CN111018961A - 白介素-2衍生物 - Google Patents

白介素-2衍生物 Download PDF

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CN111018961A
CN111018961A CN201911302355.5A CN201911302355A CN111018961A CN 111018961 A CN111018961 A CN 111018961A CN 201911302355 A CN201911302355 A CN 201911302355A CN 111018961 A CN111018961 A CN 111018961A
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CN111018961B (zh
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赵耀
彭璐佳
郭建云
朱笑婷
张建军
魏婷婷
刘慧杰
郑茜
王冀姝
张维
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Beijing Zhidao Biological Technology Co ltd
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Priority to CN202210110617.3A priority patent/CN114478743A/zh
Priority to CN202210110228.0A priority patent/CN114437198A/zh
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Abstract

本发明公开了一种IL‑2衍生物，通过在野生型IL‑2的基础上引入至少一个半胱氨酸残基，部分封闭或完全封闭IL‑2衍生物与α受体亚基的结合平面，同时基本保留对β和γ受体亚基复合物的亲和力。本发明还公开了复合物，包括IL‑2衍生物，IL‑2衍生物在野生型IL‑2的基础上引入第三半胱氨酸残基；封闭模块，封闭模块上具有或引入有第四半胱氨酸残基；IL‑2衍生物上的第三半胱氨酸残基和所述封闭模块上的第四半胱氨酸残基形成分子间二硫键，从而形成IL‑2与封闭模块的复合物，部分封闭或完全封闭IL‑2衍生物与α受体亚基的结合平面，同时基本保留对β和γ受体亚基复合物的亲和力。

Description

白介素-2衍生物
技术领域
本发明属于分子生物学领域，具体涉及一种白介素-2衍生物及其复合物。
背景技术
白介素-2(IL-2)，于1976年被发现，当时被称为T细胞生长因子(TCGF)，是一种在维持T淋巴细胞和NK细胞的正常功能中起着重要作用的球状糖蛋白。天然IL-2是一个具有133个氨基酸残基组成的多肽，分子量大约15kD，有三个半胱氨酸残基，分别位于第58、105和125位。翻译后修饰包括第3位的Thr糖基化，第58位和105位半胱氨酸残基形成二硫键，并形成其功能必不可少的主要由4个α螺旋以及一些连接序列(loop)组成的高级结构(Bazan等，Science257,410-413(1992))。
IL-2主要由活化的T细胞生成，它能促进T细胞的增殖和分化，维持T细胞活性；刺激天然杀伤(NK)细胞的生成、增殖和活化，并诱导细胞毒性T淋巴细胞(CTL)的生成以及诱导和激活淋巴因子激活的杀细胞(LAK)及肿瘤浸润淋巴细胞；促进T细胞表达细胞因子和细胞溶解分子，促进B细胞的增殖(Waldmann等,Nat Rev Immunol6,595-601(2009))；这些细胞都有或间接有杀伤外源微生物感染细胞以及癌变细胞的作用，因此IL-2有很好的抗病毒、抗癌作用和广泛的临床应用潜力。
IL-2通过结合IL-2受体(IL-2R)来介导其作用，IL-2受体由3个亚基组成，分别为α(CD25)、β(CD122)和γ(CD132)受体亚基，其中α受体主要表达在T抑制细胞(Treg)和一些内皮细胞(endothelial cells)表面，而β和γ受体亚基则高表达于效应性T细胞(Teff)和NK细胞。IL-2对不同受体亚基的复合物形式的亲和力是不同的，IL-2对α、β和γ受体亚基组成的复合体的亲和力是最高的，对由β和γ受体亚基组成的复合体的亲和力则为中等(约降低100倍)，而IL-2与两种形式的受体亚基组合结合后均能传递信号(Minami等，Annu RevImmunol 11，245-268(1993))。但是临床在IL-2低剂量的条件下，是会优先和Treg细胞表面上的高亲和力受体结合，则会产生免疫抑制，达不到治疗的效果。高剂量的IL-2会通过激活大量的效应T细胞从而中和Treg激活带来的免疫抑制，同时也会出现更多的毒副作用，以及细胞凋亡(activation induced cell apoptosis)。
基于IL-2的抗肿瘤作用，高剂量IL-2(阿地白介素)于1992年通过FDA批准用于黑色素瘤和肾细胞癌的临床治疗。但是接受高剂量IL-2治疗的患者经常经历严重的副作用，包括心血管、肺水肿、肝、胃肠、神经学和血液学等事件，大多数的这些副作用可由血管(或毛细管)渗漏综合征(VLS)的来解释，也是临床和动物实验评价IL-2治疗副作用的一项指标。而引起VLS是由于内皮细胞上表达有IL-2的高亲和力受体(α、β和γ亚基)(Krieg等，Proc Nat Acad Sci USA107,11906-11(2010))，所以减弱或消除与α受体的结合将有助于减弱IL-2促进抑制T细胞增殖活性的功能；同时还可减少对内皮细胞α受体的结合，从而降低或消除IL-2治疗引起的毒副作用。IL-2与α受体亚基的结合位点主要在第37，38，41，42，43，44，45，61，62，65，68和72位氨基酸位点(Rickert.M等(2005)Science308:1477-1480)，Merck和Roche公司或其它科研机构围绕这些与α受体亚基结合的IL-2表面氨基酸做了一些突变，如Merck公司的突变体(R38W、F42K,WO2008003473A2)，降低与α受体亚基的相互作用，以其达到效应T细胞活化以增强功效；而Roche的IL-2突变体(F42A、Y45A和L72G,US 2016/0208017A1)，其与α受体不结和，但可以正常结合β和γ受体亚基复合物，并可以发挥效应，目前正在临床中。
因此降低或消除IL-2与α受体亚基的相互作用，可能是治疗有效性并减少肿瘤患者治疗副作用的一个重要方面。
发明内容
有鉴于现有技术中的缺陷，本发明提供了一种IL-2衍生物及起复合物。
本发明的一个方面提供了一种IL-2衍生物，在一个具体实施方式中，该IL-2衍生物通过在野生型IL-2的基础上引入至少一个半胱氨酸残基，部分封闭或完全封闭IL-2衍生物与α受体亚基的结合平面，同时基本保留对β和γ受体亚基复合物的亲和力。野生型IL-2的氨基酸序列如SEQ ID NO.1所示。
进一步地，在野生型IL-2的基础上引入的至少一个半胱氨酸残基，能够：
a)形成IL-2衍生物的分子内二硫键；或者
b)使得IL-2衍生物能通过分子间二硫键与封闭模块结合。
可选地，所述引入至少一个半胱氨酸残基为通过点突变的方式引入至少一个半胱氨酸残基。
可选地，在一种方式中，在野生型IL-2的基础上通过点突变方式引入第一半胱氨酸残基和第二半胱氨酸残基；所述第一半胱氨酸残基和所述第二半胱氨酸残基中的一个或两个为野生型IL-2与α受体亚基的结合平面相关的氨基酸或其附近的氨基酸。其中，野生型IL-2与α受体亚基的结合平面相关的氨基酸位点为第37位、第38位、第41位、第42位、第43位、第44位、第45位、第61位、第62位、第65位、第68位和第72位。
进一步地，第一半胱氨酸残基为野生型IL-2的第37位、第38位、第41位、第42位、第43位、第44位、第45位、第61位、第62位氨基酸或其附近的氨基酸；第二半胱氨酸残基为野生型IL-2的第61位、第62位、第65位、第68位、第72位氨基酸或其附近的氨基酸。可选地，“附近”为：1)一级结构上相邻的1～4个氨基酸；和/或三级结构上相邻的氨基酸。
进一步地，第一半胱氨酸残基为选自下组的氨基酸点突变：K35C、L36C、R38C、M39C、L40C、T41C、F42C、K43C、F44C和E61C。
进一步地，第二半胱氨酸残基为选自下组的氨基酸点突变：V69C、E62C、P65C、T111C、Y107C、A112C、T113C、I114C、L72C和A73C。
可选地，形成分子内二硫键的第一半胱氨酸残基和第二半胱氨酸残基的组合选自下组的氨基酸点突变组合：M39C和V69C、F44C和E62C、F44C和P65C、F42C和V69C、E61C和Y107C、F42C和P65C、F42C和T111C、F42C和A112C、F42C和T113C、T41C和A112C、L40C和A112C、L40C和T113C、L40C和I114C、M39C和L72C、M39C和A73C、R38C和V69C、R38C和L72C、L36C和V69C、L36C和L72C、L36C和A73C、K35C和V69C以及K43C和A112C。
可选地，第一半胱氨酸残基和第二半胱氨酸残基的残基质心矢量距离为小于
Figure BDA0002322160500000031
可选地，在第二种方式中，在野生型IL-2的基础上通过点突变方式引入第三半胱氨酸残基；所述第三半胱氨酸残基为野生型IL-2与α受体亚基的结合平面相关的氨基酸或其附近的氨基酸。
进一步地，第三半胱氨酸残基为野生型IL-2的第37位、第38位、第41位、第42位、第43位、第44位、第45位、第61位、第62位、第65位、第68位、第72位氨基酸或其附近的氨基酸。可选地，“附近”为：1)一级结构上相邻的1～4个氨基酸；和/或三级结构上相邻的氨基酸。
进一步地，所述第三半胱氨酸残基为选自下组的氨基酸点突变：P34C、K35C、T37C、R38C、T41C、K43C、F44C、Y45C、E61C、E62C、K64C、P65C、E68C和L72C。
进一步地，封闭模块上具有或引入有第四半胱氨酸残基；IL-2衍生物上的第三半胱氨酸残基与所述封闭模块上的第四半胱氨酸残基能形成分子间二硫键。
可选地，第三半胱氨酸残基和所述第四半胱氨酸残基的残基质心矢量距离为小于
Figure BDA0002322160500000032
可选地，封闭模块为α受体亚基的胞外段。
进一步地，α受体亚基的胞外段的氨基酸序列如SEQ ID NO.24所示。
进一步地，α受体亚基的胞外段上的所述第四半胱氨酸残基位选自下组的氨基酸点突变：D4C、D5C、M25C、N27C、R35C、R36C、K38C、S39C、G40C、S41C、L42C、I118C、Y119C和H120C。
可选地，形成分子间二硫键的第三半胱氨酸残基和第四半胱氨酸残基的组合选自下组的氨基酸点突变组合：T41C和N27C、P34C和D4C、E68C和L42C、Y45C和R35C、R38C和H120C、L72C和M25C、E61C和S39C、T41C和I118C、K35C和D4C、T37C和D4C、R38C和D4C、R38C和D5C、T41C和L42C、T41C和Y119C、K43C和R35C、K43C和R36C、F44C和L42C、K43C和L42C、E61C和K38C、E62C和K38C、K64C和S39C、K64C和G40C、K64C和S41C、P65C和K38C。
无论第一种方式还是第二种方式，可选地，通过点突变的方式将野生型IL-2第125位的半胱氨酸残基转变为其他的氨基酸残基。
进一步可选地，野生型IL-2第125位的点突变为C125A。
可选地，IL-2衍生物的氨基酸序列如SEQ ID NO.3～SEQ ID NO.24所示；或如SEQID NO.26～40所示。
本发明的第二个方面提供了一种复合物，在一个具体实施方式中，该复合物包括：
1)IL-2衍生物，所述IL-2衍生物在野生型IL-2的基础上引入第三半胱氨酸残基；
2)封闭模块，所述封闭模块上具有或引入有第四半胱氨酸残基；
所述IL-2衍生物上的第三半胱氨酸残基和所述封闭模块上的第四半胱氨酸残基形成分子间二硫键，从而形成IL-2与封闭模块的复合物，部分封闭或完全封闭IL-2衍生物与α受体亚基的结合平面，同时基本保留对β和γ受体亚基复合物的亲和力。
可选地，引入至少一个半胱氨酸残基为通过点突变的方式引入至少一个半胱氨酸残基。
进一步地，所述第三半胱氨酸残基为野生型IL-2与α受体亚基的结合平面相关的氨基酸或其附近的氨基酸。
进一步地，所述第三半胱氨酸残基为野生型IL-2的第37位、第38位、第41位、第42位、第43位、第44位、第45位、第61位、第62位、第65位、第68位、第72位氨基酸或其附近的氨基酸。可选地，“附近”为：1)一级结构上相邻的1～4个氨基酸；和/或三级结构上相邻的氨基酸。
可选地，所述第三半胱氨酸残基和所述第四半胱氨酸残基的残基质心矢量距离为小于
Figure BDA0002322160500000041
进一步地，第三半胱氨酸残基为选自下组的氨基酸点突变：P34C、K35C、T37C、R38C、T41C、K43C、F44C、Y45C、E61C、E62C、K64C、P65C、E68C和L72C。
可选地，所述封闭模块为α受体亚基的胞外段，其氨基酸序列如SEQ ID NO.25所示。
进一步地，α受体亚基的胞外段上的所述第四半胱氨酸残基位选自下组的氨基酸点突变：D4C、D5C、M25C、N27C、R35C、R36C、K38C、S39C、G40C、S41C、L42C、I118C、Y119C和H120C。
可选地，形成分子间二硫键的第三半胱氨酸残基和第四半胱氨酸残基的组合选自下组的氨基酸点突变组合：T41C和N27C、P34C和D4C、E68C和L42C、Y45C和R35C、R38C和H120C、L72C和M25C、E61C和S39C、T41C和I118C、K35C和D4C、T37C和D4C、R38C和D4C、R38C和D5C、T41C和L42C、T41C和Y119C、K43C和R35C、K43C和R36C、F44C和L42C、K43C和L42C、E61C和K38C、E62C和K38C、K64C和S39C、K64C和G40C、K64C和S41C、P65C和K38C。
可选地，野生型IL-2第125位的点突变为C125A。
可选地，IL-2衍生物的氨基酸序列和所述α受体亚基的胞外段的序列组合选自下组的组合：SEQ ID NO.26和SEQ ID NO.50，SEQ ID NO.27和SEQ ID NO.51，SEQ ID NO.28和SEQ ID NO.52，SEQ ID NO.29和SEQ ID NO.53，SEQ ID NO.30和SEQ ID NO.54，SEQ IDNO.31和SEQ ID NO.55，SEQ ID NO.32和SEQ ID NO.56，SEQ ID NO.33和SEQ ID NO.57，SEQID NO.34和SEQ ID NO.58，SEQ ID NO.35和SEQ ID NO.59，SEQ ID NO.36和SEQ ID NO.60，SEQ ID NO.37和SEQ ID NO.61，SEQ ID NO.38和SEQ ID NO.62，SEQ ID NO.39和SEQ IDNO.63，SEQ ID NO.40和SEQ ID NO.64，SEQ ID NO.41和SEQ ID NO.65，SEQ ID NO.42和SEQID NO.66，SEQ ID NO.43和SEQ ID NO.67，SEQ ID NO.44和SEQ ID NO.68，SEQ ID NO.45和SEQ ID NO.69，SEQ ID NO.46和SEQ ID NO.70，SEQ ID NO.47和SEQ ID NO.71，SEQ IDNO.48和SEQ ID NO.72，SEQ ID NO.49和SEQ ID NO.73。
本发明的第三个方面提供了一种分离的多核苷酸，在一个具体实施方式中，其编码如上所述的IL-2衍生物，或者编码如上所述的复合物。
本发明的第四个方面提供了一种表达载体，在一个具体实施方式中，其包含如上所述的一种分离的多核苷酸。
本发明的第五个方面提供了一种宿主细胞，在一个具体实施方式中，包含如上所述的一种分离的多核苷酸。
本发明的第六个方面提供了一种组合物，在一个具体实施方式中，该组合物包含如上所述的IL-2衍生物或者如上所述的复合物，以及药学可接受载体。
本发明的第七个方面提供如上所述的IL-2衍生物或者如上所述的复合物用于制备用于治疗疾病的药物或制剂中的用途。
本发明的第八个方面提供如上所述的IL-2衍生物或者如上所述的复合物在制备用于刺激个体的免疫系统的组合物中的用途。
本发明的第九个方面提供了一种生成IL-2衍生物的方法，在一个具体实施方式中，该方法包括在适合于表达所述IL-2衍生物的条件下培养如上所述的宿主细胞。
本发明的第十个方面提供了一种生成复合物的方法，在一个具体实施方式中，该方法包括在适合于表达所述复合物的条件下培养如权利要求37所述的宿主细胞。
本发明的具体实施方式中的IL-2衍生物或复合物，通过形成分子内或分子间二硫键的方式，使得IL-2衍生物与α受体结合的位点被封闭，从而避免与α受体结合结构。本发明中的IL-2衍生物或复合物为减少VLS或降低或消除IL-2治疗引起的毒副作用带来的新的方向。
附图说明
图1是本发明一个实施例中纯化后的IL-2衍生物SDS-PAGE电泳；其中，还原为上样缓冲液中加入还原剂β-ME，非还原为不加还原剂。
图2是本发明一个实施例中Fortebio测试IL-2衍生物与IL2Rα结合能力测定信号图，浓度为100nM；使用的不相关的蛋白对照为HER2。其中，图2A针对IL-2wt C125A；图2B针对IL-2突变体1；图2C针对IL-2突变体2；图2D针对IL-2突变体3；图2E针对IL-2突变体4；图2F针对IL-2复合物1；图2G针对IL-2复合物2；图2H针对IL-2复合物3；图2I针对IL-2复合物4。
图3是本发明一个实施例中Fortebio测试IL-2衍生物与IL2Rβγ结合亲和力测定信号图以及Ka,kd,KD和R2，浓度范围是1.25nM-40nM。其中，图3A针对IL-2wt C125A；图3B针对IL-2突变体1；图3C针对IL-2突变体2；图3D针对IL-2突变体3；图3E针对IL-2突变体4；图3F针对IL-2复合物1；图3G针对IL-2复合物2；图3H针对IL-2复合物3；图3I针对IL-2复合物4。
图4是本发明一个实施例中CTLL-2(T细胞)的增殖实验。
具体实施方式
以下将结合实施例对本发明作进一步地说明，应理解这些实施例仅作为例证的目的，不用于限制本发明的保护范围。
下列实施例中未注明具体条件的实验方法，通常按照常规条件，例如Sambrook等分子克隆：实验室手册(New York:Cold Spring Harbor Laboratory Press,1989)中所述的条件，或按照制造厂商所建议的条件。所采用的试剂，若无特殊说明，均为市售或公开渠道可以获得的试剂。
本文中，IL-2衍生物和野生型IL-2的相关氨基酸位置，以野生型IL-2的氨基酸序列(如SEQ ID NO.1)为基准进行计算。
野生型IL-2(IL-2wt，SEQ ID NO.1)：
APTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTRMLTFKFYMPKK ATELKHLQCLEEELKPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTF MCEYADETATIVEFLNRWITFCQSIISTLT。
野生型IL-2的核苷酸序列如SEQ ID NO.146所示。
本文中，CD25-ECD的相关氨基酸位置，以SEQ ID NO.25所示的氨基酸序列为基准进行计算。
本文中，“第一”、“第二”、“第三”等，仅是为了进行区分，不是顺序的限定。例如，一个衍生物中可以仅具有“第三”半胱氨酸残基，而不必具有“第一”和“第二”半胱氨酸残基。
本文中，“IL-2衍生物”包括IL-2突变体、IL-2突变体与其他分子形成的复合物。IL-2突变体是指在野生型IL-2的基础上进行突变(比如点突变或插入突变)所形成的分子。
本文中，“IL2Rα”是指白介素-2受体α，也称为“α受体亚基”；“IL2Rβ”是指白介素-2受体β，也称为“β受体亚基”；“IL2Rγ”是指白介素-2受体γ，也称为“γ受体亚基”；“IL2Rβγ”是指白介素-2受体β和受体γ形成的复合物，也称为“β和γ受体亚基复合物”。
现有技术中，为了能够降低或消除IL-2与α受体的结合，都是对IL-2与α受体的结合表面内的氨基酸进行简单突变。
本发明的具体实施方式中，采用的策略是通过引入额外的半胱氨酸残基，使得IL-2分子内部形成新的二硫键，或者使得IL-2通过分子间二硫键与另外的封闭模块形成复合物，从而部分封闭或完全封闭IL-2与α受体的结合位点，但是也不影响与β和γ受体亚基复合物的结合。
在第一个具体实施方式中，是通过引入第一半胱氨酸残基和第二半胱氨酸残基，形成IL-2分子内二硫键，使得IL-2衍生物从结构上更加稳定，而且还可以形成屏障，破坏与α受体的结合平面。
引入第一半胱氨酸残基和第二半胱氨酸残基的方式是在野生型IL-2的基础上进行合适的点突变。关于需要引入的第一半胱氨酸残基和第二半胱氨酸残基的位置，主要通过以下方式确定：
1)第一和第二半胱氨酸残基的位置为IL-2和α受体的结合面上的氨基酸残基或其附近的氨基酸残基；IL-2和α受体的结合面上的氨基酸残基为第37，38，41，42，43，44，45，61，62，65，68和72位氨基酸位点；
2)充分考虑到IL-2的结构和原子间的距离；
