ttactaaccc cgtgcgcgcc tctgtgctgc ggtacgacga tttccacaca 120
gacgaggata agctggacac caattccgtg tatgagccct actatcactc tgatcacgcc 180
gagagctcct gggtgaaccg gggcgagtct agcaggaagg cttacgacca caacagccct 240
tatatctggc cacggaatga ctacgatggc tttctggaga acgcccacga gcaccacggc 300
gtgtataatc agggcagagg catcgactct ggcgagcggc tgatgcagcc cacccagatg 360
agcgcccagg aggatctggg cgacgataca ggcatccacg tgatccctac cctgaatggc 420
gacgataggc acaagatcgt gaacgtggat cagagacagt acggcgacgt gttcaagggc 480
gatctgaatc ccaagcctca gggccagagg ctgatcgagg tgtccgtgga ggagaaccac 540
cccttcaccc tgagagcccc tatccagcgg atctacggcg tgaggtatac cgagacatgg 600
agctttctgc catccctgac atgcaccggc gacgctgctc ctgctatcca gcacatctgc 660
ctgaagcaca ccacatgttt tcaggacgtg gtggtggacg tggattgtgc cgagaataca 720
aaggaggatc agctggctga gatctcctac cggttccagg gcaagaagga ggccgatcag 780
ccttggatcg tggtgaacac ctctacactg tttgacgagc tggagctgga tccccctgag 840
atcgagccag gcgtgctgaa ggtgctgaga accgagaagc agtacctggg cgtgtatatc 900
tggaacatgc ggggctctga cggcaccagc acatacgcta ccttcctggt cacatggaag 960
ggcgatgaga agacccggaa tccaacacct gctgtgaccc ctcagccaag gggagctgag 1020
tttcacatgt ggaactatca ctcccacgtg ttctctgtgg gcgacacctt tagcctggcc 1080
atgcacctgc aatataagat ccacgaggct cctttcgacc tgctgctgga gtggctgtat 1140
gtgcccatcg atcctacatg ccagccaatg aggctgtact ccacctgtct gtatcaccca 1200
aatgcccccc aatgcctgag ccacatgaac tccggctgta cctttacaag cccccacctg 1260
gcccagagag tggcttccac agtgtaccag aactgcgagc acgccgacaa ttacaccgct 1320
tattgtctgg gcatctctca catggagccc agcttcggcc tgatcctgca cgacggcggc 1380
accacactga agtttgtgga tacacccgag tccctgtctg gcctctacgt gttcgtggtg 1440
tacttcaacg gccacgtgga ggccgtggct tatacagtgg tgtctaccgt ggatcacttc 1500
gtgaacgcca tcgaggagag aggatttcca cctaccgctg gacagcctcc agctaccaca 1560
aagcctaagg aaatcacccc tgtcaatcct ggaacttcac ct 1602
<210> 10
<211> 1605
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> 人工序列（Artificial Sequence）
<400> 10
atgggaacag tcaacaaacc agtcgtcggc gtgctgatgg gcttcggtat tattacagga 60
actctgagga ttactaaccc cgtgcgcgcc tctgtgctgc ggtacgacga tttccacaca 120
gacgaggata agctggacac caattccgtg tatgagccct actatcactc tgatcacgcc 180
gagagctcct gggtgaaccg gggcgagtct agcaggaagg cttacgacca caacagccct 240
tatatctggc cacggaatga ctacgatggc tttctggaga acgcccacga gcaccacggc 300
gtgtataatc agggcagagg catcgactct ggcgagcggc tgatgcagcc cacccagatg 360
agcgcccagg aggatctggg cgacgataca ggcatccacg tgatccctac cctgaatggc 420
gacgataggc acaagatcgt gaacgtggat cagagacagt acggcgacgt gttcaagggc 480
gatctgaatc ccaagcctca gggccagagg ctgatcgagg tgtccgtgga ggagaaccac 540
cccttcaccc tgagagcccc tatccagcgg atctacggcg tgaggtatac cgagacatgg 600
agctttctgc catccctgac atgcaccggc gacgctgctc ctgctatcca gcacatctgc 660
ctgaagcaca ccacatgttt tcaggacgtg gtggtggacg tggattgtgc cgagaataca 720
aaggaggatc agctggctga gatctcctac cggttccagg gcaagaagga ggccgatcag 780
ccttggatcg tggtgaacac ctctacactg tttgacgagc tggagctgga tccccctgag 840
atcgagccag gcgtgctgaa ggtgctgaga accgagaagc agtacctggg cgtgtatatc 900
tggaacatgc ggggctctga cggcaccagc acatacgcta ccttcctggt cacatggaag 960
ggcgatgaga agacccggaa tccaacacct gctgtgaccc ctcagccaag gggagctgag 1020
tttcacatgt ggaactatca ctcccacgtg ttctctgtgg gcgacacctt tagcctggcc 1080
atgcacctgc aatataagat ccacgaggct cctttcgacc tgctgctgga gtggctgtat 1140
gtgcccatcg atcctacatg ccagccaatg aggctgtact ccacctgtct gtatcaccca 1200
aatgcccccc aatgcctgag ccacatgaac tccggctgta cctttacaag cccccacctg 1260
gcccagagag tggcttccac agtgtaccag aactgcgagc acgccgacaa ttacaccgct 1320
tattgtctgg gcatctctca catggagccc agcttcggcc tgatcctgca cgacggcggc 1380
accacactga agtttgtgga tacacccgag tccctgtctg gcctctacgt gttcgtggtg 1440
tacttcaacg gccacgtgga ggccgtggct tatacagtgg tgtctaccgt ggatcacttc 1500
gtgaacgcca tcgaggagag aggatttcca cctaccgctg gacagcctcc agctaccaca 1560
aagcctaagg aaatcacccc tgtcaatcct ggaacttcac ctctg 1605
<210> 11
<211> 1608
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> 人工序列（Artificial Sequence）
<400> 11
atgggaacag tcaacaaacc agtcgtcggc gtgctgatgg gcttcggtat tattacagga 60
actctgagga ttactaaccc cgtgcgcgcc tctgtgctgc ggtacgacga tttccacaca 120
gacgaggata agctggacac caattccgtg tatgagccct actatcactc tgatcacgcc 180
gagagctcct gggtgaaccg gggcgagtct agcaggaagg cttacgacca caacagccct 240
tatatctggc cacggaatga ctacgatggc tttctggaga acgcccacga gcaccacggc 300
gtgtataatc agggcagagg catcgactct ggcgagcggc tgatgcagcc cacccagatg 360
agcgcccagg aggatctggg cgacgataca ggcatccacg tgatccctac cctgaatggc 420
gacgataggc acaagatcgt gaacgtggat cagagacagt acggcgacgt gttcaagggc 480
gatctgaatc ccaagcctca gggccagagg ctgatcgagg tgtccgtgga ggagaaccac 540
cccttcaccc tgagagcccc tatccagcgg atctacggcg tgaggtatac cgagacatgg 600
agctttctgc catccctgac atgcaccggc gacgctgctc ctgctatcca gcacatctgc 660
ctgaagcaca ccacatgttt tcaggacgtg gtggtggacg tggattgtgc cgagaataca 720
aaggaggatc agctggctga gatctcctac cggttccagg gcaagaagga ggccgatcag 780
