CN117043342A - 靶向脱氨酶及使用其进行的碱基编辑 - Google Patents

靶向脱氨酶及使用其进行的碱基编辑 Download PDF

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CN117043342A
CN117043342A CN202180076508.8A CN202180076508A CN117043342A CN 117043342 A CN117043342 A CN 117043342A CN 202180076508 A CN202180076508 A CN 202180076508A CN 117043342 A CN117043342 A CN 117043342A
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fusion protein
dna
protein
nucleic acid
tale
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金晋秀
林佳映
赵诚翼
姜汎昌
李城贤
李贤智
穆荣根
李知玟
郑有眞
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Institute for Basic Science
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Abstract

本发明涉及分离形式的胞嘧啶或腺嘌呤脱氨酶或其变体，无毒全长胞嘧啶脱氨酶或其变体，包含所述脱氨酶或其变体的融合蛋白，用于碱基编辑的组合物以及用于通过使用所述脱氨酶或其变体、所述融合蛋白、所述组合物编辑碱基的方法。

Description

靶向脱氨酶及使用其进行的碱基编辑
【技术领域】
本发明涉及分离形式的胞嘧啶或腺嘌呤脱氨酶或其变体，无毒全长胞嘧啶脱氨酶或其变体，包含所述脱氨酶或其变体的融合蛋白，用于碱基编辑的组合物以及使用所述脱氨酶或其变体、所述融合蛋白、所述组合物编辑碱基的方法。
【背景技术】
其中DNA结合蛋白和脱氨酶彼此融合的融合蛋白使得能够在不产生DNA双链断裂(DSB)的情况下在基因组中实现靶向核苷酸取代或碱基编辑，纠正引起遗传障碍的点突变，或以靶向方式进行单核苷酸转化以将所需的单核苷酸突变引入原核细胞以及人和其他真核细胞中。
与在靶位点诱导小插入或缺失(插入缺失)的核酸酶如CRISPR-Cas9不同，脱氨酶融合蛋白在靶位点的几个核苷酸的窗口内转化单个碱基。因此，可以编辑在培养的细胞、动物和植物中引起遗传疾病的点突变或产生单核苷酸多态性(SNP)。
其中DNA结合蛋白和脱氨酶彼此融合的融合蛋白的例子可以包括1)碱基编辑器(BE)，其包括衍生自酿脓链球菌(S.pyogenes)的催化缺陷型Cas9(dCas9)或D10A Cas9切口酶(nCas9)和作为衍生自大鼠的胞嘧啶脱氨酶的rAPOBEC1；2)靶-AID，其包括dCas9或nCas9和PmCDA1(衍生自海七鳃鳗的活化诱导的胞苷脱氨酶(AID)直向同源物)或人AID；3)CRISPR-X，其包括MS2 RNA发夹连接的sgRNA和dCas9以募集与MS2结合蛋白融合的过度活性AID变体，等等。
已经开发了可编程基因组编辑工具，如ZFN(锌指核酸酶)、TALEN(转录激活因子样效应物核酸酶)、CRISPR(规律间隔成簇短回文重复序列)系统和由CRISPR相关蛋白9(Cas9)变体和核碱基脱氨酶蛋白构成的碱基编辑器，以用于植物遗传研究和通过改变碱基序列改善作物性状。然而，这些工具不适用于编辑植物细胞器(包括线粒体和叶绿体)的DNA序列，主要是因为难以将指导RNA递送到细胞器或在细胞器中共表达两种化合物。植物细胞器编码光合作用所需的必需基因。编辑细胞器的基因的方法或工具对于细胞器基因的功能研究或作物产量和性状的改善是必不可少的。例如，线粒体atp6基因中的靶向突变可导致雄性不育，这是种子生产的有用性状，并且叶绿体基因组的16S rRNA基因中的特定点突变可导致抗生素耐药性。
细菌毒素DddAtox是衍生自洋葱伯克霍尔德菌(Burkholderia cenocepacia)的细菌毒素的酶结构域，并且能够使双链DNA中的胞嘧啶脱氨基。作为脱氨酶的例子，DddAtox具有细胞毒性，因此，为了避免在宿主细胞中的毒性，将DddAtox分裂成两个无活性的半部分，每个半部分与DddA衍生的胞嘧啶碱基编辑器(DdCBE)中的DNA结合蛋白融合。当DNA结合蛋白将两个无活性的半部分结合在一起时，功能性脱氨酶在靶DNA位点重新组装。
原则上，该脱氨酶反应仅在两个无活性的半部分通过DNA结合蛋白紧密接近靶DNA时才被激活。因此，在两个DNA结合蛋白的结合位点之间的间隔子区诱导胞嘧啶到胸腺嘧啶(C到T)碱基编辑。与TALE(转录激活因子样效应物)DNA结合阵列融合的两个无活性形式在它们通过TALE-DNA相互作用而聚集在一起时具有功能。C到T编辑通常在两个TALE结合位点之间的14-18个碱基的区域中被诱导。然而，DddAtox分裂物系统在实验中具有许多限制。
编码全长DddAtox的基因由于毒性而无法在大肠杆菌(E.coli)中被克隆。只有当DddA抑制剂基因在大肠杆菌中共表达时，克隆才有可能。
另一方面，线粒体DNA在细胞呼吸中起非常重要的作用，这是通过线粒体氧化磷酸化(OXPHOS)机制实现的。因为OXPHOS机制对于存活是必需的，所以线粒体DNA中的突变可能在许多器官和肌肉中、特别是在高能量需求的组织中引起严重的功能障碍。在许多人类线粒体疾病中，野生型线粒体DNA与具有单碱基突变的突变型线粒体DNA共存，从而导致线粒体DNA的异质状态。突变型与野生型线粒体DNA之间的平衡决定临床症状性线粒体疾病的发展。在体外和体内，可编程核酸酶已被用于切割并由此去除突变型线粒体DNA而不切割野生型线粒体DNA。然而，这些核酸酶不能诱导或恢复线粒体中的特定突变：与细胞核中的DNA双链断裂不同，线粒体中的DNA双链断裂不能通过非同源末端连接或同源重组有效修复。
线粒体碱基编辑可用于创建各种疾病的模型或用于生产治疗这些疾病的治疗剂。在这方面，越来越需要开发高效的线粒体碱基编辑酶。
在这种技术背景下，我们已经通过证实以下事实完成了本发明：可以通过使用所需的CBE(胞嘧啶碱基编辑器)或ABE(腺嘌呤碱基编辑器)或通过使用不具有细胞毒性的新型全长脱氨酶纠正DNA，所述CBE或ABE是通过脱氨酶残基的取代来减少非选择性碱基编辑而产生的。
【发明内容】
本发明的一个目的是提供一种融合蛋白，所述融合蛋白包含DNA结合蛋白和分离形式的胞嘧啶或腺嘌呤脱氨酶或其变体或无毒全长胞嘧啶脱氨酶或其变体。
本发明的另一个目的是提供一种编码融合蛋白的核酸。
本发明的又另一个目的是提供一种用于碱基编辑的组合物，所述组合物包含融合蛋白或核酸。
本发明的又另一个目的是提供一种碱基编辑方法，所述方法包括用所述组合物处理细胞。
为了实现上述目的，本发明提供了一种融合蛋白，所述融合蛋白包含(i)DNA结合蛋白和(ii)衍生自胞嘧啶脱氨酶或其变体的第一分裂物和第二分裂物，其中所述第一分裂物和所述第二分裂物中的每一个与DNA结合蛋白融合。
此外，本发明提供了一种融合蛋白，所述融合蛋白包含(i)DNA结合蛋白和(ii)衍生自胞嘧啶脱氨酶或其变体的无毒全长胞嘧啶脱氨酶。
此外，本发明提供了一种融合蛋白，所述融合蛋白包含(i)DNA结合蛋白，(ii)胞嘧啶脱氨酶或其变体，和(iii)腺嘌呤脱氨酶，其中所述胞嘧啶脱氨酶或其变体包括(a)无毒全长胞嘧啶脱氨酶或(b)衍生自胞嘧啶脱氨酶或其变体的第一分裂物和第二分裂物，所述第一分裂物和所述第二分裂物中的每一个与所述DNA结合蛋白融合。
此外，本发明提供了一种编码融合蛋白的核酸。
此外，本发明提供了一种用于碱基编辑的组合物，所述组合物包含所述融合蛋白或所述核酸。
此外，本发明提供了一种用于在真核细胞中进行碱基编辑的组合物，所述组合物包含所述融合蛋白或所述核酸。
此外，本发明提供了一种用于在植物细胞中进行碱基编辑的组合物，所述组合物包含所述融合蛋白或所述核酸以及核定位信号(NLS)肽或编码其的核酸。
此外，本发明提供了一种用于在植物细胞中进行碱基编辑的组合物，所述组合物包含所述融合蛋白或所述核酸以及叶绿体转运肽或编码其的核酸。
此外，本发明提供了一种用于在植物细胞中进行碱基编辑的组合物，所述组合物包含所述融合蛋白或所述核酸以及线粒体靶向信号(MTS)或编码其的核酸。
在一些情况下，本发明还提供了一种用于在植物细胞中进行碱基编辑的组合物，所述组合物进一步包含核输出信号或编码其的核酸。
此外，本发明提供了一种用于在植物细胞中进行碱基编辑的方法，所述方法包括用所述组合物处理植物细胞。
此外，本发明提供了一种用于在植物细胞中进行碱基编辑的方法，所述方法包括用含有核定位信号(NLS)肽或编码其的核酸的所述融合蛋白或所述核酸处理植物细胞。
此外，本发明提供了一种用于在植物细胞中进行碱基编辑的方法，所述方法包括用含有叶绿体转运肽或编码其的核酸的所述融合蛋白或所述核酸处理植物细胞。
此外，本发明提供了一种用于在植物细胞中进行碱基编辑的方法，所述方法包括用含有线粒体靶向信号(MTS)或编码其的核酸的所述融合蛋白或所述核酸处理植物细胞。
此外，本发明提供了一种用于在动物细胞中进行碱基编辑的组合物，所述组合物包含含有核定位信号(NLS)肽或编码其的核酸的所述融合蛋白或所述核酸。
此外，本发明提供了一种用于在动物细胞中进行碱基编辑的组合物，所述组合物包含所述融合蛋白或所述核酸以及线粒体靶向信号(MTS)或编码其的核酸。
在一些情况下，本发明还提供了一种用于在动物细胞中进行碱基编辑的组合物，所述组合物进一步包含核输出信号或编码其的核酸。
此外，本发明提供了一种用于在动物细胞中进行碱基编辑的方法，所述方法包括用所述组合物处理动物细胞。
此外，本发明提供了一种用于在动物细胞中进行碱基编辑的方法，所述方法包括用含有核定位信号(NLS)肽或编码其的核酸的所述融合蛋白或所述核酸处理动物细胞。
此外，本发明提供了一种用于在动物细胞中进行碱基编辑的方法，所述方法包括用含有线粒体靶向信号(MTS)或编码其的核酸的所述融合蛋白或所述核酸处理动物细胞。
此外，本发明提供了一种用于在原核或真核细胞中进行A至G碱基编辑的组合物，所述组合物包含所述融合蛋白或编码其的核酸，其中所述DNA结合蛋白是锌指蛋白、TALE蛋白或CRISPR相关蛋白，以及胞嘧啶脱氨酶或其变体衍生自细菌并且对双链DNA具有特异性。
此外，本发明提供了一种用于在原核或真核细胞中进行A至G碱基编辑的组合物，所述组合物包含所述融合蛋白或编码其的核酸，其中所述DNA结合蛋白是锌指蛋白、TALE蛋白或CRISPR相关蛋白，所述融合蛋白的胞嘧啶脱氨酶或其变体衍生自细菌并且对双链DNA具有特异性。所述DNA结合蛋白与所述胞嘧啶脱氨酶或其变体的N末端和C末端融合。类似地，所述DNA结合蛋白也与所述融合蛋白的腺嘌呤脱氨酶的N末端和C末端融合。在包含DNA结合蛋白、胞嘧啶脱氨酶或其变体和腺嘌呤脱氨酶的融合蛋白的上下文中，所述腺嘌呤脱氨酶可以位于所述融合蛋白内胞嘧啶脱氨酶的N末端或C末端，或者可以作为独立于其他DNA结合蛋白的单独蛋白质存在。
此外，本发明提供了一种用于在原核或真核细胞中进行C至T碱基编辑的组合物，所述组合物包含所述融合蛋白或编码其的核酸和尿嘧啶糖基化酶抑制剂(UGI)，其中所述DNA结合蛋白是锌指蛋白、TALE蛋白或CRISPR相关蛋白，以及胞嘧啶脱氨酶或其变体是无毒全长胞嘧啶脱氨酶，以及所述融合蛋白中的胞嘧啶脱氨酶或其变体衍生自细菌并且对双链DNA具有特异性。
此外，本发明提供了一种用于在原核或真核细胞中进行C至T碱基编辑的组合物，所述组合物包含融合蛋白或编码其的核酸和UGI，其中所述DNA结合蛋白是锌指蛋白、TALE蛋白或CRISPR相关核酸酶，以及所述融合蛋白的胞嘧啶脱氨酶或其变体是包含第一分裂物和第二分裂物的分裂胞嘧啶脱氨酶，以及所述融合蛋白的胞嘧啶脱氨酶或其变体衍生自细菌并且对双链DNA具有特异性。
此外，本发明提供了一种用于在原核或真核细胞中进行A至G碱基编辑的方法，所述方法包括用所述融合蛋白或编码其的核酸处理原核或真核细胞，其中所述DNA结合蛋白是锌指蛋白、TALE蛋白或CRISPR相关核酸酶，以及所述融合蛋白的胞嘧啶脱氨酶或其变体衍生自细菌并且对双链DNA具有特异性。
此外，本发明提供了一种用于在原核或真核细胞中进行A至G碱基编辑的方法，所述方法包括用融合蛋白或编码其的核酸处理原核或真核细胞，其中所述DNA结合蛋白是锌指蛋白、TALE蛋白或CRISPR相关核酸酶，
所述融合蛋白的胞嘧啶脱氨酶或其变体衍生自细菌并且对双链DNA具有特异性，以及
所述融合蛋白的胞嘧啶脱氨酶或其变体衍生自细菌并且对双链DNA具有特异性。所述DNA结合蛋白与所述胞嘧啶脱氨酶或其变体的N末端和C末端融合。类似地，所述DNA结合蛋白也与所述融合蛋白的腺嘌呤脱氨酶的N末端和C末端融合。在包含DNA结合蛋白、胞嘧啶脱氨酶或其变体和腺嘌呤脱氨酶的融合蛋白的上下文中，所述腺嘌呤脱氨酶可以位于所述融合蛋白内胞嘧啶脱氨酶的N末端或C末端，或者可以作为独立于其他DNA结合蛋白的单独蛋白质存在。
此外，本发明提供了一种用于在原核或真核细胞中进行C至T碱基编辑的方法，所述方法包括用融合蛋白或编码其的核酸和UGI处理原核或真核细胞，其中所述DNA结合蛋白是锌指蛋白、TALE蛋白或CRISPR相关核酸酶，并且所述融合蛋白的胞嘧啶脱氨酶或其变体衍生自细菌并且对双链DNA具有特异性。
【附图说明】
图1显示了使用pTarget质粒优化ZFD的结果，
图1a显示了ZFD构建体，其中分裂DddAtox半部分融合到ZFP的C末端(锌指蛋白：C型)，图1b显示了使用pTarget文库的ZFD平台的优化，其中pTarget质粒包含大小为1至24bp的间隔子区(以红色表示)和ZFP DNA结合位点(以绿色表示)，并且ZFD构建体包含各种长度的AA接头(以黄色和橙色表示)和不同的DddAtox分裂位点和方向(以蓝色表示)，以及图1c和图1d显示了在pTarget文库中的靶位点处测量的ZFD活性，以检查图1b中描述的变量的作用，其中测试了在左和右ZFD中具有相同(c)或不同(d)长度的接头的ZFD对，通过对pTarget质粒的相关区域的靶向深度测序来测量碱基编辑频率，并且数据表示为来自n＝2个生物学上独立的样品的平均值±平均值的标准误差(s.e.m.)；
图2显示了证实pTarget质粒中在采用各种接头的情况下ZFD效率的结果，
图2a显示了在热图中描绘的长度为1-24bp的各种pTarget间隔子的从C/G到非C/G的编辑频率，其中测试了各种ZFD构型，包括ZFP与分裂DddAtox之间的各种类型的接头和其中DddAtox为分裂的区域，图2b显示了每个ZFD对的总体活性，其中用于x轴的命名法显示了在底部的左ZFD和在顶部的右ZFD，以及图2c显示了取决于间隔子长度的碱基编辑效率，其中“AA”表示接头中的氨基酸数目，数据表示为来自n＝2个生物学上独立的样品的平均值±平均值的标准误差(s.e.m.)；
图3显示了证实具有24AA接头和不同接头的ZFD的效率的结果，
其中显示了ZFD接头长度对热图中从C/G到非C/G的编辑效率的影响，其中ZFD对的左ZFD用24AA接头固定，并且右ZFD包含可变长度的接头，或反之亦然，误差条是n＝2个生物学上独立的样品的平均值的标准误差(s.e.m.)；
图4显示了证实ZFD在体内靶向细胞核的活性的结果，
图4a显示了靶向核DNA的ZFD的构型，其中分裂DddAtox半部分融合到ZFP的C末端(C型)或N末端(N型)，ZFD对被设计成CC或NC构型，其分别由C型左ZFD和C型右ZFD或N型左ZFD和C型右ZFD构成，图4b显示了由ZFD在HEK 293T细胞中的核DNA靶位点处诱导的碱基编辑频率，数据表示为来自n＝3个生物学上独立的样品的平均值±s.e.m.；图4c-图4f显示了HEK 293T细胞中NUMBL(c)、INPP5D-2(d)、TRAC-CC(e)和TRAC-NC(f)靶位点处间隔子内每个碱基位置处ZFD诱导的碱基编辑效率，数据表示为来自n＝3个生物学上独立的样品的平均值±s.e.m.；图4g显示了在ZFD蛋白或ZFD编码质粒的电穿孔或直接递送后K562细胞中ZFD诱导的碱基编辑频率，其中测试了具有一或四个NLS的ZFD蛋白，并等摩尔地使用左和右ZFD，使用Amaxa 4D-Nucleofector进行电穿孔，并且对于直接递送，将K562细胞与含有左和右ZFD蛋白的细胞培养基一起孵育，并以相同的方式处理细胞一次(1x)或两次(2x)，数据表示为来自n＝2个生物学上独立的样品的平均值±s.e.m.；
图5示意性地显示了靶向核DNA的ZFD的构型，图5a-图5d显示了四种可能的ZFD构型，其中NC和CN构型在结构上相同，但是左和右ZFD构型的类型不同；
图6显示了证实体内靶向细胞核的ZFD的插入缺失比率的结果，其中所有测试的ZFD以小于0.4％的频率产生插入缺失，并且数据表示为来自n＝3个生物学上独立的样品的平均值±s.e.m.；
图7显示了测试体外重组ZFD蛋白的活性的结果，
图7a显示了靶向TRAC位点的ZFD对的纯化，其中将GST标记的蛋白质使用谷胱甘肽琼脂糖珠从大肠杆菌细胞裂解物纯化，使用聚丙烯酰胺凝胶电泳监测纯化步骤，将凝胶用考马斯蓝染色，泳道1显示分子量标记物，泳道2显示细胞的样品(其蛋白质表达不用IPTG诱导的)，泳道3显示细胞的样品(其蛋白质表达是用IPTG诱导的)，泳道4显示超声处理后的可溶性级分，泳道5显示超声处理后的不溶性级分，泳道6显示柱流经级分，泳道6显示洗涤级分，以及泳道7显示洗脱级分，代表性标记物的大小显示在左边，并且红色框指示ZFD蛋白，图7b显示左和右ZFD结合位点，其中红色箭头指示ZFD诱导的脱氨的可能位点，图7c显示对于含有TRAC位点的PCR扩增子的ZFD活性，其中TRAC-NC ZFD对使胞嘧啶脱氨以产生尿嘧啶(以红色表示)，然后USER酶切割尿嘧啶以形成裂口(以红色三角形表示)，以及图7d显示了通过琼脂糖凝胶电泳分析的未处理的PCR扩增子(左)和用ZFD对处理的PCR扩增子(右)；
图8示意性地显示了靶向线粒体DNA的ZFD的各种构型，图8a-图8d显示了四种可能的mitoZFD构型，其中现有ZFD的NLS被MTS和NES代替，并且NC和CN构型在结构上是相同的，但是左和右ZFD构型的类型是不同的；
图9显示了证实mitoZFD的线粒体基因碱基编辑效率的结果，
图9a显示了HEK 293T细胞中由mitoZFD和TALE-DdCBE诱导的在mtDNA中的碱基编辑频率，数据表示为来自n＝2个生物学上独立的样品的平均值±平均值的标准误差(s.e.m.)，图9b-图9g显示了在HEK 293T细胞中ND2(b)、ND4L(c)、COX2(d)、ND6(e)和ND1(f)靶位点处间隔子内每个碱基位置处mitoZFD诱导的碱基编辑效率，和ND1(g)靶位点处TALE-DdCBE诱导的碱基编辑效率，数据表示为来自n＝2个生物学上独立的样品的平均值±平均值的标准误差(s.e.m.)，以及图9h显示了由mitoZFD和TALE-DdCBE引入的在ND1基因中的DNA变化和氨基酸变化的比较，其中通过靶向深度测序测量每个突变体等位基因的测序读段的频率(％)，并且ZFD对和TALE-DdCBE对的间隔子区表示为蓝色虚线；
图10显示了证实从用MT-ZFD处理的HEK293T细胞分离的单细胞衍生的克隆群体的碱基编辑效率的结果，
其中获得单细胞衍生的克隆以用于等位基因分析，通过靶向深度测序测定单独单细胞衍生的克隆中的C/G至非C/G编辑频率。图10a显示了用靶向ND1的mitoZFD处理的HEK293T细胞群体的单细胞衍生的克隆，图10b显示了用靶向ND2的mitoZFD处理的HEK 293T细胞群体的单细胞衍生的克隆，以及图10c显示了未处理的HEK 293T细胞群体的单细胞衍生的克隆，其中ZFP结合位点以绿色表示，并且经受mitoZFD诱导的编辑的克隆的高编辑频率以红色表示；
图11显示了证实从用MT-ZFD处理的HEK293T细胞分离的单细胞衍生的克隆群体的碱基编辑效率的结果，
其中提供了具有高碱基编辑频率的单细胞衍生的克隆的等位基因分析，表格显示了ND1中通过碱基编辑改变的氨基酸，并且在顶部参考序列中，红色字母指示间隔子，并且在等位基因中，红色字母指示氨基酸序列的改变(*指示终止密码子)；
图12a和图12b显示了证实从用MT-ZFD处理的HEK293T细胞分离的单细胞衍生的克隆群体的碱基编辑效率的结果，其中提供了具有高碱基编辑频率的单细胞衍生的克隆的等位基因分析，表格显示了ND2中通过碱基编辑改变的氨基酸，并且在顶部参考序列中，红色字母指示间隔子，并且在等位基因中，红色字母指示氨基酸序列的改变；
图13显示了证实通过ZFD和TALE-DdCBE的组合实现的碱基编辑效率的结果，
图13a显示了mitoZFD和TALE-DdCBE对的结合区的DNA序列，其中由TALE-DdCBE识别的位点以绿色突出显示，并且由mitoZFD识别的位点以蓝色突出显示，以及上部序列代表mtDNA重链，并且下部序列代表mtDNA轻链，图13b显示了通过ZFD、TALE-DdCBE和ZFD/DdCBE杂交对编辑的胞嘧啶的频率，数据是使用靶向深度测序获得的，并且数据表示为来自n＝2个生物学上独立的样品的平均值±平均值的标准误差(s.e.m.)，并且图13c显示了每个碱基位置处的碱基编辑活性的热图，红色框表示每种构型的间隔子区，并且蓝色箭头表示mtDNA的位置；
图14显示了证实在不同ZFD浓度下依赖于mRNA相对于质粒的编辑效率的结果，其中靶向ND1的mitoZFD的线粒体全基因组靶向特异性具有根据编码ZFD的mRNA或质粒的浓度而变化的特异性，显示了通过全mtDNA测序确定的中靶和脱靶碱基编辑频率，用指定浓度的编码靶向ND1的mitoZFD的质粒或mRNA转染的HEK 293T细胞的结果在图中以点绘制，红色箭头表示靶位点，红点表示靶位点的碱基编辑频率，以及灰点表示对照中也存在的SNP，数据表示为来自n＝2个生物学上独立的样品的平均值±平均值的标准误差(s.e.m.)；
图15显示了证实在不同ZFD浓度下依赖于mRNA相对于质粒的编辑效率的结果，图15a显示了在顶部ND1位点处的ZFD结合，其中ZFD结合位点以绿色表示，间隔子内的靶胞嘧啶以红色表示，并且显示了从图14中的全mtDNA测序数据确定的中靶活性，所述活性随着编码转染的mitoZFD的质粒或mRNA的量的减少而降低，图15b显示了对于每个质粒或mRNA量以>1％的频率编辑的C/G位点的数量，并且图15c显示了取决于每个质粒或mRNA浓度的线粒体基因组中的所有C/G的平均C/G至T/A编辑频率，数据表示为来自n＝2个生物学上独立的样品的平均值±平均值的标准误差(s.e.m.)；
图16显示了证实在不同ZFD浓度下依赖于mRNA相对于质粒的编辑效率的结果，其中显示了通过全mtDNA测序确定的中靶和脱靶碱基编辑频率，用指定浓度的编码靶向ND2的mitoZFD的质粒或mRNA转染的HEK 293T细胞的结果在图中以点绘制，红色箭头表示靶位点，红点表示靶位点的碱基编辑频率，以及灰点表示对照中也存在的SNP，数据表示为来自n＝2个生物学上独立的样品的平均值±平均值的标准误差(s.e.m.)；
图17显示了证实在不同ZFD浓度下依赖于mRNA相对于质粒的编辑效率的结果，
图17a显示了在顶部ND2位点处的ZFD结合，其中ZFD结合位点以绿色表示，间隔子内的靶胞嘧啶以红色表示，显示了从图16中的全mtDNA测序数据确定的中靶活性，并且所述活性随着编码转染的mitoZFD的质粒或mRNA的量的减少而降低，图17b显示了对于每个质粒或mRNA量以>1％的频率经受碱基编辑的C/G位点的数量，图17c显示了取决于每个质粒或mRNA浓度的线粒体基因组中的所有C/G的平均C/G至T/A编辑频率，数据表示为来自n＝2个生物学上独立的样品的平均值±平均值的标准误差(s.e.m.)；
图18显示了证实构建全线粒体测序/QQ变体后编辑效率的结果，
图18a显示了QQ mitoZFD变体，其在ZFD的每个锌指中包含R(-5)Q突变以消除非特异性DNA接触(如果在锌指框架的位置-5处没有R，则将附近的K或R转化为Q)，图18b显示了mitoZFD处理的细胞的全mtDNA测序，其中将位点处和位点外编辑频率分别表示为红点和黑点，数据表示为来自n＝2个生物学上独立的样品的平均值±平均值的标准误差(s.e.m.)，显示了效率>1％的从C/G到T/A的所有碱基编辑，并且图18c和图18d显示了随着所递送的编码ZFD的mRNA的容量而变化的编辑效率和特异性，图18c显示了线粒体基因组中所有C/G的平均C/G至T/A编辑频率，并且图18d显示了以>1％的碱基编辑频率编辑的C/G的数量；
图19显示了植物中碱基编辑器的Golden Gate组装系统，并示意性地显示了cp-DdCBE和mt-DdCBE构建体的Golden Gate组装，其中对于靶序列中的每个位置，TALE子阵列质粒选自424个序列的总集合(＝6×64个三部分+2×16个二部分+2×4个单部分)，并与所需载体混合以获得编码靶向特定序列的DdCBE的质粒；
图20显示了植物叶绿体和线粒体碱基编辑，图20a、图20b、图20c和图20d显示了由cp-DdCBE在16s rDNA(a，b)和psbA(c，d)中诱导的叶绿体碱基编辑的频率和模式，其中将分裂DdCBE G1333和G1397对转染到莴苣和油菜籽原生质体中，图20e和图20f显示了由mt-DdCBE在ATP6基因中诱导的线粒体碱基编辑的效率和模式，其中将分裂DdCBE G1333和G1397对转染到莴苣和油菜籽原生质体中，在图20a、图20c和图20e中，TALE结合区以蓝色表示，间隔子中的胞嘧啶以橙色表示，所有图中的误差条表示三个独立生物重复样品的平均值±标准偏差，并且在图20b、图20d和图20f中，转化的核苷酸以红色表示，编辑的等位基因％(平均值±标准偏差)是从三个独立实验获得的；
图21显示了通过DdCBE进行的植物细胞器DNA编辑，图21a示意性地显示了植物细胞器诱变，图21b显示了在不存在壮观霉素的情况下培养的cp-DdCBE转染的愈伤组织中C·G至T·A转化效率，包括代表性Sanger测序色谱图，其中转化的核苷酸在左侧用红色表示，并且箭头表示色谱图中的取代的核苷酸，图21c显示了DdCBE驱动的植物细胞器诱变，其中突变体愈伤组织显现出具有比模拟愈伤组织中的频率高得多的编辑频率，图21d显示了在将靶向16srDNA的编码cp-DdCBE的mRNA转染到莴苣原生质体中后诱导的C至T转化频率，误差条为n＝3个独立生物重复样品的平均值±s.d.，图21e显示了在2.5个月时壮观霉素耐药性愈伤组织的编辑频率和模式，图21f显示了使用代表性Sanger测序色谱图获得的在用DdCBE mRNA转染的链霉素耐药性植物中的C·G至T·A转化效率，箭头表示色谱图中的取代核苷酸，比例尺：1mm；
图22显示了DdCBE质粒转染的或DdCBE mRNA转染的莴苣原生质体中靶位点附近的脱靶活性的比较。将编码靶向叶绿体16S rRNA基因的cp-DdCBE对的质粒或mRNA转染到莴苣原生质体中。在紧邻靶位点附近检测到脱靶TC至TT编辑。转染后七天通过靶向深度测序测量编辑效率。频率(平均值±s.d.)是从三个独立实验中获得的。应用学生非配对双尾t检验。**P<0.01；*P<0.05；NS，不显著(P>0.05)；
