本申请是申请号为202110676901.2,申请日为2021年6月18日,发明名称为“一种产生广谱交叉中和活性的重组新型冠状病毒RBD三聚体蛋白疫苗、其制备方法和应用”的专利申请的分案申请。
具体实施方式
本发明公开了一种产生广谱交叉中和活性的重组新型冠状病毒RBD三聚体蛋白疫苗、其制备方法和应用,本领域技术人员可以借鉴本文内容,适当改进工艺参数实现。需要特别指出的是,所有类似的替换和改动对本领域技术人员来说是显而易见的,它们都被视为包括在本发明,并且相关人员明显能在不脱离本发明内容、精神和范围的基础上对本文所述内容进行改动或适当变更与组合,来实现和应用本发明技术。
在本发明中,除非另有说明,否则本文中使用的科学和技术名词具有本领域技术人员所通常理解的含义。除非另有其它明确表示,否则在整个说明书和权利要求书中,术语“包括”或其变换如“包含”或“包括有”等等将被理解为包括所陈述的元件或组成部分,而并未排除其它元件或其它组成部分。除非另有其它明确表示,否则在整个说明书和权利要求书中,术语“RBD”即代表新型冠状病毒的刺突蛋白的RBD结构域,其与“RBD”或“新型冠状病毒RBD区域”可以理解为能够相互替换使用。
下面就本发明中出现的部分术语作以解释。
术语“新型冠状病毒”,即SARS-CoV-2,属巢病毒目(Nidovirales)、冠状病毒科(Coronaviridae)、正冠状病毒亚科、Betacoronavirus属、Sarbecovirus亚属、类SARS病毒种、单股正链RNA病毒,有包膜,基因组全长约为29.9kb,绝大部分编码非结构蛋白,参与病毒复制和翻译等功能,少部分序列编码结构蛋白,如:S蛋白(spike protein)、M蛋白(membrane protein)、E蛋白(envelope protein)和N蛋白(nucleo protein),此外还有若干附属蛋白:3a,3b,p6,7a,7b,8b,9b和orf14,这些蛋白均参与病毒组装。S、M和E蛋白构成病毒囊膜,是病毒引起免疫反应的主要表面抗原。其中S蛋白是一种跨膜糖蛋白,分子量约为150kDa,在病毒表面形成突出的同源三聚体。S由两个功能亚基组成,在S1和S2亚基之间的边界处(S1/S2裂解点)被切割,这两个亚基在融合前构象中保持非共价结合。S2亚基也由多个结构域构成,它的功能主要是介导病毒与宿主细胞的融合。远端S1亚基在结构上分为四个不同的结构域:N端结构域(NTD)、受体结合结构域(RBD)、C端结构域1(CTD1)和C端结构域2(CTD2),其中RBD主要负责与宿主细胞表面的受体血管紧张素转换酶2(angiotensinconverting enzyme 2,ACE2)结合,从而介导病毒侵染宿主细胞,因此S蛋白及RBD均为目前基因工程疫苗研发的主要靶标。
术语“三聚体形式”,是蛋白质高级结构中的一种类型。其中含有三个蛋白质亚基即为三聚体形式。
术语“至少有一个”可以理解为在三个氨基酸序列中的两个相同或者三个氨基酸序列各不相同。
术语“一级结构”,是肽或蛋白质中氨基酸的线性序列。按照惯例,蛋白质的一级结构是指从氨基末端(N)端到羧基末端(C)端。
术语“融合蛋白”,fusion protein,是指通过DNA重组技术得到的一个、两个或多个基因重组后的表达产物。融合蛋白技术是为获得大量标准融合蛋白而进行的有目的性的基因融合和蛋白表达方法,利用融合蛋白技术,可构建和表达具有多种功能的新型目的蛋白。
术语“载体”,是可将多核苷酸插入其中的一种核酸运载工具。当载体能使插入的多核苷酸编码的蛋白质获得表达时,载体称为表达载体。载体可以通过转化,转导或者转染导入宿主细胞,使其携带的遗传物质元件在宿主细胞中获得表达。载体是本领域技术人员公知的,包括但不限于:质粒;噬菌粒;柯斯质粒;人工染色体,例如酵母人工染色体(YAC)、细菌人工染色体(BAC)或P1来源的人工染色体(PAC);噬菌体如λ噬菌体或M13噬菌体及动物病毒等。可用作载体的动物病毒包括但不限于,逆转录酶病毒(包括慢病毒)、腺病毒、腺相关病毒、疱疹病毒(如单纯疱疹病毒)、痘病毒、杆状病毒、乳头瘤病毒、乳头多瘤空泡病毒(如SV40)。一种载体可以含有多种控制表达的元件,包括但不限于,启动子序列、转录起始序列、增强子序列、选择元件及报告基因。另外,载体还可含有复制起始位点。
术语“宿主细胞”,是已经通过分子生物学技术将核酸分子引入的细胞。这些技术包括转染病毒载体,用质粒载体转化,以及通过电穿孔、脂转染、和粒子枪加速引入裸DNA。
术语“治疗”,是指减少疾病病理的可能性,减少疾病症状的发生,例如在一定程度上受试者具有更长的存活期或减少的不适。治疗可以是指当向受试者给予疗法时该疗法减少疾病症状、体征或病因的能力。治疗还指缓和或减少至少一种临床症状和/或抑制或延迟病症的进展和/或预防或延迟疾病或疾患的发作。
术语“受试者”是指接受预防、治疗、诊断的任何人或其他动物,特别是其他哺乳动物。其他哺乳动物可以包括,例如,狗、猫、牛、马、绵羊、猪、山羊、兔子、大鼠、豚鼠、小鼠等。
为了使本领域的技术人员更好地理解本发明的技术方案,下面结合具体实施例对本发明作进一步的详细说明。
实施例1:基于蛋白结构和计算生物学设计的新型冠状病毒RBD三聚体蛋白
通过对新型冠状病毒代表性突变株和原型株进行比对和分析(如表1所示),Beta(B.1.351)突变株发生了3个位点的氨基酸突变,即K417N、E484K和N501Y;Kappa(B.1.617.1)突变株发生了2个位点的氨基酸突变,即L452R和E484Q。其中,E484K被认为是导致免疫逃逸的最主要的位点突变,通过分子动力学模拟和自由能计算发现,E484K突变会导致RBD与多个中和性单抗的亲和力显著降低,WHO列出的归类为VOC和VOI的10种突变株中,有6种突变株均含有E484K突变。另外,L452R突变也已经被证实能够对中和抗体以及新冠病人恢复者血清产生逃逸,10种VOC和VOI突变株中,有3株含有L452R突变。有实验证据显示,K417N和N501Y突变能够对一部分中和抗体产生抵抗,在10种VOC和VOI突变株中,有4株含有N501Y突变。此外,通过对目前已知的120万条SARS-CoV-2序列进行了进化分析,发现E484K、L452R和N501Y是RBD中最主要的趋同进化突变,这些突变在众多不同的病毒谱系独立出现,表明这些突变在病毒进化中具有明显的选择优势,也预示着这些突变可能会在将来的突变株中再次独立或组合出现。因此研发一种具有跨流行株广谱保护能力的新冠疫苗具有重要意义。
对天然S蛋白三聚体空间结构分析(如图1所示),对RBD结构域的N端和C端的空间距离进行计算,发现在不引入外源载体或序列的情况下,可以利用RBD自身的结构特征实现三聚化,同时不会存在较大的空间位障。基于此,截取不同突变株的新型冠状病毒S蛋白RBD区序列片段(319~537位氨基酸),将三个RBD区片段首尾串联,形成一个新的融合蛋白。针对新型冠状病毒原型株S蛋白RBD区序列片段(319~537位氨基酸)序列如SEQ ID No.1所示,针对Beta(B.1.351)突变株的S蛋白RBD区序列片段(319~537位氨基酸)序列如SEQ IDNo.2所示,针对Kappa(B.1.617.1)突变株的S蛋白RBD区序列片段(319~537位氨基酸)序列如SEQ ID No.3所示。
对SEQ ID No.1、SEQ ID No.2和SEQ ID No.3进行不同组合,以达到一种三聚体蛋白可同时产生针对不同新型冠状毒株的交叉中和活性,达到广谱性疫苗研发目的,具体如下:由SEQ ID No.1,SEQ ID No.2,SEQ ID No.3顺序连接形成的三聚体蛋白A,氨基酸序列如SEQ ID No.4所示;由SEQ ID No.1,SEQ ID No.2,SEQ ID No.1顺序连接形成的三聚体蛋白B,氨基酸序列如SEQ ID No.5所示;由SEQ ID No.1,SEQ ID No.2,SEQ ID No.2顺序连接形成的三聚体蛋白C,氨基酸序列如SEQ ID No.6所示;由SEQ ID No.2,SEQ ID No.1,SEQID No.2顺序连接形成的三聚体蛋白D,氨基酸序列如SEQ ID No.7所示;由SEQ ID No.2,SEQ ID No.2,SEQ ID No.2顺序连接形成的三聚体蛋白E,氨基酸序列如SEQ ID No.8所示。
以三聚体蛋白A(氨基酸序列如SEQ ID No.4所示)为例,利用同源模建搭建了其可能的空间结构,结果如图2所示,显示该融合蛋白包含有三个独立RBD结构域,可以形成抗原构象稳定的三聚体形式,且该蛋白涵盖了关键的活跃突变位点和潜在的免疫逃逸位点,理论上推测对以此作为靶抗原的重组疫苗具备跨流行株的广谱保护能力,另外,与传统的制备多个单价疫苗,通过多价组合来实现广谱保护的策略相比,在一种抗原分子上实现了多价广谱保护效果,在疫苗制备的时间成本、经济成本以及疫苗产能方面具有明显的优势。
表1.SARS-CoV-2代表性流行株RBD区突变情况
实施例2:三聚体蛋白表达、纯化及鉴定
按照CHO细胞表达系统的密码子偏爱性,对编码蛋白A至蛋白E(氨基酸序列如SEQID NO.4至SEQ ID NO.8所示)的核苷酸序列进行密码子优化,蛋白A(氨基酸序列为SEQ IDNO.4)优化后的核苷酸序列如SEQ ID NO.9所示,蛋白B(氨基酸序列为SEQ ID NO.5)优化后的核苷酸序列如SEQ ID NO.10所示,蛋白C(氨基酸序列为SEQ ID NO.6)优化后的核苷酸序列如SEQ ID NO.11、SEQ ID NO.12、SEQ ID NO.13或SEQ ID NO.14所示,蛋白D(氨基酸序列为SEQ ID NO.7)优化后的核苷酸序列如SEQ ID NO.15所示,蛋白E(氨基酸序列为SEQ IDNO.8)优化后的核苷酸序列如SEQ ID NO.16或SEQ ID NO.17所示。
构建CHO细胞表达载体后转染至293FT细胞或CHO细胞内构建重组细胞株,通过有限稀释法筛选得到能够稳定分泌表达RBD三聚体蛋白的细胞株,最终蛋白A、蛋白B和蛋白C均成功得到表达,经系列层析纯化后蛋白A、蛋白B和蛋白C均获得纯度≥95%的三聚体蛋白,蛋白D和蛋白E表达量过低经纯化后未获得纯度较高的目的蛋白。蛋白A、蛋白B和蛋白C的SDS-PAGE检测结果如图3所示,蛋白分子量大小在70~100kD,同时可见有部分产品相关物质,如二聚体蛋白和单体蛋白等。
将纯化后蛋白A、蛋白B和蛋白C经SDS-PAGE电泳后电转至PVDF膜上,利用RBD特异性抗体(厂家:北京义翘神州科技有限公司;货号:40591-T62;稀释度:2000倍)进行Western-blot鉴定(结果如图4所示),所有蛋白可与RBD特异性抗体发生结合,具有良好的生物学活性。采用TSKgel G2500PW凝胶色谱柱对纯化后的蛋白A、蛋白B和蛋白C进行分子排阻色谱分析,所有纯化后蛋白纯度均大于90%。
实施例3:与中和性单克隆抗体结合生物学分析
将纯化后的三聚体蛋白A、蛋白B、蛋白C,以及三聚体蛋白(由3个如SEQ ID No.1所示的氨基酸片段顺序连接形成如SEQ ID No.18所示氨基酸序列的蛋白,经293FT细胞或CHO细胞重组表达、层析纯化所得)和二聚体蛋白(由2个如SEQ ID No.1所示的氨基酸片段顺序连接形成如SEQ ID No.19所示氨基酸序列的蛋白,经293FT细胞或CHO细胞重组表达、层析纯化所得)、RBD蛋白(厂家:北京义翘神州科技有限公司;货号:40592-V08B)、与Beta(B.1.351)株病毒突变位点一致的RBD蛋白(K417N、E484K、N501Y;厂家:北京义翘神州科技有限公司;货号:40592-V08H85),与Kappa(B.1.617.1)株病毒突变位点一致的RBD蛋白(L452R、E484Q;厂家:北京义翘神州科技有限公司;货号:40592-V08H85),利用包被液稀释至4μg/ml、2μg/ml、1μg/ml、0.5μg/ml、0.25μg/ml、0.125μg/ml、0.0625μg/ml、0.03125μg/ml、0.015625μg/ml、0.007813μg/ml、0.003906μg/ml、0.001953μg/ml,100μl/孔,包被至96孔酶标板上,4℃,8~12h,以空白孔为阴性对照;PBST溶液洗板后加入封闭液,37℃封闭3h;PBST溶液洗板后分别加入稀释后的MM43单克隆抗体(厂家:北京义翘神州科技有限公司;货号:40591-MM43,稀释度:2000倍),或MM57单克隆抗体(厂家:北京义翘神州科技有限公司;货号:40592-MM57;稀释度:2000倍),或R001单克隆抗体(厂家:北京义翘神州科技有限公司;货号:40592-R001;稀释度:2000倍),或R117单克隆抗体(厂家:北京义翘神州科技有限公司;货号:40592-R117;稀释度:2000倍),100μl/孔,37℃孵育1h;PBST溶液洗板后加入稀释后的辣根过氧化物酶标记的羊抗鼠或羊抗兔IgG抗体,100μl/孔,37℃孵育1h;PBST溶液洗板后先后加入显色液A和B,室温下显色5~10min,加入终止液C;酶标仪上进行双波长(OD450nm和630nm)读值,确定cutoff值,并绘制蛋白浓度-吸光度值曲线。
与MM43单克隆抗体结合活性结果如图5所示,与MM57单克隆抗体结合活性结果如图6所示,与R001单克隆抗体结合活性结果如图7所示,与R117单克隆抗体结合活性结果如图8所示,结果可见三聚体蛋白A与所有中和性单克隆抗体均呈不同程度的结合,其中与MM43抗体结合活性与三聚体蛋白基本一致,与MM57和R117单克隆抗体结合活性低于三聚体蛋白,说明三聚体蛋白既具有原型株RBD蛋白性质,又具有Beta(B.1.351)株RBD蛋白特性。
实施例4:三聚体蛋白A理化性质及活性检测
三聚体蛋白A一级序列中包含有原型株、Beta(B.1.351)突变株和Kappa(B.1.617.1)突变株的RBD区氨基酸序列,是一种突变集成的RBD三聚体蛋白(mutations-integrated trimeric form of RBD,mutI tri-RBD),进一步对纯化后的三聚体蛋白A进一步分析,利用SDS-PAGE检测和Western-blot分析,同时以三聚体蛋白为对照,结果如图9和图10所示,可见三聚体蛋白A与三聚体蛋白分子量大小基本一致,在70~100kD之间,利用MALDI-TOF MS方法检测蛋白完整分子量,三聚体蛋白A分子量约为87.836kD,较理论值(74kD)偏大,这与蛋白糖基化修饰相关,利用UPLC-MS方法检测蛋白糖基化修饰,结果显示重组表达的三聚体蛋白A具有较多位点的糖基化修饰。利用圆二色谱对三聚体蛋白A进行二级结构检测,结果显示α螺旋占比为13.5%,β折叠占比为23.3%,β转角占比为11.4%,其他为51.8%,这与理论值基本保持一致。利用UPLC-MS方法检测二硫键形成情况,结果也显示三聚体蛋白A中的每一个RBD单体都可形成4对二硫键,与RBD天然结构中二硫键配对结果一致。利用量热差示扫描方法检测三聚体蛋白A的热稳定性,结果如图11所示,Tm值为47.2℃,三聚体蛋白A具有良好的热稳定性。利用透射电镜观察蛋白形态,结果如图12所示,可见三聚体蛋白A粒径较小,约为3-5nm左右,但未观察到更为清晰的形态结构图像。另外,利用表面等离子共振技术检测三聚体蛋白A与hACE受体蛋白的亲和力,结果如图13所示,经过计算KD值为3.19×10-3nM,两者具有高亲和力,证明三聚体蛋白A具有良好的生物学活性。