ccttggatcg tggtgaacac ctctacactg tttgacgagc tggagctgga tccccctgag 840
atcgagccag gcgtgctgaa ggtgctgaga accgagaagc agtacctggg cgtgtatatc 900
tggaacatgc ggggctctga cggcaccagc acatacgcta ccttcctggt cacatggaag 960
ggcgatgaga agacccggaa tccaacacct gctgtgaccc ctcagccaag gggagctgag 1020
tttcacatgt ggaactatca ctcccacgtg ttctctgtgg gcgacacctt tagcctggcc 1080
atgcacctgc aatataagat ccacgaggct cctttcgacc tgctgctgga gtggctgtat 1140
gtgcccatcg atcctacatg ccagccaatg aggctgtact ccacctgtct gtatcaccca 1200
aatgcccccc aatgcctgag ccacatgaac tccggctgta cctttacaag cccccacctg 1260
gcccagagag tggcttccac agtgtaccag aactgcgagc acgccgacaa ttacaccgct 1320
tattgtctgg gcatctctca catggagccc agcttcggcc tgatcctgca cgacggcggc 1380
accacactga agtttgtgga tacacccgag tccctgtctg gcctctacgt gttcgtggtg 1440
tacttcaacg gccacgtgga ggccgtggct tatacagtgg tgtctaccgt ggatcacttc 1500
gtgaacgcca tcgaggagag aggatttcca cctaccgctg gacagcctcc agctaccaca 1560
aagcctaagg aaatcacccc tgtcaatcct ggaacttcac ctctgctg 1608
<210> 12
<211> 1611
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> 人工序列（Artificial Sequence）
<400> 12
atgggaacag tcaacaaacc agtcgtcggc gtgctgatgg gcttcggtat tattacagga 60
actctgagga ttactaaccc cgtgcgcgcc tctgtgctgc ggtacgacga tttccacaca 120
gacgaggata agctggacac caattccgtg tatgagccct actatcactc tgatcacgcc 180
gagagctcct gggtgaaccg gggcgagtct agcaggaagg cttacgacca caacagccct 240
tatatctggc cacggaatga ctacgatggc tttctggaga acgcccacga gcaccacggc 300
gtgtataatc agggcagagg catcgactct ggcgagcggc tgatgcagcc cacccagatg 360
agcgcccagg aggatctggg cgacgataca ggcatccacg tgatccctac cctgaatggc 420
gacgataggc acaagatcgt gaacgtggat cagagacagt acggcgacgt gttcaagggc 480
gatctgaatc ccaagcctca gggccagagg ctgatcgagg tgtccgtgga ggagaaccac 540
cccttcaccc tgagagcccc tatccagcgg atctacggcg tgaggtatac cgagacatgg 600
agctttctgc catccctgac atgcaccggc gacgctgctc ctgctatcca gcacatctgc 660
ctgaagcaca ccacatgttt tcaggacgtg gtggtggacg tggattgtgc cgagaataca 720
aaggaggatc agctggctga gatctcctac cggttccagg gcaagaagga ggccgatcag 780
ccttggatcg tggtgaacac ctctacactg tttgacgagc tggagctgga tccccctgag 840
atcgagccag gcgtgctgaa ggtgctgaga accgagaagc agtacctggg cgtgtatatc 900
tggaacatgc ggggctctga cggcaccagc acatacgcta ccttcctggt cacatggaag 960
ggcgatgaga agacccggaa tccaacacct gctgtgaccc ctcagccaag gggagctgag 1020
tttcacatgt ggaactatca ctcccacgtg ttctctgtgg gcgacacctt tagcctggcc 1080
atgcacctgc aatataagat ccacgaggct cctttcgacc tgctgctgga gtggctgtat 1140
gtgcccatcg atcctacatg ccagccaatg aggctgtact ccacctgtct gtatcaccca 1200
aatgcccccc aatgcctgag ccacatgaac tccggctgta cctttacaag cccccacctg 1260
gcccagagag tggcttccac agtgtaccag aactgcgagc acgccgacaa ttacaccgct 1320
tattgtctgg gcatctctca catggagccc agcttcggcc tgatcctgca cgacggcggc 1380
accacactga agtttgtgga tacacccgag tccctgtctg gcctctacgt gttcgtggtg 1440
tacttcaacg gccacgtgga ggccgtggct tatacagtgg tgtctaccgt ggatcacttc 1500
gtgaacgcca tcgaggagag aggatttcca cctaccgctg gacagcctcc agctaccaca 1560
aagcctaagg aaatcacccc tgtcaatcct ggaacttcac ctctgctgcg c 1611
<210> 13
<211> 1614
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> 人工序列（Artificial Sequence）
<400> 13
atgggaacag tcaacaaacc agtcgtcggc gtgctgatgg gcttcggtat tattacagga 60
actctgagga ttactaaccc cgtgcgcgcc tctgtgctgc ggtacgacga tttccacaca 120
gacgaggata agctggacac caattccgtg tatgagccct actatcactc tgatcacgcc 180
gagagctcct gggtgaaccg gggcgagtct agcaggaagg cttacgacca caacagccct 240
tatatctggc cacggaatga ctacgatggc tttctggaga acgcccacga gcaccacggc 300
gtgtataatc agggcagagg catcgactct ggcgagcggc tgatgcagcc cacccagatg 360
agcgcccagg aggatctggg cgacgataca ggcatccacg tgatccctac cctgaatggc 420
gacgataggc acaagatcgt gaacgtggat cagagacagt acggcgacgt gttcaagggc 480
gatctgaatc ccaagcctca gggccagagg ctgatcgagg tgtccgtgga ggagaaccac 540
cccttcaccc tgagagcccc tatccagcgg atctacggcg tgaggtatac cgagacatgg 600
agctttctgc catccctgac atgcaccggc gacgctgctc ctgctatcca gcacatctgc 660
ctgaagcaca ccacatgttt tcaggacgtg gtggtggacg tggattgtgc cgagaataca 720
aaggaggatc agctggctga gatctcctac cggttccagg gcaagaagga ggccgatcag 780
ccttggatcg tggtgaacac ctctacactg tttgacgagc tggagctgga tccccctgag 840
atcgagccag gcgtgctgaa ggtgctgaga accgagaagc agtacctggg cgtgtatatc 900
tggaacatgc ggggctctga cggcaccagc acatacgcta ccttcctggt cacatggaag 960
ggcgatgaga agacccggaa tccaacacct gctgtgaccc ctcagccaag gggagctgag 1020
tttcacatgt ggaactatca ctcccacgtg ttctctgtgg gcgacacctt tagcctggcc 1080
atgcacctgc aatataagat ccacgaggct cctttcgacc tgctgctgga gtggctgtat 1140
gtgcccatcg atcctacatg ccagccaatg aggctgtact ccacctgtct gtatcaccca 1200
aatgcccccc aatgcctgag ccacatgaac tccggctgta cctttacaag cccccacctg 1260
gcccagagag tggcttccac agtgtaccag aactgcgagc acgccgacaa ttacaccgct 1320
tattgtctgg gcatctctca catggagccc agcttcggcc tgatcctgca cgacggcggc 1380
accacactga agtttgtgga tacacccgag tccctgtctg gcctctacgt gttcgtggtg 1440
tacttcaacg gccacgtgga ggccgtggct tatacagtgg tgtctaccgt ggatcacttc 1500