图23显示了叶绿体和线粒体碱基编辑策略，其中cp-DdCBE和mt-DdCBE前蛋白各自含有叶绿体转运肽(CTP)或线粒体靶向信号(MTS)，因此在植物细胞中被翻译，然后转运到叶绿体和线粒体，并且前蛋白穿过细胞器的外膜和内膜，CTP和MTS分别被间质加工肽酶和线粒体加工肽酶切割，然后cp-DdCBE和mt-DdCBE(成熟蛋白)形成最终构象；
图24显示了莴苣原生质体中随时间而变的通过DdCBE质粒的编辑，其中在每个时间点收集转染的原生质体，并通过靶向深度测序分析编辑效率，频率(平均值±s.d.)是从三个独立实验中获得的；
图25显示了psbB基因的碱基编辑频率，其中将编码靶向叶绿体psbB基因的cp-DdCBE对左-G1333-N+右-G1333-C的质粒转染到油菜籽原生质体中，然后通过靶向深度测序分析间隔子中的碱基编辑效率，TALE结合区、靶胞嘧啶和转化的核苷酸分别以蓝色、橙色和红色表示，以及频率(平均值±标准偏差)是从n＝3个独立实验计算的；
图26显示了线粒体RPS14基因的碱基编辑频率，其中将编码靶向RPS14基因的mt-DdCBE对左-G1333-N+右-G1333-C的质粒转染到油菜籽原生质体中，然后通过靶向深度测序分析C至T转化效率，TALE结合区、靶胞嘧啶和转化的核苷酸分别以蓝色、橙色和红色表示，以及频率(平均值±标准偏差)是从n＝3个独立实验计算的；
图27a显示了靶向愈伤组织中叶绿体基因组的碱基编辑效率，其在培养4周后，DdCBE在莴苣和油菜籽愈伤组织中16srDNA和psbA的靶位点处的碱基编辑频率和模式，间隔子中转化的核苷酸以红色表示；图27b显示了靶向愈伤组织中线粒体基因组的碱基编辑效率，其中通过靶向深度测序证实了DdCBE在油菜籽愈伤组织中ATP6和RPS14基因的靶位点处的碱基编辑频率和模式，靶间隔子中转化的核苷酸以红色表示；
图28显示了无DNA碱基编辑，其在将DdCBE mRNA转染到莴苣原生质体中后，在16srDNA的靶位点处的叶绿体碱基编辑频率和模式，其中将原生质体培养7天，然后进行靶向深度测序，目的间隔子中转化的核苷酸以红色表示；
图29显示了凝胶电泳的结果，显示了原生质体和愈伤组织中不存在DdCBE mRNA或DNA序列(其中M是标记物)；
图30显示了16srDNA突变筛选，其中红色箭头表示链霉素耐药性绿色愈伤组织；
图31显示了抗生素耐药性愈伤组织或芽中DdCBE靶位点附近没有脱靶突变，图31(a)和图31(b)显示了通过靶深度测序分析的脱靶活性，其中TALE结合位点和间隔子区分别加绿色和红色下划线，图31(a)显示了由用DdCBE质粒转染的莴苣原生质体的培养物产生的壮观霉素耐药性愈伤组织，以及图31(b)显示了从链霉素耐药性芽获得的芽；
图32显示了在与靶向位点最同源的五个位点处的脱靶活性的分析结果，其中选择了莴苣叶绿体基因组中16s rRNA基因特异性DdCBE的前五个候选脱靶位点，包括TALE结合位点中最多九个错配，TALE结合序列和错配核苷酸分别以蓝色和红色表示，以及使用靶向深度测序测量了用DdCBE质粒或DdCBE mRNA转染的原生质体和耐药愈伤组织或芽中的脱靶突变频率，频率(平均值±标准偏差)是从三个独立实验中获得的；
图33示意性地显示了DdCBE组装和线粒体DNA编辑，图33a显示了用于有效DdCBE构建的一锅式Golden Gate组装，其中将总共424个序列(64个三部分阵列×6+16个二部分阵列×2+4个单部分阵列×2)和表达载体混合以构建左模块和右模块以用于最终质粒构建，并且图33b示意性地显示了DdCBE与小鼠线粒体DNA中靶基因ND5之间的相互作用，其中TALE结合位点以灰色表示，碱基编辑位点以黑色表示，并且相应的重复可变双残基模块以橙色、蓝色、绿色和黄色表示：“NI”、“NG”、“NN”和“HD”分别用于识别腺嘌呤、胸腺嘧啶、鸟嘌呤和胞嘧啶；
图34显示了由DdCBE碱基编辑引起的小鼠线粒体ND5点突变，图34a显示了在DdCBE脱氨酶介导的胞嘧啶至胸腺嘧啶碱基编辑靶序列和NIH3T3细胞中的效率，其中靶序列中的翻译密码子加下划线，可编辑位点以红色表示，DdCBE转染的组合表示为左或右、-G1333或-G1397、以及-N或-C，左-G1333-N+右-G1333-C、左-G1333-C+右-G1333-N、左-G1397-N+右-G1397-C、和左-G1397-C+右-G1397-N的C10突变的P值分别为0.0012、0.0003、0.0014和0.0009，以及C13突变的p值分别为0.0116、0.0076、0.0030和0.0003(*p<0.05和**p<0.01，学生双尾t检验)，图34b显示了小鼠胚泡中的碱基编辑效率，其中测序数据是从用左-G1397-N和右-G1397-C DdCBE mRNA显微注射的受精卵发育的胚泡获得的，图34c显示了新生幼崽的突变体序列的比对，其中通过从出生后立即从尾部获得的组织中以及在出生后7天和14天从脚趾获得的组织中提取基因组DNA来进行靶向深度测序，编辑的碱基以红色表示，并且指示了突变体线粒体基因组的编辑频率，并且图34d显示了成年F0小鼠(sipup-1)的各种组织中的编辑效率，其中在出生后50天从每个组织获得测序数据，并且在所有图中，深灰色条和浅灰色条表示m.C12539T(C10)和m.G12542A(C13)突变的相应编辑频率，误差条是n＝3个生物独立样品的平均值的标准误差(s.e.m.)；
图35显示了突变体线粒体DNA向生殖细胞的传递，图35a显示了在通过使雌性F0(sipup-3)小鼠与野生型C57BL6/J雄性杂交获得F1后代(101，102)后进行的靶向深度测序的结果，以观察mtDNA突变的种系传递，其中编辑的碱基以红色表示，并且指示了突变体线粒体基因组的编辑频率，并且图35b显示了使用基因组DNA的靶向深度测序获得的F1幼崽(101)的各种组织中的碱基编辑效率，其中深灰色条和浅灰色条表示m.C12539T(C10)和m.G12542A(C13)突变的相应频率，误差条是n＝3个生物独立样品的平均值的标准误差(s.e.m.)；
图36显示了由DdCBE引起的小鼠线粒体ND5 G12918A突变，图36a显示了用于在ND5蛋白中产生m.G12918A点突变从而引起D393N变化的DdCBE靶标，其中靶密码子加下划线，以及可编辑位点以红色表示，图36b显示了在NIH3T3细胞中使用DdCBE获得的胞嘧啶-胸腺嘧啶碱基编辑效率，其中指示了转染的DdCBE对的组合，误差条是n＝3个生物独立样品的s.e.m(n.s.：不显著，*p<0.05，**p<0.01，使用学生双尾t检验)，左-G1333-N+右-G1333-C、左-G1333-C+右-G1333-N、左-G1397-N+右-G1397-C、和左-G1397-C+右-G1397-N的C6突变的P值分别为0.0052、0.0099、0.0027和0.0040，以及n.s的P值为0.4971，图36c显示了m.G12918A小鼠胚泡中的点突变碱基编辑效率，其中测序数据是从胚泡获得，所述胚泡是通过将编码左-G1397-C和右-G1397-N DdCBE的mRNA显微注射到1-细胞期胚胎中然后将其培养而发育得到的，以及图36d显示了具有ND5点突变的小鼠(F0)，其中显示了具有在显微注射DdCBE mRNA后产生的ND5点突变的F0幼崽和在新生幼崽中鉴定的突变体序列阵列，编辑的碱基以红色表示，以及突变体线粒体基因的编辑频率示于右边；
图37显示了通过胞嘧啶脱氨酶介导的碱基编辑产生的小鼠线粒体ND5无义突变，图37a显示了产生m.C12336T无义突变和m.G12341A沉默突变的DdCBE靶序列，其中m.C12336T(C9)突变在ND5蛋白中产生Q199终止突变，而m.G12341A(C14)引起沉默Q200Q突变，转录三联体加下划线，以及可编辑位点以红色表示，图37b显示了在NIH3T3细胞中产生无义突变的胞嘧啶-胸腺嘧啶碱基编辑效率，其中指示了转染的DdCBE对的组合，深灰色条和浅灰色条表示m.C12336T(C9)和m.G12341A(C14)突变的相应频率，误差条表示n＝3个生物独立样品的s.e.m(n.s.：不显著，*p<0.05，**p<0.01，使用学生双尾t检验)，左-G1333-N+右-G1333-C、左-G1333-C+右-G1333-N、左-G1397-N+右-G1397-C、和左-G1397-C+右-G1397-N的C9突变的P值为0.0065、0.1143、0.0266和0.0037，以及C14突变的相应P值为0.0077、0.0144、0.0406和0.0214，图37c显示了小鼠胚泡中的编辑效率，其中测序数据是从用编码左-G1333-N和右-G1333-C DdCBE的mRNA显微注射受精卵后发育的胚泡获得的，以及深灰色条和浅灰色条分别表示C9和C14突变的频率，图37d显示了新生幼崽的突变体序列阵列，其中编辑的碱基以红色表示，以及突变体线粒体基因组的编辑频率示于右边，图37e显示了野生型和编辑的小鼠的Sanger测序色谱图，并且红色箭头表示取代的核苷酸；
图38示意性地显示了用于产生DdCBE构建体的Golden Gate克隆，其中所有反应在一个管中同时发生，箭头未指示连续反应，空表达载体和模块载体使用BsaI酶切割以消除线性化骨架和TALE模块插入物，包括相容的粘性末端，骨架和六个模块插入物通过T4 DNA连接酶连接以产生最终DdCBE构建体，使用八个DdCBE克隆骨架质粒，并且对于SOD2MTS，提供了左-G1333-N、左-G1333-C、左-G1397-N和左-G1397-C，并且对于COX8AMTS，提供了右-G1333-N、右-G1333-C、右-G1397-N和右-G1397-C；
图39显示了ND5突变体小鼠(F0)，图39a显示了ND5沉默突变体小鼠，图39b显示了ND5 G12918A突变体小鼠，以及图39c显示了由DdCBEmRNA显微注射产生的ND5无义突变体小鼠；
图40：图40a示意性地显示了含有DdCBE-NES和NES序列的载体，图40b显示了细胞核的4号染色体中小鼠m.G12918 ND5基因和ND5样基因的序列、细胞核的5号染色体中线粒体TrnA的序列、以及细胞核的6号染色体中线粒体Rnr2的序列，图40c显示了使用NIH3T3细胞系通过DdCBE和DdCBE-NES实现的在ND5基因中的编辑效率，图40d显示了使用NIH3T3细胞系通过DdCBE和DdCBE-NES实现的在TrnA基因中的编辑效率，图40e显示了使用NIH3T3细胞系通过DdCBE和DdCBE-NES实现的在Rnr2基因中的编辑效率，其中橙色图显示DdCBE的编辑效率，以及灰色图显示DdCBE-NES的编辑效率，图40f显示了mitoTALEN TALE阵列的DNA识别序列，以及图40g显示了在用mitoTALEN处理或不处理的实验组中的DdCBE碱基编辑效率，所有图为n＝2并且误差条为平均值的标准误差；
图41显示了使用DdCBE-NES和mitoTALEN实现的在小鼠胚胎和小鼠中改进的编辑效率，图41a显示了使用DdCBE和DdCBE-NES实现的在胚泡中对各种线粒体DNA靶标(mtND5、mtTrnA和mtRNR2)的碱基编辑效率，图41b显示了在有或没有mitoTALEN情况下使用DdCBE和DdCBE-NES实现的m.G12918A碱基编辑效率的比较，以及图41c显示了小鼠中m.G12918A碱基编辑效率的比较，所有的图为n>＝3，以及误差条为平均值的标准误差(n.s.：不显著，*p<0.05，**p<0.01，***p<0.001，使用学生双尾t检验获得的)；
图42：图42a示意性地显示了DdCBE蛋白的改进，图42b和图42c显示了DddAtox脱氨酶的晶体结构，其中分裂物二聚体界面处的残基表示为棒状，图42b显示了分别以紫色和浅蓝色表示的G1397-N分裂物和G1397-C分裂物，图42c显示了分别以橙色和绿色表示的G1333-N分裂物和G1333-C分裂物，以及图42d和图42e显示了G1397-N和G1397-C(d)以及G1333-N和G1333-C(e)的氨基酸序列，以及界面残基以红色表示；
图43：图43a显示了G1397界面突变体的碱基编辑效率的图，其中编辑范围和靶胞嘧啶表示在顶部，并且将突变体和野生型/无TALE的DddAtox蛋白共转染，如下面所示，并且对于左-DdCBE，无TALE的DddAtox蛋白是G1397-N，并且对于右-DdCBE，无TALE的DddAtox蛋白是G1397-C，并且图43b显示了DdCBE和突变体的靶胞嘧啶-胸腺嘧啶(鸟嘌呤-腺嘌呤)碱基编辑效率的热图；
图44：图44a显示了G1333界面突变体的碱基编辑效率的图，其中编辑范围和靶胞嘧啶表示在顶部，并且将突变体和野生型/无TALE的DddAtox蛋白共转染，如下面所示，并且对于左-DdCBE，无TALE的DddAtox蛋白是G1333-N，并且对于右-DdCBE，无TALE的DddAtox蛋白是G1333-C，并且图44b显示了DdCBE和突变体的靶胞嘧啶-胸腺嘧啶(鸟嘌呤-腺嘌呤)碱基编辑效率的热图；
图45显示了野生型和新型全长DddA的氨基酸序列的比较结果；
图46显示了其中将全长DddA递送到动物或植物细胞的构象；
图47显示了证实人细胞基因组背景ROR1位点(a)、HEK3位点(b)和TYRO3位点(c)中TC基序中胞嘧啶向胸腺嘧啶转化的活性的结果；
图48显示了全长DddA的优点；
图49显示了在人细胞基因组背景TRAC位点1(a)、TRAC位点2(b)、FANCF(c)和HBB(d)中测量全长DddA活性的结果；
图50显示了使用DddA-dCas9(D10A,H840A)-UGI在人细胞基因组背景TYRO3(a)、ROR1(b)、HEK3(c)、EMX1位点2(d)、TRAC位点1(e)和HBB(f)中测量DddA活性的结果；
图51显示了HEK293T细胞中全长DddAtox的碱基编辑效率，图51a示意性地显示了以基于结构的方式筛选全长DddAtox，红色丙氨酸指示带正电荷的氨基酸残基被丙氨酸取代，图51b显示了被丙氨酸取代的DddA变体的大肠杆菌转化体，E1347A用作对照中的活性位点突变体，图51c显示了在TYRO3位点处DddA AAAAA和CBE的编辑频率和插入缺失，以及图51d显示了TYRO3位点的等位基因频率，C至T转化以红色表示，原型间隔子以蓝色表示，以及原型间隔子邻近基序(PAM)以橙色表示；
图52显示了无毒DddA GSVG，图52a示意性地显示了基于易错PCR筛选无毒全长DddAtox变体，以及图52b和图52c显示了与Cas9、nCas9(D10A)、nCas9(H840A)和dCas9(D10A，H840A)的N末端和C末端融合的基因(b)和等位基因(c)的编辑频率，原型间隔子以蓝色表示，以及原型间隔子邻近基序(PAM)以橙色表示；
图53显示了在nCas9(D10A)的N末端处DddAtox变体的编辑频率，其中TYRO3位点(a)、ROR1位点1(b)和HEK3位点(c)中的带正电荷的氨基酸残基被丙氨酸取代，原型间隔子以蓝色表示，以及原型间隔子邻近基序(PAM)以橙色表示；
图54显示了在几个位点处的编辑频率。图54a、图54b、图54c、图54d、图54e、图54f、图54g和图54h至图54j分别显示了ROR1位点1、ROR1位点2、ROR1位点3、FANCF位点、HBB位点、HEK3位点、TRAC5位点1和EMX1位点，其中原型间隔子以蓝色表示，原型间隔子邻近基序(PAM)以橙色表示，C至T转化以红色表示，以及DddA的靶窗口通过计数原型间隔子的5'上游而指示为负数；
图55a、图55b、图55c、图55d、图55e、图55f、图55g、图55h、图55i和图55j分别显示了HeLa细胞中AAAAA和E1347A中TYRO3位点、ROR1位点1、ROR1位点2、ROR1位点3、FANCF位点、HBB位点、HEK3位点、TRAC5位点1、TRAC5位点2和EMX1位点2处的编辑频率，其中原型间隔子以蓝色表示，原型间隔子邻近基序(PAM)以橙色表示，DddA的靶窗口通过计数原型间隔子的5'上游而指示为负数，以及靶胞嘧啶以红色表示；
图56显示了TYRO3位点(a)和ROR1位点1(b)处的AAAAA和E1347A的时间依赖性碱基编辑和插入缺失率；
图57a、图57b、图57c、图57d、图57e、图57f、图57g和图57h分别显示了在EMX1位点2、FANCF位点、TRAC5位点1、TRAC5位点2、ROR1位点1、ROR1位点2、ROR1位点3和HBB位点处与nCas9(D10A)、nCas9(H840A)和dCas9的N末端融合的GSVG的编辑、插入-缺失和等位基因频率，其中原型间隔子以蓝色表示，原型间隔子邻近基序(PAM)以橙色表示，C至T转化以红色表示，以及GSVG的靶窗口通过计数原型间隔子的5'上游而指示为负数；
图58a、图58b、图58c和图58d分别显示了在EMX1位点2、EMX1位点4、ROR1位点2和HBB位点处与nCas9(D10A)、nCas9(H840A)和dCas9的C末端融合的GSVG的编辑、插入-缺失和等位基因频率，其中原型间隔子以蓝色表示，原型间隔子邻近基序(PAM)以橙色表示，G至A转化以红色表示，以及GSVG的靶窗口通过从原型间隔子的位置1的3'下游计数来指示；
图59显示了在TYRO3(a)和EMX1位点2(b)处与nCas9(H840A)的C末端融合的E1347A、GSVG、SSVG、GSAG和GSVS的时间依赖性编辑和插入-缺失频率；
图60显示了HEK293T细胞中mDdCBE的线粒体碱基编辑，图60a和图60b分别显示了ND4和ND6的编辑效率，其中靶胞嘧啶和TALE结合位点分别以红色和灰色表示，图60c至图60f显示了当仅一半DddAtox与TALE阵列融合且剩余一半不含TALE时ND4(c，d)和ND6(e，f)的编辑效率，其中左TALE阵列和右TALE阵列分别表示为L和R，并且ND6TALE阵列与参考基因组的错配加紫色下划线；
图61：
图61a示意性地显示了使用常规ZFP DNA结合蛋白的锌指胞嘧啶脱氨酶(ZFD)，
图61b显示了腺嘌呤脱氨酶插入ZFD中的位置(其中红色箭头是插入位点)，
图61c显示了构建的ZF-DdABE在核DNA Trac位点处的碱基编辑效率(C至T)，
图61d显示了构建的ZF-DdABE在核DNA Trac位点处的碱基编辑效率(A至G)
(其中WT-ZFD是在不存在腺嘌呤脱氨酶的情况下具有单独的分裂DddAtox的C至T脱氨酶)，
图61e显示了靶向线粒体DNA的ZF-DdABE在ND1位点处的效率(C至T)，以及图61f显示了靶向线粒体DNA的ZF-DdABE在ND1位点处的效率(A至G)；
图62：
图62a示意性地显示了使用TALE和分裂DddAtox的DdABE(其中组分包括分裂DddAtox、腺嘌呤脱氨酶和TALE阵列)，
图62b显示了将单独的腺嘌呤脱氨酶连接到靶向线粒体ND4位点的TALE时的碱基编辑效率，
图62c显示了将腺嘌呤脱氨酶连接到靶向线粒体ND1位点的TALE-分裂DddAtox时的碱基编辑效率，
图62d显示了当在左边使用DdCBE对并且在右边将腺嘌呤脱氨酶连接到TALE-分裂DddAtox时(其中绿色框是TALE所连接的部分)，在单个核苷酸单元中的碱基编辑效率，以及
图62e显示了当在左边将腺嘌呤脱氨酶连接到TALE-分裂DddAtox并且在右边使用DdCBE对时(其中绿色框是TALE所连接的部分)，在单个核苷酸单元中的碱基编辑效率；
图63：
图63a显示了在不存在或存在UGI的情况下靶向线粒体ND1位点的DdABE的C至T和A至G碱基编辑效率(其中红色框指示腺嘌呤脱氨酶)，
图63b显示了在不存在或存在UGI的情况下靶向线粒体ND4位点的DdABE的C至T和A至G碱基编辑效率(其中红色框指示腺嘌呤脱氨酶)，
图63c显示了靶向线粒体ND1位点的DdABE构型中在单个核苷酸单元中具有最高效率的构型(其中绿色框是TALE所连接的部分)，以及
图63d显示了靶向线粒体ND4位点的DdABE构型中在单个核苷酸单元中具有最高效率的构型(其中绿色框是TALE所连接的部分)；
图64：
图64a在顶部示意性地显示了在一个TALE模块中具有所有构建体的单个TALE模块(其中组分包括全长DddAtox、腺嘌呤脱氨酶和TALE阵列)，以及
在底部还显示了使用两个TALE模块的双TALE模块(其中组分包括在一侧的全长DddAtox和TALE阵列以及在另一侧的腺嘌呤脱氨酶和TALE阵列)，
图64b显示了靶向线粒体ND1位点的单模块和双模块DdABE的碱基编辑效率，以及图64c显示了靶向线粒体ND4位点的单模块和双模块DdABE的碱基编辑效率；
图65显示了证实靶向ND1位点的单模块的碱基编辑效率的结果(其中组分包括TALE阵列、腺嘌呤脱氨酶(AD)和全长DddAtox(GSVG、AAAA和E1347A是变体))；
图66显示了证实靶向ND1位点的双模块的碱基编辑效率的结果(其中组分包括TALE阵列、腺嘌呤脱氨酶(AD)和全长DddAtox(GSVG、AAAA和E1347A是变体))；
图67：图67a显示了当将单独的TadA(AD)腺嘌呤脱氨酶连接到靶向ND1位点的TALE结合蛋白时的碱基编辑效率，以及
图67b显示了当将单独的TadA(AD)腺嘌呤脱氨酶连接到靶向ND4位点的TALE结合蛋白时的碱基编辑效率；以及
图68显示了靶向ND1位点的TALE中双模块、单模块和分裂DddA-AD的腺嘌呤和胞嘧啶碱基编辑效率，其中(从最下面开始)当将UGI在没有AD的情况下连接到两侧时称为DdCBE，仅发生胞嘧啶碱基编辑，或当AD被UGI的任一侧替代时，胞嘧啶碱基编辑和腺嘌呤碱基编辑均发生，或当UGI不存在时，仅选择性地发生腺嘌呤碱基编辑，以及类似地，甚至在双模块和单模块中仅选择性地发生腺嘌呤碱基编辑。
【具体实施方式】
除非另外定义，否则本文使用的所有技术和科学术语均具有与本发明所属领域的技术人员通常所理解的含义相同的含义。一般来说，本文使用的命名法是本领域熟知的，并且是典型的。
如本文所用，术语“编辑”可与“校正”互换使用，并且是指改变细胞中特定基因组靶位点处的核酸序列的方法。这种特定基因组靶标包括但不限于染色体区域、基因、启动子、开放阅读框或任何核酸序列。
如本文所用，术语“单碱基”是指核酸序列中的仅一个核苷酸。当在单碱基编辑的上下文中使用时，它是指核酸序列中特定位置的碱基被不同的碱基取代。这种取代可以通过多种机制发生，包括取代或修饰，但不限于此。
如本文所用，术语“靶标”或“靶位点”是指任何组成和/或长度的先前鉴定的核酸序列。这些靶位点包括但不限于染色体区域、基因、启动子、开放阅读框或任何核酸序列。
如本文所用，术语“中靶”是指特定基因组靶标的子序列，其被可编程DNA结合蛋白结合或可与单个指导RNA序列完全互补。
如本文所用，术语“脱靶”是指特定基因组靶标的亚序列，其可以与可编程DNA结合区识别的中靶序列和/或单个指导RNA序列部分互补。
1.分裂胞嘧啶脱氨酶
根据本发明的一方面的融合蛋白包括胞嘧啶脱氨酶或其变体，其中所述胞嘧啶脱氨酶或其变体包含衍生自胞嘧啶脱氨酶或其变体的第一分裂物和第二分裂物，并且所述第一分裂物和所述第二分裂物中的每一个与DNA结合蛋白融合。
胞嘧啶脱氨酶是一种去除胞嘧啶碱基中的氨基并能够将胞嘧啶(C)转化为尿苷(U)的酶。
它可以是胞嘧啶脱氨酶。胞嘧啶脱氨酶的例子可以包括APOBEC1(载脂蛋白B编辑复合物1)和AID(活化诱导的脱氨酶)，但是大多数DNA脱氨酶可能仅作用于单链DNA，并且可能不适于通过与DNA结合蛋白连接进行碱基编辑。具体地，胞嘧啶脱氨酶可以衍生自双链DNA脱氨酶(DddA)或其直系同源物。更具体地，胞嘧啶脱氨酶可以是双链DNA特异性细菌胞嘧啶脱氨酶。
胞嘧啶脱氨酶以分裂形式提供，胞嘧啶脱氨酶包含第一分裂物和第二分裂物，并且第一分裂物和第二分裂物中的每一个不具有脱氨酶活性。
可以包括对应于全长胞嘧啶脱氨酶中的DddAtox分裂物的SEQ ID NO:1的序列。胞嘧啶脱氨酶包含第一分裂物和第二分裂物，并且第一分裂物和第二分裂物中的每一个不具有脱氨酶活性。
[SEQ ID NO:1]
GSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLESKVFSSGGPTPYPNYANAGHVEGQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGAIPVKRGATGETKVFTGNSNSPKSPTKGGC
在一个实施方案中，所述胞嘧啶脱氨酶的第一分裂物或第二分裂物可以包含SEQID NO:1序列中从N末端到选自G33、G44、A54、N68、G82、N98和G108中的至少一个的序列。胞嘧啶脱氨酶的第一分裂物或第二分裂物可以包含SEQ ID NO:1序列中从选自G34、P45、G55、N69、T83、A99和A109中的至少一个到C末端的序列。
具体地，胞嘧啶脱氨酶可以包含SEQ ID NO:23的第一分裂物(G1333-N)和SEQ IDNO:24的第二分裂物(G1333-C)、SEQ ID NO:25的第一分裂物(G1397-N)和SEQ ID NO:26的第二分裂物(G1397-C)、SEQ ID NO:23的第一分裂物(G1333-N)和SEQ ID NO:26的第二分裂物(G1397-C)、或SEQ ID NO:25的第一分裂物(G1397-N)和SEQ ID NO:24的第二分裂物(G1333-C)。
(SEQ ID NO:23)野生型DddAtox G1333-N
GSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLESKVFSSGG
(SEQ ID NO:27)
GGCTCTGGTTCCTACGCCCTGGGTCCATATCAGATTAGTGCTCCCCAACTCCCCGCCTACAACGGTCAGACAGTGGGGACCTTTTACTATGTCAACGACGCCGGGGGATTGGAATCCAAGGTTTTCTCTAGCGGTGGG
(SEQ ID NO:24)野生型G1333-C
PTPYPNYANAGHVEGQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGAIPVKRGATGETKVFTGNSNSPKSPTKGGC
(SEQ ID NO:28)
CCAACACCTTATCCTAACTACGCTAACGCCGGGCACGTCGAGGGGCAGTCAGCTCTTTTTATGAGAGATAACGGCATTAGCGAAGGGCTTGTGTTCCATAATAATCCTGAGGGCACCTGTGGCTTCTGTGTAAATATGACCGAAACACTTCTGCCTGAGAACGCTAAAATGACTGTCGTACCACCCGAAGGCGCAATCCCAGTTAAACGGGGCGCAACCGGCGAAACCAAAGTATTCACCGGAAACAGCAATAGTCCAAAGTCCCCCACCAAGGGAGGTTGC