3)通过生物信息学和蛋白质工程学设计，在上述氨基酸残基周围根据残基质心的矢量距离判据，寻找两个合适的位点，其能形成分子内二硫键，但是不容易或不会形成IL-2分子间二硫键。
确定合适的位点后，通过突变将原氨基酸残基突变成第一半胱氨酸残基和第二半胱氨酸残基。突变后的IL-2衍生物(及IL-2突变体)通过正常的转录和翻译后形成分子内的二硫键。
在一些实施例中，将IL-2第125位原有的自由半胱氨酸突变掉，防止其干扰分子间二硫键的形成。
在一些实施例中，设计获得的IL-2突变体的氨基酸序列如表1所示：
表1设计获得的IL-2突变体的氨基酸序列
Figure BDA0002322160500000081
Figure BDA0002322160500000091
Figure BDA0002322160500000101
表达宿主可以是E.Coli或哺乳动物细胞。
在第二个具体实施方式中，IL-2通过分子间二硫键与封闭模块形成复合物，从而全部或部分封闭IL-2与α受体的结合平面，进而阻断IL-2与内源性α受体的结合。IL-2与封闭模块之间的二硫键，是通过在野生型IL-2上引入第三半胱氨酸残基，在封闭模块上具有或引入第四半胱氨酸残基，由第三半胱氨酸残基和第四半胱氨酸残基之间形成的。
在一个实施例中，封闭模块是α受体的胞外段。野生型α受体的胞外段的氨基酸序列如SEQ ID NO.25所示。在天然状态下，α受体与IL-2的结合并不稳定(高解离系数Kd)，不能形成稳定的异源二聚体，必须与β和γ受体亚基共同构成IL-2的高亲和力的受体。因此，无法通过共表达两种野生型的分子而形成稳定的复合物。但是，在野生型IL-2上引入第三半胱氨酸残基，在α受体的胞外段(CD25-ECD)上引入第四半胱氨酸残基，通过第三半胱氨酸残基和第四半胱氨酸残基之间形成的二硫键，使IL-2衍生物和α受体的胞外段(CD25-ECD)形成复合物，阻断IL-2与内源性α受体的结合。
野生型α受体的胞外段的氨基酸序列如SEQ ID NO.25所示：
ELCDDDPPEIPHATFKAMAYKEGTMLNCECKRGFRRIKSGSLYMLCTGNSSHSSWDNQCQCTSSATRNTTKQVTPQPEEQKERKTTEMQSPMQPVDQASLPGHCREPPPWENEATERIYHFVVGQMVYYQCVQGYRALHRGPAESVCKMTHGKTRWTQPQLICTG；
野生型α受体的胞外段的核苷酸序列如SEQ ID NO.145所示。
关于需要引入的第三半胱氨酸残基和第四半胱氨酸残基的位置，主要通过以下方式确定：
1)第三半胱氨酸残基的位置为IL-2和α受体的结合面上的氨基酸残基或其附近的氨基酸残基；IL-2和α受体的结合面上的氨基酸残基为第37，38，41，42，43，44，45，61，62，65，68和72位氨基酸位点；
2)通过生物信息学和蛋白质工程学设计，分别在IL-2和α受体胞外区(CD25-ECD)根据残基质心的矢量距离判据，寻找合适的结合面位点，该突变不影响蛋白质的结构。
确定合适的位点后，通过突变将IL-2上的原氨基酸残基突变成第三半胱氨酸残基，将α受体胞外区(CD25-ECD)上的原氨基酸残基突变成第四半胱氨酸残基。IL-2和α受体胞外区的突变体进行共表达之后，通过转录和翻译后，可以在IL-2与α受体胞外区之间形成二硫键，形成一个全新的分子IL-2/CD25-ECD异源二聚体(heterodimer)。该复合物无法与体内的内源性α受体结合，但是可以与β和γ受体亚基复合物结合，从而达到不激活Treg的目的。
在一个实施例中，将IL-2第125位原有的自由半胱氨酸突变掉，防止其在与带有半胱氨酸突变的α受体的胞外段及IL-2形成额外的二硫键，影响α受体与IL-2二聚体的形成。
在一些实施例中，设计获得的IL-2突变体和CD25-ECD突变体的氨基酸序列如表2所示：
表2设计获得的IL-2突变体和CD25-ECD突变体的氨基酸序列
Figure BDA0002322160500000121
Figure BDA0002322160500000131
Figure BDA0002322160500000141
Figure BDA0002322160500000151
Figure BDA0002322160500000161
Figure BDA0002322160500000171
Figure BDA0002322160500000181
Figure BDA0002322160500000191
Figure BDA0002322160500000201
表达宿主为哺乳动物细胞(HEK293或CHO)。
实施例1 IL-2(C125A)、IL-2突变体和IL-2复合物的制备
本实施例中挑选IL-2wt(C125A)，IL-2突变体1～4和IL-2复合物1～4进行分别表达，并依靠分子C端带有的HPC4标签进行纯化和制备。
1.1表达质粒构建
委托苏州金唯智生物科技有限公司合成带有IL-2wt C125A(SEQ ID NO.74)、IL-2突变体1～4、IL-2复合物1～4(IL-2复合物中IL-2Pair 1～4和CD25-ECD Pair 1～4)的基因，然后或按照《分子克隆》提到的操作方法，进行重叠PCR得到目的片段。然后进行片段和pTT5通用载体的重组连接、转化DH10B、测序和保菌，从而得到需要的IL-2wt(C125A)，IL-2突变体1～4和IL-2复合物1～4的质粒(IL-2复合物中IL-2Pair 1～4的质粒和CD25-ECDPair 1～4的质粒)；或按照《Agilent QuikChange Lightning Site-DirectedMutagenesis Kit》提到的操作方法，进行PCR、DpnI消化、转化DH10B、测序和保菌，从而得到需要的IL-2wt(C125A)，IL-2突变体1～4和IL-2复合物1～4的质粒。
1.2质粒提取及HEK293细胞准备
1.2.1质粒提取
按照《Qiagen Mini-prep Kit》和《Qiagen Endofree Maxi-prep Kit》提到的操作方法，进行IL-2wt(C125A)，IL-2突变体1～4和IL-2复合物1～4的质粒的制备。
1.2.2 HEK293细胞准备
新鲜传代的密度为1-1.2×10^6/ml的HEK293细胞(National Research Council,Canada)用于瞬时表达。
1.3 HEK293瞬时表达
1.3.1试剂制备
A)G418溶液：称取250mg GeneticinTM加入4.5ml超纯水溶解，超纯水定容至5ml，0.22um滤膜过滤，-20℃保存；
B)PEI溶液：称取50mgPEI加入45ml超纯水溶解，1M NaOH调节pH至7.0，超纯水定容至50ml，0.22um滤膜过滤，-20℃保存；
C)培养基：在1L FreeStyleTM 293Expression Medium中加入10ml PluronicdTMF-68和500ul G418；
D)质粒提前准备在2ml去内毒素离心管中；
E)根据转染需要体积准备好新鲜传代至1-1.2×106个/ml的细胞悬液。
1.3.2配制转染试剂-质粒复合物
A液：质粒1ug/ml+Opti-MEMTM 33.3ul/ml
B液：PEI 2ug/ml+Opti-MEMTM 33.3ul/ml
将B液倒入A液混匀孵育10min后，加入细胞悬液。
转染中，IL-2wt(C125A)，IL-2突变体1～4的质粒分别单独转染；IL-2复合物1～4的质粒，是将两种质粒混合后，进行转染。
1.3.3换液
在115rpm，36.8℃，5％CO2培养4h后，800g 5min离心，换成未添加F68和G418的FreeStyleTM 293Expression Medium
1.3.4表达培养和收获
在115rpm，36.8℃，5％CO2培养5天后，离心8500rpm 15min，收集细胞上清。
1.4纯化制备
所有IL-2相关的衍生物C端都带有HPC4标签，因此可以用偶联有HPC4抗体的填料进行亲和纯化，然后再经过凝胶过滤层析(superdex200)进行进一步纯化得到纯度较高的蛋白。按照《分子克隆》描述的方法进行SDS-PAGE分析。
结果如图1所示，表明质粒转染细胞后，都生成了IL-2wt(C125A)，IL-2突变体1～4和IL-2复合物1～4相应的蛋白和复合物，质粒构建及蛋白表达及纯化成功。
实施例2通过生物膜干涉技术(biolayer interferomeory，BLI)测定IL-2wt(C125A)，IL-2突变体1～4和IL-2复合物1～4分别与IL2Rβγ和IL2Rα的亲和力
1、实验材料
实验所用蛋白均为北京志道生物科技有限公司生产，IL2Rα-his(购自北京志道生物科技有限公司)，IL2Rβγ-Fc(购自北京志道生物科技有限公司)以及IL2突变体通过HEK293瞬时表达以及亲和纯化后获得。缓冲液配方：10mM HEPES，150mM氯化钠，3mM EDTA，0.1％BSA和0.05％吐温20；ProA传感器(Pall Fortebio公司，货号#18-5010)；HISIK传感器(Pall Fortebio公司，货号#18-5120)；BLI设备为Pall Fortebio公司生产的Octet RED96；数据获取和分析工作分别采用Data acquisition 11.0和Data analysis 11.0软件进行。
2、实验方法
1)IL2Rβγ-Fc准备
用缓冲液稀释IL2Rβγ-Fc至浓度10ug/ml，加入96孔测定板第2列，控制程序中设为Loading，600s。
2)IL2Rα-his准备
用缓冲液稀释IL2Rα-his至浓度10ug/ml，加入96孔测定板第3列，控制程序中设为Loading，600s.
3)样品准备
用缓冲液将IL-2衍生物稀释到100nM，然后1：1向下系列稀释6个梯度(共7个梯度)至浓度为1.625nM以及0浓度，分别加入96孔测定板5-9列，控制程序中设定为Association，200s。在96孔测定板第1，4，10，11列加入缓冲液，12列加入PH1.7的甘氨酸，上述样品和溶液的加样量均为200ul。
4)检测与IL2Rβγ-Fc的亲和力
取8个ProA传感器分别置于传感器支架第1列的A-H，在Data acquisition11.0软件中将检测条件设置如下：1预湿：Baseline,60s,位置：第1列。2循环检测：第2列：loading，600s；第4列：Baseline1，60s；样品5-9列：Association，200s，第10列：Dissociation，600s；第11列：中和；第12列：再生。
5)检测与IL2Rα的结合
取8个HISIK传感器分别置于传感器支架第2列的A-H，在Data acquisition11.0软件中将检测条件设置如下：1预湿：Baseline,60s,位置：第1列。2循环检测：第3列：loading，600s；第4列：Baseline1，60s；样品5-9列：Association，200s，第10列：Dissociation，600s；第11列：中和；第12列：再生。
6)数据分析
使用Data analysis 11.0软件对数据进行分析。以0浓度为对照扣减背景，通过Fitting curve计算KD值。
3、结果
如图2所示，从IL-2衍生物与IL2Rα受体结合曲线上看，除了IL-2突变体1大大减弱了与IL2Rα受体的结合外，其余IL-2衍生物几乎都不与其结合；如图3所示，从IL-2衍生物与IL2Rβγ受体结合曲线上看，和IL-2wt C125A相比，都没有显著变化(图3)。
实施例3促进T细胞的增殖实验
CTLL-2(T细胞)的增殖实验是普遍应用的细胞水平测定白介素刺激免疫细胞活性的实验。因此，此处通过CTLL-2细胞的增殖实验来查看IL-2衍生物的生物活性。
1)CTLL-2细胞准备：将细胞用含有FBS和Rat-T-Stim的培养液重悬起来。
2)加样：将细胞接种于96孔培养板，每孔0.1ml。同时将待测样品IL-2衍生物11、18、21和28的蛋白(即实施例1中制备的蛋白)分别做倍比稀释，每孔加入0.1ml，每个稀释浓度均设3复孔。另设培养液对照孔(100ul细胞+100ul培养液)。37度，5％CO2孵育72小时。
3)MTS加入：每孔加入20μl CellTiter
Figure BDA0002322160500000231
AQueous One Solution Reagent，37度，5％CO2孵育2～4小时。
4)测定：用酶标测定仪于波长490nm测吸光值(A)并计算EC50值。
5)结果：挑选的具有代表性的IL-2复合物2和IL-2突变体4，他们都具有CTLL-2(T细胞)的增殖活性，说明没有严重影响β和γ受体亚基复合物的信号传导功能(图4)。
序列表
<110> 北京志道生物科技有限公司
<120> 白介素-2衍生物
<160> 146
<170> SIPOSequenceListing 1.0
<210> 1
<211> 133
<212> PRT
<213> Homo sapiens
<400> 1
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 2
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 2
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 3
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 3
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Cys Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Cys Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 4
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 4
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Cys Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Cys Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 5
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 5
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Cys Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Cys Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 6
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 6
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Cys Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Cys Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 7
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 7
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Cys Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Cys Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 8
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 8
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Cys Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Cys Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 9
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 9
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Cys Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Cys Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 10
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 10
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Cys Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Cys
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 11
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 11
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Cys Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Cys Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 12
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 12
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Cys Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Cys
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 13
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 13
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Cys Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Cys
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 14
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 14
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Cys Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Cys Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 15
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 15
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Cys Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Cys Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 16
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 16
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Cys Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Cys Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 17
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 17
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Cys Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Cys Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 18
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 18
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Cys Met Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Cys Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 19