gtgaacgcca tcgaggagag aggatttcca cctaccgctg gacagcctcc agctaccaca 1560
aagcctaagg aaatcacccc tgtcaatcct ggaacttcac ctctgctgcg ctac 1614
<210> 14
<211> 1617
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> 人工序列（Artificial Sequence）
<400> 14
atgggaacag tcaacaaacc agtcgtcggc gtgctgatgg gcttcggtat tattacagga 60
actctgagga ttactaaccc cgtgcgcgcc tctgtgctgc ggtacgacga tttccacaca 120
gacgaggata agctggacac caattccgtg tatgagccct actatcactc tgatcacgcc 180
gagagctcct gggtgaaccg gggcgagtct agcaggaagg cttacgacca caacagccct 240
tatatctggc cacggaatga ctacgatggc tttctggaga acgcccacga gcaccacggc 300
gtgtataatc agggcagagg catcgactct ggcgagcggc tgatgcagcc cacccagatg 360
agcgcccagg aggatctggg cgacgataca ggcatccacg tgatccctac cctgaatggc 420
gacgataggc acaagatcgt gaacgtggat cagagacagt acggcgacgt gttcaagggc 480
gatctgaatc ccaagcctca gggccagagg ctgatcgagg tgtccgtgga ggagaaccac 540
cccttcaccc tgagagcccc tatccagcgg atctacggcg tgaggtatac cgagacatgg 600
agctttctgc catccctgac atgcaccggc gacgctgctc ctgctatcca gcacatctgc 660
ctgaagcaca ccacatgttt tcaggacgtg gtggtggacg tggattgtgc cgagaataca 720
aaggaggatc agctggctga gatctcctac cggttccagg gcaagaagga ggccgatcag 780
ccttggatcg tggtgaacac ctctacactg tttgacgagc tggagctgga tccccctgag 840
atcgagccag gcgtgctgaa ggtgctgaga accgagaagc agtacctggg cgtgtatatc 900
tggaacatgc ggggctctga cggcaccagc acatacgcta ccttcctggt cacatggaag 960
ggcgatgaga agacccggaa tccaacacct gctgtgaccc ctcagccaag gggagctgag 1020
tttcacatgt ggaactatca ctcccacgtg ttctctgtgg gcgacacctt tagcctggcc 1080
atgcacctgc aatataagat ccacgaggct cctttcgacc tgctgctgga gtggctgtat 1140
gtgcccatcg atcctacatg ccagccaatg aggctgtact ccacctgtct gtatcaccca 1200
aatgcccccc aatgcctgag ccacatgaac tccggctgta cctttacaag cccccacctg 1260
gcccagagag tggcttccac agtgtaccag aactgcgagc acgccgacaa ttacaccgct 1320
tattgtctgg gcatctctca catggagccc agcttcggcc tgatcctgca cgacggcggc 1380
accacactga agtttgtgga tacacccgag tccctgtctg gcctctacgt gttcgtggtg 1440
tacttcaacg gccacgtgga ggccgtggct tatacagtgg tgtctaccgt ggatcacttc 1500
gtgaacgcca tcgaggagag aggatttcca cctaccgctg gacagcctcc agctaccaca 1560
aagcctaagg aaatcacccc tgtcaatcct ggaacttcac ctctgctgcg ctacgca 1617
<210> 15
<211> 1620
<212> DNA
<213> 人工序列（Artificial Sequence）
<220>
<223> 人工序列（Artificial Sequence）
<400> 15
atgggaacag tcaacaaacc agtcgtcggc gtgctgatgg gcttcggtat tattacagga 60
actctgagga ttactaaccc cgtgcgcgcc tctgtgctgc ggtacgacga tttccacaca 120
gacgaggata agctggacac caattccgtg tatgagccct actatcactc tgatcacgcc 180
gagagctcct gggtgaaccg gggcgagtct agcaggaagg cttacgacca caacagccct 240
tatatctggc cacggaatga ctacgatggc tttctggaga acgcccacga gcaccacggc 300
gtgtataatc agggcagagg catcgactct ggcgagcggc tgatgcagcc cacccagatg 360
agcgcccagg aggatctggg cgacgataca ggcatccacg tgatccctac cctgaatggc 420
gacgataggc acaagatcgt gaacgtggat cagagacagt acggcgacgt gttcaagggc 480
gatctgaatc ccaagcctca gggccagagg ctgatcgagg tgtccgtgga ggagaaccac 540
cccttcaccc tgagagcccc tatccagcgg atctacggcg tgaggtatac cgagacatgg 600
agctttctgc catccctgac atgcaccggc gacgctgctc ctgctatcca gcacatctgc 660
ctgaagcaca ccacatgttt tcaggacgtg gtggtggacg tggattgtgc cgagaataca 720
aaggaggatc agctggctga gatctcctac cggttccagg gcaagaagga ggccgatcag 780
ccttggatcg tggtgaacac ctctacactg tttgacgagc tggagctgga tccccctgag 840
atcgagccag gcgtgctgaa ggtgctgaga accgagaagc agtacctggg cgtgtatatc 900
tggaacatgc ggggctctga cggcaccagc acatacgcta ccttcctggt cacatggaag 960
ggcgatgaga agacccggaa tccaacacct gctgtgaccc ctcagccaag gggagctgag 1020
tttcacatgt ggaactatca ctcccacgtg ttctctgtgg gcgacacctt tagcctggcc 1080
atgcacctgc aatataagat ccacgaggct cctttcgacc tgctgctgga gtggctgtat 1140
gtgcccatcg atcctacatg ccagccaatg aggctgtact ccacctgtct gtatcaccca 1200
aatgcccccc aatgcctgag ccacatgaac tccggctgta cctttacaag cccccacctg 1260
gcccagagag tggcttccac agtgtaccag aactgcgagc acgccgacaa ttacaccgct 1320
tattgtctgg gcatctctca catggagccc agcttcggcc tgatcctgca cgacggcggc 1380
accacactga agtttgtgga tacacccgag tccctgtctg gcctctacgt gttcgtggtg 1440
tacttcaacg gccacgtgga ggccgtggct tatacagtgg tgtctaccgt ggatcacttc 1500
gtgaacgcca tcgaggagag aggatttcca cctaccgctg gacagcctcc agctaccaca 1560
aagcctaagg aaatcacccc tgtcaatcct ggaacttcac ctctgctgcg ctacgcagcc 1620
1620
<210> 16
<211> 500
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> 人工序列（Artificial Sequence）
<400> 16
Met Gly Thr Val Asn Lys Pro Val Val Gly Val Leu Met Gly Phe Gly
1 5 10 15
Ile Ile Thr Gly Thr Leu Arg Ile Thr Asn Pro Val Arg Ala Ser Val
20 25 30
Leu Arg Tyr Asp Asp Phe His Thr Asp Glu Asp Lys Leu Asp Thr Asn
35 40 45
Ser Val Tyr Glu Pro Tyr Tyr His Ser Asp His Ala Glu Ser Ser Trp
50 55 60
Val Asn Arg Gly Glu Ser Ser Arg Lys Ala Tyr Asp His Asn Ser Pro
65 70 75 80
Tyr Ile Trp Pro Arg Asn Asp Tyr Asp Gly Phe Leu Glu Asn Ala His
85 90 95
Glu His His Gly Val Tyr Asn Gln Gly Arg Gly Ile Asp Ser Gly Glu
100 105 110