(SEQ ID NO:25)野生型DddAtox G1397-N
GSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLESKVFSSGGPTPYPNYANAGHVEGQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEG
(SEQ ID NO:29)
GGTAGCTACGCACTTGGTCCTTACCAGATTAGCGCACCCCAACTCCCCGCCTATAATGGTCAAACCGTCGGGACCTTTTACTACGTAAACGATGCTGGTGGGCTGGAATCCAAAGTATTCTCCTCAGGGGGCCCTACACCCTACCCCAACTACGCCAATGCTGGTCATGTAGAAGGGCAGTCAGCACTGTTTATGCGCGATAATGGTATAAGCGAGGGGTTGGTCTTCCATAACAACCCAGAGGGTACTTGTGGCTTCTGTGTGAATATGACTGAAACCCTTCTGCCCGAAAATGCCAAGATGACTGTCGTCCCACCTGAAGGC
(SEQ ID NO:26)野生型DddAtox G1397-C
AIPVKRGATGETKVFTGNSNSPKSPTKGGC
(SEQ ID NO:30)
GCCATACCTGTGAAGCGGGGAGCAACAGGGGAGACAAAGGTGTTCACAGGCAACTCTAACAGTCCAAAGAGCCCCACCAAAGGCGGGTGT
组合的G1333N、G1333C、G1397N和G1397C可用作分裂形式的脱氨酶。具体地，可以使用左-G1333-N+右-G133-C、左-G1397-N+右-G1397-C、左-G1397-N+右-G1333-C或左-G1333-N+右-G1397-C的形式。
2.变体
本申请的发明人试图通过其中氨基酸残基被取代的DdCBE突变来抑制不需要的碱基编辑。提出了一种能够降低DdCBE脱靶效应的高精度DddA衍生的胞嘧啶碱基编辑器。这种脱靶碱基编辑效应是由DddAtox脱氨酶分裂物的自发组装引起的现象，与TALE和DNA之间的相互作用无关。
这里，突变的氨基酸残基是位于DddAtox分裂物二聚体彼此相互作用的表面上的接触位点。通过用丙氨酸取代位于DddAtox分裂物之间表面上的氨基酸残基来构建高保真度DdCBE(HF-DdCBE)。HF-DdCBE防止了一对与TALE连接的两个分裂物脱氨酶半部分在不与DNA结合时正常发挥功能。通过全线粒体基因组分析，证实了HF-DdCBE是非常有效和精确的，这与在人线粒体DNA中引起许多不希望的脱靶C至T转化的常规DdCBE不同。
对于DddAtox，原则上只有当两个分裂物二聚体被募集到DNA的靶位点时才可诱导碱基编辑。基于实际实验的结果，即使当使用DdCBE时，也发生靶向碱基编辑，其中的一半结合DNA，而剩余的一半不结合。为了解决这个问题，通过取代分裂物二聚体彼此相互作用的蛋白质表面上的残基来防止DdCBE对在不需要的位置结合。
基于此，本发明涉及通过取代胞嘧啶脱氨酶DddAtox的氨基酸残基来减少非选择性碱基编辑的新变体。
胞嘧啶脱氨酶包含SEQ ID NO:23的第一分裂物(G1333-N)和SEQ ID NO:24的第二分裂物(G1333-C)、或SEQ ID NO:25的第一分裂物(G1397-N)和SEQ ID NO:26的第二分裂物(G1397-C)。
胞嘧啶脱氨酶的变体可以被配置为使得SEQ ID NO:23的第一分裂物中的选自位置3、5、10、11、13、14、15、16、17、18、19、28、30和31处的氨基酸的至少一个氨基酸被不同的氨基酸取代，或者还使得SEQ ID NO:24的第二分裂物中的选自位置13、16、17、20、21、28、29、30、31、32、33、56、57、58和60处的氨基酸的至少一个氨基酸被不同的氨基酸取代。
根据本发明的变体可以被配置为使得SEQ ID NO:25的第一分裂物中的选自位置87、88、91、92、95、100、101、102和103处的氨基酸的至少一个氨基酸或SEQ ID NO:26的第二分裂物中的选自位置13、14、15和16处的氨基酸的至少一个氨基酸被不同的氨基酸取代。
这里，“不同的氨基酸”可以是丙氨酸、异亮氨酸、亮氨酸、甲硫氨酸、苯丙氨酸、脯氨酸、色氨酸、缬氨酸、天冬酰胺、半胱氨酸、谷氨酰胺、甘氨酸、丝氨酸、苏氨酸、酪氨酸、天冬氨酸、谷氨酸、精氨酸、组氨酸或赖氨酸，并且可以指选自从上述氨基酸的所有已知变体中排除野生型蛋白质中原始突变位置处的氨基酸的氨基酸中的氨基酸。在一个示例性实施方案中，“不同的氨基酸”可以是丙氨酸。
具体地，可以包括SEQ ID NO:23的第一分裂物中选自Y3A、L5A、I10A、S11A、V13A、G14A、T15A、F16A、Y17A、Y18A、V19A、K28A、F30A和S31A的至少一个的氨基酸取代(分别对应于Y1292A、L1294A、I1299A、S1300A、V1312A、G1313A、T1314A、F1315A、Y1316A、Y1317A、V1318A、K1327A、F1329A和S1330A)。
此外，可以包括SEQ ID NO:24的第二分裂物中选自V13A、Q16A、S17A、F20A、M21A、E28A、G29A、L30A、V31A、F32A、H33A、K56A、M57A、T58A和V60A的至少一个的氨基酸取代(分别对应于V1346A、Q1349A、S1350A、F1353A、M1354A、E1361A、G1362A、L1363A、V1364A、F1365A、H1366A、K1389A、M1390A、T1391A和V1393A)。
具体地，可以包括SEQ ID NO:25的第一分裂物中选自C87A、V88A、T91A、E92A、L95A、K100A、M101A、T102A和V103A的至少一个的氨基酸取代(分别对应于C1376A、V1377A、T1380A、E1381A、L1384A、K1389A、M1390A、T1391A和V1392A)。
此外，可以包括SEQ ID NO:26的第二分裂物中选自K13A、V14A、F15A和T16A的至少一个的氨基酸取代(分别对应于K1410A、V1411A、F1412A和T1413A)。
[DddAtox G1333-N变体]
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[DddAtox G1333-C变体]
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[DddAtox G1397-N变体]
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[DddAtox G1397-C变体]
根据本发明的胞嘧啶脱氨酶变体可以包括选自上表中所述的氨基酸序列的至少一个序列。根据本发明的胞嘧啶脱氨酶变体显示出对非特异性靶位点中各种碱基减少不希望的编辑的可能性。
3.全长胞嘧啶脱氨酶
本申请的发明人开发了使用全长DddA的新型可编程胞嘧啶脱氨酶，其通过修饰野生型胞嘧啶脱氨酶DddAtox的带正电荷的氨基酸而制备，所述新型可编程胞嘧啶脱氨酶由于其细胞毒性而以分裂形式使用。
本发明涉及一种融合蛋白，所述融合蛋白包含(i)DNA结合蛋白和(ii)胞嘧啶脱氨酶或其变体，其中所述胞嘧啶脱氨酶或其变体是无毒的全长胞嘧啶脱氨酶。
在DddAtox的C末端，带正电荷的氨基酸(KRKKK)是特异性簇集的。由于DNA是带负电荷的，因此它与蛋白质的带正电荷的氨基酸结合。通过用不带电荷的氨基酸取代带正电荷的氨基酸，减弱了DddAtox与DNA的结合力，从而降低或消除了细胞毒性。特别地，通过用非极性氨基酸取代带正电荷的氨基酸而产生的无毒组合使得能够使用大肠杆菌进行克隆以提供全长DddA。
野生型DddAtox由于其细胞毒性而以两种分裂形式使用，这在实验中具有许多限制。特别地，当使用Cas9时，使用识别其他PAM的正交Cas9变体。因此，由于PAM的存在是有限的，因此通常难以在所需位置将胞嘧啶准确地脱氨基为胸腺嘧啶。此外，具有最高活性的靶窗口是彼此结合的两个Cas9变体之间的40bp长的区域，并且该区域中不需要的胞嘧啶也被脱氨基。然而，全长DddA不受PAM的约束，因为它不是分离的。此外，它在距离靶位置10bp内的TC基序中最有活性，导致高精度。
已证实，Cas9使通过与靶位点结合形成的R环中的ACA、GC和CC基序中的胞嘧啶脱氨基为胸腺嘧啶。这是尚未以分离形式鉴定的活性。
在全长DddA中，可以使用TALE模块或锌指蛋白以及Cas9在所需位置用胸腺嘧啶取代胞嘧啶。现有的DddAtox必须以分离的形式成对递送，但全长DddA可仅使用TALE模块或锌指蛋白中的一个模块，使得可无限制地选择靶位点。此外，通过靶向线粒体、植物叶绿体或质体中的DNA以及基因组位点，可以将特定DNA的胞嘧啶转化为胸腺嘧啶。
此外，可以将所有的构建体插入到AAV中，AAV由于其小尺寸而是用于基因治疗的病毒载体。现有的CBE(胞嘧啶碱基编辑器)在通过Cas9与靶位点结合形成的R环中用胸腺嘧啶取代胞嘧啶，但本文发明的全长DddA使R环外的胞嘧啶脱氨基。因此，可以在限制使用常规CBE进行编辑的位置将胞嘧啶转化为胸腺嘧啶。
基于此，在无毒的全长胞嘧啶脱氨酶中，SEQ ID NO:1的野生型脱氨酶的至少一个、至少两个、至少三个、至少四个或至少五个氨基酸可以被不同的氨基酸取代。
这里，“不同的氨基酸”可以是丙氨酸、异亮氨酸、亮氨酸、甲硫氨酸、苯丙氨酸、脯氨酸、色氨酸、缬氨酸、天冬酰胺、半胱氨酸、谷氨酰胺、甘氨酸、丝氨酸、苏氨酸、酪氨酸、天冬氨酸、谷氨酸、精氨酸、组氨酸或赖氨酸，并且可以指选自从上述氨基酸的所有已知变体中排除野生型蛋白质中原始突变位置处的氨基酸的氨基酸中的氨基酸。在一个示例性实施方案中，“不同的氨基酸”可以是丙氨酸。
根据其类型，无毒全长DddA可以包括选自SEQ ID NO:12至SEQ ID NO:18的序列。
野生型(SEQ ID No:1)
GSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLESKVFSSGGPTPYPNYANAGHVEGQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGAIPVKRGATGETKVFTGNSNSPKSPTKGGC
A1341D KRKKA(SEQ ID No:12)
GSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLESKVFSSGGPTPYPNYDNAGHVEGQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGAIPVKRGATGETKVFTGNSNSPKSPTAGGC
AAAAA(SEQ ID No:13)
GSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLESKVFSSGGPTPYPNYANAGHVEGQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGAIPVAAGATGETAVFTGNSNSPASPTAGGC
AAAAK(SEQ ID No:14)
GSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLESKVFSSGGPTPYPNYANAGHVEGQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGAIPVAAGATGETAVFTGNSNSPASPTKGGC
AAKAA(SEQ ID No:15)
GSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLESKVFSSGGPTPYPNYANAGHVEGQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGAIPVAAGATGETKVFTGNSNSPASPTAGGC
AAKAK(SEQ ID No:16)
GSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLESKVFSSGGPTPYPNYANAGHVEGQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGAIPVAAGATGETKVFTGNSNSPASPTKGGC
KAAAA(SEQ ID No:17)
GSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLESKVFSSGGPTPYPNYANAGHVEGQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGAIPVKAGATGETAVFTGNSNSPASPTAGGC
E1347A(SEQ ID No:18)
GSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLESKVFSSGGPTPYPNYANAGHVAGQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGAIPVKRGATGETKVFTGNSNSPKSPTKGGC
全长脱氨酶变体可以在SEQ ID NO:1的氨基酸序列中包括至少一个选自以下的取代：
位置37处用G取代S；
位置59处用S取代G；
位置109处用V取代A；以及
位置129处用G取代S。
在一个实施方案中，全长脱氨酶变体可包括SEQ ID NO:19的序列，包括在SEQ IDNO:1的氨基酸序列中位置37处用G取代S，位置59处用S取代G，位置109处用V取代A，以及位置129处用G取代S。
GSVG(SEQ ID No:19)
GSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLEGKVFSSGGPTPYPNYANAGHVESQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGVIPVKRGATGETKVFTGNSNGPKSPTKGGC
全长DddA GSVG能够使用通用大肠杆菌进行克隆。在人细胞基因组背景中证实，全长DddA GSVG使靶部位处TC基序的胞嘧啶脱氨基为胸腺嘧啶。可将全长DddA GSVG克隆到Cas9的N末端和C末端中的每一个。与Cas9的N末端连接的DddA GSVG在同一靶位点可以用胸腺嘧啶取代胞嘧啶。在人细胞中证实，与Cas9的C末端连接的DddA GSVG诱导TC基序中的胞嘧啶到胸腺嘧啶的取代(互补序列中的鸟嘌呤到腺嘌呤的取代)。
在一个实施方案中，全长脱氨酶变体可包括选自SEQ ID NO:20至22的序列。
SSVG(SEQ ID No:20)
GSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLESKVFSSGGPTPYPNYANAGHVESQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGVIPVKRGATGETKVFTGNSNGPKSPTKGGC
GSAG(SEQ ID No:21)
GSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLEGKVFSSGGPTPYPNYANAGHVESQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGAIPVKRGATGETKVFTGNSNGPKSPTKGGC
GSVS(SEQ ID No:22)
GSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLEGKVFSSGGPTPYPNYANAGHVESQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGVIPVKRGATGETKVFTGNSNSPKSPTKGGC
4.DNA结合蛋白
DNA结合蛋白可以是例如锌指蛋白、TALE(转录激活因子样效应物)蛋白、CRISPR相关核酸酶或其两种或更多种的组合。
锌指蛋白的锌指基序具有DNA结合结构域，并且指的c末端部分特异性识别DNA序列。含有3至6个锌指基序的DNA结合蛋白识别DNA序列。
在一个实施方案中，胞嘧啶脱氨酶的第一分裂物和第二分裂物中的每一个可以与锌指蛋白的N末端或C末端融合。
锌指蛋白的C末端(ZF-左)与胞嘧啶脱氨酶的第一分裂物的N末端融合，以及锌指蛋白的C末端(ZF-右)与胞嘧啶脱氨酶的第二分裂物的N末端融合(CC构型)。
锌指蛋白的N末端(ZF-左)与胞嘧啶脱氨酶的第一分裂物的C末端融合，以及锌指蛋白的C末端(ZF-右)与胞嘧啶脱氨酶的第二分裂物的N末端融合(NC构型)。
锌指蛋白的C末端(ZF-左)与胞嘧啶脱氨酶的第一分裂物的N末端融合，以及锌指蛋白的N末端(ZF-右)与胞嘧啶脱氨酶的第二分裂物的C末端融合(CN构型)。
锌指蛋白的N末端(ZF-左)与胞嘧啶脱氨酶的第一分裂物的C末端融合，以及锌指蛋白的N末端(ZF-右)与胞嘧啶脱氨酶的第二分裂物的C末端融合(NN构型)。
ZF-左可包括以下SEQ ID NO:2的序列：
[SEQ ID NO:2]
GIHGVPAAMAERPFQCRICMRNFSDRSNLSRHIRTHTGEKPFACDICGRKFAISSNLNSHTKIHTGSQKPFQCRICMRNFSRSDNLARHIRTHTGEKPFACDICGRKFATSGNLTRHTKIHLR。
ZF-右可包括以下SEQ ID NO:3的序列：
[SEQ ID NO:3]
GIHGVPAAMAERPFQCRICMRNFSRSDNLSVHIRTHTGEKPFACDICGRKFAQKINLQVHTKIHTGEKPFQCRICMRNFSRSDVLSEHIRTHTGEKPFACDICGRKFAQRNHRTTHTKIHLR。
ZF的序列可以根据DNA靶标而变化。ZF可以根据DNA靶序列进行定制。由于ZF识别3-bp DNA，因此可以通过连接3至6个ZF构建识别9-18bp DNA的ZF组合。例如，它可以使用包括识别GNN、TNN、CNN或ANN的模块的库来产生。
在一些情况下，锌指蛋白可通过接头与脱氨酶连接。接头可以是包括2至40个氨基酸残基的肽接头。接头可以是例如长度为2aa、5aa、10aa、16aa、24aa或32aa的接头，但不限于此。
在一个实施方案中，接头可以包括：
2a.a接头：GS
5a.a接头：TGEKQ(SEQ ID NO:8)
10a.a接头：SGAQGSTLDF(SEQ ID NO:9)
16a.a接头：SGSETPGTSESATPES(SEQ ID NO:10)；
24a.a接头：SGTPHEVGVYTLSGTPHEVGVYTL(SEQ ID NO:115)；或
32a.a接头：GSGGSSGGSSGSETPGTSESATPESSGGSSGGS(SEQ ID NO:11)。
在根据本发明的一个具体实施方案中，分裂物脱氨酶和锌指蛋白可以通过接头连接，锌指蛋白与包括第一分裂物的分裂半脱氨酶的N末端融合，以及锌指蛋白与包括第二分裂物的半脱氨酶的N末端融合。这里，C至T碱基转化可以发生在左和右ZFP结合位点之间的间隔子中。当通过24a.a接头分别连接到包括第一分裂物的半脱氨酶和包括第二分裂物的半脱氨酶时，左和右ZFP都被证实显示出高编辑效率。
TAL效应物(TALE)被配置为使得33-34个氨基酸序列被重复，并且约九个RVD(重复变体结构域)被重复。根据第12-13个氨基酸序列(HD->胞嘧啶、NI->腺嘌呤、NG->胸腺嘧啶、NN->鸟嘌呤)，它能够识别每个结构域一个核苷酸并且可以结合特定的DNA序列。TAL效应物(TALE)识别靶位点内的单链DNA。靶位点之间的距离可以是12-14个核苷酸。
TALE结构域是通过至少一个TALE-重复序列组合以序列特异性方式与核苷酸结合的蛋白质结构域。它包括至少一个TALE-重复序列，特别是1至30个TALE-重复，但不限于此。TALE-重复序列是识别TALE结构域中的特定核苷酸序列的结构域。
TALE结构域包括含有TALE的N末端的区域和含有TALE的C末端的区域作为骨架结构。包括TALE的N末端的第一TALE可以由SEQ ID NO:4或5编码。包括TALE的C末端的第二TALE可以由SEQ ID NO:6或7编码。
根据基于切割位点的TALE结构域结合的位置，单个TALE阵列或第一TALE阵列和第二TALE阵列中的每一个可以与其结合。
第一TALE(左TALE)可以与胞嘧啶脱氨酶的第一分裂物融合，以及第二TALE(右TALE)可以与胞嘧啶脱氨酶的第二分裂物融合。相应的构建体可以被描述为N’-TALE-第一分裂物-C’和N’-TALE-第二分裂物-C’。
当胞嘧啶脱氨酶是全长时，单个模块TALE可以与胞嘧啶脱氨酶的N末端结合。在N-C方向上包括单个TALE模块和胞嘧啶脱氨酶。可以包括双模块TALE，其中第一TALE可以与全长胞嘧啶脱氨酶的N末端融合，以及第二TALE可以单独包括。在N-C方向上包括第一TALE模块和胞嘧啶脱氨酶，并且提供了N’-TALE-胞嘧啶脱氨酶-C’和N’-TALE-C’的构建体。
TALE阵列可以根据靶DNA序列进行定制。TALE阵列被配制为使得由33至35个氨基酸残基构成的模块被重复排列。这些衍生于植物病原体黄单胞菌属(Xanthomonas)，并且模块识别碱基A、C、G和T中的每一个，然后结合DNA。每个模块的碱基特异性由第12和第13个氨基酸残基决定，所谓的重复可变双残基(RVD)。例如，其中RVD是NN的模块识别G，NI识别A，HD识别C，以及NG识别T。TALE阵列可由至少14至18个模块构成，并可被设计为识别15-20bp长的靶DNA序列。
关于CRISPR相关的核酸酶，在CRISPR阵列中编码两种RNA，一种是crRNA(CRISPRRNA)，另一种是tracrRNA(反式激活CRISPR RNA)。此外，crRNA在原型间隔子位点被转录并结合tracrRNA以形成三级结构。两种类型的RNA有助于识别和切割外来DNA。
Cas蛋白可包括但不限于Cas1、Cas1B、Cas2、Cas3、Cas4、Cas5、Cas6、Cas7、Cas8、Cas9、Cas10、Cas12a、Cas12b、Cas12c、Cas12d、Cas12e、Cas12g、Cas12h、Cas12i、Cas12j、Cas13a、Cas13b、Cas13c、Cas13d、Cas14、Csy1、Csy2、Csy3、Cse1、Cse2、Csc1、Csc2、Csa5、Csn2、CsMT2、Csm3、Csm4、Csm5、Csm6、Cmr1、Cmr3、Cmr4、Cmr5、Cmr6、Csb1、Csb2、Csb3、Csx17、Csx14、Csx10、Csx16、CsaX、Csx3、Csx1、Csx15、Csf1、Csf2、Csf3或Csf4核酸内切酶。
Cas蛋白可以衍生自含有Cas蛋白直向同源物的微生物属，其选自棒杆菌属(Corynebacter)、萨特氏菌属(Sutterella)、军团菌属(Legionella)、密螺旋体属(Treponema)、产丝菌属(Filifactor)、真杆菌属(Eubacterium)、链球菌属(Streptococcus)(酿脓链球菌(Streptococcus pyogenes))、乳杆菌属(Lactobacillus)、支原体属(Mycoplasma)、拟杆菌属(Bacteroides)、黄沃拉菌属(Flaviivola)、黄杆菌属(Flavobacterium)、固氮螺菌属(Azospirillum)、葡糖醋杆菌属(Gluconacetobacter)、奈瑟菌属(Neisseria)、罗氏菌属(Roseburia)、细小棒菌属(Parvibaculum)、葡萄球菌属(Staphylococcus)(金黄色葡萄球菌(Staphylococcus aureus))、Nitratifractor、棒杆菌属(Corynebacterium)和弯曲杆菌属(Campylobacter)，并且可以简单地从其中分离或重组。
Cas蛋白可以以突变形式被包含在内，其可能丧失核酸内切酶活性。举例说明了选自被突变以丧失核酸内切酶活性并具有切口酶活性的突变靶特异性核酸酶以及被突变以丧失核酸内切酶活性和切口酶活性的形式中的至少一种。
当具有切口酶活性时，与通过胞嘧啶脱氨酶的碱基转化(例如胞嘧啶到尿苷的转化)同时地或不管顺序如何顺序地，可以将切口引入到发生碱基转化的链或相对的链(例如与发生碱基转化的链相对的链)中(例如在与PAM所在的链相对的链上，在PAM序列的5'末端的方向上在第3个核苷酸与第4个核苷酸之间的位置处引入切口)。这样的突变(例如氨基酸取代等)可以发生在催化活性结构域(例如Cas9中的RuvC催化结构域)中。另外，酿脓链球菌衍生的Cas9可包括其中选自催化活性天冬氨酸残基(位置10的天冬氨酸(D10)等)、位置762的谷氨酸(E762)、位置840的组氨酸(H840)、位置854的天冬酰胺(N854)、位置863的天冬酰胺(N863)、位置986的天冬氨酸(D986)等的至少一个被任何不同氨基酸取代的突变。这里，被取代的任何不同的氨基酸可以是丙氨酸，但不限于此。
在一些情况下，酿脓链球菌衍生的Cas9蛋白可被突变以识别不同于野生型Cas9的PAM序列(NGG)的NGA(其中N是选自A、T、G和C中的任何碱基)，这是通过用不同的氨基酸取代选自位置1135的天冬氨酸(D1135)、位置1335的精氨酸(R1335)和位置1337的苏氨酸(T1337)中的至少，例如全部三个。
例如，在酿脓链球菌衍生的Cas9蛋白的氨基酸序列中，氨基酸取代可以发生在：
(1)D10、H840或D10+H840；
(2)D1135、R1335、T1337或D1135+R1335+T1337；或
(3)残基(1)和(2)。
这里，“不同的氨基酸”可以是丙氨酸、异亮氨酸、亮氨酸、甲硫氨酸、苯丙氨酸、脯氨酸、色氨酸、缬氨酸、天冬酰胺、半胱氨酸、谷氨酰胺、甘氨酸、丝氨酸、苏氨酸、酪氨酸、天冬氨酸、谷氨酸、精氨酸、组氨酸或赖氨酸，并且可以指选自从上述氨基酸的所有已知变体中排除野生型蛋白质中原始突变位置处的氨基酸的氨基酸中的氨基酸。在一个示例性实施方案中，“不同的氨基酸”可以是丙氨酸、缬氨酸、谷氨酰胺或精氨酸。
在一些情况下，可以进一步包括指导RNA。指导RNA可以是例如选自CRISPR RNA(crRNA)、反式激活crRNA(tracrRNA)和单指导RNA(sgRNA)中的至少一种。具体地，它可以是其中crRNA和tracrRNA彼此结合的双链crRNA:tracrRNA复合物，或其中crRNA或其一部分和tracrRNA或其一部分通过寡核苷酸接头连接的单链指导RNA(sgRNA)。
5.腺嘌呤脱氨酶的添加