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 19
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Cys Met Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Cys Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 20
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 20
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Cys Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Cys Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 21
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 21
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Cys Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Cys Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 22
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 22
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Cys Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Cys Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 23
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 23
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Cys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Cys Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 24
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 24
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Cys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Cys
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 25
<211> 165
<212> PRT
<213> Artificial Sequence
<400> 25
Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys
1 5 10 15
Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg
20 25 30
Gly Phe Arg Arg Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly
35 40 45
Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser
50 55 60
Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln
65 70 75 80
Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp
85 90 95
Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn
100 105 110
Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr
115 120 125
Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu
130 135 140
Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln
145 150 155 160
Leu Ile Cys Thr Gly
165
<210> 26
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 26
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Cys Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 27
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 27
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Cys Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 28
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 28
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Cys Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 29
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 29
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Cys Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 30
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 30
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Cys Met Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 31
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 31
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Cys Met Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Cys Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 32
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 32
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Cys Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 33
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 33
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Cys Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 34
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 34
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Cys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 35
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 35
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Cys Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 36
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 36
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Cys Met Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 37
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 37
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Cys Met Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 38
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 38
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Cys Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 39
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 39
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Cys Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 40
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 40
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Cys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 41
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 41
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Cys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 42
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 42
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Cys Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 43
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 43
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Cys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 44
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 44
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Cys Glu Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 45
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 45
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Cys Leu Lys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 46
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 46
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Cys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 47
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 47
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Cys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 48
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 48
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Cys
50 55 60
Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 49
<211> 133
<212> PRT
<213> Artificial Sequence
<400> 49
Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His
1 5 10 15
Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys
20 25 30
Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys
35 40 45
Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys
50 55 60
Cys Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu
65 70 75 80
Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu
85 90 95
Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala
100 105 110
Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ala Gln Ser Ile
115 120 125
Ile Ser Thr Leu Thr
130
<210> 50
<211> 165
<212> PRT
<213> Artificial Sequence
<400> 50
Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys
1 5 10 15
Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Cys Cys Glu Cys Lys Arg
20 25 30
Gly Phe Arg Arg Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly
35 40 45
Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser
50 55 60
Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln
65 70 75 80
Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp
85 90 95
Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn
100 105 110
Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr
115 120 125
Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu
130 135 140
Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln
145 150 155 160
Leu Ile Cys Thr Gly
165
<210> 51
<211> 165
<212> PRT
<213> Artificial Sequence
<400> 51
Glu Leu Cys Cys Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys
1 5 10 15
Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg
20 25 30
Gly Phe Arg Arg Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly
35 40 45
Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser
50 55 60
Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln
65 70 75 80
Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp
85 90 95
Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn
100 105 110
Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr
115 120 125
Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu
130 135 140
Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln
145 150 155 160
Leu Ile Cys Thr Gly
165
<210> 52
<211> 165
<212> PRT
<213> Artificial Sequence
<400> 52
Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys
1 5 10 15
Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg
20 25 30
Gly Phe Arg Arg Ile Lys Ser Gly Ser Cys Tyr Met Leu Cys Thr Gly
35 40 45
Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser
50 55 60
Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln
65 70 75 80
Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp
85 90 95
Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn
100 105 110
Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr
115 120 125
Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu
130 135 140
Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln
145 150 155 160
Leu Ile Cys Thr Gly
165
<210> 53
<211> 165
<212> PRT
<213> Artificial Sequence
<400> 53
Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys
1 5 10 15
Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg
20 25 30
Gly Phe Cys Arg Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly
35 40 45
Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser
50 55 60
Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln
65 70 75 80
Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp
85 90 95
Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn
100 105 110
Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr
115 120 125
Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu
130 135 140
Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln
145 150 155 160
Leu Ile Cys Thr Gly
165
<210> 54
<211> 165
<212> PRT
<213> Artificial Sequence
<400> 54
Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys
1 5 10 15
Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg
20 25 30
Gly Phe Arg Arg Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly
35 40 45
Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser
50 55 60
Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln
65 70 75 80
Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp
85 90 95
Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn
100 105 110
Glu Ala Thr Glu Arg Ile Tyr Cys Phe Val Val Gly Gln Met Val Tyr
115 120 125
Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu
130 135 140
Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln
145 150 155 160
Leu Ile Cys Thr Gly
165
<210> 55
<211> 165
<212> PRT
<213> Artificial Sequence
<400> 55
Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys
1 5 10 15
Ala Met Ala Tyr Lys Glu Gly Thr Cys Leu Asn Cys Glu Cys Lys Arg
20 25 30
Gly Phe Arg Arg Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly
35 40 45
Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser
50 55 60
Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln
65 70 75 80
Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp
85 90 95
Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn
100 105 110
Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr
115 120 125
Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu
130 135 140
Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln
145 150 155 160
Leu Ile Cys Thr Gly
165
<210> 56
<211> 165
<212> PRT
<213> Artificial Sequence
<400> 56
Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys
1 5 10 15
Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg
20 25 30
Gly Phe Arg Arg Ile Lys Cys Gly Ser Leu Tyr Met Leu Cys Thr Gly
35 40 45
Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser
50 55 60
Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln
65 70 75 80
Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp
85 90 95
Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn
100 105 110
Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr
115 120 125
Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu
130 135 140
Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln
145 150 155 160
Leu Ile Cys Thr Gly
165
<210> 57
<211> 165
<212> PRT
<213> Artificial Sequence
<400> 57
Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys
1 5 10 15
Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg
20 25 30
Gly Phe Arg Arg Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly
35 40 45
Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser
50 55 60
Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln
65 70 75 80
Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp
85 90 95
Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn
100 105 110
Glu Ala Thr Glu Arg Cys Tyr His Phe Val Val Gly Gln Met Val Tyr
115 120 125
Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu
130 135 140
Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln
145 150 155 160
Leu Ile Cys Thr Gly
165
<210> 58
<211> 165
<212> PRT
<213> Artificial Sequence
<400> 58
Glu Leu Cys Cys Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys
1 5 10 15
Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg
20 25 30
Gly Phe Arg Arg Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly
35 40 45
Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser
50 55 60
Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln
65 70 75 80
Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp
85 90 95
Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn
100 105 110
Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr
115 120 125
Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu
130 135 140
Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln
145 150 155 160
Leu Ile Cys Thr Gly
165
<210> 59
<211> 165
<212> PRT
<213> Artificial Sequence
<400> 59
Glu Leu Cys Cys Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys
1 5 10 15
Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg
20 25 30
Gly Phe Arg Arg Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly
35 40 45
Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser
50 55 60
Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln
65 70 75 80
Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp
85 90 95
Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn
100 105 110
Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr
115 120 125
Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu
130 135 140
Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln
145 150 155 160
Leu Ile Cys Thr Gly
165
<210> 60
<211> 165
<212> PRT
<213> Artificial Sequence
<400> 60
Glu Leu Cys Cys Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys
1 5 10 15
Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg
20 25 30
Gly Phe Arg Arg Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly
35 40 45
Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser
50 55 60
Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln
65 70 75 80
Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp
85 90 95
Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn
100 105 110
Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr
115 120 125
Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu
130 135 140
Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln
145 150 155 160
Leu Ile Cys Thr Gly
165
<210> 61
<211> 165
<212> PRT
<213> Artificial Sequence
<400> 61
Glu Leu Cys Asp Cys Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys
1 5 10 15
Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg
20 25 30
Gly Phe Arg Arg Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly
35 40 45
Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser
50 55 60
Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln
65 70 75 80
Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp
85 90 95
Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn
100 105 110
Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr
115 120 125
Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu
130 135 140
Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln
145 150 155 160
Leu Ile Cys Thr Gly
165
<210> 62
<211> 165
<212> PRT
<213> Artificial Sequence
<400> 62
Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys
1 5 10 15
Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg
20 25 30
Gly Phe Arg Arg Ile Lys Ser Gly Ser Cys Tyr Met Leu Cys Thr Gly
35 40 45
Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser
50 55 60
Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln
65 70 75 80
Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp
85 90 95
Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn
100 105 110
Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr
115 120 125
Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu
130 135 140
Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln
145 150 155 160
Leu Ile Cys Thr Gly
165
<210> 63
<211> 165
<212> PRT
<213> Artificial Sequence
<400> 63
Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys
1 5 10 15
Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg
20 25 30
Gly Phe Arg Arg Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly
35 40 45
Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser
50 55 60
Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln
65 70 75 80
Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp
85 90 95
Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn
100 105 110
Glu Ala Thr Glu Arg Ile Cys His Phe Val Val Gly Gln Met Val Tyr
115 120 125
Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu
130 135 140
Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln
145 150 155 160
Leu Ile Cys Thr Gly
165
<210> 64
<211> 165
<212> PRT
<213> Artificial Sequence
<400> 64
Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys
1 5 10 15
Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg
20 25 30
Gly Phe Cys Arg Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly
35 40 45
Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser
50 55 60
Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln
65 70 75 80
Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp
85 90 95
Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn
100 105 110
Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr
115 120 125
Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu
130 135 140
Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln
145 150 155 160
Leu Ile Cys Thr Gly
165
<210> 65
<211> 165
<212> PRT
<213> Artificial Sequence
<400> 65
Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys
1 5 10 15
Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg
20 25 30