实施例5:重组新型冠状病毒疫苗制备
将纯化后的蛋白稀释至2倍目标抗原浓度,与1.2mg/ml氢氧化铝佐剂按1:1比例(w/w)混合吸附,并于磁力搅拌器上搅拌40~120min,转速为200~300rpm,获得疫苗半成品,上清残留蛋白含量应低于总蛋白含量的10%,半成品每瓶按0.5ml装量无菌分装后即为疫苗成品。
实施例6:重组新型冠状病毒疫苗免疫学效果评价
将制备的疫苗(如三聚体蛋白A或三聚体蛋白),分别经腹腔注射免疫BALB/c小鼠(购自北京维通利华实验动物技术有限公司,SPF级,雌性,6-8周龄),2μg/剂/只,分别于0w、3w免疫2针,5w采血分离血清。利用假病毒微量中和试验检测免后小鼠血清针对多种假病毒(如表2所示)的中和活性,结果如图14所示,血清抗体GMT值如表3所示,可见三聚体蛋白A可产生针对多种假病毒的中和活性,对目前引起免疫逃逸突变株以及将来可能出现的突变株具有一定的保护潜力,预期可产生广谱保护能力。
表2.SARS-CoV-2假病毒信息列表
表3.中和抗体GMT值(假病毒微量中和试验)
假病毒微量中和试验:利用微量中和试验检测免后血清针对多种假病毒(如表2所示)的中和抗体滴度,假病毒报告基因均为萤火虫荧光素酶,假病毒工作浓度为(1~2)×104TCID50/ml,使用Reed-Muench法计算50%感染抑制率时血清稀释倍数,即为该血清样品的中和抗体滴度。
以上所述仅是本发明的优选实施方式,应当指出,对于本技术领域的普通技术人员来说,在不脱离本发明原理的前提下,还可以做出若干改进和润饰,这些改进和润饰也应视为本发明的保护范围。
序列表
<110> 国药中生生物技术研究院有限公司
<120> 一种产生广谱交叉中和活性的重组新型冠状病毒RBD三聚体蛋白疫苗、其制备方法和应用
<130> /
<160> 19
<170> SIPOSequenceListing 1.0
<210> 1
<211> 219
<212> PRT
<213> Virus
<400> 1
Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn
1 5 10 15
Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val
20 25 30
Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser
35 40 45
Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val
50 55 60
Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp
65 70 75 80
Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln
85 90 95
Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr
100 105 110
Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly
115 120 125
Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys
130 135 140
Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr
145 150 155 160
Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser
165 170 175
Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val
180 185 190
Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly
195 200 205
Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys
210 215
<210> 2
<211> 219
<212> PRT
<213> Virus
<400> 2
Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn
1 5 10 15
Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val
20 25 30
Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser
35 40 45
Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val
50 55 60
Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp
65 70 75 80
Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln
85 90 95
Thr Gly Asn Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr
100 105 110
Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly
115 120 125
Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys
130 135 140
Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr
145 150 155 160
Pro Cys Asn Gly Val Lys Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser
165 170 175
Tyr Gly Phe Gln Pro Thr Tyr Gly Val Gly Tyr Gln Pro Tyr Arg Val
180 185 190
Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly
195 200 205
Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys
210 215
<210> 3
<211> 219
<212> PRT
<213> Virus
<400> 3
Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn
1 5 10 15
Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val
20 25 30
Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser
35 40 45
Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val
50 55 60
Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp
65 70 75 80
Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln
85 90 95
Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr
100 105 110
Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly
115 120 125
Gly Asn Tyr Asn Tyr Arg Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys
130 135 140
Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr
145 150 155 160
Pro Cys Asn Gly Val Gln Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser
165 170 175
Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val
180 185 190
Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly
195 200 205
Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys
210 215
<210> 4
<211> 658
<212> PRT
<213> Artificial
<400> 4
Met Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr
1 5 10 15
Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser
20 25 30
Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr
35 40 45
Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly
50 55 60
Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala
65 70 75 80
Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly
85 90 95
Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe
100 105 110
Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val
115 120 125
Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu
130 135 140
Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser
145 150 155 160
Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln
165 170 175
Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg
180 185 190
Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys
195 200 205
Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Arg Val Gln Pro
210 215 220
Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe
225 230 235 240
Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn
245 250 255
Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn
260 265 270
Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys
275 280 285
Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile
290 295 300
Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly Asn Ile
305 310 315 320
Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile
325 330 335
Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn
340 345 350
Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg
355 360 365
Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn Gly
370 375 380
Val Lys Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe Gln
385 390 395 400
Pro Thr Tyr Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser
405 410 415
Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser
420 425 430
Thr Asn Leu Val Lys Asn Lys Arg Val Gln Pro Thr Glu Ser Ile Val
435 440 445
Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn
450 455 460
Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser
465 470 475 480
Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser
485 490 495
Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys
500 505 510
Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val
515 520 525
Arg Gln Ile Ala Pro Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr
530 535 540
Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn
545 550 555 560
Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn Tyr Arg Tyr Arg Leu
565 570 575
Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu
580 585 590
Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn Gly Val Gln Gly Phe Asn
595 600 605
Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val
610 615 620
Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu His
625 630 635 640
Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys
645 650 655
Asn Lys
<210> 5
<211> 658
<212> PRT
<213> Artificial
<400> 5
Met Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr
1 5 10 15
Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser
20 25 30
Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr
35 40 45
Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly
50 55 60
Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala
65 70 75 80
Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly
85 90 95
Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe
100 105 110
Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val
115 120 125
Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu
130 135 140
Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser
145 150 155 160
Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln
165 170 175
Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg
180 185 190
Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys
195 200 205
Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Arg Val Gln Pro
210 215 220
Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe
225 230 235 240
Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn
245 250 255
Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn
260 265 270
Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys
275 280 285
Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile
290 295 300
Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly Asn Ile
305 310 315 320
Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile
325 330 335
Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn
340 345 350
Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg
355 360 365
Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn Gly
370 375 380
Val Lys Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe Gln
385 390 395 400
Pro Thr Tyr Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser
405 410 415
Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser
420 425 430
Thr Asn Leu Val Lys Asn Lys Arg Val Gln Pro Thr Glu Ser Ile Val
435 440 445
Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn
450 455 460
Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser
465 470 475 480
Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser
485 490 495
Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys
500 505 510
Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val
515 520 525
Arg Gln Ile Ala Pro Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr
530 535 540
Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn
545 550 555 560
Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu
565 570 575
Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu
580 585 590
Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn
595 600 605
Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val
610 615 620
Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu His
625 630 635 640
Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys
645 650 655
Asn Lys
<210> 6
<211> 658
<212> PRT
<213> Artificial
<400> 6
Met Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr
1 5 10 15
Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser
20 25 30
Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr
35 40 45
Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly
50 55 60
Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala
65 70 75 80
Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly
85 90 95
Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe
100 105 110
Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val
115 120 125
Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu
130 135 140
Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser
145 150 155 160
Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln
165 170 175
Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg
180 185 190
Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys
195 200 205
Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Arg Val Gln Pro
210 215 220
Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe
225 230 235 240
Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn
245 250 255
Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn
260 265 270
Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys
275 280 285
Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile
290 295 300
Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly Asn Ile
305 310 315 320
Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile
325 330 335
Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn
340 345 350
Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg
355 360 365
Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn Gly
370 375 380
Val Lys Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe Gln
385 390 395 400
Pro Thr Tyr Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser
405 410 415
Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser
420 425 430
Thr Asn Leu Val Lys Asn Lys Arg Val Gln Pro Thr Glu Ser Ile Val
435 440 445
Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn
450 455 460
Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser
465 470 475 480
Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser
485 490 495
Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys
500 505 510
Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val
515 520 525
Arg Gln Ile Ala Pro Gly Gln Thr Gly Asn Ile Ala Asp Tyr Asn Tyr
530 535 540
Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn
545 550 555 560
Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu
565 570 575
Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu
580 585 590
Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn Gly Val Lys Gly Phe Asn
595 600 605
Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe Gln Pro Thr Tyr Gly Val
610 615 620
Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu His
625 630 635 640
Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys
645 650 655
Asn Lys
<210> 7