本发明涉及包含三种组分的融合蛋白：DNA结合蛋白，胞嘧啶脱氨酶或其变体，以及腺嘌呤脱氨酶。胞嘧啶脱氨酶或其变体被分裂成两部分，称为“分裂物”，其衍生自无毒全长胞嘧啶脱氨酶或胞嘧啶脱氨酶或其变体。两个分裂物都与DNA结合蛋白融合。
本申请的发明人通过将能够引起A至G转化的腺嘌呤脱氨酶与DddAtox胞嘧啶脱氨酶连接到能够结合DNA的TALE或ZFP蛋白，构建能够编辑碱基A的碱基编辑器。
使用现有DddAtox的脱氨酶(DdCBE)是使用TALE重复序列作为DNA结合模块的胞嘧啶脱氨酶。与仅引起C至T转化的DdCBE不同，DdABE可诱导A至G转化，因此可产生其他突变模式。
由于DdABE自身识别双链DNA并引起脱氨，因此不存在另外的组分如RNA。对于线粒体或叶绿体，RNA的递送机制尚不清楚，因此不能应用Crispr系统。然而，没有这种组分的DdABE不仅能够靶向细胞中的基因组DNA，而且能够靶向细胞器如线粒体或叶绿体中的DNA，诱导特定DNA的A至G转化。
目前，DdCBE仅是靶向线粒体或细胞器的基因编辑技术。因此，可通过所有常规技术引入的突变仅包括C至T转化，但DdABE可诱导A至G转化，因此可引入突变的范围变得更加多样化。这使得有可能产生或治疗迄今为止还不可能的线粒体疾病模型。
现有的DdCBE需要两个TALE模块(连接到左边和右边)，因此，它不能装载在AAV上，AAV是在基因治疗中具有低基因容量的病毒载体。然而，由于DdABE可以用作能够仅使用一个TALE模块的单个模块，因此它可以装载在AAV上并且可用于基因治疗。
DdABE具有高度兼容性，因为它能够根据需要使用分裂DddAtox或全长DddAtox变体。
腺嘌呤脱氨酶可选自例如APOBEC1(载脂蛋白B编辑复合物1)、AID(活化诱导的脱氨酶)和tadA(tRNA特异性腺苷脱氨酶)，并且可以特别是tadA(tRNA特异性腺苷脱氨酶)。腺嘌呤脱氨酶可以是例如作为大肠杆菌TadA的变体的脱氧腺嘌呤脱氨酶。
在其中胞嘧啶脱氨酶被分裂，DNA结合蛋白是锌指蛋白，锌指蛋白的N末端(ZF-左)与胞嘧啶脱氨酶的第一分裂物的C末端融合，并且锌指蛋白的C末端(ZF-右)与胞嘧啶脱氨酶的第二分裂物的N末端融合的构建体(NC构型)中，腺嘌呤脱氨酶可以与锌指蛋白的C末端(ZF-左)，胞嘧啶脱氨酶的第一分裂物的N末端或C末端，锌指蛋白的N末端(ZF-右)，或胞嘧啶脱氨酶的第二分裂物的N末端或C末端融合。
此外，腺嘌呤脱氨酶可以与锌指蛋白的C末端(ZF-左)，胞嘧啶脱氨酶的第一分裂物的N末端或C末端，锌指蛋白的N末端(ZF-右)，或胞嘧啶脱氨酶的第二分裂物的N末端或C末端融合，即使在其中锌指蛋白的C末端(ZF-左)与胞嘧啶脱氨酶的第一分裂物的N末端融合并且锌指蛋白的C末端(ZF-右)与胞嘧啶脱氨酶的第二分裂物的N末端融合(CC构型)；锌指蛋白的C末端(ZF-左)与胞嘧啶脱氨酶的第一分裂物的N末端融合并且锌指蛋白的N末端(ZF-右)与胞嘧啶脱氨酶的第二分裂物的C末端融合(CN构型)；或者锌指蛋白的N末端(ZF-左)与胞嘧啶脱氨酶的第一分裂物的C末端融合并且锌指蛋白的N末端(ZF-右)与胞嘧啶脱氨酶的第二分裂物的C末端融合(NN构型)的构建体中。
当包括分裂形式的胞嘧啶脱氨酶并且DNA结合蛋白是TALE时，第一TALE可以与胞嘧啶脱氨酶的第一分裂物融合，并且第二TALE可以与胞嘧啶脱氨酶的第二分裂物融合，并且相应的构建体可以被描述为N’-TALE-第一分裂DddA-C’和N’-TALE-第二分裂DddA-C’。腺嘌呤脱氨酶可以与胞嘧啶脱氨酶的第一分裂物的N末端或C末端或与胞嘧啶脱氨酶的第二分裂物的N末端或C末端融合。
当包括全长形式的胞嘧啶脱氨酶并且DNA结合蛋白是TALE时，单个TALE模块可以是N’-TALE-全长DDDA-C’。这里，腺嘌呤脱氨酶可以与胞嘧啶脱氨酶的N末端或C末端融合。这里，腺嘌呤脱氨酶可以与单个TALE模块的C末端方向融合或与胞嘧啶脱氨酶的N末端或C末端融合。
当包括全长形式的胞嘧啶脱氨酶并且DNA结合蛋白是TALE时，可以包括双TALE模块，可以在N-C方向上包括第一TALE模块和胞嘧啶脱氨酶(N’-TALE-全长DDDA-C’)，并且还可以包括腺嘌呤脱氨酶和包含第二TALE的第二分裂物(N’-TALE-腺嘌呤脱氨酶-C’)。这里，腺嘌呤脱氨酶可以与TALE的N末端或C末端融合，例如在N’-TALE-胞嘧啶脱氨酶-C’和N’-TALE-腺嘌呤脱氨酶-C’的构建体中。
在一些情况下，可以另外包括能够增加碱基编辑效率的UGI(尿嘧啶DNA糖基化酶抑制剂)。UGI能够通过抑制UDG(尿嘧啶DNA糖基化酶)的活性来增加碱基编辑效率，UDG是一种通过从DNA中去除U来修复突变DNA的酶。
本发明涉及一种用于在原核或真核细胞中进行A至G碱基编辑的方法，所述方法包括融合蛋白或编码其的核酸，其中所述DNA结合蛋白是锌指蛋白、TALE蛋白或CRISPR相关核酸酶，以及所述融合蛋白的胞嘧啶脱氨酶或其变体衍生自细菌并且对双链DNA具有特异性。
本发明涉及一种用于在原核或真核细胞中进行A至G碱基编辑的组合物，其包含融合蛋白或编码其的核酸，其中DNA结合蛋白是锌指蛋白、TALE蛋白或CRISPR相关蛋白。
所述融合蛋白的胞嘧啶脱氨酶或其变体衍生自细菌并且对双链DNA具有特异性。所述DNA结合蛋白与所述胞嘧啶脱氨酶或其变体的N末端和C末端融合。类似地，所述DNA结合蛋白也与所述融合蛋白的腺嘌呤脱氨酶的N末端和C末端融合。在包含DNA结合蛋白、胞嘧啶脱氨酶或其变体和腺嘌呤脱氨酶的融合蛋白的上下文中，所述腺嘌呤脱氨酶可以位于所述融合蛋白内胞嘧啶脱氨酶的N末端或C末端，或者可以作为独立于其他DNA结合蛋白的单独蛋白质存在。
本发明涉及一种用于在原核或真核细胞中进行C至T碱基编辑的组合物，所述组合物包含所述融合蛋白或编码其的核酸和UGI(尿嘧啶糖基化酶抑制剂)，其中所述DNA结合蛋白是锌指蛋白、TALE蛋白或CRISPR相关核酸酶，并且所述融合蛋白的胞嘧啶脱氨酶或其变体衍生自细菌并且对双链DNA具有特异性。
具体地，本发明涉及一种用于在原核和真核细胞中进行A至G碱基编辑(无UGI)的组合物，所述组合物包含1)DNA结合蛋白，2)全长双链DNA特异性细菌胞嘧啶脱氨酶或其变体，和3)衍生自大肠杆菌TadA的脱氧腺嘌呤脱氨酶，其中所述DNA结合蛋白是锌指蛋白(ZFP)、转录激活因子样效应物(TALE)阵列、或催化缺陷型CRISPR-Cas9(nCas9或dCas9)或Cas12a，并且所述全长双链DNA特异性细菌胞嘧啶脱氨酶是衍生自洋葱伯克霍尔德菌(Burkholderia cenocepacia)的DddAtox。
本发明涉及一种用于在原核和真核细胞中进行A至G碱基编辑(无UGI)的组合物，所述组合物包含1)与全长双链DNA特异性细菌胞嘧啶脱氨酶或其变体可操作地连接的左DNA结合蛋白，和2)与衍生自大肠杆菌TadA的脱氧腺嘌呤脱氨酶可操作地连接的右DNA结合蛋白，其中左或右DNA结合蛋白是锌指蛋白(ZFP)、转录激活因子样效应物(TALE)阵列、或催化缺陷型CRISPR-Cas9(nCas9或dCas9)或Cas12a，并且所述全长双链DNA特异性细菌胞嘧啶脱氨酶是衍生自洋葱伯克霍尔德菌的DddAtox。融合蛋白中左右组分的顺序可以互换。
本发明还涉及一种用于在原核和真核细胞中进行A至G和C至T碱基编辑的组合物，所述组合物包含1)DNA结合蛋白，2)全长双链DNA特异性细菌胞嘧啶脱氨酶或其变体，3)衍生自大肠杆菌TadA的脱氧腺嘌呤脱氨酶，和4)UGI(尿嘧啶糖基化酶抑制剂)，其中所述DNA结合蛋白是锌指蛋白(ZFP)、转录激活因子样效应物(TALE)阵列、或催化缺陷型CRISPR-Cas9(nCas9或dCas9)或Cas12a，并且所述全长双链DNA特异性细菌胞嘧啶脱氨酶是衍生自洋葱伯克霍尔德菌的DddAtox。
本发明还涉及一种用于在原核和真核细胞中进行A至G碱基编辑(无UGI)的组合物，所述组合物包含1)DNA结合蛋白，2)分裂物双链DNA特异性细菌胞嘧啶脱氨酶或其变体，和3)衍生自大肠杆菌TadA的脱氧腺嘌呤脱氨酶，其中所述DNA结合蛋白是锌指蛋白(ZFP)、转录激活因子样效应物(TALE)阵列、或催化缺陷型CRISPR-Cas9(nCas9或dCas9)或Cas12a，并且所述分裂物双链DNA特异性细菌胞嘧啶脱氨酶是衍生自洋葱伯克霍尔德菌的DddAtox。
本发明还涉及一种用于在原核和真核细胞中进行A至G和C至T碱基编辑的组合物，所述组合物包含1)DNA结合蛋白，2)分裂物双链DNA特异性细菌胞嘧啶脱氨酶或其变体，3)衍生自大肠杆菌TadA的脱氧腺嘌呤脱氨酶，和4)UGI(尿嘧啶糖基化酶抑制剂)，其中所述DNA结合蛋白是锌指蛋白(ZFP)、转录激活因子样效应物(TALE)阵列、或催化缺陷型CRISPR-Cas9(nCas9或dCas9)或Cas12a，并且所述分裂物双链DNA特异性细菌胞嘧啶脱氨酶是衍生自洋葱伯克霍尔德菌的DddAtox。
本发明涉及一种用于在原核或真核细胞中进行A至G碱基编辑的方法，所述方法包括用融合蛋白或编码其的核酸处理原核细胞或真核细胞，其中DNA结合蛋白是锌指蛋白、TALE蛋白或CRISPR相关核酸酶，以及所述融合蛋白的胞嘧啶脱氨酶或其变体衍生自细菌并且对双链DNA具有特异性。
本发明涉及一种用于在原核或真核细胞中进行A至G碱基编辑的方法，所述方法包括用融合蛋白或编码其的核酸处理原核细胞或真核细胞，其中DNA结合蛋白是锌指蛋白、TALE蛋白或CRISPR相关核酸酶，所述融合蛋白的胞嘧啶脱氨酶或其变体衍生自细菌并且对双链DNA具有特异性。所述DNA结合蛋白与所述胞嘧啶脱氨酶或其变体的N末端和C末端融合。类似地，所述DNA结合蛋白也与所述融合蛋白的腺嘌呤脱氨酶的N末端和C末端融合。在包含DNA结合蛋白、胞嘧啶脱氨酶或其变体和腺嘌呤脱氨酶的融合蛋白的上下文中，所述腺嘌呤脱氨酶可以位于所述融合蛋白内胞嘧啶脱氨酶的N末端或C末端，或者可以作为独立于其他DNA结合蛋白的单独蛋白质存在。
本发明涉及一种用于在原核或真核细胞中进行C至T碱基编辑的方法，所述方法包括用融合蛋白或编码其的核酸和UGI(尿嘧啶糖基化酶抑制剂)处理原核细胞或真核细胞，其中DNA结合蛋白是锌指蛋白、TALE蛋白或CRISPR相关核酸酶，以及所述融合蛋白的胞嘧啶脱氨酶或其变体衍生自细菌并且对双链DNA具有特异性。
包含在根据本发明的组合物或方法中的组分的具体序列如下。
ND1-ZFP-右-1397C-AD(图61f-61g：SEQ ID No:410)
MLGFVGRVAAAPASGALRRLTPSASLPPAQLLLRAAPTAVHPVRDYAAQDYKDDDDKVDEMTKKFGTLTIHDTEKAAEFGIRIPGEKPFQCRICMRNFSDSGNLRVHIRTHTGEKPYKCPDCGKSFSQSSSLIRHQRTHTGEKPYECDHCGKSFSQSSHLNVHKRTHTGEKPYRCKYCDRSFSISSNLQRHVRNIHLRSGTPHEVGVYTLSGTPHEVGVYTLAIPVKRGATGETKVFTGNSNSPKSPTKGGCSGSETPGTSESATPESSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFGVRNSKRGAAGSLMNVLNYPGMNHRVEITEGILADECAALLCDFYRMPRQVFNAQKKAQSSIN
ND1-ZFP-左-1397C-UGI(图61f-61g：SEQ ID No:411)
MLGFVGRVAAAPASGALRRLTPSASLPPAQLLLRAAPTAVHPVRDYAAQDYKDDDDKVDEMTKKFGTLTIHDTEKAAEFGIRIPGEKPFQCRICMRNFSDSGNLRVHIRTHTGEKPYKCPDCGKSFSQSSSLIRHQRTHTGEKPYECDHCGKSFSQSSHLNVHKRTHTGEKPYRCKYCDRSFSISSNLQRHVRNIHLRSGTPHEVGVYTLSGTPHEVGVYTLAIPVKRGATGETKVFTGNSNSPKSPTKGGCSGGSTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTAYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML
ND1-ZFP-右-1397N-UGI(图61f-61g：SEQ ID No:412)
MLGFVGRVAAAPASGALRRLTPSASLPPAQLLLRAAPTAVHPVRDYAAQYPYDVPDYAVDEMTKKFGTLTIHDTEKAAEFGIHGVPAAMGGSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLESKVFSSGGPTPYPNYANAGHVEGQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGSGTPHEVGVYTLSGTPHEVGVYTLYKCPECGKSFSTKNSLTEHQRTHTGEKPYKCPECGKSFSSKKALTEHQRTHTGEKPYECNYCGKTFSVSSTLIRHQRIHTGEKPYRCKYCDRSFSISSNLQRHVRNIHLRSGGSTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTAYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML
Trac-ZFP-左-1397N-UGI(图61d：SEQ ID No:413)
MAPKKKRKVGIHGVPAAMGGSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLESKVFSSGGPTPYPNYANAGHVEGQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGSGTPHEVGVYTLSGTPHEVGVYTLFQCRICMRKFATSGSLTRHTKIHTGEKPFQCRICMRNFSRSDHLSTHIRTHTGEKPFACDICGRKFATSSNRTKHTKIHTHPRAPIPKPFQCRICMRNFSRSDNLSEHIRTHTGEKPFACDICGRKFAWHSSLRVHTKIHLRSGGSTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTAYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML
Trac-ZFP-右-1397C-AD(图61d：SEQ ID No:414)
MAPKKKRKVGIHGVPAAMAERPFQCRICMRNFSRSDHLSTHIRTHTGEKPFACDICGRKFADRSHLARHTKIHTGSQKPFQCRICMRKFALKQHLNEHTKIHTGEKPFQCRICMRNFSQSGNLARHIRTHTGEKPFACDICGRKFAHNSSLKDHTKIHLRSGTPHEVGVYTLSGTPHEVGVYTLAIPVKRGATGETKVFTGNSNSPKSPTKGGCSGSETPGTSESATPESSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFGVRNSKRGAAGSLMNVLNYPGMNHRVEITEGILADECAALLCDFYRMPRQVFNAQKKAQSSIN
ND1-左-TALE-1397N-UGI(图62：SEQ ID No:415)
MALSRAVCGTSRQLAPVLGYLGSRQKHSLPDYPYDVPDYAGYPYDVPDYAGYPYDVPDYAGIRIQDLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGHGFTHAHIVALSQHPAALGTVAVKYQDMIAALPEATHEAIVGVGKQWSGARALEALLTVAGELRGPPLQLDTGQLLKIAKRGGVTAVEAVHAWRNALTGAPLNLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQDHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALESIVAQLSRPDPALAALLVGSGSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLESKVFSSGGPTPYPNYANAGHVEGQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGSGGSTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTAYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML
ND1-左-TALE-1397C-UGI(图62：SEQ ID No:416)
MALSRAVCGTSRQLAPVLGYLGSRQKHSLPDYPYDVPDYAGYPYDVPDYAGYPYDVPDYAGIRIQDLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGHGFTHAHIVALSQHPAALGTVAVKYQDMIAALPEATHEAIVGVGKQWSGARALEALLTVAGELRGPPLQLDTGQLLKIAKRGGVTAVEAVHAWRNALTGAPLNLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQDHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALESIVAQLSRPDPALAALLVGSAIPVKRGATGETKVFTGNSNSPKSPTKGGCSGGSTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTAYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML
ND1-右-TALE-1397N-UGI(图62：SEQ ID No:417)
MASVLTPLLLRGLTGSARRLPVPRAKIHSLDYKDHDGDYKDHDIDYKDDDDKGIRIQDLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGHGFTHAHIVALSQHPAALGTVAVKYQDMIAALPEATHEAIVGVGKQWSGARALEALLTVAGELRGPPLQLDTGQLLKIAKRGGVTAVEAVHAWRNALTGAPLNLTPDQVVAIASNNGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNNGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALESIVAQLSRPDPALAALLVGSGSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLESKVFSSGGPTPYPNYANAGHVEGQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGSGGSTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTAYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML
ND1-右-TALE-1397C-UGI(图62：SEQ ID No:418)
MASVLTPLLLRGLTGSARRLPVPRAKIHSLDYKDHDGDYKDHDIDYKDDDDKGIRIQDLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGHGFTHAHIVALSQHPAALGTVAVKYQDMIAALPEATHEAIVGVGKQWSGARALEALLTVAGELRGPPLQLDTGQLLKIAKRGGVTAVEAVHAWRNALTGAPLNLTPDQVVAIASNNGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNNGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALESIVAQLSRPDPALAALLVGSAIPVKRGATGETKVFTGNSNSPKSPTKGGCSGGSTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTAYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML
ND4-左-TALE-AD(图62b：SEQ ID No:419)
MALSRAVCGTSRQLAPVLGYLGSRQKHSLPDYPYDVPDYAGYPYDVPDYAGYPYDVPDYAMDIADLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGHGFTHAHIVALSQHPAALGTVAVKYQDMIAALPEATHEAIVGVGKQWSGARALEALLTVAGELRGPPLQLDTGQLLKIAKRGGVTAVEAVHAWRNALTGAPLNLTPEQVVAIASNNGGKQALETVQALLPVLCQAHGLTPQQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPQQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNIGGKQALETVQALLPVLCQAHGLTPEQVVAIASNNGGKQALETVQALLPVLCQAHGLTPEQVVAIASNGGGKQALETVQALLPVLCQAHGLTPEQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNIGGKQALETVQALLPVLCQAHGLTPEQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASHDGGKQALETVQALLPVLCQAHGLTPEQVVAIASNIGGKQALETVQALLPVLCQAHGLTPEQVVAIASHDGGKQALETVQALLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPQQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALETVQALLPVLCQAHGLTPEQVVAIASHDGGKQALETVQALLPVLCQAHGLTPQQVVAIASNGGGRPALESIVAQLSRPDPALAALTNDHLVALACLGGRPALDAVKKGLGGSSGSETPGTSESATPESSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFGVRNSKRGAAGSLMNVLNYPGMNHRVEITEGILADECAALLCDFYRMPRQVFNAQKKAQSSIN
ND4-右-TALE-AD(图62b：SEQ ID No:420)
MASVLTPLLLRGLTGSARRLPVPRAKIHSLDYKDHDGDYKDHDIDYKDDDDKMDIADLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGHGFTHAHIVALSQHPAALGTVAVKYQDMIAALPEATHEAIVGVGKQWSGARALEALLTVAGELRGPPLQLDTGQLLKIAKRGGVTAVEAVHAWRNALTGAPLNLTPEQVVAIASHDGGKQALETVQALLPVLCQAHGLTPQQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPQQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQALLPVLCQAHGLTPEQVVAIASNGGGKQALETVQALLPVLCQAHGLTPEQVVAIASNIGGKQALETVQALLPVLCQAHGLTPEQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQALLPVLCQAHGLTPEQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQALLPVLCQAHGLTPEQVVAIASNNGGKQALETVQALLPVLCQAHGLTPEQVVAIASNIGGKQALETVQALLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPQQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPQQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPQQVVAIASNGGGRPALESIVAQLSRPDPALAALTNDHLVALACLGGRPALDAVKKGLGGSSGSETPGTSESATPESSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFGVRNSKRGAAGSLMNVLNYPGMNHRVEITEGILADECAALLCDFYRMPRQVFNAQKKAQSSIN
ND1-左-TALE-1333N-AD(图63：SEQ ID No:421)
MALSRAVCGTSRQLAPVLGYLGSRQKHSLPDYPYDVPDYAGYPYDVPDYAGYPYDVPDYAGIRIQDLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGHGFTHAHIVALSQHPAALGTVAVKYQDMIAALPEATHEAIVGVGKQWSGARALEALLTVAGELRGPPLQLDTGQLLKIAKRGGVTAVEAVHAWRNALTGAPLNLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQDHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALESIVAQLSRPDPALAALLVGSGSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLESKVFSSGGSGSETPGTSESATPESSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFGVRNSKRGAAGSLMNVLNYPGMNHRVEITEGILADECAALLCDFYRMPRQVFNAQKKAQSSIN