Arg Leu Met Gln Pro Thr Gln Met Ser Ala Gln Glu Asp Leu Gly Asp
115 120 125
Asp Thr Gly Ile His Val Ile Pro Thr Leu Asn Gly Asp Asp Arg His
130 135 140
Lys Ile Val Asn Val Asp Gln Arg Gln Tyr Gly Asp Val Phe Lys Gly
145 150 155 160
Asp Leu Asn Pro Lys Pro Gln Gly Gln Arg Leu Ile Glu Val Ser Val
165 170 175
Glu Glu Asn His Pro Phe Thr Leu Arg Ala Pro Ile Gln Arg Ile Tyr
180 185 190
Gly Val Arg Tyr Thr Glu Thr Trp Ser Phe Leu Pro Ser Leu Thr Cys
195 200 205
Thr Gly Asp Ala Ala Pro Ala Ile Gln His Ile Cys Leu Lys His Thr
210 215 220
Thr Cys Phe Gln Asp Val Val Val Asp Val Asp Cys Ala Glu Asn Thr
225 230 235 240
Lys Glu Asp Gln Leu Ala Glu Ile Ser Tyr Arg Phe Gln Gly Lys Lys
245 250 255
Glu Ala Asp Gln Pro Trp Ile Val Val Asn Thr Ser Thr Leu Phe Asp
260 265 270
Glu Leu Glu Leu Asp Pro Pro Glu Ile Glu Pro Gly Val Leu Lys Val
275 280 285
Leu Arg Thr Glu Lys Gln Tyr Leu Gly Val Tyr Ile Trp Asn Met Arg
290 295 300
Gly Ser Asp Gly Thr Ser Thr Tyr Ala Thr Phe Leu Val Thr Trp Lys
305 310 315 320
Gly Asp Glu Lys Thr Arg Asn Pro Thr Pro Ala Val Thr Pro Gln Pro
325 330 335
Arg Gly Ala Glu Phe His Met Trp Asn Tyr His Ser His Val Phe Ser
340 345 350
Val Gly Asp Thr Phe Ser Leu Ala Met His Leu Gln Tyr Lys Ile His
355 360 365
Glu Ala Pro Phe Asp Leu Leu Leu Glu Trp Leu Tyr Val Pro Ile Asp
370 375 380
Pro Thr Cys Gln Pro Met Arg Leu Tyr Ser Thr Cys Leu Tyr His Pro
385 390 395 400
Asn Ala Pro Gln Cys Leu Ser His Met Asn Ser Gly Cys Thr Phe Thr
405 410 415
Ser Pro His Leu Ala Gln Arg Val Ala Ser Thr Val Tyr Gln Asn Cys
420 425 430
Glu His Ala Asp Asn Tyr Thr Ala Tyr Cys Leu Gly Ile Ser His Met
435 440 445
Glu Pro Ser Phe Gly Leu Ile Leu His Asp Gly Gly Thr Thr Leu Lys
450 455 460
Phe Val Asp Thr Pro Glu Ser Leu Ser Gly Leu Tyr Val Phe Val Val
465 470 475 480
Tyr Phe Asn Gly His Val Glu Ala Val Ala Tyr Thr Val Val Ser Thr
485 490 495
Val Asp His Phe
500
<210> 17
<211> 505
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> 人工序列（Artificial Sequence）
<400> 17
Met Gly Thr Val Asn Lys Pro Val Val Gly Val Leu Met Gly Phe Gly
1 5 10 15
Ile Ile Thr Gly Thr Leu Arg Ile Thr Asn Pro Val Arg Ala Ser Val
20 25 30
Leu Arg Tyr Asp Asp Phe His Thr Asp Glu Asp Lys Leu Asp Thr Asn
35 40 45
Ser Val Tyr Glu Pro Tyr Tyr His Ser Asp His Ala Glu Ser Ser Trp
50 55 60
Val Asn Arg Gly Glu Ser Ser Arg Lys Ala Tyr Asp His Asn Ser Pro
65 70 75 80
Tyr Ile Trp Pro Arg Asn Asp Tyr Asp Gly Phe Leu Glu Asn Ala His
85 90 95
Glu His His Gly Val Tyr Asn Gln Gly Arg Gly Ile Asp Ser Gly Glu
100 105 110
Arg Leu Met Gln Pro Thr Gln Met Ser Ala Gln Glu Asp Leu Gly Asp
115 120 125
Asp Thr Gly Ile His Val Ile Pro Thr Leu Asn Gly Asp Asp Arg His
130 135 140
Lys Ile Val Asn Val Asp Gln Arg Gln Tyr Gly Asp Val Phe Lys Gly
145 150 155 160
Asp Leu Asn Pro Lys Pro Gln Gly Gln Arg Leu Ile Glu Val Ser Val
165 170 175
Glu Glu Asn His Pro Phe Thr Leu Arg Ala Pro Ile Gln Arg Ile Tyr
180 185 190
Gly Val Arg Tyr Thr Glu Thr Trp Ser Phe Leu Pro Ser Leu Thr Cys
195 200 205
Thr Gly Asp Ala Ala Pro Ala Ile Gln His Ile Cys Leu Lys His Thr
210 215 220
Thr Cys Phe Gln Asp Val Val Val Asp Val Asp Cys Ala Glu Asn Thr
225 230 235 240
Lys Glu Asp Gln Leu Ala Glu Ile Ser Tyr Arg Phe Gln Gly Lys Lys
245 250 255
Glu Ala Asp Gln Pro Trp Ile Val Val Asn Thr Ser Thr Leu Phe Asp
260 265 270
Glu Leu Glu Leu Asp Pro Pro Glu Ile Glu Pro Gly Val Leu Lys Val
275 280 285
Leu Arg Thr Glu Lys Gln Tyr Leu Gly Val Tyr Ile Trp Asn Met Arg
290 295 300
Gly Ser Asp Gly Thr Ser Thr Tyr Ala Thr Phe Leu Val Thr Trp Lys
305 310 315 320
Gly Asp Glu Lys Thr Arg Asn Pro Thr Pro Ala Val Thr Pro Gln Pro
325 330 335
Arg Gly Ala Glu Phe His Met Trp Asn Tyr His Ser His Val Phe Ser
340 345 350
Val Gly Asp Thr Phe Ser Leu Ala Met His Leu Gln Tyr Lys Ile His
355 360 365
Glu Ala Pro Phe Asp Leu Leu Leu Glu Trp Leu Tyr Val Pro Ile Asp
370 375 380
Pro Thr Cys Gln Pro Met Arg Leu Tyr Ser Thr Cys Leu Tyr His Pro
385 390 395 400
Asn Ala Pro Gln Cys Leu Ser His Met Asn Ser Gly Cys Thr Phe Thr
405 410 415
Ser Pro His Leu Ala Gln Arg Val Ala Ser Thr Val Tyr Gln Asn Cys
420 425 430
Glu His Ala Asp Asn Tyr Thr Ala Tyr Cys Leu Gly Ile Ser His Met
435 440 445
Glu Pro Ser Phe Gly Leu Ile Leu His Asp Gly Gly Thr Thr Leu Lys
450 455 460
Phe Val Asp Thr Pro Glu Ser Leu Ser Gly Leu Tyr Val Phe Val Val
465 470 475 480
Tyr Phe Asn Gly His Val Glu Ala Val Ala Tyr Thr Val Val Ser Thr
485 490 495
Val Asp His Phe Val Asn Ala Ile Glu
500 505
<210> 18
<211> 510
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> 人工序列（Artificial Sequence）
<400> 18
Met Gly Thr Val Asn Lys Pro Val Val Gly Val Leu Met Gly Phe Gly
1 5 10 15
Ile Ile Thr Gly Thr Leu Arg Ile Thr Asn Pro Val Arg Ala Ser Val
20 25 30
Leu Arg Tyr Asp Asp Phe His Thr Asp Glu Asp Lys Leu Asp Thr Asn
35 40 45
Ser Val Tyr Glu Pro Tyr Tyr His Ser Asp His Ala Glu Ser Ser Trp
50 55 60
Val Asn Arg Gly Glu Ser Ser Arg Lys Ala Tyr Asp His Asn Ser Pro