<211> 658
<212> PRT
<213> Artificial
<400> 7
Met Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr
1 5 10 15
Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser
20 25 30
Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr
35 40 45
Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly
50 55 60
Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala
65 70 75 80
Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly
85 90 95
Gln Thr Gly Asn Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe
100 105 110
Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val
115 120 125
Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu
130 135 140
Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser
145 150 155 160
Thr Pro Cys Asn Gly Val Lys Gly Phe Asn Cys Tyr Phe Pro Leu Gln
165 170 175
Ser Tyr Gly Phe Gln Pro Thr Tyr Gly Val Gly Tyr Gln Pro Tyr Arg
180 185 190
Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys
195 200 205
Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Arg Val Gln Pro
210 215 220
Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe
225 230 235 240
Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn
245 250 255
Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn
260 265 270
Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys
275 280 285
Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile
290 295 300
Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly Lys Ile
305 310 315 320
Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile
325 330 335
Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn
340 345 350
Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg
355 360 365
Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn Gly
370 375 380
Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe Gln
385 390 395 400
Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser
405 410 415
Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser
420 425 430
Thr Asn Leu Val Lys Asn Lys Arg Val Gln Pro Thr Glu Ser Ile Val
435 440 445
Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn
450 455 460
Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser
465 470 475 480
Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser
485 490 495
Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys
500 505 510
Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val
515 520 525
Arg Gln Ile Ala Pro Gly Gln Thr Gly Asn Ile Ala Asp Tyr Asn Tyr
530 535 540
Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn
545 550 555 560
Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu
565 570 575
Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu
580 585 590
Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn Gly Val Lys Gly Phe Asn
595 600 605
Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe Gln Pro Thr Tyr Gly Val
610 615 620
Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu His
625 630 635 640
Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys
645 650 655
Asn Lys
<210> 8
<211> 658
<212> PRT
<213> Artificial
<400> 8
Met Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr
1 5 10 15
Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser
20 25 30
Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr
35 40 45
Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly
50 55 60
Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala
65 70 75 80
Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly
85 90 95
Gln Thr Gly Asn Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe
100 105 110
Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val
115 120 125
Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu
130 135 140
Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser
145 150 155 160
Thr Pro Cys Asn Gly Val Lys Gly Phe Asn Cys Tyr Phe Pro Leu Gln
165 170 175
Ser Tyr Gly Phe Gln Pro Thr Tyr Gly Val Gly Tyr Gln Pro Tyr Arg
180 185 190
Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys
195 200 205
Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Arg Val Gln Pro
210 215 220
Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe
225 230 235 240
Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn
245 250 255
Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn
260 265 270
Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys
275 280 285
Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile
290 295 300
Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly Asn Ile
305 310 315 320
Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile
325 330 335
Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn
340 345 350
Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg
355 360 365
Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn Gly
370 375 380
Val Lys Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe Gln
385 390 395 400
Pro Thr Tyr Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser
405 410 415
Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser
420 425 430
Thr Asn Leu Val Lys Asn Lys Arg Val Gln Pro Thr Glu Ser Ile Val
435 440 445
Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn
450 455 460
Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser
465 470 475 480
Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser
485 490 495
Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys
500 505 510
Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val
515 520 525
Arg Gln Ile Ala Pro Gly Gln Thr Gly Asn Ile Ala Asp Tyr Asn Tyr
530 535 540
Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn
545 550 555 560
Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu
565 570 575
Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu
580 585 590
Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn Gly Val Lys Gly Phe Asn
595 600 605
Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe Gln Pro Thr Tyr Gly Val
610 615 620
Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu His
625 630 635 640
Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys
645 650 655
Asn Lys
<210> 9
<211> 1977
<212> DNA
<213> Artificial
<400> 9
atgagagtgc agcctaccga gagcattgtc agattcccta acatcaccaa tctgtgccct 60
ttcggcgagg tgttcaacgc tacaagattt gctagcgtgt acgcttggaa cagaaagaga 120
atcagcaatt gcgtcgccga ctacagcgtg ctgtacaaca gcgcctcctt cagcaccttc 180
aagtgctacg gcgtcagccc tacaaaactg aatgacctgt gctttacaaa tgtgtatgcc 240
gatagcttcg tcatcagagg cgacgaggtg aggcaaatcg ctcctggcca aaccggcaag 300
atcgccgact acaattacaa gctgcctgac gatttcaccg gatgcgtgat tgcctggaac 360