ND1-左-TALE-1333C-AD(图63：SEQ ID No:422)
MALSRAVCGTSRQLAPVLGYLGSRQKHSLPDYPYDVPDYAGYPYDVPDYAGYPYDVPDYAGIRIQDLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGHGFTHAHIVALSQHPAALGTVAVKYQDMIAALPEATHEAIVGVGKQWSGARALEALLTVAGELRGPPLQLDTGQLLKIAKRGGVTAVEAVHAWRNALTGAPLNLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQDHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALESIVAQLSRPDPALAALLVGSPTPYPNYANAGHVEGQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGAIPVKRGATGETKVFTGNSNSPKSPTKGGCSGSETPGTSESATPESSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFGVRNSKRGAAGSLMNVLNYPGMNHRVEITEGILADECAALLCDFYRMPRQVFNAQKKAQSSIN
ND1-右-TALE-1333N-AD(图63：SEQ ID No:423)
MASVLTPLLLRGLTGSARRLPVPRAKIHSLDYKDHDGDYKDHDIDYKDDDDKGIRIQDLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGHGFTHAHIVALSQHPAALGTVAVKYQDMIAALPEATHEAIVGVGKQWSGARALEALLTVAGELRGPPLQLDTGQLLKIAKRGGVTAVEAVHAWRNALTGAPLNLTPDQVVAIASNNGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNNGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALESIVAQLSRPDPALAALLVGSGSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLESKVFSSGGSGSETPGTSESATPESSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFGVRNSKRGAAGSLMNVLNYPGMNHRVEITEGILADECAALLCDFYRMPRQVFNAQKKAQSSIN
ND1-右-TALE-1333C-AD(图63：SEQ ID No:424)
MASVLTPLLLRGLTGSARRLPVPRAKIHSLDYKDHDGDYKDHDIDYKDDDDKGIRIQDLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGHGFTHAHIVALSQHPAALGTVAVKYQDMIAALPEATHEAIVGVGKQWSGARALEALLTVAGELRGPPLQLDTGQLLKIAKRGGVTAVEAVHAWRNALTGAPLNLTPDQVVAIASNNGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNNGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALESIVAQLSRPDPALAALLVGSPTPYPNYANAGHVEGQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGAIPVKRGATGETKVFTGNSNSPKSPTKGGCSGSETPGTSESATPESSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFGVRNSKRGAAGSLMNVLNYPGMNHRVEITEGILADECAALLCDFYRMPRQVFNAQKKAQSSIN
ND1-左-TALE-1397C-AD(图62-63：SEQ ID No:425)
MALSRAVCGTSRQLAPVLGYLGSRQKHSLPDYPYDVPDYAGYPYDVPDYAGYPYDVPDYAGIRIQDLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGHGFTHAHIVALSQHPAALGTVAVKYQDMIAALPEATHEAIVGVGKQWSGARALEALLTVAGELRGPPLQLDTGQLLKIAKRGGVTAVEAVHAWRNALTGAPLNLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQDHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALESIVAQLSRPDPALAALTNDHLVALACLGGRPALDAVKKGLLVGSAIPVKRGATGETKVFTGNSNSPKSPTKGGCSGSETPGTSESATPESSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFGVRNSKRGAAGSLMNVLNYPGMNHRVEITEGILADECAALLCDFYRMPRQVFNAQKKAQSSIN
ND1-右-TALE-1397C-AD(图62-63：SEQ ID No:426)
MASVLTPLLLRGLTGSARRLPVPRAKIHSLDYKDHDGDYKDHDIDYKDDDDKGIRIQDLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGHGFTHAHIVALSQHPAALGTVAVKYQDMIAALPEATHEAIVGVGKQWSGARALEALLTVAGELRGPPLQLDTGQLLKIAKRGGVTAVEAVHAWRNALTGAPLNLTPDQVVAIASNNGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNNGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALESIVAQLSRPDPALAALTNDHLVALACLGGRPALDAVKKGLGGSGSAIPVKRGATGETKVFTGNSNSPKSPTKGGCSGSETPGTSESATPESSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFGVRNSKRGAAGSLMNVLNYPGMNHRVEITEGILADECAALLCDFYRMPRQVFNAQKKAQSSIN
ND1-左-TALE-1333N(图63：SEQ ID No:427)
MALSRAVCGTSRQLAPVLGYLGSRQKHSLPDYPYDVPDYAGYPYDVPDYAGYPYDVPDYAGIRIQDLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGHGFTHAHIVALSQHPAALGTVAVKYQDMIAALPEATHEAIVGVGKQWSGARALEALLTVAGELRGPPLQLDTGQLLKIAKRGGVTAVEAVHAWRNALTGAPLNLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQDHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALESIVAQLSRPDPALAALTNDHLVALACLGGRPALDAVKKGLGGSGSGSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLESKVFSSGG
ND1-左-TALE-1333C(图63：SEQ ID No:428)
MALSRAVCGTSRQLAPVLGYLGSRQKHSLPDYPYDVPDYAGYPYDVPDYAGYPYDVPDYAGIRIQDLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGHGFTHAHIVALSQHPAALGTVAVKYQDMIAALPEATHEAIVGVGKQWSGARALEALLTVAGELRGPPLQLDTGQLLKIAKRGGVTAVEAVHAWRNALTGAPLNLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQDHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALESIVAQLSRPDPALAALTNDHLVALACLGGRPALDAVKKGLGGSGSPTPYPNYANAGHVEGQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGAIPVKRGATGETKVFTGNSNSPKSPTKGGC
ND1-左-TALE-1397N(图62-63：SEQ ID No:429)
MALSRAVCGTSRQLAPVLGYLGSRQKHSLPDYPYDVPDYAGYPYDVPDYAGYPYDVPDYAGIRIQDLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGHGFTHAHIVALSQHPAALGTVAVKYQDMIAALPEATHEAIVGVGKQWSGARALEALLTVAGELRGPPLQLDTGQLLKIAKRGGVTAVEAVHAWRNALTGAPLNLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQDHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALESIVAQLSRPDPALAALTNDHLVALACLGGRPALDAVKKGLGGSGSGSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLESKVFSSGGPTPYPNYANAGHVEGQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEG
ND1-右-TALE-1333N(图63：SEQ ID No:430)
MASVLTPLLLRGLTGSARRLPVPRAKIHSLDYKDHDGDYKDHDIDYKDDDDKGIRIQDLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGHGFTHAHIVALSQHPAALGTVAVKYQDMIAALPEATHEAIVGVGKQWSGARALEALLTVAGELRGPPLQLDTGQLLKIAKRGGVTAVEAVHAWRNALTGAPLNLTPDQVVAIASNNGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNNGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALESIVAQLSRPDPALAALLVGSGSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLESKVFSSGG
ND1-右-TALE-1333C(图63：SEQ ID No:431)
MASVLTPLLLRGLTGSARRLPVPRAKIHSLDYKDHDGDYKDHDIDYKDDDDKGIRIQDLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGHGFTHAHIVALSQHPAALGTVAVKYQDMIAALPEATHEAIVGVGKQWSGARALEALLTVAGELRGPPLQLDTGQLLKIAKRGGVTAVEAVHAWRNALTGAPLNLTPDQVVAIASNNGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNNGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALESIVAQLSRPDPALAALLVGSPTPYPNYANAGHVEGQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGAIPVKRGATGETKVFTGNSNSPKSPTKGGC
ND1-右-TALE-1397N(图62-63：SEQ ID No:432)
MASVLTPLLLRGLTGSARRLPVPRAKIHSLDYKDHDGDYKDHDIDYKDDDDKGIRIQDLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGHGFTHAHIVALSQHPAALGTVAVKYQDMIAALPEATHEAIVGVGKQWSGARALEALLTVAGELRGPPLQLDTGQLLKIAKRGGVTAVEAVHAWRNALTGAPLNLTPDQVVAIASNNGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNNGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALESIVAQLSRPDPALAALLVGSGSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLESKVFSSGGPTPYPNYANAGHVEGQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEG
ND4-左-TALE-1333N-AD(图63：SEQ ID No:433)
MALSRAVCGTSRQLAPVLGYLGSRQKHSLPDYPYDVPDYAGYPYDVPDYAGYPYDVPDYAMDIADLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGHGFTHAHIVALSQHPAALGTVAVKYQDMIAALPEATHEAIVGVGKQWSGARALEALLTVAGELRGPPLQLDTGQLLKIAKRGGVTAVEAVHAWRNALTGAPLNLTPEQVVAIASNNGGKQALETVQALLPVLCQAHGLTPQQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPQQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNIGGKQALETVQALLPVLCQAHGLTPEQVVAIASNNGGKQALETVQALLPVLCQAHGLTPEQVVAIASNGGGKQALETVQALLPVLCQAHGLTPEQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNIGGKQALETVQALLPVLCQAHGLTPEQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASHDGGKQALETVQALLPVLCQAHGLTPEQVVAIASNIGGKQALETVQALLPVLCQAHGLTPEQVVAIASHDGGKQALETVQALLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPQQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALETVQALLPVLCQAHGLTPEQVVAIASHDGGKQALETVQALLPVLCQAHGLTPQQVVAIASNGGGRPALESIVAQLSRPDPALAALTNDHLVALACLGGRPALDAVKKGLGGSGSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLESKVFSSGGSGSETPGTSESATPESSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFGVRNSKRGAAGSLMNVLNYPGMNHRVEITEGILADECAALLCDFYRMPRQVFNAQKKAQSSIN
ND4-左-TALE-1333C-AD(图63：SEQ ID No:434)
MALSRAVCGTSRQLAPVLGYLGSRQKHSLPDYPYDVPDYAGYPYDVPDYAGYPYDVPDYAMDIADLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGHGFTHAHIVALSQHPAALGTVAVKYQDMIAALPEATHEAIVGVGKQWSGARALEALLTVAGELRGPPLQLDTGQLLKIAKRGGVTAVEAVHAWRNALTGAPLNLTPEQVVAIASNNGGKQALETVQALLPVLCQAHGLTPQQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPQQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNIGGKQALETVQALLPVLCQAHGLTPEQVVAIASNNGGKQALETVQALLPVLCQAHGLTPEQVVAIASNGGGKQALETVQALLPVLCQAHGLTPEQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNIGGKQALETVQALLPVLCQAHGLTPEQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASHDGGKQALETVQALLPVLCQAHGLTPEQVVAIASNIGGKQALETVQALLPVLCQAHGLTPEQVVAIASHDGGKQALETVQALLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPQQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALETVQALLPVLCQAHGLTPEQVVAIASHDGGKQALETVQALLPVLCQAHGLTPQQVVAIASNGGGRPALESIVAQLSRPDPALAALTNDHLVALACLGGRPALDAVKKGLGGSPTPYPNYANAGHVEGQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGAIPVKRGATGETKVFTGNSNSPKSPTKGGCSGSETPGTSESATPESSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFGVRNSKRGAAGSLMNVLNYPGMNHRVEITEGILADECAALLCDFYRMPRQVFNAQKKAQSSIN
ND4-左-TALE-1397C-AD(图63：SEQ ID No:435)
MALSRAVCGTSRQLAPVLGYLGSRQKHSLPDYPYDVPDYAGYPYDVPDYAGYPYDVPDYAMDIADLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGHGFTHAHIVALSQHPAALGTVAVKYQDMIAALPEATHEAIVGVGKQWSGARALEALLTVAGELRGPPLQLDTGQLLKIAKRGGVTAVEAVHAWRNALTGAPLNLTPEQVVAIASNNGGKQALETVQALLPVLCQAHGLTPQQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPQQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNIGGKQALETVQALLPVLCQAHGLTPEQVVAIASNNGGKQALETVQALLPVLCQAHGLTPEQVVAIASNGGGKQALETVQALLPVLCQAHGLTPEQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNIGGKQALETVQALLPVLCQAHGLTPEQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASHDGGKQALETVQALLPVLCQAHGLTPEQVVAIASNIGGKQALETVQALLPVLCQAHGLTPEQVVAIASHDGGKQALETVQALLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPQQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALETVQALLPVLCQAHGLTPEQVVAIASHDGGKQALETVQALLPVLCQAHGLTPQQVVAIASNGGGRPALESIVAQLSRPDPALAALTNDHLVALACLGGRPALDAVKKGLGGSGSAIPVKRGATGETKVFTGNSNSPKSPTKGGCSGSETPGTSESATPESSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFGVRNSKRGAAGSLMNVLNYPGMNHRVEITEGILADECAALLCDFYRMPRQVFNAQKKAQSSIN
ND4-右-TALE-1333N-AD(图63：SEQ ID No:436)
MASVLTPLLLRGLTGSARRLPVPRAKIHSLDYKDHDGDYKDHDIDYKDDDDKMDIADLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGHGFTHAHIVALSQHPAALGTVAVKYQDMIAALPEATHEAIVGVGKQWSGARALEALLTVAGELRGPPLQLDTGQLLKIAKRGGVTAVEAVHAWRNALTGAPLNLTPEQVVAIASHDGGKQALETVQALLPVLCQAHGLTPQQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPQQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQALLPVLCQAHGLTPEQVVAIASNGGGKQALETVQALLPVLCQAHGLTPEQVVAIASNIGGKQALETVQALLPVLCQAHGLTPEQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQALLPVLCQAHGLTPEQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQALLPVLCQAHGLTPEQVVAIASNNGGKQALETVQALLPVLCQAHGLTPEQVVAIASNIGGKQALETVQALLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPQQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPQQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPQQVVAIASNGGGRPALESIVAQLSRPDPALAALTNDHLVALACLGGRPALDAVKKGLGGSGSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLESKVFSSGGSGSETPGTSESATPESSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFGVRNSKRGAAGSLMNVLNYPGMNHRVEITEGILADECAALLCDFYRMPRQVFNAQKKAQSSIN
ND4-右-TALE-1333C-AD(图63：SEQ ID No:437)
MASVLTPLLLRGLTGSARRLPVPRAKIHSLDYKDHDGDYKDHDIDYKDDDDKMDIADLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGHGFTHAHIVALSQHPAALGTVAVKYQDMIAALPEATHEAIVGVGKQWSGARALEALLTVAGELRGPPLQLDTGQLLKIAKRGGVTAVEAVHAWRNALTGAPLNLTPEQVVAIASHDGGKQALETVQALLPVLCQAHGLTPQQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPQQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQALLPVLCQAHGLTPEQVVAIASNGGGKQALETVQALLPVLCQAHGLTPEQVVAIASNIGGKQALETVQALLPVLCQAHGLTPEQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQALLPVLCQAHGLTPEQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQALLPVLCQAHGLTPEQVVAIASNNGGKQALETVQALLPVLCQAHGLTPEQVVAIASNIGGKQALETVQALLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPQQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPQQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPQQVVAIASNGGGRPALESIVAQLSRPDPALAALTNDHLVALACLGGRPALDAVKKGLGGSPTPYPNYANAGHVEGQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGAIPVKRGATGETKVFTGNSNSPKSPTKGGCSGSETPGTSESATPESSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFGVRNSKRGAAGSLMNVLNYPGMNHRVEITEGILADECAALLCDFYRMPRQVFNAQKKAQSSIN