65 70 75 80
Tyr Ile Trp Pro Arg Asn Asp Tyr Asp Gly Phe Leu Glu Asn Ala His
85 90 95
Glu His His Gly Val Tyr Asn Gln Gly Arg Gly Ile Asp Ser Gly Glu
100 105 110
Arg Leu Met Gln Pro Thr Gln Met Ser Ala Gln Glu Asp Leu Gly Asp
115 120 125
Asp Thr Gly Ile His Val Ile Pro Thr Leu Asn Gly Asp Asp Arg His
130 135 140
Lys Ile Val Asn Val Asp Gln Arg Gln Tyr Gly Asp Val Phe Lys Gly
145 150 155 160
Asp Leu Asn Pro Lys Pro Gln Gly Gln Arg Leu Ile Glu Val Ser Val
165 170 175
Glu Glu Asn His Pro Phe Thr Leu Arg Ala Pro Ile Gln Arg Ile Tyr
180 185 190
Gly Val Arg Tyr Thr Glu Thr Trp Ser Phe Leu Pro Ser Leu Thr Cys
195 200 205
Thr Gly Asp Ala Ala Pro Ala Ile Gln His Ile Cys Leu Lys His Thr
210 215 220
Thr Cys Phe Gln Asp Val Val Val Asp Val Asp Cys Ala Glu Asn Thr
225 230 235 240
Lys Glu Asp Gln Leu Ala Glu Ile Ser Tyr Arg Phe Gln Gly Lys Lys
245 250 255
Glu Ala Asp Gln Pro Trp Ile Val Val Asn Thr Ser Thr Leu Phe Asp
260 265 270
Glu Leu Glu Leu Asp Pro Pro Glu Ile Glu Pro Gly Val Leu Lys Val
275 280 285
Leu Arg Thr Glu Lys Gln Tyr Leu Gly Val Tyr Ile Trp Asn Met Arg
290 295 300
Gly Ser Asp Gly Thr Ser Thr Tyr Ala Thr Phe Leu Val Thr Trp Lys
305 310 315 320
Gly Asp Glu Lys Thr Arg Asn Pro Thr Pro Ala Val Thr Pro Gln Pro
325 330 335
Arg Gly Ala Glu Phe His Met Trp Asn Tyr His Ser His Val Phe Ser
340 345 350
Val Gly Asp Thr Phe Ser Leu Ala Met His Leu Gln Tyr Lys Ile His
355 360 365
Glu Ala Pro Phe Asp Leu Leu Leu Glu Trp Leu Tyr Val Pro Ile Asp
370 375 380
Pro Thr Cys Gln Pro Met Arg Leu Tyr Ser Thr Cys Leu Tyr His Pro
385 390 395 400
Asn Ala Pro Gln Cys Leu Ser His Met Asn Ser Gly Cys Thr Phe Thr
405 410 415
Ser Pro His Leu Ala Gln Arg Val Ala Ser Thr Val Tyr Gln Asn Cys
420 425 430
Glu His Ala Asp Asn Tyr Thr Ala Tyr Cys Leu Gly Ile Ser His Met
435 440 445
Glu Pro Ser Phe Gly Leu Ile Leu His Asp Gly Gly Thr Thr Leu Lys
450 455 460
Phe Val Asp Thr Pro Glu Ser Leu Ser Gly Leu Tyr Val Phe Val Val
465 470 475 480
Tyr Phe Asn Gly His Val Glu Ala Val Ala Tyr Thr Val Val Ser Thr
485 490 495
Val Asp His Phe Val Asn Ala Ile Glu Glu Arg Gly Phe Pro
500 505 510
<210> 19
<211> 515
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> 人工序列（Artificial Sequence）
<400> 19
Met Gly Thr Val Asn Lys Pro Val Val Gly Val Leu Met Gly Phe Gly
1 5 10 15
Ile Ile Thr Gly Thr Leu Arg Ile Thr Asn Pro Val Arg Ala Ser Val
20 25 30
Leu Arg Tyr Asp Asp Phe His Thr Asp Glu Asp Lys Leu Asp Thr Asn
35 40 45
Ser Val Tyr Glu Pro Tyr Tyr His Ser Asp His Ala Glu Ser Ser Trp
50 55 60
Val Asn Arg Gly Glu Ser Ser Arg Lys Ala Tyr Asp His Asn Ser Pro
65 70 75 80
Tyr Ile Trp Pro Arg Asn Asp Tyr Asp Gly Phe Leu Glu Asn Ala His
85 90 95
Glu His His Gly Val Tyr Asn Gln Gly Arg Gly Ile Asp Ser Gly Glu
100 105 110
Arg Leu Met Gln Pro Thr Gln Met Ser Ala Gln Glu Asp Leu Gly Asp
115 120 125
Asp Thr Gly Ile His Val Ile Pro Thr Leu Asn Gly Asp Asp Arg His
130 135 140
Lys Ile Val Asn Val Asp Gln Arg Gln Tyr Gly Asp Val Phe Lys Gly
145 150 155 160
Asp Leu Asn Pro Lys Pro Gln Gly Gln Arg Leu Ile Glu Val Ser Val
165 170 175
Glu Glu Asn His Pro Phe Thr Leu Arg Ala Pro Ile Gln Arg Ile Tyr
180 185 190
Gly Val Arg Tyr Thr Glu Thr Trp Ser Phe Leu Pro Ser Leu Thr Cys
195 200 205
Thr Gly Asp Ala Ala Pro Ala Ile Gln His Ile Cys Leu Lys His Thr
210 215 220
Thr Cys Phe Gln Asp Val Val Val Asp Val Asp Cys Ala Glu Asn Thr
225 230 235 240
Lys Glu Asp Gln Leu Ala Glu Ile Ser Tyr Arg Phe Gln Gly Lys Lys
245 250 255
Glu Ala Asp Gln Pro Trp Ile Val Val Asn Thr Ser Thr Leu Phe Asp
260 265 270
Glu Leu Glu Leu Asp Pro Pro Glu Ile Glu Pro Gly Val Leu Lys Val
275 280 285
Leu Arg Thr Glu Lys Gln Tyr Leu Gly Val Tyr Ile Trp Asn Met Arg
290 295 300
Gly Ser Asp Gly Thr Ser Thr Tyr Ala Thr Phe Leu Val Thr Trp Lys
305 310 315 320
Gly Asp Glu Lys Thr Arg Asn Pro Thr Pro Ala Val Thr Pro Gln Pro
325 330 335
Arg Gly Ala Glu Phe His Met Trp Asn Tyr His Ser His Val Phe Ser
340 345 350
Val Gly Asp Thr Phe Ser Leu Ala Met His Leu Gln Tyr Lys Ile His
355 360 365
Glu Ala Pro Phe Asp Leu Leu Leu Glu Trp Leu Tyr Val Pro Ile Asp
370 375 380
Pro Thr Cys Gln Pro Met Arg Leu Tyr Ser Thr Cys Leu Tyr His Pro
385 390 395 400
Asn Ala Pro Gln Cys Leu Ser His Met Asn Ser Gly Cys Thr Phe Thr
405 410 415
Ser Pro His Leu Ala Gln Arg Val Ala Ser Thr Val Tyr Gln Asn Cys
420 425 430
Glu His Ala Asp Asn Tyr Thr Ala Tyr Cys Leu Gly Ile Ser His Met
435 440 445
Glu Pro Ser Phe Gly Leu Ile Leu His Asp Gly Gly Thr Thr Leu Lys
450 455 460
Phe Val Asp Thr Pro Glu Ser Leu Ser Gly Leu Tyr Val Phe Val Val
465 470 475 480
Tyr Phe Asn Gly His Val Glu Ala Val Ala Tyr Thr Val Val Ser Thr
485 490 495
Val Asp His Phe Val Asn Ala Ile Glu Glu Arg Gly Phe Pro Pro Thr
500 505 510
Ala Gly Gln
515
<210> 20
<211> 520
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> 人工序列（Artificial Sequence）
<400> 20
Met Gly Thr Val Asn Lys Pro Val Val Gly Val Leu Met Gly Phe Gly