agcaacaacc tggacagcaa ggtgggcggc aactacaact acctgtacag actgtttaga 420
aaaagcaacc tgaagccttt cgagagagac atcagcaccg agatctacca agccggcagc 480
acaccttgca acggcgtgga gggattcaac tgctatttcc ctctgcagag ctacggcttt 540
cagcctacca acggcgtggg ctatcagcct tatagagtcg tggtgctcag cttcgaactc 600
ctgcacgccc ctgctacagt gtgtggccct aagaagtcca ccaacctggt gaagaataag 660
agagtgcaac ctaccgagag catcgtgaga ttccccaata ttaccaacct gtgccccttt 720
ggcgaagtgt tcaacgccac aagattcgct agcgtgtacg cctggaacag aaagagaatc 780
agcaactgcg tggccgacta cagcgtgctc tacaactccg ctagcttcag cacctttaag 840
tgctacggcg tgagccctac caagctgaac gacctgtgct ttaccaacgt ctatgctgac 900
agctttgtga tcagaggcga cgaggtgaga cagatcgctc ccggacagac cggcaatatc 960
gccgattaca actacaagct gcccgacgat ttcaccggat gcgtgatcgc ctggaacagc 1020
aacaatctgg actccaaggt gggaggcaat tacaattacc tgtacagact gtttagaaaa 1080
tccaacctga agcctttcga gagagacatt tccaccgaga tctaccaagc cggctccaca 1140
ccttgcaatg gagtgaaggg cttcaactgc tacttccctc tgcagagcta cggctttcag 1200
cctacatacg gagtcggcta tcagccttac agagtcgtcg tcctgagctt cgagctcctg 1260
cacgcccccg ccaccgtctg cggacctaag aagtccacaa acctcgtcaa aaacaagaga 1320
gtgcagccta ccgagagcat cgtgaggttt cctaacatca ccaacctgtg ccctttcggc 1380
gaggtcttta acgccacaag attcgctagc gtctacgcct ggaacagaaa gaggattagc 1440
aattgtgtcg ccgactacag cgtgctgtac aacagcgcta gcttcagcac cttcaagtgc 1500
tacggcgtga gccctaccaa gctgaacgac ctgtgcttta caaacgtgta tgccgacagc 1560
ttcgtgatta ggggcgacga ggtgaggcaa atcgctcctg gccaaaccgg caagatcgcc 1620
gactacaact acaagctgcc tgacgatttc accggctgcg tgatcgcctg gaacagcaac 1680
aacctggata gcaaggtggg cggcaattac aactacagat atagactgtt cagaaagagc 1740
aacctgaagc ctttcgagag agacatctcc accgagatct atcaagccgg cagcacacct 1800
tgcaacggag tgcaaggctt caactgctat ttccctctgc aatcctacgg ctttcagcct 1860
accaacggag tgggctatca gccttacaga gtcgtcgtgc tgagcttcga gctgctgcac 1920
gcccctgcca cagtgtgcgg acctaaaaag agcacaaatc tggtgaagaa caagtga 1977
<210> 10
<211> 1977
<212> DNA
<213> Artificial
<400> 10
atgagagtgc agcctacaga gagcattgtg agattcccta acatcacaaa cctgtgccct 60
ttcggcgagg tgttcaacgc cacaagattc gctagcgtgt acgcctggaa cagaaaaaga 120
attagcaatt gcgtcgccga ttacagcgtg ctgtataaca gcgcttcctt cagcaccttc 180
aagtgctacg gcgtcagccc tacaaagctg aacgacctgt gcttcacaaa cgtgtatgcc 240
gacagcttcg tgatcagagg cgacgaggtg aggcaaatcg ctcctggcca aaccggcaag 300
atcgccgact acaattacaa gctgcctgat gacttcaccg gatgcgtgat tgcctggaac 360
agcaacaacc tggacagcaa agtcggagga aactacaact acctgtacag actgttcaga 420
aagagcaacc tgaagccttt cgagagagac atcagcaccg agatctacca agccggctcc 480
acaccttgca acggcgtgga gggattcaac tgctacttcc ctctgcagag ctacggcttt 540
cagcccacca acggcgtggg ctatcagcct tacagagtgg tcgtcctgag ctttgagctg 600
ctccacgccc ctgccaccgt ctgcggcccc aaaaaaagca ccaatctggt gaagaacaag 660
agggtgcagc ctacagagag catcgtgaga ttccctaaca ttaccaacct gtgccctttc 720
ggagaagtgt ttaacgccac aagattcgcc tccgtctacg cttggaatag aaagaggatc 780
agcaactgcg tggccgacta cagcgtgctg tacaattccg cctccttcag caccttcaag 840
tgttacggcg tgagccctac caagctgaac gacctgtgct tcaccaacgt gtacgccgat 900
agcttcgtga ttagaggcga cgaggtgagg cagatcgccc ctggacaaac cggcaacatt 960
gctgactaca attacaagct gcctgacgac ttcaccggct gcgtgatcgc ctggaacagc 1020
aacaacctgg acagcaaggt cggaggcaat tacaattacc tctatagact gttcagaaaa 1080
agcaatctca agcctttcga aagagacatt agcaccgaga tctaccaagc cggcagcacc 1140
ccctgcaacg gcgtgaaagg attcaactgc tacttccccc tgcagagcta cggctttcag 1200
cccacctacg gagtgggcta tcaaccctac agagtggtcg tgctctcctt cgagctgctg 1260
catgcccctg ccacagtgtg cggacctaaa aagtccacca acctcgtgaa gaacaagaga 1320
gtgcagccta cagagagcat cgtgaggttc cccaacatca ccaacctgtg ccctttcggc 1380
gaggtgttta acgctacaag attcgctagc gtgtacgctt ggaacagaaa gagaatctcc 1440
aattgcgtgg ccgactacag cgtgctgtac aacagcgcta gcttcagcac cttcaagtgc 1500
tacggagtgt cccctaccaa gctgaacgac ctgtgcttca ccaacgtgta cgccgatagc 1560
ttcgtgatta gaggagacga agtgagacag attgctcctg gacagaccgg caagatcgcc 1620
gactacaatt acaagctgcc tgatgacttt accggctgtg tgattgcctg gaacagcaac 1680
aacctggaca gcaaggtggg cggcaactac aactatctgt acagactctt cagaaagagc 1740
aatctgaagc cttttgaaag ggacatcagc accgagatct atcaagccgg cagcacccct 1800
tgcaacggag tcgagggctt caactgctac tttcctctgc agagctatgg ctttcagcct 1860
accaacggcg tcggatatca gccttacaga gtcgtggtgc tgagcttcga gctgctccac 1920
gcccctgcca ccgtctgcgg ccctaagaaa agcaccaacc tggtcaagaa caaatga 1977
<210> 11
<211> 1977
<212> DNA
<213> Artificial
<400> 11
atgagagtgc agcctacaga gagcattgtg agattcccta acatcaccaa tctgtgccct 60
ttcggcgagg tgttcaacgc cacaagattc gctagcgtgt acgcctggaa cagaaagagg 120
atctccaatt gcgtggctga ctatagcgtg ctgtacaata gcgctagctt cagcaccttc 180
aagtgctacg gcgtcagccc tacaaagctg aacgacctgt gcttcacaaa cgtgtatgcc 240
gacagcttcg tgatcagagg cgacgaggtg aggcaaatcg ctcctggcca aaccggcaag 300
atcgccgact ataattataa gctgcctgat gacttcaccg gctgcgtcat cgcctggaac 360
agcaataatc tggacagcaa ggtcggaggc aactacaact acctgtacag actgttcaga 420
aagagcaacc tgaagccttt cgagagagac atcagcaccg agatttacca agccggctcc 480
accccttgca acggcgtgga aggcttcaac tgctacttcc ctctgcagtc ctacggcttt 540
cagcctacaa acggcgtggg ctaccaaccc tacagagtcg tggtgctcag cttcgagctg 600
ctgcatgccc ctgctaccgt gtgcggcccc aaaaagagca ccaacctcgt gaagaataag 660
agagtgcagc ctaccgagag catcgtcaga ttccctaaca tcaccaatct ctgccccttc 720
ggcgaggtgt tcaacgccac aagattcgct agcgtctacg cttggaatag aaagagaatc 780
agcaactgtg tggccgacta tagcgtgctg tacaacagcg ctagctttag cacctttaag 840
tgctacggcg tgtcccctac caagctgaac gacctgtgct tcacaaatgt gtacgccgac 900
agcttcgtga tcagaggaga cgaggtgagg cagatcgccc ctggacaaac cggcaatatt 960
gccgactaca actacaagct gcccgacgac ttcaccggct gcgtgatcgc ctggaacagc 1020
aacaacctcg acagcaaagt gggaggcaat tacaactacc tgtatagact gtttagaaag 1080
agcaacctga agcctttcga gagggacatc tccaccgaga tctaccaagc cggcagcacc 1140
ccttgtaacg gcgtgaaggg cttcaactgt tacttccctc tgcagagcta tggctttcag 1200
cctacatacg gagtgggcta tcagccttac agagtggtgg tcctctcctt tgaactcctg 1260
catgcccctg ccacagtgtg cggacctaaa aagagcacca acctcgtgaa gaataagaga 1320
gtgcagccta ccgagagcat cgtgagattc cccaatatca ccaacctgtg tcctttcggc 1380
gaggtgttca atgccacaag attcgctagc gtctatgcct ggaacagaaa gagaatctcc 1440
aattgcgtgg ccgactacag cgtgctgtac aacagcgcta gcttcagcac cttcaagtgc 1500
tacggagtga gccctaccaa gctgaacgac ctctgcttta ccaatgtgta cgccgatagc 1560
ttcgtcatta gaggagacga ggtgagacag attgctcctg gacagaccgg caacatcgcc 1620
gactacaact acaagctgcc tgacgatttt accggctgtg tgatcgcctg gaacagcaac 1680
aacctggaca gcaaggtcgg cggcaactac aactatctgt acagactgtt tagaaagagc 1740
aacctgaagc ctttcgaaag ggacatcagc accgagatct atcaagccgg ctccacccct 1800