ND4-右-TALE-1397C-AD(图63：SEQ ID No:438)
MASVLTPLLLRGLTGSARRLPVPRAKIHSLDYKDHDGDYKDHDIDYKDDDDKMDIADLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGHGFTHAHIVALSQHPAALGTVAVKYQDMIAALPEATHEAIVGVGKQWSGARALEALLTVAGELRGPPLQLDTGQLLKIAKRGGVTAVEAVHAWRNALTGAPLNLTPEQVVAIASHDGGKQALETVQALLPVLCQAHGLTPQQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPQQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQALLPVLCQAHGLTPEQVVAIASNGGGKQALETVQALLPVLCQAHGLTPEQVVAIASNIGGKQALETVQALLPVLCQAHGLTPEQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQALLPVLCQAHGLTPEQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQALLPVLCQAHGLTPEQVVAIASNNGGKQALETVQALLPVLCQAHGLTPEQVVAIASNIGGKQALETVQALLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPQQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPQQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPQQVVAIASNGGGRPALESIVAQLSRPDPALAALTNDHLVALACLGGRPALDAVKKGLGGSGSAIPVKRGATGETKVFTGNSNSPKSPTKGGCSGSETPGTSESATPESSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFGVRNSKRGAAGSLMNVLNYPGMNHRVEITEGILADECAALLCDFYRMPRQVFNAQKKAQSSIN
ND1-左-TALE-AD-GSVG(图64：SEQ ID No:439)
MALSRAVCGTSRQLAPVLGYLGSRQKHSLPDYPYDVPDYAGYPYDVPDYAGYPYDVPDYAGIRIQDLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGHGFTHAHIVALSQHPAALGTVAVKYQDMIAALPEATHEAIVGVGKQWSGARALEALLTVAGELRGPPLQLDTGQLLKIAKRGGVTAVEAVHAWRNALTGAPLNLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQDHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALESIVAQLSRPDPALAALLVSGSETPGTSESATPESSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFGVRNSKRGAAGSLMNVLNYPGMNHRVEITEGILADECAALLCDFYRMPRQVFNAQKKAQSSINLVGSGSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLEGKVFSSGGPTPYPNYANAGHVESQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGVIPVKRGATGETKVFTGNSNGPKSPTKGGC
ND1-左-TALE-AD-E1347A(图64：SEQ ID No:440)
MALSRAVCGTSRQLAPVLGYLGSRQKHSLPDYPYDVPDYAGYPYDVPDYAGYPYDVPDYAGIRIQDLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGHGFTHAHIVALSQHPAALGTVAVKYQDMIAALPEATHEAIVGVGKQWSGARALEALLTVAGELRGPPLQLDTGQLLKIAKRGGVTAVEAVHAWRNALTGAPLNLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQDHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALESIVAQLSRPDPALAALLVSGSETPGTSESATPESSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFGVRNSKRGAAGSLMNVLNYPGMNHRVEITEGILADECAALLCDFYRMPRQVFNAQKKAQSSINLVGSGSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLESKVFSSGGPTPYPNYANAGHVAGQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGAIPVKRGATGETKVFTGNSNSPKSPTKGGC
ND1-左-TALE-AD-AAAAA(图64：SEQ ID No:441)
MALSRAVCGTSRQLAPVLGYLGSRQKHSLPDYPYDVPDYAGYPYDVPDYAGYPYDVPDYAGIRIQDLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGHGFTHAHIVALSQHPAALGTVAVKYQDMIAALPEATHEAIVGVGKQWSGARALEALLTVAGELRGPPLQLDTGQLLKIAKRGGVTAVEAVHAWRNALTGAPLNLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQDHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALESIVAQLSRPDPALAALLVSGSETPGTSESATPESSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFGVRNSKRGAAGSLMNVLNYPGMNHRVEITEGILADECAALLCDFYRMPRQVFNAQKKAQSSINLVGSGSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLESKVFSSGGPTPYPNYANAGHVEGQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGAIPVAAGATGETAVFTGNSNSPASPTAGGC
ND1-右-TALE-AD(图64：SEQ ID No:442)
MASVLTPLLLRGLTGSARRLPVPRAKIHSLDYKDHDGDYKDHDIDYKDDDDKGIRIQDLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGHGFTHAHIVALSQHPAALGTVAVKYQDMIAALPEATHEAIVGVGKQWSGARALEALLTVAGELRGPPLQLDTGQLLKIAKRGGVTAVEAVHAWRNALTGAPLNLTPDQVVAIASNNGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNNGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALESIVAQLSRPDPALAALLVSGSETPGTSESATPESSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFGVRNSKRGAAGSLMNVLNYPGMNHRVEITEGILADECAALLCDFYRMPRQVFNAQKKAQSSIN
ND1-左-TALE-GSVG(图64：SEQ ID No:443)
MALSRAVCGTSRQLAPVLGYLGSRQKHSLPDYPYDVPDYAGYPYDVPDYAGYPYDVPDYAGIRIQDLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGHGFTHAHIVALSQHPAALGTVAVKYQDMIAALPEATHEAIVGVGKQWSGARALEALLTVAGELRGPPLQLDTGQLLKIAKRGGVTAVEAVHAWRNALTGAPLNLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQDHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALESIVAQLSRPDPALAALLVGSGSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLEGKVFSSGGPTPYPNYANAGHVESQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGVIPVKRGATGETKVFTGNSNGPKSPTKGGC
ND1-左-TALE-E1347A(图64：SEQ ID No:444)
MALSRAVCGTSRQLAPVLGYLGSRQKHSLPDYPYDVPDYAGYPYDVPDYAGYPYDVPDYAGIRIQDLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGHGFTHAHIVALSQHPAALGTVAVKYQDMIAALPEATHEAIVGVGKQWSGARALEALLTVAGELRGPPLQLDTGQLLKIAKRGGVTAVEAVHAWRNALTGAPLNLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQDHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALESIVAQLSRPDPALAALLVGSGSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLESKVFSSGGPTPYPNYANAGHVAGQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGAIPVKRGATGETKVFTGNSNSPKSPTKGGC
ND1-左-TALE-AAAAA(图64SEQ ID No:445)
MALSRAVCGTSRQLAPVLGYLGSRQKHSLPDYPYDVPDYAGYPYDVPDYAGYPYDVPDYAGIRIQDLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGHGFTHAHIVALSQHPAALGTVAVKYQDMIAALPEATHEAIVGVGKQWSGARALEALLTVAGELRGPPLQLDTGQLLKIAKRGGVTAVEAVHAWRNALTGAPLNLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQDHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALESIVAQLSRPDPALAALLVGSGSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLESKVFSSGGPTPYPNYANAGHVEGQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGAIPVAAGATGETAVFTGNSNSPASPTAGGC
ND1-TALE-左(SEQ ID No:446)
DLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGHGFTHAHIVALSQHPAALGTVAVKYQDMIAALPEATHEAIVGVGKQWSGARALEALLTVAGELRGPPLQLDTGQLLKIAKRGGVTAVEAVHAWRNALTGAPLNLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQDHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALESIVAQLSRPDPALAAL
ND1-TALE-右(SEQ ID No:447)
DLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGHGFTHAHIVALSQHPAALGTVAVKYQDMIAALPEATHEAIVGVGKQWSGARALEALLTVAGELRGPPLQLDTGQLLKIAKRGGVTAVEAVHAWRNALTGAPLNLTPDQVVAIASNNGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPAQVVAIASNNGGKQALETVQRLLPVLCQDHGLTPAQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPDQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPDQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPDQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQDHGLTPAQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALESIVAQLSRPDPALAAL
ND4-TALE-左(SEQ ID No:448)
MDIADLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGHGFTHAHIVALSQHPAALGTVAVKYQDMIAALPEATHEAIVGVGKQWSGARALEALLTVAGELRGPPLQLDTGQLLKIAKRGGVTAVEAVHAWRNALTGAPLNLTPEQVVAIASNNGGKQALETVQALLPVLCQAHGLTPQQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPQQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNIGGKQALETVQALLPVLCQAHGLTPEQVVAIASNNGGKQALETVQALLPVLCQAHGLTPEQVVAIASNGGGKQALETVQALLPVLCQAHGLTPEQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNIGGKQALETVQALLPVLCQAHGLTPEQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPEQVVAIASHDGGKQALETVQALLPVLCQAHGLTPEQVVAIASNIGGKQALETVQALLPVLCQAHGLTPEQVVAIASHDGGKQALETVQALLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPQQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNGGGKQALETVQALLPVLCQAHGLTPEQVVAIASHDGGKQALETVQALLPVLCQAHGLTPQQVVAIASNGGGRPALESIVAQLSRPDPALAALTNDHLVALACLGGRPALDAVKKGLG
ND4-TALE-右(SEQ ID No:449)
MDIADLRTLGYSQQQQEKIKPKVRSTVAQHHEALVGHGFTHAHIVALSQHPAALGTVAVKYQDMIAALPEATHEAIVGVGKQWSGARALEALLTVAGELRGPPLQLDTGQLLKIAKRGGVTAVEAVHAWRNALTGAPLNLTPEQVVAIASHDGGKQALETVQALLPVLCQAHGLTPQQVVAIASHDGGKQALETVQRLLPVLCQAHGLTPQQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQALLPVLCQAHGLTPEQVVAIASNGGGKQALETVQALLPVLCQAHGLTPEQVVAIASNIGGKQALETVQALLPVLCQAHGLTPEQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQALLPVLCQAHGLTPEQVVAIASNGGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPEQVVAIASNNGGKQALETVQALLPVLCQAHGLTPEQVVAIASNNGGKQALETVQALLPVLCQAHGLTPEQVVAIASNIGGKQALETVQALLPVLCQAHGLTPEQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPQQVVAIASNIGGKQALETVQRLLPVLCQAHGLTPQQVVAIASNNGGKQALETVQRLLPVLCQAHGLTPQQVVAIASNGGGRPALESIVAQLSRPDPALAALTNDHLVALACLGGRPALDAVKKGLG
TRAC-ZFP-左(SEQ ID No:450)
FQCRICMRKFATSGSLTRHTKIHTGEKPFQCRICMRNFSRSDHLSTHIRTHTGEKPFACDICGRKFATSSNRTKHTKIHTHPRAPIPKPFQCRICMRNFSRSDNLSEHIRTHTGEKPFACDICGRKFAWHSSLRVHTKIHLR
TRAC-ZFP-右(SEQ ID No:451)
FQCRICMRNFSRSDHLSTHIRTHTGEKPFACDICGRKFADRSHLARHTKIHTGSQKPFQCRICMRKFALKQHLNEHTKIHTGEKPFQCRICMRNFSQSGNLARHIRTHTGEKPFACDICGRKFAHNSSLKDHTKIHLR
ND1-ZFP-左(SEQ ID No:452)
FQCRICMRNFSDSGNLRVHIRTHTGEKPYKCPDCGKSFSQSSSLIRHQRTHTGEKPYECDHCGKSFSQSSHLNVHKRTHTGEKPYRCKYCDRSFSISSNLQRHVRNIHLR
ND1-ZFP-右(SEQ ID No:453)
YKCPECGKSFSTKNSLTEHQRTHTGEKPYKCPECGKSFSSKKALTEHQRTHTGEKPYECNYCGKTFSVSSTLIRHQRIHTGEKPYRCKYCDRSFSISSNLQRHVRNIHLR
G1333N-DddAtox(SEQ ID No:454)
GSGSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLESKVFSSGG
G1333C-DddAtox(SEQ ID No:455)
GSPTPYPNYANAGHVEGQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGAIPVKRGATGETKVFTGNSNSPKSPTKGGC
G1397N-DddAtox(SEQ ID No:456)
GSGSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLESKVFSSGGPTPYPNYANAGHVEGQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEG
G1397C-DddAtox(SEQ ID No:457)
GSAIPVKRGATGETKVFTGNSNSPKSPTKGGC
腺嘌呤脱氨酶(AD:ABE 8e)(SEQ ID No:458)
SEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFGVRNSKRGAAGSLMNVLNYPGMNHRVEITEGILADECAALLCDFYRMPRQVFNAQKKAQSSIN
全长DddAtox变体GSVG(SEQ ID No:459)
GSGSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLEGKVFSSGGPTPYPNYANAGHVESQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGVIPVKRGATGETKVFTGNSNGPKSPTKGGC
全长DddAtox变体E1347A(SEQ ID No:460)
GSGSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLESKVFSSGGPTPYPNYANAGHVAGQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGAIPVKRGATGETKVFTGNSNSPKSPTKGGC
全长DddAtox变体AAAAA(SEQ ID No:461)
GSGSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLESKVFSSGGPTPYPNYANAGHVEGQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGAIPVAAGATGETAVFTGNSNSPASPTAGGC
UGI(SEQ ID No:462)
TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTAYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML
单个模块
(TALE)-接头-AD-GSVG(SEQ ID No:463)
SGSETPGTSESATPESSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFGVRNSKRGAAGSLMNVLNYPGMNHRVEITEGILADECAALLCDFYRMPRQVFNAQKKAQSSINLVGSGSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLEGKVFSSGGPTPYPNYANAGHVESQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGVIPVKRGATGETKVFTGNSNGPKSPTKGGC
(TALE)-接头-AD-E1347A(SEQ ID No:464)
SGSETPGTSESATPESSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFGVRNSKRGAAGSLMNVLNYPGMNHRVEITEGILADECAALLCDFYRMPRQVFNAQKKAQSSINLVGSGSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLESKVFSSGGPTPYPNYANAGHVAGQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGAIPVKRGATGETKVFTGNSNSPKSPTKGGC
(TALE)-接头-AD-AAAAA(SEQ ID No:465)
SGSETPGTSESATPESSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFGVRNSKRGAAGSLMNVLNYPGMNHRVEITEGILADECAALLCDFYRMPRQVFNAQKKAQSSINLVGSGSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLESKVFSSGGPTPYPNYANAGHVEGQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGAIPVAAGATGETAVFTGNSNSPASPTAGGC
双模块
(TALE)-接头-AD(SEQ ID No:466)
SGSETPGTSESATPESSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFGVRNSKRGAAGSLMNVLNYPGMNHRVEITEGILADECAALLCDFYRMPRQVFNAQKKAQSSIN
(TALE)-GSVG(SEQ ID No:467)
GSGSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLEGKVFSSGGPTPYPNYANAGHVESQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGVIPVKRGATGETKVFTGNSNGPKSPTKGGC