1 5 10 15
Ile Ile Thr Gly Thr Leu Arg Ile Thr Asn Pro Val Arg Ala Ser Val
20 25 30
Leu Arg Tyr Asp Asp Phe His Thr Asp Glu Asp Lys Leu Asp Thr Asn
35 40 45
Ser Val Tyr Glu Pro Tyr Tyr His Ser Asp His Ala Glu Ser Ser Trp
50 55 60
Val Asn Arg Gly Glu Ser Ser Arg Lys Ala Tyr Asp His Asn Ser Pro
65 70 75 80
Tyr Ile Trp Pro Arg Asn Asp Tyr Asp Gly Phe Leu Glu Asn Ala His
85 90 95
Glu His His Gly Val Tyr Asn Gln Gly Arg Gly Ile Asp Ser Gly Glu
100 105 110
Arg Leu Met Gln Pro Thr Gln Met Ser Ala Gln Glu Asp Leu Gly Asp
115 120 125
Asp Thr Gly Ile His Val Ile Pro Thr Leu Asn Gly Asp Asp Arg His
130 135 140
Lys Ile Val Asn Val Asp Gln Arg Gln Tyr Gly Asp Val Phe Lys Gly
145 150 155 160
Asp Leu Asn Pro Lys Pro Gln Gly Gln Arg Leu Ile Glu Val Ser Val
165 170 175
Glu Glu Asn His Pro Phe Thr Leu Arg Ala Pro Ile Gln Arg Ile Tyr
180 185 190
Gly Val Arg Tyr Thr Glu Thr Trp Ser Phe Leu Pro Ser Leu Thr Cys
195 200 205
Thr Gly Asp Ala Ala Pro Ala Ile Gln His Ile Cys Leu Lys His Thr
210 215 220
Thr Cys Phe Gln Asp Val Val Val Asp Val Asp Cys Ala Glu Asn Thr
225 230 235 240
Lys Glu Asp Gln Leu Ala Glu Ile Ser Tyr Arg Phe Gln Gly Lys Lys
245 250 255
Glu Ala Asp Gln Pro Trp Ile Val Val Asn Thr Ser Thr Leu Phe Asp
260 265 270
Glu Leu Glu Leu Asp Pro Pro Glu Ile Glu Pro Gly Val Leu Lys Val
275 280 285
Leu Arg Thr Glu Lys Gln Tyr Leu Gly Val Tyr Ile Trp Asn Met Arg
290 295 300
Gly Ser Asp Gly Thr Ser Thr Tyr Ala Thr Phe Leu Val Thr Trp Lys
305 310 315 320
Gly Asp Glu Lys Thr Arg Asn Pro Thr Pro Ala Val Thr Pro Gln Pro
325 330 335
Arg Gly Ala Glu Phe His Met Trp Asn Tyr His Ser His Val Phe Ser
340 345 350
Val Gly Asp Thr Phe Ser Leu Ala Met His Leu Gln Tyr Lys Ile His
355 360 365
Glu Ala Pro Phe Asp Leu Leu Leu Glu Trp Leu Tyr Val Pro Ile Asp
370 375 380
Pro Thr Cys Gln Pro Met Arg Leu Tyr Ser Thr Cys Leu Tyr His Pro
385 390 395 400
Asn Ala Pro Gln Cys Leu Ser His Met Asn Ser Gly Cys Thr Phe Thr
405 410 415
Ser Pro His Leu Ala Gln Arg Val Ala Ser Thr Val Tyr Gln Asn Cys
420 425 430
Glu His Ala Asp Asn Tyr Thr Ala Tyr Cys Leu Gly Ile Ser His Met
435 440 445
Glu Pro Ser Phe Gly Leu Ile Leu His Asp Gly Gly Thr Thr Leu Lys
450 455 460
Phe Val Asp Thr Pro Glu Ser Leu Ser Gly Leu Tyr Val Phe Val Val
465 470 475 480
Tyr Phe Asn Gly His Val Glu Ala Val Ala Tyr Thr Val Val Ser Thr
485 490 495
Val Asp His Phe Val Asn Ala Ile Glu Glu Arg Gly Phe Pro Pro Thr
500 505 510
Ala Gly Gln Pro Pro Ala Thr Thr
515 520
<210> 21
<211> 525
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> 人工序列（Artificial Sequence）
<400> 21
Met Gly Thr Val Asn Lys Pro Val Val Gly Val Leu Met Gly Phe Gly
1 5 10 15
Ile Ile Thr Gly Thr Leu Arg Ile Thr Asn Pro Val Arg Ala Ser Val
20 25 30
Leu Arg Tyr Asp Asp Phe His Thr Asp Glu Asp Lys Leu Asp Thr Asn
35 40 45
Ser Val Tyr Glu Pro Tyr Tyr His Ser Asp His Ala Glu Ser Ser Trp
50 55 60
Val Asn Arg Gly Glu Ser Ser Arg Lys Ala Tyr Asp His Asn Ser Pro
65 70 75 80
Tyr Ile Trp Pro Arg Asn Asp Tyr Asp Gly Phe Leu Glu Asn Ala His
85 90 95
Glu His His Gly Val Tyr Asn Gln Gly Arg Gly Ile Asp Ser Gly Glu
100 105 110
Arg Leu Met Gln Pro Thr Gln Met Ser Ala Gln Glu Asp Leu Gly Asp
115 120 125
Asp Thr Gly Ile His Val Ile Pro Thr Leu Asn Gly Asp Asp Arg His
130 135 140
Lys Ile Val Asn Val Asp Gln Arg Gln Tyr Gly Asp Val Phe Lys Gly
145 150 155 160
Asp Leu Asn Pro Lys Pro Gln Gly Gln Arg Leu Ile Glu Val Ser Val
165 170 175
Glu Glu Asn His Pro Phe Thr Leu Arg Ala Pro Ile Gln Arg Ile Tyr
180 185 190
Gly Val Arg Tyr Thr Glu Thr Trp Ser Phe Leu Pro Ser Leu Thr Cys
195 200 205
Thr Gly Asp Ala Ala Pro Ala Ile Gln His Ile Cys Leu Lys His Thr
210 215 220
Thr Cys Phe Gln Asp Val Val Val Asp Val Asp Cys Ala Glu Asn Thr
225 230 235 240
Lys Glu Asp Gln Leu Ala Glu Ile Ser Tyr Arg Phe Gln Gly Lys Lys
245 250 255
Glu Ala Asp Gln Pro Trp Ile Val Val Asn Thr Ser Thr Leu Phe Asp
260 265 270
Glu Leu Glu Leu Asp Pro Pro Glu Ile Glu Pro Gly Val Leu Lys Val
275 280 285
Leu Arg Thr Glu Lys Gln Tyr Leu Gly Val Tyr Ile Trp Asn Met Arg
290 295 300
Gly Ser Asp Gly Thr Ser Thr Tyr Ala Thr Phe Leu Val Thr Trp Lys
305 310 315 320
Gly Asp Glu Lys Thr Arg Asn Pro Thr Pro Ala Val Thr Pro Gln Pro
325 330 335
Arg Gly Ala Glu Phe His Met Trp Asn Tyr His Ser His Val Phe Ser
340 345 350
Val Gly Asp Thr Phe Ser Leu Ala Met His Leu Gln Tyr Lys Ile His
355 360 365
Glu Ala Pro Phe Asp Leu Leu Leu Glu Trp Leu Tyr Val Pro Ile Asp
370 375 380
Pro Thr Cys Gln Pro Met Arg Leu Tyr Ser Thr Cys Leu Tyr His Pro
385 390 395 400
Asn Ala Pro Gln Cys Leu Ser His Met Asn Ser Gly Cys Thr Phe Thr
405 410 415
Ser Pro His Leu Ala Gln Arg Val Ala Ser Thr Val Tyr Gln Asn Cys
420 425 430
Glu His Ala Asp Asn Tyr Thr Ala Tyr Cys Leu Gly Ile Ser His Met
435 440 445
Glu Pro Ser Phe Gly Leu Ile Leu His Asp Gly Gly Thr Thr Leu Lys
450 455 460
Phe Val Asp Thr Pro Glu Ser Leu Ser Gly Leu Tyr Val Phe Val Val