tgcaacggcg tcaagggctt taactgctac ttccctctgc agagctacgg ctttcagcct 1860
acctacggcg tcggatatca gccttataga gtggtcgtgc tgagcttcga gctgctccac 1920
gcccctgcca ccgtctgcgg ccctaaaaag agcaccaacc tggtcaagaa caaatga 1977
<210> 12
<211> 1977
<212> DNA
<213> Artificial
<400> 12
atgagagtgc agcccacaga gagcattgtg agattcccca acatcaccaa tctgtgcccc 60
ttcggcgagg tgttcaacgc cacaagattc gctagcgtgt acgcctggaa cagaaagcgg 120
atctccaatt gcgtggctga ctatagcgtg ctgtacaata gcgctagctt cagcaccttc 180
aagtgctacg gcgtcagccc cacaaagctg aacgacctgt gcttcacaaa cgtgtatgcc 240
gacagcttcg tgatcagagg cgacgaggtg cggcaaatcg ctcccggcca aaccggcaag 300
atcgccgact ataattataa gctgcccgat gacttcaccg gctgcgtcat cgcctggaac 360
agcaataatc tggacagcaa ggtcgggggc aactacaact acctgtacag actgttcaga 420
aagagcaacc tgaagccctt cgagagagac atcagcaccg agatttacca agccggctcc 480
accccctgca acggcgtgga aggcttcaac tgctacttcc ccctgcagtc ctacggcttt 540
cagcccacaa acggcgtggg ctaccaacct tacagagtcg tggtgctcag cttcgagctg 600
ctgcatgccc ccgctaccgt gtgcggccct aaaaagagca ccaacctcgt gaagaataag 660
agagtgcagc ccaccgagag catcgtcaga ttccccaaca tcaccaatct ctgccctttc 720
ggcgaggtgt tcaacgccac aagattcgct agcgtctacg cttggaaccg gaagagaatc 780
agcaactgtg tggccgacta tagcgtgctg tacaacagcg ctagctttag cacctttaag 840
tgctacggcg tgtcccccac caagctgaac gacctgtgct tcacaaatgt gtacgccgac 900
agcttcgtga tcagagggga cgaggtgcgg cagatcgccc ccgggcaaac cggcaatatt 960
gccgactaca actacaagct gcctgacgac ttcaccggct gcgtgatcgc ctggaacagc 1020
aacaacctcg acagcaaagt ggggggcaat tacaactacc tgtatagact gtttagaaag 1080
agcaacctga agcccttcga gcgggacatc tccaccgaga tctaccaagc cggcagcacc 1140
ccctgtaacg gcgtgaaggg cttcaactgt tacttccccc tgcagagcta tggctttcag 1200
cccacatacg gggtgggcta tcagccctac agagtggtgg tcctctcctt tgaactcctg 1260
catgcccccg ccacagtgtg cgggcccaaa aagagcacca acctcgtgaa gaataagaga 1320
gtgcagccca ccgagagcat cgtgagattc cctaatatca ccaacctgtg tcccttcggc 1380
gaggtgttca atgccacaag attcgctagc gtctatgcct ggaacagaaa gagaatctcc 1440
aattgcgtgg ccgactacag cgtgctgtac aacagcgcta gcttcagcac cttcaagtgc 1500
tacggggtga gccccaccaa gctgaacgac ctctgcttta ccaatgtgta cgccgatagc 1560
ttcgtcatcc ggggggacga ggtgagacag attgctcccg ggcagaccgg caacatcgcc 1620
gactacaact acaagctgcc cgacgatttt accggctgtg tgatcgcctg gaacagcaac 1680
aacctggaca gcaaggtcgg cggcaactac aactatctgt acagactgtt tagaaagagc 1740
aacctgaagc ccttcgaacg ggacatcagc accgagatct atcaagccgg ctccaccccc 1800
tgcaacggcg tcaagggctt taactgctac ttccccctgc agagctacgg ctttcagccc 1860
acctacggcg tcgggtatca gccctaccgg gtggtcgtgc tgagcttcga gctgctccac 1920
gcccccgcca ccgtctgcgg ccccaaaaag agcaccaacc tggtcaagaa caaatga 1977
<210> 13
<211> 1977
<212> DNA
<213> Artificial
<400> 13
atgagagtgc agcctaccga gagcatcgtg agatttccta atatcaccaa tctctgcccc 60
ttcggcgagg tgtttaacgc cacaagattt gctagcgtgt acgcctggaa cagaaaaaga 120
atctccaact gcgtggccga ctacagcgtc ctgtacaata gcgctagctt cagcaccttc 180
aagtgctacg gcgtctcccc tacaaagctg aatgacctgt gttttaccaa cgtgtatgcc 240
gacagcttcg tgattagagg cgatgaggtg aggcagatcg cccctggaca gaccggcaag 300
atcgctgatt acaactacaa gctgcctgac gacttcaccg gctgcgtgat cgcctggaac 360
agcaacaacc tggatagcaa ggtgggcggc aactacaact atctgtacag actgttcaga 420
aaaagcaacc tgaagccttt tgagagagac atcagcacag agatctacca agccggcagc 480
accccttgca acggcgtgga aggatttaac tgctatttcc ctctgcagag ctatggcttc 540
caacctacca acggagtggg ctatcagccc tacagagtgg tcgtgctgag cttcgagctg 600
ctgcacgctc ctgccaccgt gtgcggcccc aaaaagtcca ccaacctggt gaagaataaa 660
agagtgcagc ctaccgaatc catcgtgaga ttccctaaca tcaccaacct gtgccctttc 720
ggcgaggtct tcaacgccac aagatttgct agcgtgtacg cctggaacag aaagagaatc 780
agcaactgcg tggccgacta cagcgtcctg tacaacagcg cctccttcag caccttcaaa 840
tgctacggcg tgtcccctac caagctgaat gacctgtgct ttaccaacgt gtacgccgac 900
agctttgtga tcagaggcga cgaggtgaga cagattgctc ctggacagac cggcaacatt 960
gccgattaca attacaagct gcctgatgat ttcaccggct gtgtgatcgc ctggaacagc 1020
aacaacctgg acagcaaggt gggcggcaac tataattacc tgtatagact gttcagaaaa 1080
agcaacctga agcctttcga gagagacatc tccaccgaaa tttaccaagc cggatccaca 1140
ccttgcaacg gcgtgaaggg cttcaattgt tacttccctc tgcaaagcta cggctttcag 1200
cctacatacg gagtgggcta ccaaccctac agagtggtcg tgctgagctt cgagctgctc 1260
cacgcccctg ccaccgtgtg cggccctaaa aagagcacca acctggtgaa gaataagaga 1320
gtgcaaccca cagagagcat tgtgagattc cctaacatca caaatctgtg ccctttcggc 1380
gaggtgttca acgccacaag attcgctagc gtgtacgctt ggaacagaaa gagaatcagc 1440
aactgtgtgg ctgactacag cgtgctctac aacagcgctt ccttcagcac ctttaaatgc 1500
tacggcgtga gccccacaaa gctcaacgac ctgtgcttca ccaacgtcta cgccgacagc 1560
ttcgtgatta gaggagacga agtgagacag atcgctcctg gacagaccgg caatatcgcc 1620
gactacaact acaaactgcc tgacgacttc accggctgcg tgatcgcctg gaacagcaat 1680
aatctcgaca gcaaggtcgg cggcaactac aattacctgt atagactgtt tagaaagagc 1740
aacctgaagc ccttcgagag agacatctcc accgagatct accaagccgg ctccacacct 1800
tgcaacggcg tgaagggctt caactgctac ttccctctgc agagctacgg cttccaacct 1860
acctacggag tgggctatca gccttataga gtggtcgtgc tgagcttcga actgctgcac 1920
gcccccgcca ccgtgtgcgg ccccaaaaag agcacaaacc tggtgaagaa caagtga 1977
<210> 14
<211> 1977
<212> DNA
<213> Artificial
<400> 14
atgagagtgc agcctaccga gagcatcgtg agattcccta acatcacaaa cctctgccct 60
tttggagagg tcttcaacgc cacaagattc gcctccgtgt atgcctggaa cagaaagaga 120
atcagcaatt gcgtggctga ctacagcgtg ctgtacaact ccgctagctt tagcaccttc 180
aagtgctacg gcgtgagccc taccaaactc aacgatctgt gctttaccaa cgtctatgcc 240
gacagctttg tgatcagagg cgacgaggtc agacaaatcg cccccggaca gaccggcaag 300
atcgccgact acaactataa actgcctgac gacttcaccg gctgcgtgat cgcctggaac 360
agcaacaacc tggactccaa ggtgggcggc aactacaact acctgtacag actcttcaga 420
aagagcaacc tgaaaccttt cgaaagagac atcagcaccg aaatctacca agccggcagc 480
accccttgca acggcgtgga gggctttaat tgctactttc ctctgcaaag ctacggcttt 540
cagcccacca acggagtggg ctatcagcct tatagagtcg tcgtgctgag ctttgagctg 600
ctgcacgccc ctgccaccgt ctgcggccct aaaaagagca caaatctggt caagaacaag 660
agagtgcagc ctaccgagag cattgtgaga tttcctaaca tcaccaatct gtgccctttc 720
ggcgaggtgt tcaatgccac aagatttgct agcgtctacg cctggaacag aaaaagaatc 780
agcaactgcg tggccgacta cagcgtgctg tacaactccg ctagcttcag cacctttaag 840
tgctacggcg tcagccctac caaactgaac gacctctgct ttaccaacgt gtatgccgac 900
agcttcgtga tcagaggaga cgaggtgaga cagatcgccc ctggacagac cggcaacatc 960
gccgattaca attacaaact gcccgacgac ttcaccggct gtgtgattgc ttggaactcc 1020
aacaacctcg acagcaaggt gggcggcaac tacaactacc tgtatagact gttcagaaag 1080
agcaacctga agcctttcga gagagatatc agcaccgaga tctaccaagc cggcagcaca 1140
ccttgtaacg gcgtgaaagg cttcaattgt tatttccccc tgcagagcta cggctttcag 1200