Gly Phe Arg Cys Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly
35 40 45
Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser
50 55 60
Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln
65 70 75 80
Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp
85 90 95
Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn
100 105 110
Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr
115 120 125
Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu
130 135 140
Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln
145 150 155 160
Leu Ile Cys Thr Gly
165
<210> 66
<211> 165
<212> PRT
<213> Artificial Sequence
<400> 66
Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys
1 5 10 15
Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg
20 25 30
Gly Phe Arg Arg Ile Lys Ser Gly Ser Cys Tyr Met Leu Cys Thr Gly
35 40 45
Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser
50 55 60
Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln
65 70 75 80
Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp
85 90 95
Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn
100 105 110
Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr
115 120 125
Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu
130 135 140
Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln
145 150 155 160
Leu Ile Cys Thr Gly
165
<210> 67
<211> 165
<212> PRT
<213> Artificial Sequence
<400> 67
Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys
1 5 10 15
Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg
20 25 30
Gly Phe Arg Arg Ile Lys Ser Gly Ser Cys Tyr Met Leu Cys Thr Gly
35 40 45
Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser
50 55 60
Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln
65 70 75 80
Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp
85 90 95
Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn
100 105 110
Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr
115 120 125
Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu
130 135 140
Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln
145 150 155 160
Leu Ile Cys Thr Gly
165
<210> 68
<211> 165
<212> PRT
<213> Artificial Sequence
<400> 68
Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys
1 5 10 15
Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg
20 25 30
Gly Phe Arg Arg Ile Cys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly
35 40 45
Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser
50 55 60
Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln
65 70 75 80
Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp
85 90 95
Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn
100 105 110
Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr
115 120 125
Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu
130 135 140
Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln
145 150 155 160
Leu Ile Cys Thr Gly
165
<210> 69
<211> 165
<212> PRT
<213> Artificial Sequence
<400> 69
Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys
1 5 10 15
Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg
20 25 30
Gly Phe Arg Arg Ile Cys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly
35 40 45
Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser
50 55 60
Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln
65 70 75 80
Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp
85 90 95
Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn
100 105 110
Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr
115 120 125
Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu
130 135 140
Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln
145 150 155 160
Leu Ile Cys Thr Gly
165
<210> 70
<211> 165
<212> PRT
<213> Artificial Sequence
<400> 70
Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys
1 5 10 15
Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg
20 25 30
Gly Phe Arg Arg Ile Lys Cys Gly Ser Leu Tyr Met Leu Cys Thr Gly
35 40 45
Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser
50 55 60
Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln
65 70 75 80
Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp
85 90 95
Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn
100 105 110
Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr
115 120 125
Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu
130 135 140
Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln
145 150 155 160
Leu Ile Cys Thr Gly
165
<210> 71
<211> 165
<212> PRT
<213> Artificial Sequence
<400> 71
Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys
1 5 10 15
Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg
20 25 30
Gly Phe Arg Arg Ile Lys Ser Cys Ser Leu Tyr Met Leu Cys Thr Gly
35 40 45
Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser
50 55 60
Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln
65 70 75 80
Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp
85 90 95
Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn
100 105 110
Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr
115 120 125
Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu
130 135 140
Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln
145 150 155 160
Leu Ile Cys Thr Gly
165
<210> 72
<211> 165
<212> PRT
<213> Artificial Sequence
<400> 72
Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys
1 5 10 15
Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg
20 25 30
Gly Phe Arg Arg Ile Lys Ser Gly Cys Leu Tyr Met Leu Cys Thr Gly
35 40 45
Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser
50 55 60
Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln
65 70 75 80
Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp
85 90 95
Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn
100 105 110
Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr
115 120 125
Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu
130 135 140
Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln
145 150 155 160
Leu Ile Cys Thr Gly
165
<210> 73
<211> 165
<212> PRT
<213> Artificial Sequence
<400> 73
Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys
1 5 10 15
Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg
20 25 30
Gly Phe Arg Arg Ile Cys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly
35 40 45
Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser
50 55 60
Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln
65 70 75 80
Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp
85 90 95
Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn
100 105 110
Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr
115 120 125
Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu
130 135 140
Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln
145 150 155 160
Leu Ile Cys Thr Gly
165
<210> 74
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 74
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120
accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 75
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 75
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagatgcctc 120
accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctga agcctctgga agagtgtctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 76
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 76
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120
accttcaagt gctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gagtgtctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 77
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 77
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120
accttcaagt gctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctga agtgtctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 78
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 78
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120
acctgcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctga agcctctgga agagtgtctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 79
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 79
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120
accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
tgcgagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagtg tgccgatgag accgccacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 80
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 80
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120
acctgcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctga agtgtctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 81
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 81
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120
acctgcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag tgtgccacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 82
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 82
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120
acctgcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag acctgtacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 83
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 83
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120
acctgcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgcctgta tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 84
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 84
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120
tgcttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag acctgtacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 85
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 85
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgtgc 120
accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag acctgtacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 86
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 86
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgtgc 120
accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgcctgta tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 87
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 87
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgtgc 120
accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgccacct gtgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 88
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 88
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagatgcctc 120
accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctga agcctctgga agaggtgctg aattgtgccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 89
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 89
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagatgcctc 120
accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctga agcctctgga agaggtgctg aatctgtgtc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 90
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 90
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac atgcatgctc 120
accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctga agcctctgga agagtgtctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 91
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 91
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac atgcatgctc 120
accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctga agcctctgga agaggtgctg aattgtgccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 92
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 92
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagtgcac aagaatgctc 120
accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctga agcctctgga agagtgtctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 93
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 93
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagtgcac aagaatgctc 120
accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctga agcctctgga agaggtgctg aattgtgccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 94
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 94
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagtgcac aagaatgctc 120
accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctga agcctctgga agaggtgctg aatctgtgtc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 95
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 95
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc cctgcctgac aagaatgctc 120
accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctga agcctctgga agagtgtctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 96
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 96
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120
accttctgct tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag acctgtacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 97
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 97
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120
tgcttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 98
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 98
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatt gcaagctgac aagaatgctc 120
accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 99
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 99
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120
accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctga agcctctgga atgcgtgctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 100
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 100
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120
accttcaagt tctgcatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 101
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 101
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac atgcatgctc 120
accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 102
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 102
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120
accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctga agcctctgga agaggtgctg aattgcgccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 103
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 103
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120
accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
tgcgagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 104
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 104
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120
tgcttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 105
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 105
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc cctgcctgac aagaatgctc 120
accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 106
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 106
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgtg cagaatgctc 120
accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 107
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 107
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac atgcatgctc 120
accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 108
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 108
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac atgcatgctc 120
accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 109
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 109
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120
tgcttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 110
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 110
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120
tgcttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 111
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 111
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120
accttctgct tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 112
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 112
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120
accttctgct tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 113
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 113
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120
accttcaagt gctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 114
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 114
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120
accttctgct tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 115
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 115
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120
accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
tgcgagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 116
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 116
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120
accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gagtgcctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 117
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 117
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120
accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctgt gccctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 118
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 118
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120
accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctgt gccctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 119
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 119
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120
accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctgt gccctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 120
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 120
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120
accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctga agtgcctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360
tggatcacct tcgcccagag catcatcagc acactgacc 399
<210> 121
<211> 495
<212> DNA
<213> Artificial Sequence
<400> 121
gagctctgcg acgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60
aaggagggaa ccatgctctg ttgcgagtgc aagaggggat ttaggaggat caagagcggc 120
tctctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180
tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240
aaggaaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300
cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360
tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420
ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480
ctgatttgta ccggc 495
<210> 122
<211> 495
<212> DNA
<213> Artificial Sequence
<400> 122
gagctctgct gtgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60
aaggagggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggaggat caagagcggc 120
tctctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180
tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240
aaggaaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300
cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360
tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420
ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480
ctgatttgta ccggc 495
<210> 123
<211> 495
<212> DNA
<213> Artificial Sequence
<400> 123
gagctctgcg acgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60
aaggagggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggaggat caagagcggc 120
tcttgttaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180
tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240
aaggaaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300
cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360
tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420
ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480
ctgatttgta ccggc 495
<210> 124
<211> 495
<212> DNA
<213> Artificial Sequence
<400> 124
gagctctgcg acgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60
aaggagggaa ccatgctcaa ctgcgagtgc aagaggggat tttgtaggat caagagcggc 120
tctctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180
tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240
aaggaaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300
cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360
tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420
ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480
ctgatttgta ccggc 495
<210> 125
<211> 495
<212> DNA
<213> Artificial Sequence
<400> 125
gagctctgcg acgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60
aaggagggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggaggat caagagcggc 120
tctctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180
tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240
aaggaaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300
cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctactgt 360
tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420
ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480
ctgatttgta ccggc 495
<210> 126
<211> 495
<212> DNA
<213> Artificial Sequence
<400> 126
gagctctgcg acgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60
aaggagggaa cctgtctcaa ctgcgagtgc aagaggggat ttaggaggat caagagcggc 120
tctctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180
tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240
aaggaaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300
cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360
tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420
ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480
ctgatttgta ccggc 495
<210> 127
<211> 495
<212> DNA
<213> Artificial Sequence
<400> 127
gagctctgcg acgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60
aaggagggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggaggat caagtgtggc 120
tctctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180
tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240
aaggaaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300
cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360
tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420
ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480
ctgatttgta ccggc 495
<210> 128
<211> 495
<212> DNA
<213> Artificial Sequence
<400> 128
gagctctgcg acgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60
aaggagggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggaggat caagagcggc 120
tctctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180
tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240
aaggaaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300
cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gtgttaccac 360
tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420
ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480
ctgatttgta ccggc 495
<210> 129
<211> 495
<212> DNA
<213> Artificial Sequence
<400> 129
gagctctgct gtgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60
aaggagggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggaggat caagagcggc 120
tctctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180
tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240
aaggaaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300
cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360
tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420
ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480
ctgatttgta ccggc 495
<210> 130
<211> 495
<212> DNA
<213> Artificial Sequence
<400> 130
gagctctgct gtgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60
aaggagggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggaggat caagagcggc 120
tctctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180
tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240
aaggaaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300
cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360
tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420
ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480
ctgatttgta ccggc 495
<210> 131
<211> 495
<212> DNA
<213> Artificial Sequence
<400> 131
gagctctgct gtgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60
aaggagggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggaggat caagagcggc 120
tctctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180
tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240
aaggaaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300
cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360
tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420
ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480
ctgatttgta ccggc 495
<210> 132
<211> 495
<212> DNA
<213> Artificial Sequence
<400> 132
gagctctgcg actgtgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60
aaggagggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggaggat caagagcggc 120
tctctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180
tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240
aaggaaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300
cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360
tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420
ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480
ctgatttgta ccggc 495
<210> 133
<211> 495
<212> DNA
<213> Artificial Sequence
<400> 133
gagctctgcg acgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60
aaggagggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggaggat caagagcggc 120
tcttgttaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180
tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240
aaggaaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300
cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360
tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420
ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480
ctgatttgta ccggc 495
<210> 134
<211> 495
<212> DNA
<213> Artificial Sequence
<400> 134
gagctctgcg acgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60
aaggagggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggaggat caagagcggc 120
tctctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180
tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240
aaggaaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300
cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctgtcac 360
tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420
ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480
ctgatttgta ccggc 495
<210> 135
<211> 495
<212> DNA
<213> Artificial Sequence
<400> 135
gagctctgcg acgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60
aaggagggaa ccatgctcaa ctgcgagtgc aagaggggat tttgtaggat caagagcggc 120
tctctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180
tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240
aaggaaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300
cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360
tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420
ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480
ctgatttgta ccggc 495
<210> 136
<211> 495
<212> DNA
<213> Artificial Sequence
<400> 136
gagctctgcg acgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60
aaggagggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggtgtat caagagcggc 120
tctctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180
tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240
aaggaaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300
cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360
tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420
ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480
ctgatttgta ccggc 495
<210> 137
<211> 495
<212> DNA
<213> Artificial Sequence
<400> 137
gagctctgcg acgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60
aaggagggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggaggat caagagcggc 120
tcttgttaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180
tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240
aaggaaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300
cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360
tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420
ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480
ctgatttgta ccggc 495
<210> 138
<211> 495
<212> DNA
<213> Artificial Sequence
<400> 138
gagctctgcg acgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60
aaggagggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggaggat caagagcggc 120
tcttgttaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180
tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240
aaggaaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300
cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360
tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420
ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480
ctgatttgta ccggc 495
<210> 139
<211> 495
<212> DNA
<213> Artificial Sequence
<400> 139
gagctctgcg acgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60
aaggagggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggaggat ctgtagcggc 120
tctctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180
tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240
aaggaaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300
cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360
tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420
ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480
ctgatttgta ccggc 495
<210> 140
<211> 495
<212> DNA
<213> Artificial Sequence
<400> 140
gagctctgcg acgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60
aaggagggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggaggat ctgtagcggc 120
tctctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180
tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240
aaggaaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300
cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360
tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420
ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480
ctgatttgta ccggc 495
<210> 141
<211> 495
<212> DNA
<213> Artificial Sequence
<400> 141
gagctctgcg acgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60
aaggagggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggaggat caagtgtggc 120
tctctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180
tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240
aaggaaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300
cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360
tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420
ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480
ctgatttgta ccggc 495
<210> 142
<211> 495
<212> DNA
<213> Artificial Sequence
<400> 142
gagctctgcg acgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60
aaggagggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggaggat caagagctgt 120
tctctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180
tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240
aaggaaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300
cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360
tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420
ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480
ctgatttgta ccggc 495
<210> 143
<211> 495
<212> DNA
<213> Artificial Sequence
<400> 143
gagctctgcg acgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60
aaggagggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggaggat caagagcggc 120
tgtctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180
tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240
aaggaaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300
cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360
tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420
ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480
ctgatttgta ccggc 495
<210> 144
<211> 495
<212> DNA
<213> Artificial Sequence
<400> 144
gagctctgcg acgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60
aaggagggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggaggat ctgtagcggc 120
tctctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180
tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240
aaggaaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300
cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360
tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420
ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480
ctgatttgta ccggc 495
<210> 145
<211> 495
<212> DNA
<213> Artificial Sequence
<400> 145
gagctctgcg acgatgatcc ccccgaaatc ccccacgcca cattcaaggc catggcctac 60
aaggagggaa ccatgctcaa ctgcgagtgc aagaggggat ttaggaggat caagagcggc 120
tctctgtaca tgctgtgcac cggcaacagc agccacagca gctgggataa tcagtgccag 180
tgcaccagca gcgctacaag aaacaccaca aaacaagtga ccccccagcc cgaggagcaa 240
aaggaaagaa agaccaccga aatgcagtcc cccatgcagc ccgtggatca agcctctctg 300
cccggccact gtagagagcc ccctccttgg gaaaatgagg ccacagagag gatctaccac 360
tttgtggtgg gacagatggt ctactaccag tgtgtccaag gctatagagc tctccacaga 420
ggccccgccg agagcgtctg caaaatgaca cacggcaaga caagatggac ccaaccccag 480
ctgatttgta ccggc 495
<210> 146
<211> 399
<212> DNA
<213> Artificial Sequence
<400> 146
gcccccacaa gctccagcac caagaagaca cagctgcagc tggaacatct gctgctggac 60
ctccagatga ttctgaacgg catcaacaac tacaagaatc ccaagctgac aagaatgctc 120
accttcaagt tctacatgcc caagaaggcc accgagctga agcatctgca gtgtctggag 180
gaggagctga agcctctgga agaggtgctg aatctggccc agtccaagaa cttccatctg 240
aggcctagag atctgatcag caacatcaac gtcatcgtgc tggagctgaa gggcagcgag 300
accaccttca tgtgcgagta cgccgatgag accgccacca tcgtggagtt tctgaataga 360
tggatcacct tctgccagag catcatcagc acactgacc 399

Claims (35)

1.IL-2衍生物，其特征在于，所述IL-2衍生物通过在野生型IL-2的基础上引入至少一个半胱氨酸残基，部分封闭或完全封闭IL-2衍生物与α受体亚基的结合平面，同时基本保留对β和γ受体亚基复合物的亲和力。
2.如权利要求了1所述的IL-2衍生物，其特征在于，在野生型IL-2的基础上引入的至少一个半胱氨酸残基，能够：
a)形成IL-2衍生物的分子内二硫键；或者
b)使得IL-2衍生物能通过分子间二硫键与封闭模块结合。
3.如权利要求了2所述的IL-2衍生物，其特征在于，所述引入至少一个半胱氨酸残基为通过点突变的方式引入至少一个半胱氨酸残基。
4.如权利要求3所述的IL-2衍生物，其特征在于，在野生型IL-2的基础上通过点突变方式引入第一半胱氨酸残基和第二半胱氨酸残基；所述第一半胱氨酸残基和所述第二半胱氨酸残基中的一个或两个为野生型IL-2与α受体亚基的结合平面相关的氨基酸或其附近的氨基酸。
5.如权利要求4所述的IL-2衍生物，其特征在于，所述第一半胱氨酸残基为野生型IL-2的第37位、第38位、第41位、第42位、第43位、第44位、第45位、第61位、第62位氨基酸或其附近的氨基酸；所述第二半胱氨酸残基为野生型IL-2的第61位、第62位、第65位、第68位、第72位氨基酸或其附近的氨基酸。
6.如权利要求4所述的IL-2衍生物，其特征在于，所述第一半胱氨酸残基为选自下组的氨基酸点突变：K35C、L36C、R38C、M39C、L40C、T41C、F42C、K43C、F44C和E61C。
7.如权利要求4所述的IL-2衍生物，其特征在于，所述第二半胱氨酸残基为选自下组的氨基酸点突变：V69C、E62C、P65C、T111C、Y107C、A112C、T113C、I114C、L72C和A73C。
8.如权利要求4所述的IL-2衍生物，其特征在于，形成分子内二硫键的第一半胱氨酸残基和第二半胱氨酸残基的组合选自下组的氨基酸点突变组合：M39C和V69C、F44C和E62C、F44C和P65C、F42C和V69C、E61C和Y107C、F42C和P65C、F42C和T111C、F42C和A112C、F42C和T113C、T41C和A112C、L40C和A112C、L40C和T113C、L40C和I114C、M39C和L72C、M39C和A73C、R38C和V69C、R38C和L72C、L36C和V69C、L36C和L72C、L36C和A73C、K35C和V69C以及K43C和A112C。
9.如权利要求4所述的IL-2衍生物，其特征在于，所述第一半胱氨酸残基和所述第二半胱氨酸残基的残基质心矢量距离为小于
Figure FDA0002322160490000021
10.如权利要求3所述的IL-2衍生物，其特征在于，在野生型IL-2的基础上通过点突变方式引入第三半胱氨酸残基；所述第三半胱氨酸残基为野生型IL-2与α受体亚基的结合平面相关的氨基酸或其附近的氨基酸。
11.如权利要求10所述的IL-2衍生物，其特征在于，所述第三半胱氨酸残基为野生型IL-2的第37位、第38位、第41位、第42位、第43位、第44位、第45位、第61位、第62位、第65位、第68位、第72位氨基酸或其附近的氨基酸。
12.如权利要求10所述的IL-2衍生物，其特征在于，所述第三半胱氨酸残基为选自下组的氨基酸点突变：P34C、K35C、T37C、R38C、T41C、K43C、F44C、Y45C、E61C、E62C、K64C、P65C、E68C和L72C。
13.如权利要求10所述的IL-2衍生物，其特征在于，所述封闭模块上具有或引入有第四半胱氨酸残基；所述IL-2衍生物上的第三半胱氨酸残基与所述封闭模块上的第四半胱氨酸残基能形成分子间二硫键。
14.如权利要求13所述的IL-2衍生物，其特征在于，所述第三半胱氨酸残基和所述第四半胱氨酸残基的残基质心矢量距离为小于
Figure FDA0002322160490000022
15.如权利要求13所述的IL-2衍生物，其特征在于，所述封闭模块为α受体亚基的胞外段。
16.如权利要求3所述的IL-2衍生物，其特征在于，通过点突变的方式将野生型IL-2第125位的半胱氨酸残基转变为其他的氨基酸残基。
17.如权利要求16所述的IL-2衍生物，其特征在于，野生型IL-2第125位的点突变为C125A。
18.如权利要求3所述的IL-2衍生物，其特征在于，所述IL-2衍生物的氨基酸序列如SEQID NO.3～SEQ ID NO.24所示；或如SEQ ID NO.26～40所示。
19.一种复合物，其特征在于，包括：
1)IL-2衍生物，所述IL-2衍生物在野生型IL-2的基础上引入第三半胱氨酸残基；
2)封闭模块，所述封闭模块上具有或引入有第四半胱氨酸残基；
所述IL-2衍生物上的第三半胱氨酸残基和所述封闭模块上的第四半胱氨酸残基形成分子间二硫键，从而形成IL-2与封闭模块的复合物，部分封闭或完全封闭IL-2衍生物与α受体亚基的结合平面，同时基本保留对β和γ受体亚基复合物的亲和力。
20.如权利要求19所述的复合物，其特征在于，所述第三半胱氨酸残基为野生型IL-2与α受体亚基的结合平面相关的氨基酸或其附近的氨基酸。
21.如权利要求20所述的复合物，其特征在于，所述第三半胱氨酸残基和所述第四半胱氨酸残基的残基质心矢量距离为小于
Figure FDA0002322160490000031
22.如权利要求19所述的复合物，其特征在于，所述第三半胱氨酸残基为选自下组的氨基酸点突变：P34C、K35C、T37C、R38C、T41C、K43C、F44C、Y45C、E61C、E62C、K64C、P65C、E68C和L72C。
23.如权利要求19所述的复合物，其特征在于，所述封闭模块为α受体亚基的胞外段，其氨基酸序列如SEQ ID NO.25所示。
24.如权利要求23所述的复合物，其特征在于，α受体亚基的胞外段上的所述第四半胱氨酸残基位选自下组的氨基酸点突变：D4C、D5C、M25C、N27C、R35C、R36C、K38C、S39C、G40C、S41C、L42C、I118C、Y119C和H120C。
25.如权利要求23所述的复合物，其特征在于，形成分子间二硫键的第三半胱氨酸残基和第四半胱氨酸残基的组合选自下组的氨基酸点突变组合：T41C和N27C、P34C和D4C、E68C和L42C、Y45C和R35C、R38C和H120C、L72C和M25C、E61C和S39C、T41C和I118C、K35C和D4C、T37C和D4C、R38C和D4C、R38C和D5C、T41C和L42C、T41C和Y119C、K43C和R35C、K43C和R36C、F44C和L42C、K43C和L42C、E61C和K38C、E62C和K38C、K64C和S39C、K64C和G40C、K64C和S41C、P65C和K38C。
26.如权利要求19所述的复合物，其特征在于，野生型IL-2第125位的点突变为C125A。
27.如权利要求23所述的复合物，其特征在于，所述IL-2衍生物的氨基酸序列和所述α受体亚基的胞外段的序列组合选自下组的组合：SEQ ID NO.26和SEQ ID NO.50，SEQ IDNO.27和SEQ ID NO.51，SEQ ID NO.28和SEQ ID NO.52，SEQ ID NO.29和SEQ ID NO.53，SEQID NO.30和SEQ ID NO.54，SEQ ID NO.31和SEQ ID NO.55，SEQ ID NO.32和SEQ ID NO.56，SEQ ID NO.33和SEQ ID NO.57，SEQ ID NO.34和SEQ ID NO.58，SEQ ID NO.35和SEQ IDNO.59，SEQ ID NO.36和SEQ ID NO.60，SEQ ID NO.37和SEQ ID NO.61，SEQ ID NO.38和SEQID NO.62，SEQ ID NO.39和SEQ ID NO.63，SEQ ID NO.40和SEQ ID NO.64，SEQ ID NO.41和SEQ ID NO.65，SEQ ID NO.42和SEQ ID NO.66，SEQ ID NO.43和SEQ ID NO.67，SEQ IDNO.44和SEQ ID NO.68，SEQ ID NO.45和SEQ ID NO.69，SEQ ID NO.46和SEQ ID NO.70，SEQID NO.47和SEQ ID NO.71，SEQ ID NO.48和SEQ ID NO.72，SEQ ID NO.49和SEQ ID NO.73。
28.一种分离的多核苷酸，其特征在于，其编码如权利要求1～18中任一项所述的IL-2衍生物，或者编码如权利要求19～27中任一项所述的复合物。
29.一种表达载体，其特征在于，包含如权利要求28所述的一种分离的多核苷酸。
30.一种宿主细胞，其特征在于，包含如权利要求28所述的一种分离的多核苷酸。
31.一种组合物，其特征在于，包含如权利要求1～18中任一项所述的IL-2衍生物或者如权利要求19～27中任一项所述的复合物，以及药学可接受载体。
32.如权利要求1～18中任一项所述的IL-2衍生物或者如权利要求19～27中任一项所述的复合物用于制备用于治疗疾病的药物或制剂中的用途。
33.如权利要求1～18中任一项所述的IL-2衍生物或者如权利要求19～27中任一项所述的复合物在制备用于刺激个体的免疫系统的组合物中的用途。
34.一种生成IL-2衍生物的方法，其特征在于，所述方法包括在适合于表达所述IL-2衍生物的条件下培养如权利要求30所述的宿主细胞。
35.一种生成复合物的方法，其特征在于，所述方法包括在适合于表达所述复合物的条件下培养如权利要求30所述的宿主细胞。
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