465 470 475 480
Tyr Phe Asn Gly His Val Glu Ala Val Ala Tyr Thr Val Val Ser Thr
485 490 495
Val Asp His Phe Val Asn Ala Ile Glu Glu Arg Gly Phe Pro Pro Thr
500 505 510
Ala Gly Gln Pro Pro Ala Thr Thr Lys Pro Lys Glu Ile
515 520 525
<210> 22
<211> 530
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> 人工序列（Artificial Sequence）
<400> 22
Met Gly Thr Val Asn Lys Pro Val Val Gly Val Leu Met Gly Phe Gly
1 5 10 15
Ile Ile Thr Gly Thr Leu Arg Ile Thr Asn Pro Val Arg Ala Ser Val
20 25 30
Leu Arg Tyr Asp Asp Phe His Thr Asp Glu Asp Lys Leu Asp Thr Asn
35 40 45
Ser Val Tyr Glu Pro Tyr Tyr His Ser Asp His Ala Glu Ser Ser Trp
50 55 60
Val Asn Arg Gly Glu Ser Ser Arg Lys Ala Tyr Asp His Asn Ser Pro
65 70 75 80
Tyr Ile Trp Pro Arg Asn Asp Tyr Asp Gly Phe Leu Glu Asn Ala His
85 90 95
Glu His His Gly Val Tyr Asn Gln Gly Arg Gly Ile Asp Ser Gly Glu
100 105 110
Arg Leu Met Gln Pro Thr Gln Met Ser Ala Gln Glu Asp Leu Gly Asp
115 120 125
Asp Thr Gly Ile His Val Ile Pro Thr Leu Asn Gly Asp Asp Arg His
130 135 140
Lys Ile Val Asn Val Asp Gln Arg Gln Tyr Gly Asp Val Phe Lys Gly
145 150 155 160
Asp Leu Asn Pro Lys Pro Gln Gly Gln Arg Leu Ile Glu Val Ser Val
165 170 175
Glu Glu Asn His Pro Phe Thr Leu Arg Ala Pro Ile Gln Arg Ile Tyr
180 185 190
Gly Val Arg Tyr Thr Glu Thr Trp Ser Phe Leu Pro Ser Leu Thr Cys
195 200 205
Thr Gly Asp Ala Ala Pro Ala Ile Gln His Ile Cys Leu Lys His Thr
210 215 220
Thr Cys Phe Gln Asp Val Val Val Asp Val Asp Cys Ala Glu Asn Thr
225 230 235 240
Lys Glu Asp Gln Leu Ala Glu Ile Ser Tyr Arg Phe Gln Gly Lys Lys
245 250 255
Glu Ala Asp Gln Pro Trp Ile Val Val Asn Thr Ser Thr Leu Phe Asp
260 265 270
Glu Leu Glu Leu Asp Pro Pro Glu Ile Glu Pro Gly Val Leu Lys Val
275 280 285
Leu Arg Thr Glu Lys Gln Tyr Leu Gly Val Tyr Ile Trp Asn Met Arg
290 295 300
Gly Ser Asp Gly Thr Ser Thr Tyr Ala Thr Phe Leu Val Thr Trp Lys
305 310 315 320
Gly Asp Glu Lys Thr Arg Asn Pro Thr Pro Ala Val Thr Pro Gln Pro
325 330 335
Arg Gly Ala Glu Phe His Met Trp Asn Tyr His Ser His Val Phe Ser
340 345 350
Val Gly Asp Thr Phe Ser Leu Ala Met His Leu Gln Tyr Lys Ile His
355 360 365
Glu Ala Pro Phe Asp Leu Leu Leu Glu Trp Leu Tyr Val Pro Ile Asp
370 375 380
Pro Thr Cys Gln Pro Met Arg Leu Tyr Ser Thr Cys Leu Tyr His Pro
385 390 395 400
Asn Ala Pro Gln Cys Leu Ser His Met Asn Ser Gly Cys Thr Phe Thr
405 410 415
Ser Pro His Leu Ala Gln Arg Val Ala Ser Thr Val Tyr Gln Asn Cys
420 425 430
Glu His Ala Asp Asn Tyr Thr Ala Tyr Cys Leu Gly Ile Ser His Met
435 440 445
Glu Pro Ser Phe Gly Leu Ile Leu His Asp Gly Gly Thr Thr Leu Lys
450 455 460
Phe Val Asp Thr Pro Glu Ser Leu Ser Gly Leu Tyr Val Phe Val Val
465 470 475 480
Tyr Phe Asn Gly His Val Glu Ala Val Ala Tyr Thr Val Val Ser Thr
485 490 495
Val Asp His Phe Val Asn Ala Ile Glu Glu Arg Gly Phe Pro Pro Thr
500 505 510
Ala Gly Gln Pro Pro Ala Thr Thr Lys Pro Lys Glu Ile Thr Pro Val
515 520 525
Asn Pro
530
<210> 23
<211> 543
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> 人工序列（Artificial Sequence）
<400> 23
Met Gly Thr Val Asn Lys Pro Val Val Gly Val Leu Met Gly Phe Gly
1 5 10 15
Ile Ile Thr Gly Thr Leu Arg Ile Thr Asn Pro Val Arg Ala Ser Val
20 25 30
Leu Arg Tyr Asp Asp Phe His Thr Asp Glu Asp Lys Leu Asp Thr Asn
35 40 45
Ser Val Tyr Glu Pro Tyr Tyr His Ser Asp His Ala Glu Ser Ser Trp
50 55 60
Val Asn Arg Gly Glu Ser Ser Arg Lys Ala Tyr Asp His Asn Ser Pro
65 70 75 80
Tyr Ile Trp Pro Arg Asn Asp Tyr Asp Gly Phe Leu Glu Asn Ala His
85 90 95
Glu His His Gly Val Tyr Asn Gln Gly Arg Gly Ile Asp Ser Gly Glu
100 105 110
Arg Leu Met Gln Pro Thr Gln Met Ser Ala Gln Glu Asp Leu Gly Asp
115 120 125
Asp Thr Gly Ile His Val Ile Pro Thr Leu Asn Gly Asp Asp Arg His
130 135 140
Lys Ile Val Asn Val Asp Gln Arg Gln Tyr Gly Asp Val Phe Lys Gly
145 150 155 160
Asp Leu Asn Pro Lys Pro Gln Gly Gln Arg Leu Ile Glu Val Ser Val
165 170 175
Glu Glu Asn His Pro Phe Thr Leu Arg Ala Pro Ile Gln Arg Ile Tyr
180 185 190
Gly Val Arg Tyr Thr Glu Thr Trp Ser Phe Leu Pro Ser Leu Thr Cys
195 200 205
Thr Gly Asp Ala Ala Pro Ala Ile Gln His Ile Cys Leu Lys His Thr
210 215 220
Thr Cys Phe Gln Asp Val Val Val Asp Val Asp Cys Ala Glu Asn Thr
225 230 235 240
Lys Glu Asp Gln Leu Ala Glu Ile Ser Tyr Arg Phe Gln Gly Lys Lys
245 250 255
Glu Ala Asp Gln Pro Trp Ile Val Val Asn Thr Ser Thr Leu Phe Asp
260 265 270
Glu Leu Glu Leu Asp Pro Pro Glu Ile Glu Pro Gly Val Leu Lys Val
275 280 285
Leu Arg Thr Glu Lys Gln Tyr Leu Gly Val Tyr Ile Trp Asn Met Arg
290 295 300
Gly Ser Asp Gly Thr Ser Thr Tyr Ala Thr Phe Leu Val Thr Trp Lys
305 310 315 320
Gly Asp Glu Lys Thr Arg Asn Pro Thr Pro Ala Val Thr Pro Gln Pro
325 330 335
Arg Gly Ala Glu Phe His Met Trp Asn Tyr His Ser His Val Phe Ser
340 345 350
Val Gly Asp Thr Phe Ser Leu Ala Met His Leu Gln Tyr Lys Ile His
355 360 365
Glu Ala Pro Phe Asp Leu Leu Leu Glu Trp Leu Tyr Val Pro Ile Asp
370 375 380