cctacctatg gcgtgggata ccaaccttac agagtggtcg tgctgagctt cgaactgctg 1260
cacgcccctg ccaccgtgtg cggccccaaa aagagcacca atctggtgaa gaacaaaaga 1320
gtgcaaccta ccgagagcat cgtgagattc cccaatatta caaacctgtg cccctttggc 1380
gaggtgttca acgccacaag attcgctagc gtgtacgcct ggaacagaaa gagaatcagc 1440
aactgcgtgg ctgactacag cgtgctgtac aacagcgcct ccttcagcac attcaagtgc 1500
tacggagtca gccctaccaa gctgaatgat ctgtgtttca caaacgtgta cgccgacagc 1560
ttcgtgatca gaggcgacga agtgagacag attgcccctg gacagaccgg caacatcgcc 1620
gactacaatt acaagctgcc tgacgatttc accggctgcg tcatcgcctg gaacagcaac 1680
aacctggact ccaaagtggg cggcaactac aattacctgt atagactgtt cagaaagtcc 1740
aatctcaagc ccttcgagag agacatcagc accgagattt atcaagccgg cagcacccct 1800
tgcaatggag tgaaaggctt caactgctac ttccctctgc agagctacgg atttcagcct 1860
acctacggag tgggctatca gccttacaga gtggtcgtgc tgagctttga gctgctgcac 1920
gctcctgcca ccgtgtgcgg ccctaaaaag agcaccaacc tggtgaagaa caagtga 1977
<210> 15
<211> 1977
<212> DNA
<213> Artificial
<400> 15
atgagagtgc agcctacaga gagcattgtg agattcccta acatcacaaa cctgtgccct 60
ttcggcgagg tgttcaacgc cacaagattc gctagcgtgt acgcctggaa cagaaaaaga 120
attagcaatt gcgtcgccga ttacagcgtg ctgtataaca gcgcttcctt cagcaccttc 180
aagtgctacg gcgtcagccc tacaaagctg aacgacctgt gcttcacaaa cgtgtatgcc 240
gacagcttcg tgatcagagg cgacgaggtg aggcaaatcg ctcctggcca aaccggcaac 300
atcgccgact acaattacaa gctgcctgat gacttcaccg gatgcgtgat tgcctggaac 360
agcaacaacc tggacagcaa agtcggagga aactacaact acctgtacag actgttcaga 420
aagagcaacc tgaagccttt cgagagagac atcagcaccg agatctacca agccggctcc 480
acaccttgca acggcgtgaa gggattcaac tgctacttcc ctctgcagag ctacggcttt 540
cagcccacct acggcgtggg ctatcagcct tacagagtgg tcgtcctgag ctttgagctg 600
ctccacgccc ctgccaccgt ctgcggcccc aaaaaaagca ccaatctggt gaagaacaag 660
agggtgcagc ctacagagag catcgtgaga ttccctaaca ttaccaacct gtgccctttc 720
ggagaagtgt ttaacgccac aagattcgcc tccgtctacg cttggaatag aaagaggatc 780
agcaactgcg tggccgacta cagcgtgctg tacaattccg cctccttcag caccttcaag 840
tgttacggcg tgagccctac caagctgaac gacctgtgct tcaccaacgt gtacgccgat 900
agcttcgtga ttagaggcga cgaggtgagg cagatcgccc ctggacaaac cggcaagatt 960
gctgactaca attacaagct gcctgacgac ttcaccggct gcgtgatcgc ctggaacagc 1020
aacaacctgg acagcaaggt cggaggcaat tacaattacc tctatagact gttcagaaaa 1080
agcaatctca agcctttcga aagagacatt agcaccgaga tctaccaagc cggcagcacc 1140
ccctgcaacg gcgtggaagg attcaactgc tacttccccc tgcagagcta cggctttcag 1200
cccaccaacg gagtgggcta tcaaccctac agagtggtcg tgctctcctt cgagctgctg 1260
catgcccctg ccacagtgtg cggacctaaa aagtccacca acctcgtgaa gaacaagaga 1320
gtgcagccta cagagagcat cgtgaggttc cccaacatca ccaacctgtg ccctttcggc 1380
gaggtgttta acgctacaag attcgctagc gtgtacgctt ggaacagaaa gagaatctcc 1440
aattgcgtgg ccgactacag cgtgctgtac aacagcgcta gcttcagcac cttcaagtgc 1500
tacggagtgt cccctaccaa gctgaacgac ctgtgcttca ccaacgtgta cgccgatagc 1560
ttcgtgatta gaggagacga agtgagacag attgctcctg gacagaccgg caacatcgcc 1620
gactacaatt acaagctgcc tgatgacttt accggctgtg tgattgcctg gaacagcaac 1680
aacctggaca gcaaggtggg cggcaactac aactatctgt acagactctt cagaaagagc 1740
aatctgaagc cttttgaaag ggacatcagc accgagatct atcaagccgg cagcacccct 1800
tgcaacggag tcaagggctt caactgctac tttcctctgc agagctatgg ctttcagcct 1860
acctacggcg tcggatatca gccttacaga gtcgtggtgc tgagcttcga gctgctccac 1920
gcccctgcca ccgtctgcgg ccctaagaaa agcaccaacc tggtcaagaa caaatga 1977
<210> 16
<211> 1977
<212> DNA
<213> Artificial
<400> 16
atgagggtgc agcccacaga gagcatcgtc agattcccta acatcacaaa cctgtgtcct 60
tttggcgagg tcttcaacgc cacaagattc gctagcgtgt acgcctggaa tagaaaaagg 120
attagcaact gtgtggctga ctacagcgtc ctgtataaca gcgcttcctt ttccaccttt 180
aagtgttatg gcgtgtcccc cacaaagctg aacgacctgt gttttaccaa tgtgtacgcc 240
gacagcttcg tcatcagagg agacgaggtg agacaaatcg cccctggaca gaccggcaac 300
atcgccgact acaattataa gctgcctgac gactttaccg gctgcgtgat tgcttggaac 360
tccaacaacc tggatagcaa ggtgggagga aactacaact acctctacag actcttcaga 420
aagagcaatc tgaagccttt cgagagagat atctccacag agatctatca agccggcagc 480
accccttgta acggagtgaa gggattcaat tgttatttcc ctctgcaaag ctatggattt 540
cagcctacct acggagtggg atatcagccc tatagagtgg tcgtgctgag cttcgagctc 600
ctgcacgccc ctgctacagt ctgtggccct aagaagtcca caaacctcgt gaagaacaag 660
agagtgcagc ctaccgagag catcgtgagg ttccctaaca ttaccaacct ctgtcctttt 720
ggagaagtct ttaatgccac aagatttgct agcgtgtacg cttggaatag aaagaggatt 780
tccaattgcg tggccgacta ctccgtcctg tacaacagcg ctagcttcag caccttcaaa 840
tgctacggag tgagccctac caagctcaac gacctgtgct ttaccaacgt gtacgctgac 900
agcttcgtca ttaggggaga cgaggtgagg cagatcgctc ctggacagac cggcaacatc 960
gccgactaca actacaagct gcccgacgat ttcaccggct gcgtcattgc ctggaatagc 1020
aacaatctgg acagcaaggt gggaggaaac tataattacc tgtacagact gtttaggaaa 1080
agcaacctca aacccttcga aagagacatt agcacagaga tctaccaagc cggctccacc 1140
ccctgcaatg gcgtgaaggg atttaattgt tacttccccc tgcagagcta tggctttcaa 1200
cctacctacg gcgtgggcta tcagccctat agggtggtgg tcctcagctt tgaactcctg 1260
cacgctcctg ccaccgtgtg cggccccaaa aagagcacca atctggtcaa gaacaagaga 1320
gtgcagccca cagagtccat cgtgagattt cctaatatta ccaacctgtg ccctttcgga 1380
gaggtgttta atgctacaag atttgctagc gtctatgcct ggaacagaaa gagaatcagc 1440
aactgcgtgg ccgattacag cgtcctgtac aatagcgctt ccttctccac ctttaaatgc 1500
tacggcgtga gccccaccaa gctgaatgac ctctgtttca ccaacgtgta tgccgactcc 1560
tttgtgatta gaggagatga ggtgagacag atcgcccctg gacaaaccgg caacattgcc 1620
gattacaatt acaagctgcc tgatgacttc accggctgtg tgattgcctg gaacagcaat 1680
aacctggaca gcaaggtggg cggcaattat aactacctgt atagactgtt cagaaagagc 1740
aacctgaagc cttttgagag agacatcagc accgagattt accaagccgg ctccacccct 1800
tgcaacggcg tgaagggctt caattgctac ttccctctgc agtcctacgg ctttcagcct 1860
acatacggcg tgggatatca gccttataga gtcgtggtgc tcagcttcga gctgctgcac 1920
gcccccgcta cagtgtgtgg acctaaaaag tccaccaatc tcgtcaagaa taagtga 1977
<210> 17
<211> 1977
<212> DNA
<213> Artificial
<400> 17
atgagagtgc agcctaccga gagcatcgtg agattcccta acatcaccaa cctctgccct 60
ttcggagagg tcttcaacgc cacaagattt gctagcgtgt acgcctggaa cagaaaaaga 120
atctccaact gcgtggccga ctacagcgtc ctgtacaata gcgctagctt cagcaccttc 180
aagtgctacg gcgtctcccc tacaaagctg aatgacctgt gcttcaccaa cgtgtatgcc 240
gacagcttcg tgattagagg cgatgaggtg aggcagatcg cccctggaca aaccggcaac 300
atcgccgatt ataactacaa gctgcctgac gacttcaccg gctgcgtgat cgcctggaac 360
agcaacaacc tggacagcaa ggtcggcggc aactacaact acctctacag actgttcaga 420
aagtccaacc tgaagccttt cgagagggac atcagcaccg agatttacca agccggcagc 480
accccttgca acggcgtgaa gggatttaac tgctatttcc ctctgcagag ctacggcttc 540
caacctacct atggagtggg ctaccaaccc tacagagtgg tggtcctgag cttcgaactg 600
ctgcacgccc ctgccaccgt gtgtggccct aaaaagagca ccaatctcgt gaaaaacaag 660