(TALE)-E1347A(SEQ ID No:468)
GSGSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLESKVFSSGGPTPYPNYANAGHVAGQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGAIPVKRGATGETKVFTGNSNSPKSPTKGGC
(TALE)-AAAAA(SEQ ID No:469)
GSGSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLESKVFSSGGPTPYPNYANAGHVEGQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGAIPVAAGATGETAVFTGNSNSPASPTAGGC
(TALE)-1397C-AD(SEQ ID No:470)
GSAIPVKRGATGETKVFTGNSNSPKSPTKGGCSGSETPGTSESATPESSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFGVRNSKRGAAGSLMNVLNYPGMNHRVEITEGILADECAALLCDFYRMPRQVFNAQKKAQSSIN
(TALE)-1333N-AD(SEQ ID No:471)
GSGSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLESKVFSSGGSGSETPGTSESATPESSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFGVRNSKRGAAGSLMNVLNYPGMNHRVEITEGILADECAALLCDFYRMPRQVFNAQKKAQSSIN(TALE)-1333C-AD(SEQ ID No:472)
GSPTPYPNYANAGHVEGQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGAIPVKRGATGETKVFTGNSNSPKSPTKGGCSGSETPGTSESATPESSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFGVRNSKRGAAGSLMNVLNYPGMNHRVEITEGILADECAALLCDFYRMPRQVFNAQKKAQSSIN
接头(AA代表氨基酸)
8AA：SGGGLGST(SEQ ID No:473)
16AA：SGSETPGTSESATPES(SEQ ID No:474)
32AA：SGGSSGGSSGSETPGTSESATPESSGGSSGGS(SEQ ID No:475)
SOD2 MTS-3xHA:
MALSRAVCGTSRQLAPVLGYLGSRQKHSLPDYPYDVPDYAGYPYDVPDYAGYPYDVPDYA(SEQ IDNo:476)
COX8A MTS-3xFLAG:
MASVLTPLLLRGLTGSARRLPVPRAKIHSLDYKDHDGDYKDHDIDYKDDDDK(SEQ ID No:477)
6.递送
根据本发明的融合蛋白可通过本领域已知的各种方法递送至细胞，例如微注射、电穿孔、DEAE-葡聚糖处理、脂质转染、纳米颗粒介导的转染、蛋白转导结构域介导的引入和PEG介导的转染，但本发明不限于此。
本发明的另一方面涉及一种编码融合蛋白的核酸。
核酸可与“多核苷酸”、“核苷酸”、“核苷酸序列”和“寡核苷酸”互换使用。它可以包括聚合形式的任何长度的核苷酸，脱氧核糖核苷酸或核糖核苷酸或其类似物。多核苷酸可以具有任何三维结构，并且可以执行已知或未知的任何功能。多核苷酸可以包括至少一个修饰的核苷酸，例如甲基化的核苷酸和核苷酸类似物。对核苷酸结构的修饰可以在聚合物组装之前或之后进行。
多核苷酸可以具有RNA序列、DNA序列或其组合(RNA-DNA组合序列)。
为了表达融合蛋白，可以使用已知的表达载体，例如质粒载体、粘粒载体、噬菌体载体等，并且本领域技术人员可以根据任何已知的方法使用DNA重组技术容易地构建载体。
载体可以是质粒载体或病毒载体，特别地，病毒载体的例子可以包括但不限于腺病毒、腺相关病毒、慢病毒和逆转录病毒载体。
重组表达载体可以含有适于在宿主细胞中表达核酸的核酸形式，并且可以包含至少一个在宿主细胞基础上选择的调节元件使得重组表达载体用于表达，即所述调节元件与待表达的核酸序列可操作地连接。
在重组表达载体中，“可操作地连接”意指目的核苷酸序列与调节元件以允许所述核苷酸序列表达(例如在体外转录/翻译系统中或将载体引入宿主细胞时在宿主细胞中)的方式连接。
重组表达载体可以以适合信使RNA合成的形式提供，包括T7启动子，这意指包括至少一个调节元件以使得能够进行体外mRNA合成，即通过T7聚合酶进行信使RNA合成。
“调节元件”可包括启动子、增强子、内部核糖体进入位点(IRES)和其他表达控制元件(例如转录终止信号，诸如聚腺苷酸化信号和多聚U序列)。调节元件包括指导核苷酸序列在许多类型的宿主细胞中诱导或组成型表达的元件，和指导核苷酸序列仅在某些宿主细胞中表达的元件(例如组织特异性调节序列)。组织特异性启动子可指导主要在所需的感兴趣组织如肌肉、神经元、骨骼、皮肤、血液、特定器官(例如肝脏、胰腺)或特定细胞类型(例如淋巴细胞)中的表达。调节元件还可以以瞬时依赖性方式指导表达，例如以细胞周期或发育阶段依赖性方式，其可能对组织或细胞具有特异性或不具有。
在一些情况下，载体包括至少一个pol III启动子、至少一个pol II启动子、至少一个pol I启动子或其组合。pol III启动子的例子包括但不限于U6和H1启动子。pol II启动子的例子包括但不限于逆转录病毒劳斯肉瘤病毒(RSV)LTR启动子(任选地具有RSV增强子)、巨细胞病毒(CMV)启动子(任选地具有CMV增强子)(例如Boshart等人al(1985)Cell41:521-530)、SV40启动子、二氢叶酸还原酶启动子、β-肌动蛋白启动子、磷酸甘油激酶(PGK)启动子和EF1α启动子。
“调节元件”可包括增强子如WPRE；CMV增强子；HTLV-I的LTR中的R-U5’片段；SV40增强子；以及兔β-珠蛋白的外显子2和3之间的内含子序列。本领域技术人员应当理解，表达载体的设计可以取决于多种因素，例如待转化的宿主细胞的选择、期望的表达水平等。可以将载体引入宿主细胞中以形成包括由本文所述的核酸编码的融合蛋白或肽的转录物、蛋白质或肽(例如，规律间隔成簇短回文重复序列(CRISPR)转录物、蛋白质、酶、其突变体、其融合蛋白等)。有用的载体可以包括慢病毒和腺相关病毒载体，并且也可以选择这些类型的载体以靶向某些类型的细胞。
载体可以通过微注射(例如，直接注射到病变或靶位点)、电穿孔、脂质转染、病毒载体、纳米颗粒、PTD(蛋白质易位结构域)融合蛋白方法等体内递送或递送到细胞中。
核酸可以以核糖核酸的形式注射，例如信使核糖核酸mRNA，从而可以不受限制地对细胞如动物细胞或植物细胞进行基因碱基编辑。
根据本发明的核酸可以是mRNA的形式，并且当以mRNA的形式递送时，与以使用DNA的载体形式递送相比，转录到mRNA过程是不必要的，并且因此基因编辑可以快速启动。瞬时蛋白质表达的可能性很高。
本申请的发明人发现，当将胞嘧啶碱基编辑器以核糖核酸(例如信使核糖核酸)的形式注射到植物细胞中用于植物细胞器基因编辑时，与用质粒递送相比，脱靶效应降低。在植物细胞器基因编辑中，当将胞嘧啶碱基编辑器以mRNA的形式转化到植物细胞中时，首次证明与质粒相比，脱靶效应具有优势。
mRNA可以直接或通过载体递送。在一些情况下，核酸酶和/或切割因子的mRNA可以被化学修饰或以合成的自我复制RNA的形式直接递送。
考虑了在体外或体内将mRNA分子递送到细胞中的方法，包括在体内将mRNA递送到细胞的方法或将mRNA递送到生物体如人或动物的细胞的方法。例如，可以使用脂质(例如脂质体、胶束等)、纳米颗粒或纳米管、或阳离子化合物(例如聚乙烯亚胺或PEI)将mRNA分子递送到细胞中。在一些情况下，可以使用生物发射方法如基因枪或生物发射颗粒递送系统将mRNA递送到细胞中。
载体的例子可包括但不限于细胞穿透肽(CPP)、纳米颗粒和聚合物。
CPP是促进细胞摄取各种分子货物(从纳米级颗粒到小化学分子和大DNA片段)的短肽。
关于纳米颗粒，根据本发明的组合物可通过聚合物纳米颗粒、金属纳米颗粒、金属/无机纳米颗粒或脂质纳米颗粒递送。聚合物纳米颗粒可以是例如通过滚环扩增合成的DNA纳米线或线状DNA纳米颗粒。DNA纳米线或线状DNA纳米颗粒可以负载mRNA并用PEI包被以改善内体逃逸。这些复合物与细胞膜结合、被内化，然后通过内体逃逸递送到细胞核。
关于金属纳米颗粒，金颗粒可与阳离子内体破坏性聚合物连接和复合，并因此递送到细胞。阳离子内体破坏性聚合物可包括例如聚乙烯亚胺、聚(精氨酸)、聚(赖氨酸)、聚(组氨酸)、聚-[2-{(2-氨基乙基)氨基}-乙基-天冬酰胺(pAsp(DET))、聚(乙二醇)(PEG)和聚(精氨酸)的嵌段共聚物、PEG和聚(赖氨酸)的嵌段共聚物、或PEG和聚{N-[N-(2-氨基乙基)-2-氨基乙基]天冬酰胺}(PEG-pAsp(DET))的嵌段共聚物。
关于金属/无机纳米颗粒，mRNA可以通过例如沸石咪唑酯骨架-8(ZIF-8)包封。
在一些情况下，带负电荷的mRNA可以与阳离子材料偶联以形成纳米颗粒，其可以通过受体介导的内吞作用或吞噬作用穿透细胞。
阳离子聚合物的例子可以包括聚烯丙胺(PAH)；聚乙烯亚胺(PEI)；聚(L-赖氨酸)(PLL)；聚(L-精氨酸)(PLA)；聚乙烯胺均聚物或共聚物；聚(乙烯基苄基-三-C1-C4-烷基铵盐)；脂族或脂环族二卤化物和脂族N,N,N’,N’-四-C1-C4-烷基-亚烷基二胺的聚合物；聚(乙烯基吡啶)或聚(乙烯基吡啶鎓盐)；聚(N,N-二烯丙基-N,N-二-C1-C4-烷基-卤化铵)；季铵化的丙烯酸或甲基丙烯酸二-C1-C4-烷基-氨基乙酯的均聚物或共聚物；POLYQUADTM；聚氨基酰胺等。
阳离子脂质可包括阳离子脂质体配制品。脂质体的脂质双层可以保护包封的核酸免于降解，并且可以防止能够结合核酸的抗体的特异性中和。在内体成熟期间，内体膜和脂质体融合，使得阳离子脂质-核酸酶能够有效地内体逃逸。阳离子脂质的例子可以包括聚乙烯亚胺、星放射状聚酰胺胺(PAMAM)树枝状大分子、Lipofectin(DOTMA和DOPE的组合)、脂质体酶、(例如/>2000、/>3000、RNAiMAX、/>LTX)、SAINT-RED(Synvolux Therapeutics，荷兰格罗宁根)、DOPE、Cytofectin(Gilead Sciences，加利福尼亚州福斯特城)和Eufectin(JBL，加利福尼亚州圣路易斯奥比斯波)。代表性的阳离子脂质体可以由N-[1-(2,3-二油酰氧基)-丙基]-N,N,N-三甲基氯化铵(DOTMA)、N-[1-(2,3-二油酰氧基)-丙基]-N,N,N-三甲基甲基硫酸铵(DOTAP)、3β-[N-(N’,N’-二甲基氨基乙烷)氨基甲酰基]胆固醇(DC-Chol)、2,3-二油酰氧基-N-[2(精胺甲酰氨基)乙基]-N,N-二甲基-1-三氟乙酸丙胺酯(DOSPA)、1,2-二肉豆蔻酰氧基丙基-3-二甲基-羟乙基溴化铵或双十八烷基二甲基溴化铵(DDAB)制备。
关于脂质纳米颗粒，它们可以使用脂质体作为载体进行递送。脂质体是由包围内部水性隔室的单层或多层脂质双层和相对不可渗透的外部亲脂性磷脂双层构成的球形囊泡结构。脂质体配制品可主要含有天然磷脂和脂质如1,2-二硬脂酰-sn-甘油-3-磷脂酰胆碱(DSPC)、鞘磷脂、磷脂酰胆碱或单唾液神经节苷酯。在一些情况下，可以将胆固醇或1,2-二油酰基-sn-甘油-3-磷酸乙醇胺(DOPE)添加至脂质膜中以解决血浆中的不稳定性。胆固醇的添加降低了包封的生物活性化合物向血浆中的快速释放，或者1,2-二油酰基-sn-甘油-3-磷酸乙醇胺(DOPE)增加了稳定性。
7.碱基编辑
本发明的又另一方面涉及一种用于碱基编辑的组合物，其包含融合蛋白或核酸。
本发明的又另一方面涉及一种碱基编辑方法，其包括用所述组合物处理细胞。
在DNA结合蛋白如TALE或ZFP(锌指蛋白)与靶DNA结合后，融合蛋白的胞嘧啶脱氨酶水解胞嘧啶的氨基，从而将其转化为尿嘧啶。由于尿嘧啶可与腺嘌呤形成碱基对，因此在细胞内DNA复制期间，胞嘧啶-鸟嘌呤碱基对可最终经由尿嘧啶-腺嘌呤碱基对编辑为胸腺嘧啶-腺嘌呤碱基对。此外，融合蛋白的腺嘌呤脱氨酶水解腺嘌呤的氨基，以将其转化为次黄嘌呤。由于次黄嘌呤可与胞嘧啶形成碱基对，类似地，在细胞内DNA复制期间，腺嘌呤-胸腺嘧啶碱基对可经由次黄嘌呤-胞嘧啶碱基对编辑为鸟嘌呤-胞嘧啶碱基对。
细胞可以是真核细胞(例如真菌如酵母、真核动物和/或真核植物衍生的细胞(例如胚胎细胞、干细胞、体细胞、生殖细胞等)等)、真核动物(例如灵长类动物如人、猴、狗、猪、牛、绵羊、山羊、小鼠、大鼠等)或真核植物(例如藻类如绿藻、玉米、大豆、小麦、水稻等)，但不限于此。
(1)植物细胞DNA的碱基编辑
本发明涉及一种用于植物细胞DNA的碱基编辑的组合物或方法。用于植物细胞中碱基编辑的组合物包含融合蛋白或编码其的核酸；以及核定位信号(NLS)肽、叶绿体转运肽、线粒体靶向信号(MTS)、核输出信号或编码其的核酸。
本发明还提供了一种用于在植物细胞中进行碱基编辑的组合物，所述组合物包含融合蛋白或核酸；以及核定位信号(NLS)肽或编码其的核酸。
本发明还提供了一种用于在植物细胞中进行碱基编辑的组合物，所述组合物包含融合蛋白或核酸；以及叶绿体转运肽或编码其的核酸。
本发明还提供了一种用于在植物细胞中进行碱基编辑的组合物，所述组合物包含融合蛋白或核酸；以及线粒体靶向信号(MTS)或编码其的核酸。
在一些情况下，本发明还提供了一种用于在植物细胞中进行碱基编辑的组合物，所述组合物进一步包含核输出信号或编码其的核酸。
具体地，本发明涉及一种用于植物细胞中核DNA、线粒体DNA或叶绿体DNA的碱基编辑的组合物或方法。
具体地，融合蛋白可以通过以下递送到植物细胞：
使用基因枪注射(轰击)；
PEG介导的原生质体转染；
通过电穿孔进行原生质体转染；或
通过显微注射进行原生质体注射。
编码根据本发明的融合蛋白的多核苷酸序列可以是RNA序列、DNA序列或其组合(RNA-DNA组合序列)。
编码融合蛋白的多核苷酸可通过以下递送到植物细胞：
使用土壤杆菌属如根癌土壤杆菌、发根土壤杆菌等进行转化。
-二元载体，
-病毒载体：双联病毒、烟草脆裂病毒(TRV)、番茄花叶病毒(ToMV)、:狗尾草花叶病毒(FoMV)、大麦黄条点花叶病毒(BYSMV)、苦苣菜黄网弹状病毒(SYNV)等；
使用病毒进行转染；
注射(轰击、基因枪)；
PEG介导的原生质体转染；
通过电穿孔进行原生质体转染；或
通过显微注射进行原生质体注射。
病毒的例子可包括病毒载体中的双联病毒、烟草脆裂病毒(TRV)、番茄花叶病毒(ToMV)、:狗尾草花叶病毒(FoMV)、大麦黄条点花叶病毒(BYSMV)、苦苣菜黄网弹状病毒(SYNV)等。
载体可以通过微注射(例如，直接注射到病变或靶位点)、电穿孔、脂质转染、病毒载体、纳米颗粒、PTD(蛋白质易位结构域)融合蛋白方法等递送到细胞中。
关于转运到植物细胞器的蛋白质或编码其的核酸，植物细胞器可以是线粒体、叶绿体或质体(白色体、色质体)。
转运到植物细胞器的蛋白质可以是例如叶绿体转运肽或线粒体靶向信号(MTS)。
例如，叶绿体转运肽(CTP)或线粒体靶向信号(MTS)结合，然后递送到植物细胞中的叶绿体或线粒体中。当递送到叶绿体或线粒体时，除了N末端CTP或MTS之外的剩余部分以前体蛋白的形式递送到叶绿体或线粒体中。在进入叶绿体或线粒体内部的过程中，递送的蛋白质部分被分离，并且靶向叶绿体或线粒体以诱导位点特异性碱基编辑。
除了融合蛋白或编码其的核酸之外，叶绿体转运肽(CTP)或编码其的核酸或线粒体靶向信号(MTS)或编码其的核酸可以融合并被递送到植物细胞，使得能够对植物线粒体、叶绿体、色质体或白色体DNA进行碱基编辑。
当核输出信号在线粒体基因编辑过程中与碱基编辑蛋白连接时，可以以更高的效率实现碱基编辑。核输出信号可以衍生自例如MVM(小鼠微小病毒)，但本发明不限于此。核输出信号可包括例如SEQ ID NO:31的氨基酸序列，但不限于此。
VDEMTKKFGTLTIHDTEK(SEQ ID NO:31)
本发明还包括TAL(转录激活因子样)效应物(TALE)-FokI核酸酶或编码其的核酸，所述核酸酶切割野生型DNA序列但不切割编辑的碱基序列；或ZFN(锌指核酸酶)或编码其的核酸，特别是线粒体核酸酶mitoTALEN(线粒体TALE核酸酶)或编码其的核酸；或ZFN(锌指核酸酶)或编码其的核酸，从而即使当同时使用线粒体序列切割蛋白时，也预期以更高效率进行线粒体碱基编辑。
(2)动物细胞DNA的碱基编辑
本发明涉及一种用于动物细胞DNA的碱基编辑的组合物或方法。用于动物细胞中进行碱基编辑的组合物包含融合蛋白或编码其的核酸；以及核定位信号(NLS)肽、线粒体靶向信号(MTS)、核输出信号或编码其的核酸。
本发明还提供了一种用于在动物细胞中进行碱基编辑的组合物，所述组合物包含融合蛋白或核酸；以及核定位信号(NLS)肽或编码其的核酸。
本发明还提供了一种用于在动物细胞中进行碱基编辑的组合物，所述组合物包含融合蛋白或核酸；以及线粒体靶向信号(MTS)或编码其的核酸。
在一些情况下，本发明还提供了一种用于在动物细胞中进行碱基编辑的组合物，所述组合物进一步包含核输出信号或编码其的核酸。
动物细胞是非人动物细胞，并且用核输出信号或编码其的核酸和/或线粒体靶向信号(MTS)或编码其的核酸处理使得能够在非人动物细胞中对线粒体DNA进行碱基编辑。
例如，除了融合蛋白或编码其的核酸之外，线粒体靶向信号(MTS)结合并递送到线粒体。当递送到线粒体时，除了N末端MTS之外的剩余部分以前体蛋白的形式递送到线粒体中。在进入线粒体内部的过程中，递送的蛋白质部分被分离，并且靶向线粒体DNA以诱导位点特异性碱基编辑。
本发明涉及一种用于在非人动物细胞中对线粒体DNA进行碱基编辑的组合物或方法，其中核输出信号(NES)或编码其的核酸与包括线粒体靶向信号、TAL效应物和胞嘧啶脱氨酶(DddAtox)的TALE-DdCBE(TALE DddA衍生的胞嘧啶碱基编辑器)或编码其的核酸融合。在融合蛋白中添加核输出信号可以减少在具有相似DNA序列的位点处的核DNA碱基编辑。
根据本发明，通过包括核输出信号或编码其的核酸可以实现更有效的动物线粒体DNA碱基编辑。此外，对线粒体核样序列的核DNA碱基编辑可由于核输出信号而减少，从而可以仅编辑线粒体DNA。
核输出信号可以衍生自例如MVM(小鼠微小病毒)，但本发明不限于此。核输出信号可包括例如VDEMTKKFGTLTIHDTEK(SEQ ID NO:31)的氨基酸序列，但不限于此。
在与(1)核输出信号或编码其的核酸和(2)DNA结合蛋白、脱氨酶或其变体或编码其的核酸同时或顺序编辑之前，可以将切割野生型DNA碱基序列但不切割编辑的碱基序列或编码其的核酸或ZFN(锌指核酸酶)或编码其的核酸的TAL(转录激活因子样)效应物(TALE)-FokI核酸酶注射到真核细胞中。
特别地，关于真核细胞中线粒体基因的碱基编辑，可以包括核输出信号或编码其的核酸和/或线粒体靶向信号(MTS)或编码其的核酸。
根据本发明，当核输出信号在动物线粒体基因编辑过程中与碱基编辑蛋白连接时，碱基编辑可能具有更高的效率，并且在动物胚胎中，细胞核中同源序列的非特异性碱基编辑也被抑制。
在本发明中，通过进一步包括线粒体核酸酶，即mitoTALEN(线粒体TALE核酸酶)或编码其的核酸，即使当同时使用线粒体核酸酶时，也可以以更高的效率实现线粒体碱基编辑。可以使用线粒体DNA核酸酶mitoTALEN(线粒体TALE核酸酶)切割线粒体DNA，并且可以切割野生型线粒体基因组以在动物中以高效率获得碱基编辑的基因组。
在本发明中，通过进一步包括线粒体核酸酶mitoTALEN(线粒体TALE核酸酶)或编码其的核酸，即使当同时使用线粒体核酸酶时，也可以以更高的效率实现线粒体碱基编辑。具体地，可以包括融合蛋白(mitoTALEN)，其包括连接线粒体靶向信号和FokI核酸酶的TAL效应物结构域或ZFN或编码其的核酸，或编码其的核酸。
可以使用线粒体DNA核酸酶mitoTALEN(线粒体TALE核酸酶)切割线粒体DNA，并且可以切割野生型线粒体基因组以在动物中以高效率获得碱基编辑的基因组。
在一些情况下，可以另外包括能够增加碱基编辑效率的UGI(尿嘧啶DNA糖基化酶抑制剂)。UGI能够通过抑制UDG(尿嘧啶DNA糖基化酶)的活性来增加碱基编辑效率，UDG是一种通过催化从DNA中去除U来修复突变DNA的酶。
具体地，由分裂细菌间毒素DddAtox、设计为结合DNA的转录激活因子样效应物(TALE)和尿嘧啶糖基化酶抑制剂(UGI)构成的DddA衍生的胞嘧啶碱基编辑器(DdCBE)使得能够在线粒体DNA中进行靶向胞嘧啶-胸腺嘧啶碱基编辑。根据实施方案，高效的线粒体DNA编辑在小鼠胚胎中是可能的。在线粒体基因中，靶向编码催化NADH脱水和电子转移到泛醌的NADH脱氢酶的亚基的MT-ND5(ND5)，引起与人线粒体疾病相关的突变，例如m.G12918A，和产生早期终止密码子的突变，例如m.C12336T。因此，可以在小鼠中构建线粒体疾病模型，这表明治疗线粒体疾病的可能性。
(2)DNA结合蛋白、脱氨酶或其变体或编码其的核酸可以与(1)核输出信号或编码其的核酸连接，以及(3)核酸酶mitoTALEN(线粒体TALE核酸酶)或编码其的核酸可以与(1)连接。为了递送(1)至(3)，单个递送载体或多个递送载体可以与相同或不同的构型组合使用。
(1)可以包括在第一递送载体中，(2)可以包括在第二递送载体中，以及(3)可以包括在第三递送载体中。这些单独的递送系统可以同时是病毒递送载体，可以是病毒和非病毒递送载体，或者可以同时是非病毒递送载体。
可以将(1)至(3)的核输出信号、DdCBE和mitoTALEN混合并递送。
选自(1)至(3)中的至少一种可递送至核输出信号、DdCBE或mitoTALEN，并且一些可通过将编码(1)至(3)的DNA序列定位在载体上来递送。
编码上述(1)至(3)的DNA序列可位于同一载体上并通过一个载体同时递送，或可位于不同载体上并递送。
根据本发明的动物可以包括人或非人动物。非人转基因动物的例子可以是昆虫、环节动物、软体动物、腕足动物、线虫、腔肠动物、海绵、脊索动物和脊椎动物，脊椎动物可以是鱼类、两栖动物、爬行动物、鸟类或哺乳动物，昆虫可以是果蝇，线虫可以是秀丽隐杆线虫，鱼类可以是斑马鱼，哺乳动物可以是灵长类动物、食肉类动物、食虫类动物、啮齿类动物、偶蹄类动物、奇蹄类动物或长鼻类动物，啮齿类动物可以包括大鼠或小鼠。
可以通过将根据本发明的组合物引入人或非人动物的胚胎中，将胚胎转移到雌性老鼠中，并怀上胚胎来产生碱基编辑的动物。可以将根据本发明的组合物引入动物的受精卵中并进行培养。
可以将由此获得的受精卵转移到雌性老鼠并分娩。可以进一步包括证实非人转基因动物在递送后是否是转基因的。可以将非人转基因动物交配以产生后代转基因动物。
“后代”是指由非人转基因动物交配产生的所有有活力的转基因动物后代，更具体地，可以是通过将转基因动物作为亲本彼此交配或将其与正常动物交配产生的F1代，通过将F1代的动物与正常动物交配产生的F2代，以及随后的几代，但本发明不限于此。
交配的特征可在于转基因动物或正常动物的交配。本发明可以包括从转基因动物或后代转基因动物分离的细胞、组织和副产物。副产物可以包括衍生自转基因兔的任何材料，并且优选地选自血液、血清、尿液、粪便、唾液、器官和皮肤，但不限于此。
实施例
通过以下实施例可以获得对本发明的更好理解。这些实施例仅用于说明本发明，而不应解释为限制本发明的范围，如本领域技术人员将清楚的。
实施例1.锌指脱氨酶(ZFD)
核DNA或线粒体DNA的碱基编辑广泛用于生物医学研究、医学和生物技术。ZFD平台包含DNA结合蛋白、分裂细菌间毒素脱氨酶DDDAtox和尿嘧啶糖基化酶抑制剂(UGI)。这里，ZFD催化靶向的C至T碱基转化，而不会在人细胞中诱导不希望的小插入和缺失(插入缺失)。使用公开可用的锌指资源，构建编码ZFD的质粒，从而在核DNA中实现频率高达60％的碱基编辑以及在线粒体DNA中实现频率为30％的碱基编辑。与基于CRISPR的碱基编辑不同，ZFD不通过DNA切割产生单链或双链断裂，使得在靶位点处不产生由易错的非同源末端连接引起的不希望的插入和缺失(插入缺失)。此外，从大肠杆菌纯化的重组ZFD蛋白自发地穿透人细胞以诱导靶向的碱基转化。这证明了无基因的基因疗法的原理论证。
用于真核细胞和生物体中的基因组编辑的技术包括锌指核酸酶(ZFN)、转录激活因子样效应物(TALE)核酸酶(TALEN)、TALE连接的分裂细菌间脱氨酶毒素DddA衍生的胞嘧啶碱基编辑器(又称DdCBE)、CRISPR-Cas9和不具有切割活性的Cas9连接的脱氨酶(又称碱基编辑器)，但不限于此。这些工具原则上由两个功能单元构成，即DNA结合部分和催化部分。因此，锌指阵列或TALE阵列用作DNA结合部分，而核酸酶(ZFN和TALEN中的FokI)或脱氨酶(DdCBE中的分裂DddAtox和CBE中的APOBEC1)用作催化单元。Crispr-cas9具有核酸酶功能和RNA指导的DNA结合蛋白功能。定制设计的可编程核酸酶如ZFN、TALEN和Cas9切割DNA，从而产生双链断裂，它的修复以靶向方式引起基因敲除和敲入。然而，可编程核酸酶诱导的双链断裂可引起靶位点处不希望的大基因缺失、p53活化和在中靶和脱靶位点的两个并行DSB修复期间的染色体重排。相比之下，包括胞嘧啶和腺嘌呤碱基编辑器(CBE和ABE)在内的可编程碱基编辑器不产生DSB，从而避免细胞中这些不希望的事件，并且有效地催化单核苷酸转化而不需要修复模板或供体DNA。然而，含有Cas9切口酶变体的CBE或ABE切割靶DNA链以产生切口或单链断裂，导致在基因靶位点处不希望的插入缺失。
CBE催化细胞中核DNA和线粒体DNA中的C至T碱基转化。我们使用定制设计的DdCBE来证明小鼠中的线粒体DNA编辑和植物中的叶绿体DNA编辑。我们还通过将分裂DddAtox与定制设计的锌指蛋白连接产生了锌指脱氨酶(ZFD)，以用于在人和其他真核细胞中进行无插入缺失的精确碱基编辑。由于锌指阵列(2×0.3-0.6k碱基对)的大小小于TALE阵列(2×1.7-2k碱基对)或酿脓链球菌Cas9(4.1k碱基对)，因此编码ZFD的基因可以容易地包装在具有有限货物空间的病毒载体中，例如用于体内研究和基因治疗应用的AAV中。与TALE阵列不同，锌指阵列在C末端或N末端缺少庞大的结构域，使得它们在工程化上是友好的。分裂DddAtox半部分可以与锌指蛋白的C末端或N末端融合。此外，具有穿透细胞的内在能力的锌指蛋白能够在人细胞中进行无核酸的基因编辑。这些性质使锌指蛋白成为用于在核或其他细胞器中进行碱基编辑的DNA结合模块的理想平台。
1-1.材料和方法
质粒构建