Pro Thr Cys Gln Pro Met Arg Leu Tyr Ser Thr Cys Leu Tyr His Pro
385 390 395 400
Asn Ala Pro Gln Cys Leu Ser His Met Asn Ser Gly Cys Thr Phe Thr
405 410 415
Ser Pro His Leu Ala Gln Arg Val Ala Ser Thr Val Tyr Gln Asn Cys
420 425 430
Glu His Ala Asp Asn Tyr Thr Ala Tyr Cys Leu Gly Ile Ser His Met
435 440 445
Glu Pro Ser Phe Gly Leu Ile Leu His Asp Gly Gly Thr Thr Leu Lys
450 455 460
Phe Val Asp Thr Pro Glu Ser Leu Ser Gly Leu Tyr Val Phe Val Val
465 470 475 480
Tyr Phe Asn Gly His Val Glu Ala Val Ala Tyr Thr Val Val Ser Thr
485 490 495
Val Asp His Phe Val Asn Ala Ile Glu Glu Arg Gly Phe Pro Pro Thr
500 505 510
Ala Gly Gln Pro Pro Ala Thr Thr Lys Pro Lys Glu Ile Thr Pro Val
515 520 525
Asn Pro Gly Thr Ser Pro Leu Leu Arg Tyr Ala Ala Trp Thr Gly
530 535 540
<210> 24
<211> 546
<212> PRT
<213> 人工序列（Artificial Sequence）
<220>
<223> 人工序列（Artificial Sequence）
<400> 24
Met Gly Thr Val Asn Lys Pro Val Val Gly Val Leu Met Gly Phe Gly
1 5 10 15
Ile Ile Thr Gly Thr Leu Arg Ile Thr Asn Pro Val Arg Ala Ser Val
20 25 30
Leu Arg Tyr Asp Asp Phe His Thr Asp Glu Asp Lys Leu Asp Thr Asn
35 40 45
Ser Val Tyr Glu Pro Tyr Tyr His Ser Asp His Ala Glu Ser Ser Trp
50 55 60
Val Asn Arg Gly Glu Ser Ser Arg Lys Ala Tyr Asp His Asn Ser Pro
65 70 75 80
Tyr Ile Trp Pro Arg Asn Asp Tyr Asp Gly Phe Leu Glu Asn Ala His
85 90 95
Glu His His Gly Val Tyr Asn Gln Gly Arg Gly Ile Asp Ser Gly Glu
100 105 110
Arg Leu Met Gln Pro Thr Gln Met Ser Ala Gln Glu Asp Leu Gly Asp
115 120 125
Asp Thr Gly Ile His Val Ile Pro Thr Leu Asn Gly Asp Asp Arg His
130 135 140
Lys Ile Val Asn Val Asp Gln Arg Gln Tyr Gly Asp Val Phe Lys Gly
145 150 155 160
Asp Leu Asn Pro Lys Pro Gln Gly Gln Arg Leu Ile Glu Val Ser Val
165 170 175
Glu Glu Asn His Pro Phe Thr Leu Arg Ala Pro Ile Gln Arg Ile Tyr
180 185 190
Gly Val Arg Tyr Thr Glu Thr Trp Ser Phe Leu Pro Ser Leu Thr Cys
195 200 205
Thr Gly Asp Ala Ala Pro Ala Ile Gln His Ile Cys Leu Lys His Thr
210 215 220
Thr Cys Phe Gln Asp Val Val Val Asp Val Asp Cys Ala Glu Asn Thr
225 230 235 240
Lys Glu Asp Gln Leu Ala Glu Ile Ser Tyr Arg Phe Gln Gly Lys Lys
245 250 255
Glu Ala Asp Gln Pro Trp Ile Val Val Asn Thr Ser Thr Leu Phe Asp
260 265 270
Glu Leu Glu Leu Asp Pro Pro Glu Ile Glu Pro Gly Val Leu Lys Val
275 280 285
Leu Arg Thr Glu Lys Gln Tyr Leu Gly Val Tyr Ile Trp Asn Met Arg
290 295 300
Gly Ser Asp Gly Thr Ser Thr Tyr Ala Thr Phe Leu Val Thr Trp Lys
305 310 315 320
Gly Asp Glu Lys Thr Arg Asn Pro Thr Pro Ala Val Thr Pro Gln Pro
325 330 335
Arg Gly Ala Glu Phe His Met Trp Asn Tyr His Ser His Val Phe Ser
340 345 350
Val Gly Asp Thr Phe Ser Leu Ala Met His Leu Gln Tyr Lys Ile His
355 360 365
Glu Ala Pro Phe Asp Leu Leu Leu Glu Trp Leu Tyr Val Pro Ile Asp
370 375 380
Pro Thr Cys Gln Pro Met Arg Leu Tyr Ser Thr Cys Leu Tyr His Pro
385 390 395 400
Asn Ala Pro Gln Cys Leu Ser His Met Asn Ser Gly Cys Thr Phe Thr
405 410 415
Ser Pro His Leu Ala Gln Arg Val Ala Ser Thr Val Tyr Gln Asn Cys
420 425 430
Glu His Ala Asp Asn Tyr Thr Ala Tyr Cys Leu Gly Ile Ser His Met
435 440 445
Glu Pro Ser Phe Gly Leu Ile Leu His Asp Gly Gly Thr Thr Leu Lys
450 455 460
Phe Val Asp Thr Pro Glu Ser Leu Ser Gly Leu Tyr Val Phe Val Val
465 470 475 480
Tyr Phe Asn Gly His Val Glu Ala Val Ala Tyr Thr Val Val Ser Thr
485 490 495
Val Asp His Phe Val Asn Ala Ile Glu Glu Arg Gly Phe Pro Pro Thr
500 505 510
Ala Gly Gln Pro Pro Ala Thr Thr Lys Pro Lys Glu Ile Thr Pro Val
515 520 525
Asn Pro Gly Thr Ser Pro Leu Leu Arg Tyr Ala Ala Trp Thr Gly Gly
530 535 540
Leu Ala
545

Claims (12)

1.一种水痘带状疱疹病毒表面蛋白抗原变体，包括一变体，该变体是水痘带状疱疹病毒表面蛋白(gE)抗原中选自第525位至第543位个氨基酸残基的任何一个氨基酸残基的羧基末端的截断，其氨基酸序列如SEQ ID NO:1所示。
2.如权利要求1所述的水痘带状疱疹病毒表面蛋白抗原变体，其中SEQ ID NO:1所示氨基酸序列中的第40个氨基酸残基是苏氨酸。
3.如权利要求1所述的水痘带状疱疹病毒表面蛋白抗原变体，其中SEQ ID NO:1所示氨基酸序列中的第536个氨基酸残基是亮氨酸。
4.如权利要求1所述的水痘带状疱疹病毒表面蛋白抗原变体，其中所述抗原变体由SEQID NO:2至8和SEQ ID NO:21至23所示的任一氨基酸序列表示。
5.一种基因，其编码如权利要求1-4中任一项所述的水痘带状疱疹病毒表面蛋白抗原变体。
6.如权利要求5所述的基因，其中，所述基因由SEQ ID NO:9至15所示的任一核苷酸序列所示。
7.一种重组载体，其包含：如权利要求5所述的基因。
8.一种宿主细胞，其中转化有如权利要求7所述的重组载体。
9.一种用于预防或治疗水痘或带状疱疹的疫苗组合物，其包含活性成分：如权利要求1-4中任一项所述的水痘带状疱疹病毒表面蛋白抗原变体。
10.一种预防或治疗水痘或带状疱疹的方法，包括：
向患者施用疫苗组合物的步骤，所述疫苗组合物包含如权利要求1-4中任一项所述的水痘带状疱疹病毒表面蛋白抗原变体作为活性成分。
11.一种疫苗组合物的用途，所述疫苗组合物包含如权利要求1-4中任一项所述的水痘带状疱疹病毒表面蛋白抗原变体作为活性成分，用于预防或治疗水痘或带状疱疹。
12.一种疫苗组合物的用途，所述疫苗组合物包含权利要求1-4中任一项的水痘带状疱疹病毒表面蛋白抗原变体作为活性成分，用于制备预防或治疗水痘或带状疱疹的药物。
CN201980034772.8A 2018-05-23 2019-05-22 水痘带状疱疹病毒的抗原变体及其用途 Pending CN112189017A (zh)

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CN113683704A (zh) * 2021-07-28 2021-11-23 安徽智飞龙科马生物制药有限公司 一种水痘-带状疱疹病毒r-gE融合蛋白、重组水痘-带状疱疹疫苗及其制备方法和应用
WO2023056912A1 (en) * 2021-10-08 2023-04-13 Suzhou Abogen Biosciences Co., Ltd. Nucleic acid vaccines for vzv
CN116284455A (zh) * 2023-04-17 2023-06-23 北京康乐卫士生物技术股份有限公司 一种带状疱疹病毒疫苗的嵌合抗原及其应用

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* Cited by examiner, † Cited by third party
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