agagtgcagc ctacagaaag cattgtgaga tttcctaata tcaccaacct gtgccctttc 720
ggcgaggtgt tcaacgctac aagattcgct agcgtgtacg cctggaatag aaagagaatc 780
agcaactgcg tggccgacta cagcgtgctg tacaacagcg cttcctttag caccttcaag 840
tgttacggcg tgagccccac caagctcaac gacctgtgct tcacaaacgt gtacgctgac 900
agcttcgtga tcagaggcga tgaggtgaga cagatcgccc ctggccaaac cggcaacatc 960
gccgattaca actataagct ccctgatgac ttcaccggct gcgtgatcgc ttggaacagc 1020
aacaacctgg acagcaaggt gggcggcaac tacaactacc tctatagact gttcagaaaa 1080
tccaacctga agcctttcga gagagacatc tccaccgaga tttatcaagc cggaagcacc 1140
ccctgcaatg gcgtgaaggg cttcaactgc tacttccctc tccaatccta cggcttccaa 1200
cctacctacg gagtgggcta tcagccctac agagtggtcg tgctgagctt cgagctcctg 1260
cacgcccctg ccaccgtgtg cggccctaaa aagagcacca acctggtcaa aaataagaga 1320
gtgcagccca ccgagagcat cgtgagattc cctaatatca ccaacctgtg cccttttgga 1380
gaggtgttca acgccacaag attcgctagc gtgtacgcct ggaatagaaa gaggatcagc 1440
aactgcgtgg ccgactacag cgtgctctac aacagcgcct ccttcagcac ctttaagtgt 1500
tacggcgtct cccctaccaa gctgaacgat ctctgcttta ccaacgtgta cgccgacagc 1560
tttgtgatca gaggcgacga agtgaggcaa atcgctcctg gacagaccgg aaacatcgcc 1620
gactacaact acaagctgcc tgacgacttc accggctgcg tcatcgcctg gaatagcaac 1680
aacctggata gcaaagtcgg aggcaactac aactacctgt acagactctt cagaaagagc 1740
aacctcaaac ctttcgagag agacatcagc acagagatct accaagccgg cagcacccct 1800
tgcaacggag tcaagggctt caactgctac tttcctctgc agagctacgg ctttcagcct 1860
acctacggcg tcggctatca gccttacaga gtggtcgtgc tcagcttcga actcctgcac 1920
gcccccgcca ccgtgtgcgg acctaaaaag agcaccaatc tggtgaagaa caaatga 1977
<210> 18
<211> 658
<212> PRT
<213> Artificial
<400> 18
Met Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr
1 5 10 15
Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser
20 25 30
Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr
35 40 45
Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly
50 55 60
Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala
65 70 75 80
Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly
85 90 95
Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe
100 105 110
Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val
115 120 125
Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu
130 135 140
Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser
145 150 155 160
Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln
165 170 175
Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg
180 185 190
Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys
195 200 205
Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Arg Val Gln Pro
210 215 220
Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe
225 230 235 240
Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn
245 250 255
Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn
260 265 270
Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys
275 280 285
Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile
290 295 300
Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly Lys Ile
305 310 315 320
Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile
325 330 335
Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn
340 345 350
Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg
355 360 365
Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn Gly
370 375 380
Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe Gln
385 390 395 400
Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser
405 410 415
Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser
420 425 430
Thr Asn Leu Val Lys Asn Lys Arg Val Gln Pro Thr Glu Ser Ile Val
435 440 445
Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn
450 455 460
Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser
465 470 475 480
Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser
485 490 495
Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys
500 505 510
Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val
515 520 525
Arg Gln Ile Ala Pro Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr
530 535 540
Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn
545 550 555 560
Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu
565 570 575
Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu
580 585 590
Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn
595 600 605
Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val
610 615 620
Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu His
625 630 635 640
Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys
645 650 655
Asn Lys
<210> 19
<211> 439
<212> PRT
<213> Artificial
<400> 19
Met Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr
1 5 10 15
Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser
20 25 30
Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr
35 40 45
Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly
50 55 60
Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala
65 70 75 80
Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly
85 90 95
Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe
100 105 110
Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val
115 120 125
Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu
130 135 140
Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser
145 150 155 160
Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln
165 170 175
Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg
180 185 190
Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys
195 200 205
Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Arg Val Gln Pro
210 215 220
Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe
225 230 235 240
Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn
245 250 255
Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn
260 265 270
Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys
275 280 285
Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile
290 295 300
Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly Lys Ile
305 310 315 320
Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile
325 330 335
Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn
340 345 350
Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg
355 360 365
Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn Gly
370 375 380
Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe Gln
385 390 395 400
Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser
405 410 415
Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser
420 425 430
Thr Asn Leu Val Lys Asn Lys
435