通过在用HindIII和XhoI(NEB)酶消化后修饰p3s-ABE7.10质粒(Addgene,#113128)产生用于哺乳动物表达的p3s-ZFD质粒。使用HiFi DNA组装试剂盒(NEB)组装经消化的p3s质粒和合成的插入DNA。所有编码MTS、ZFP(Toolgen、Sangamo和Barbas模块)、分裂DddA或UGI的插入DNA由IDT合成。设计pTarget质粒以确定对于ZFD活性的间隔子序列的最佳长度。通过将ZFP结合序列和间隔子序列插入用两种酶(EcoRI和BamHI，NEB)消化的pRGS-CCR5-NHEJ报告质粒中，构建含有两个ZFP结合位点的每个pTarget质粒，在所述两个ZFP结合位点之间具有不同长度的间隔子。通过在用NcoI和XhoI(NEB)酶消化后修饰pET-Hisx6-rAPOBEC1-XTEN-nCas9-UGI-NLS质粒(Addgene,#89508)产生用于在大肠杆菌中产生蛋白质的pET-ZFD质粒。使用PCR从p3s-ZFD质粒扩增ZFD序列，并且合成Hisx6标签和GST标签序列作为寡核苷酸(Macrogen)。使用HiFi DNA组装试剂盒(NEB)产生所有质粒以将编码ZFD和用于蛋白质纯化的标签的序列插入经消化的pET质粒中。使用化学感受态DH5ɑ大肠杆菌细胞进行质粒的转化，并根据制造商的方案用AccuPrep质粒微型提取试剂盒(Bioneer)纯化质粒。用Sanger测序鉴定整个序列后，选择所需质粒。
HEK293T细胞培养和转染
将HEK 293T细胞(ATCC CRL-11268)在补充有10％胎牛血清(Welgene)和1％抗生素-抗真菌溶液(Welgene)的杜尔贝科改良伊格尔培养基(Welgene)中培养。将HEK 293T细胞(7.5×104)接种到48孔板中。18-24小时后，使用Lipofectamine 2000(1.5μl，Invitrogen)将细胞用编码左和右ZFD的质粒(各500ng)或连同pTarget质粒(10ng)以70％-80％汇合度转染细胞。转染后96小时收获细胞，然后将所述细胞通过在补充有5μl蛋白酶K(Qiagen)的100μl细胞裂解缓冲液(50mM Tris-HCl(pH 8.0)(Sigma-Aldrich)、1mM EDTA(Sigma-Aldrich)、0.005％十二烷基硫酸钠(Sigma-Aldrich))中在55℃下孵育1小时，然后在95℃下孵育10分钟来裂解。对于全mtDNA测序，用连续稀释浓度的编码靶向ND1或ND2的mitoZFD对的质粒或mRNA转染HEK 293T细胞。示出了每7.5×104个细胞递送的构建体的量(ng)。转染后96小时，从细胞中分离mtDNA。
K562细胞培养和转染
将K562细胞在补充有10％胎牛血清(Welgene)和1％抗生素-抗真菌溶液(Welgene)的RPMI 1640中培养。为了通过电穿孔将ZFD递送到K562细胞中，使用具有程序FF-120(Lonza)的Amaxa 4D-NucleofectorTMX单元系统。当使用16孔NucleocuvetteTMStrip时，添加至每个样品中的底物溶液的最大体积为2μl。将K562细胞(1×105)用220pmol(对于最大容量)或110pmol(对于最大容量的一半)的每种左和右ZFD蛋白或500ng编码左和右ZFD的质粒转染。处理后96小时，通过以100g离心5分钟收获细胞，并将所述细胞通过在补充有5μl蛋白酶K(Qiagen)的100μl细胞裂解缓冲液(50mM Tris-HCl(pH 8.0)(Sigma-Aldrich)、1M EDTA(Sigma-Aldrich)、0.005％十二烷基硫酸钠(Sigma-Aldrich))中在55℃下孵育1小时，然后在95℃下孵育10分钟来裂解。为了将ZFD或编码ZFD的质粒直接递送到K562细胞中，参考以前用于直接递送ZFN的方法。用pH 7.4的含有100mM L-精氨酸和90μM ZnCl2的无血清培养基将左和右ZFD蛋白的混合物(终浓度为50μM)或编码左和右ZFD的质粒的混合物(各500ng)稀释至终体积20μl。将K562细胞(1×105)以100g离心5分钟，并弃去上清液。然后将细胞重悬于稀释的ZFD溶液中并在37℃下孵育1小时。孵育后，将细胞以100g离心5分钟，然后重悬于新鲜培养基中。将细胞在30℃(对于瞬时低温条件)或37℃保持18小时，然后使其在37℃再生长两天。将一些细胞根据上述程序处理两次。处理后96小时分析细胞。
ZFD蛋白表达和纯化
将编码每对ZFD(每一个具有C末端GST标签)的质粒转化到Rosetta(DE3)感受态细胞中，然后将其在含有卡那霉素的LB琼脂板中培养。培养过夜后，挑取单个菌落并在37℃下在含有50μg/ml卡那霉素和100μM ZnCl2的液体培养基中培养过夜(预培养)。第二天，将一部分预培养物转移到大体积的液体培养基中，随后在37℃下培养直至A600 nm吸光度为约0.5-0.70。将培养物置于冰上约1小时，然后通过添加0.5mM异丙基β-D-1-硫代吡喃半乳糖苷(IPTG；GoldBio)诱导ZFD蛋白表达，并将培养物在18℃下孵育14小时。
在蛋白质纯化过程中，将细胞重悬于裂解缓冲液(50mM Tris-HCl(Sigma-Aldrich)、500mM NaCl(Sigma-Aldrich)、1mM MgCl2(Sigma-Aldrich)、10mM 1,4-二硫苏糖醇(DTT；GoldBio)、1％Triton X-10(Sigma-Aldrich)、10％甘油、1mM苯甲基磺酰氟(Sigma-Aldrich)、1mg/ml来自鸡蛋清的溶菌酶(Sigma-Aldrich)、100μM ZnCl2(Sigma-Aldrich)、100mM精氨酸(Sigma-Aldrich)，pH 8.0)中，然后超声处理(总共3min，开启5s，关闭10s)以用于进一步裂解。此后，将溶液离心(13,000rpm)以仅提取上清液。通过向其中添加谷胱甘肽琼脂糖4B(GE Healthcare)将上清液孵育1小时。孵育后，将树脂-裂解物混合物置于柱中，随后用洗涤缓冲液(50mM Tris-HCl(Sigma-Aldrich)、500mM NaCl(Sigma-Aldrich)、10mM DTT(GoldBio)、1mM MgCl2(Sigma-Aldrich)、100μM ZnCl2(Sigma-Aldrich)、10％甘油、100mM精氨酸(Sigma-Aldrich)，pH 8.0)洗涤三次。使用洗脱缓冲液(50mM Tris-HCl(Sigma-Aldrich)、500mM NaCl(Sigma-Aldrich)、1mM MgCl2(Sigma-Aldrich)、40mM谷胱甘肽(Sigma-Aldrich)、10％甘油、1mM DTT(GoldBio)、100μM ZnCl2(Sigma-Aldrich)、100mM精氨酸(Sigma-Aldrich)，pH 8.0)从树脂洗脱附着于树脂的蛋白质。最后，将洗脱的蛋白质浓缩至约15ng/μl的浓度(200-240pmol/μl，取决于蛋白质大小)。
通过ZFD实现对PCR扩增子的体外脱氨
使用PCR制备含有TRAC位点的扩增子。将8μg扩增子与2μg每种ZFD蛋白(左-G1397N和右-G1397C)在含有100μM ZnCl2的NEB3.1缓冲液中在37℃下孵育1-2小时。反应后，通过与4μl蛋白酶K溶液(Qiagen)在55℃下孵育30分钟去除ZFD蛋白，并使用PCR纯化试剂盒(MGmed)纯化扩增子。将1μg纯化的扩增子与2个单位的USER酶(NEB)在37℃下孵育1小时。然后将扩增子与4μl蛋白酶K溶液(Qiagen)孵育并使用PCR纯化试剂盒(MGmed)再次纯化。将纯化的PCR产物在琼脂糖凝胶上进行电泳并成像。
靶向深度测序
为了分析中靶和脱靶位点的碱基编辑比率，使用GXL DNA聚合酶(TAKARA)、使用含有TruSeq HT双索引的引物，对靶位点进行重叠的一级PCR、二级PCR扩增和三级PCR，从而产生深度测序文库。使用Illumina MiniSeq对文库进行配对末端测序。
mRNA制备
使用正向和反向引物(正向：5’-CATCAATGGGCGTGGATAG-3’SEQ ID No:116，反向：5’-CATCAATGGGCGTGGATAG-3’SEQ ID No:117，反向：5’-GACACCTACTCAGACAATGC-3’SEQ IDNo:118)通过PCR产生在ZFD序列上游含有T7 RNA聚合酶启动子的DNA模板。然后，使用mMESSAGE mMACHINETMT7 ULTRA转录试剂盒(Thermo Fisher)体外合成mRNA。使用MEGAclearTM转录清除试剂盒(Thermo Fisher)根据制造商的方案纯化体外转录的mRNA。
全线粒体基因组测序
对于全线粒体基因组测序，需要三个步骤。1.从分离的线粒体中提取mtDNA：在用靶向ND1或ND2的mitoZFD对转染后96小时，将3×105个HEK293T细胞胰蛋白酶化并通过离心(500g，4分钟，4℃)收集。然后用磷酸盐缓冲盐水(Welgene)洗涤细胞并再次离心收集。去除上清液，并根据制造商的方案，使用用于培养细胞的线粒体分离试剂盒(Thermo Fisher)的基于试剂的方法从培养细胞中分离线粒体。然后，使用DNeasy血液和组织试剂盒(Qiagen)从分离的线粒体中提取mtDNA。2.NGS文库的产生：使用具有NexteraTMDNA CD索引的Illumina DNA制备试剂盒(Illumina)从提取的mtDNA中产生NGS文库。3.NGS：将文库合并并加载到MiniSeq测序仪(Illumina)上。平均测序深度>50。
线粒体全基因组DNA编辑分析
为了分析来自全线粒体基因组测序的NGS数据，使用BWA将Fastq文件与GRCh38.p13(v102版)参考基因组比对，并且通过固定读取的配对信息和标记产生具有SAMtools(v.1.9)的BAM文件。然后，使用来自REDItools(v.1.2.1)的REDItoolDenovo.py脚本来鉴定线粒体基因组中所有胞嘧啶和鸟嘌呤中碱基编辑率为1％或更高的位置。碱基编辑率为50％或更高的位置被认为是细胞系中的单核苷酸变异，并从所有样品中排除。对于脱靶分析，排除每个ZFD的靶位点。将编辑频率≥1％的剩余位置视为脱靶位点，并计数编辑的C/G核苷酸的数量。为了计算线粒体基因组中所有C/G的平均C/G至T/A碱基编辑频率，将脱靶位点中的编辑速率取平均值。通过将平均中靶编辑频率除以平均脱靶编辑频率来计算特异性比率。通过绘制中靶和脱靶位点的碱基编辑率来产生线粒体全基因组图。
1-2.ZFD构建体的优化
为了开发用于在人和其他真核细胞中进行碱基编辑的ZFD，优化了连接到分裂DddAtox半部分的锌指蛋白(ZFP)的氨基酸接头和间隔子的长度。在左与右ZFP结合位点之间的间隔子中诱导C至T碱基转化。选择良好表征的靶向人CCR5基因的ZFN对。使用相同的方法，制备具有2、5、10、16、24和32个氨基酸的各种接头的ZFD，并构建一系列具有长度范围为1至24个碱基对的各种间隔子以及ZFD的左和右ZFP结合位点和重复的TC序列的靶质粒(图1a和图1b和表1)。
[表1]pTarget文库序列
DddAtox可以在两个位置(G1333和G1397)分裂，并且每一半可以融合到左或右ZFP。对于具有间隔子的24个靶质粒中的每一个，测量所得到的24个(＝6个接头×2个分裂位置×ZFP融合位置(左或右))ZFD构建体的碱基编辑效率。在Hek293T细胞中转染的第4天用深度测序进行测量。
[构建体]
*左-ZFD：SV40 NLS-ZFP(S162-左)-接头-DddAtox半部分-4aa接头-UGI
*右-ZFD：SV40 NLS-ZFP(S162-右)-接头-DddAtox半部分-4aa接头-UGI
-SV40 NLS:
PKKKRKV(SEQ ID No:478)
-ZFP(S162-左)
GIHGVPAAMAERPFQCRICMRNFSDRSNLSRHIRTHTGEKPFACDICGRKFAISSNLNSHTKIHTGSQKPFQCRICMRNFSRSDNLARHIRTHTGEKPFACDICGRKFATSGNLTRHTKIHLR(SEQ ID NO:2)
-ZFP(S162-右)
GIHGVPAAMAERPFQCRICMRNFSRSDNLSVHIRTHTGEKPFACDICGRKFAQKINLQVHTKIHTGEKPFQCRICMRNFSRSDVLSEHIRTHTGEKPFACDICGRKFAQRNHRTTHTKIHLR(SEQ ID NO:3)
-锌指蛋白与DddAtox半部分之间的接头：
-2aa：GS
-5aa：TGEKP(SEQ ID No:479)
-10aa：SGAQGSTLDF(SEQ ID No:9)
-16aa：SGSETPGTSESATPES(SEQ ID No:10)
-24aa：SGTPHEVGVYTLSGTPHEVGVYTL(SEQ ID No:115)
-32aa：GSGGSSGGSSGSETPGTSESATPESSGGSSGGS(SEQ ID No:11)
-分裂DddAtox G1333-N
GSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLESKVFSSGG(SEQ ID No:27)
-分裂DddAtox G1333-C
PTPYPNYANAGHVEGQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEGAIPVKRGATGETKVFTGNSNSPKSPTKGGC(SEQ ID No:272)
-分裂DddAtox G1397-N
GSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLESKVFSSGGPTPYPNYANAGHVEGQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEG(SEQ ID No:273)
-分裂DddAtox G1397-C
AIPVKRGATGETKVFTGNSNSPKSPTKGGC(SEQ ID No:26)
-4aa接头
SGGS(SEQ ID NO:480)
-UGI
TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTAYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML(SEQ ID NO:481)
具有短接头(2和5个氨基酸(AA)接头)的ZFD具有低效率或没有效率。相比之下，在具有10AA或更多的接头的ZFD中，在4个碱基对或更多的间隔子中的C至T碱基编辑效率在1％至24％的范围内(图1c、图2a和图2c)。在ZFD对中，具有24AA接头的ZFD对显示出最高的编辑效率。为了确定最佳接头组合，构建其中ZFD对的左ZFD用24AA接头固定并且各种长度的接头用于右ZFD的ZFD组合，或反之亦然，并且测量其编辑效率(图1d和图3)。发现在两个位点使用相同的24AA接头是最有效的。还发现G1397处的DddAtox分裂物比G1333处的DddAtox分裂物更有效(图1c、图2a和图2b)。因此，通过这些最有效的ZFD对编辑胞嘧啶，其中在长度为7-21bp的间隔子区中具有>6.7％的最高效率(图1c、图2a和图2c)。
1-3.体内核DNA靶标中的碱基编辑研究了人细胞中具有24AA接头的ZFD是否能够在体内催化染色体靶位置处的C至T碱基编辑。构建了22对靶向总共八个基因中的11个位点的ZFD(每个位点一对两个ZFD)(图4)。其中，使用公开可用的锌指资源组装14对ZFD。通过调整先前表征的ZFN(对CCR5和TRAC具有特异性)产生另外8对ZFD。原因是ZFN在长度为5-7bp的间隔子中切割靶DNA，而ZFD在至少7bp的间隔子中起作用，从而产生能够通过将一个或两个锌指连接到ZFN对或从ZFN对分离而起作用的ZFD。由于具有四种不同构型的ZFN可以通过将FokI核酸酶与ZFP的N末端或C末端融合来构建，因此构建了两对具有不同构型的ZFD(图4b中的Trac-NC)，并且测试了分裂DddAtox半部分是否可以与ZFP的N末端以及现有ZFP的C末端融合(图4a和图5中所示的NC构型)。
[构建体]
*C型：SV40 NLS-锌指蛋白-24aa接头-DddAtox半部分-4aa接头-UGI
*N型：SV40 NLS-DddAtox半部分-24aa接头-锌指蛋白-4aa接头-UGI
-SV40 NLS
PKKKRKV(SEQ ID No:478)
-24aa接头
SGTPHEVGVYTLSGTPHEVGVYTL
-分裂DddAtox G1397-N
GSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLESKVFSSGGPTPYPNYANAGHVEGQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEG
-分裂DddAtox G1397-C
AIPVKRGATGETKVFTGNSNSPKSPTKGGC
-4aa接头
SGGS
-UGI
TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTAYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML
-ZFP
CCR5-1左(C型)[S162 ZFN-左]
GIHGVPAAMAERPFQCRICMRNFSDRSNLSRHIRTHTGEKPFACDICGRKFAISSNLNSHTKIHTGSQKPFQCRICMRNFSRSDNLARHIRTHTGEKPFACDICGRKFATSGNLTRHTKIHLR
CCR5-1右(C型)[S162 ZFN-右]
GIHGVPAAMAERPFQCRICMRNFSRSDNLSVHIRTHTGEKPFACDICGRKFAQKINLQVHTKIHTGEKPFQCRICMRNFSRSDVLSEHIRTHTGEKPFACDICGRKFAQRNHRTTHTKIHLR
CCR5-2左(C型)[S162 ZFN-左]
GIHGVPAAMAERPFQCRICMRNFSDRSNLSRHIRTHTGEKPFACDICGRKFAISSNLNSHTKIHTGSQKPFQCRICMRNFSRSDNLARHIRTHTGEKPFACDICGRKFATSGNLTRHTKIHLR(SEQ ID NO:2)
CCR5-2右(C型)[使用Barbas锌指模块组通过另外的ZF修饰S162 ZFN-右]
GIHGVPAAMAERPFQCRICMRNFSQSGDLRRHIRTHTGEKPFACDICGRKFARSDNLSVHTKIHTGSQKPFQCRICMRNFSQKINLQVHIRTHTGEKPFACDICGRKFARSDVLSEHTKIHLR(SEQ ID NO:482)
TRAC-左(C型)[改编自Paschon,D.E.等人,2019]
GIHGVPAAMAERPFQCRICMRNFSDQSNLRAHIRTHTGEKPFACDICGRKFATSSNRKTHTKIHTGSQKPFQCRICMRNFSLQQTLADHIRTHTGEKPFACDICGRKFAQSGNLARHTKIHLR(SEQ ID NO:483)
TRAC-左(N型)[改编自Paschon,D.E.等人,2019]
FQCRICMRKFATSGSLTRHTKIHTGEKPFQCRICMRNFSRSDHLSTHIRTHTGEKPFACDICGRKFAT SSNRTKHTKIHTHPRAPIPKPFQCRICMRNFSRSDNLSEHIRTHTGEKPFACDICGRKFAWHSSLRVHTKIHLR(SEQ ID NO:484)
TRAC-右(C型)[来自Paschon,D.E.等人,2019]
GIHGVPAAMAERPFQCRICMRNFSRSDHLSTHIRTHTGEKPFACDICGRKFADRSHLARHTKIHTGSQKPFQCRICMRKFALKQHLNEHTKIHTGEKPFQCRICMRNFSQSGNLARHIRTHTGEKPFACDICGRKFAHNSSLKDHTKIHLR(SEQ ID NO:485)
MFAP1左(C型)[使用Toolgen锌指模块组进行重新设计]
GIRIPGEKPYSCGICGKSFSDSSAKRRHCILHTGEKPYTCSDCGKAFRDKSCLNRHRRTHTGEKPYKCMECGKAFNRRSHLTRHQRIHTGEKPYECNYCGKTFSVSSTLIRHQRIHLR(SEQ ID NO:486)
MFAP1右(C型)[使用Barbas锌指模块组通过另外的ZF修饰S162 ZFN-右]
GIRERPYACPVESCDRRFSTSGSLVRHIRIHTGQKPFQCRICMRNFSRSDELTRHIRTHTGEKPFACDICGRKFARSDHLTTHTKIHTGEKPFQCRICMRKFAQSSNLVRHTKIHLR(SEQ ID NO:487)
CCDC28B左(C型)[使用Barbas锌指模块组通过另外的ZF修饰S162 ZFN-右]
GIRERPYACPVESCDRRFSDPGHLVRHIRIHTGQKPFQCRICMRNFSRSDELTRHIRTHTGEKPFACDICGRKFARSDHLTTHTKIHTGEKPFQCRICMRKFARSDKLVRHTKIHLR(SEQ ID NO:488)
CCDC28B右(C型)[使用Toolgen锌指模块组进行重新设计]
GIRIPGEKPYECNYCGKTFSVSSTLIRHQRIHTGEKPYHCDWDGCGWKFARSDELTRHYRKHTGEKPFECKDCGKAFIQKSNLIRHQRTHTGEKPYHCDWDGCGWKFARSDELTRHYRKHLR(SEQ ID NO:489)
KDM4B左(C型)[使用Barbas锌指模块组通过另外的ZF修饰S162 ZFN-右]
GIRERPYACPVESCDRRFSDCRDLARHIRIHTGQKPFQCRICMRNFSRSDELTRHIRTHTGEKPFACDICGRKFARSDHLTTHTKIHTGEKPFQCRICMRKFARSDKLVRHTKIHLR(SEQ ID NO:490)
KDM4B右(C型)[使用Toolgen锌指模块组进行重新设计]
GIRIPGEKPFECKDCGKAFIQKSNLIRHQRTHTGEKPYRCEECGKAFRWPSNLTRHKRIHTGEKPYRCEECGKAFRWPSNLTRHKRIHTGEKPYSCGICGKSFSDSSAKRRHCILHLR(SEQ ID NO:491)
NUMBL左(C型)[使用Barbas锌指模块组通过另外的ZF修饰S162 ZFN-右]
GIRERPYACPVESCDRRFSDCRDLARHIRIHTGQKPFQCRICMRNFSRSDELTRHIRTHTGEKPFACDICGRKFARSDHLTTHTKIHTGEKPFQCRICMRKFARSDKLVRHTKIHLR(SEQ ID NO:492)
NUMBL右(C型)[使用Toolgen锌指模块组进行重新设计]
GIRIPGEKPYTCSDCGKAFRDKSCLNRHRRTHTGEKPYKCGQCGKFYSQVSHLTRHQKIHTGEKPFECKDCGKAFIQKSNLIRHQRTHTGEKPYHCDWDGCGWKFARSDELTRHYRKHLR(SEQ ID NO:493)
INPP5D-1左(C型)[使用Barbas锌指模块组通过另外的ZF修饰S162 ZFN-右]
GIRERPYACPVESCDRRFSRSDKLVRHIRIHTGQKPFQCRICMRNFSRSDELTRHIRTHTGEKPFACDICGRKFARSDHLTTHTKIHTGEKPFQCRICMRKFARSDKLVRHTKIHLR(SEQ ID NO:494)
INPP5D-1右(C型)[使用Toolgen锌指模块组进行重新设计]
GIRIPGEKPYTCSDCGKAFRDKSCLNRHRRTHTGEKPYECNYCGKTFSVSSTLIRHQRIHTGEKPYTCSDCGKAFRDKSCLNRHRRTHTGEKPYTCSDCGKAFRDKSCLNRHRRTHLR(SEQ ID NO:495)
INPP5D-2左(C型)[使用Barbas锌指模块组通过另外的ZF修饰S162 ZFN-右]
GIRERPYACPVESCDRRFSRSDKLVRHIRIHTGQKPFQCRICMRNFSRSDELTRHIRTH TGEKPFACDICGRKFARSDHLTTHTKIHTGEKPFQCRICMRKFARSDKLVRHTKIHLR(SEQ ID NO:496)
INPP5D-2右(C型)[使用Toolgen锌指模块组进行重新设计]
GIRIPGEKPYECNYCGKTFSVSSTLIRHQRIHTGEKPYTCSDCGKAFRDKSCLNRHRRTHTGEKPYTCSDCGKAFRDKSCLNRHRRTHTGEKPYHCDWDGCGWKFARSDELTRHYRKH(SEQ ID NO:497)
DVL3左(C型)[使用Barbas锌指模块进行重新设计]
GIHGVPAAMAERPFQCRICMRNFSTSGHLVRHIRTHTGEKPFACDICGRKFATSGHLVRHTKIHTGEKPFQCRICMRNFSTSGELVRHIRTHTGEKPFACDICGRKFAQSSNLVRHTKIHLR(SEQ ID NO:498)
DVL3右(C型)[S162 ZFN-左]
GIHGVPAAMAERPFQCRICMRNFSDRSNLSRHIRTHTGEKPFACDICGRKFAISSNLNSHTKIHTGSQKPFQCRICMRNFSRSDNLARHIRTHTGEKPFACDICGRKFATSGNLTRHTKIHLR(SEQ ID NO:499)
在Hek 293T细胞中，ZFD(包括具有NC构型的ZFD)的C至T碱基编辑效率为1.0％至60％。另一方面，插入-缺失(插入缺失)<0.4％，因此很少发生(图4b和图6)。如上述基于质粒的实验中所见，具有5bp长间隔子的靶向CCR5的ZFD效率非常低。在具有至少7bp间隔子的靶标中，其他20个ZFD对显示12.0％±3.4％的平均编辑效率，其与cas9衍生的碱基编辑器2的8.3％±2.2％的平均值是可比较的。另外，C至T碱基编辑不仅发生在TC背景中，而且发生在AC或GCC中(图4c-图4f)。NUMBL的C6处的AC和INPP5D-2的C7处的GCC的相应C碱基编辑效率为4.58％和1.85％。
1-4.纯化的ZFD蛋白向人细胞的直接递送
将纯化的基因编辑蛋白递送到细胞中，而不是递送编码基因编辑蛋白的质粒DNA，能够降低脱靶效应，避免外来DNA的先天免疫反应，并防止外来质粒DNA在体内插入基因组中。其他组显示ZFP可在体外和体内自发通过哺乳动物细胞。为了证实蛋白质介导的ZFD的碱基编辑，选择高效靶向TRAC基因的ZFD对，并从大肠杆菌中纯化具有一个或四个核定位信号(NLS)的重组ZFD蛋白。首先，使用具有TRAC位点的PCR扩增子体外测试ZFD蛋白的碱基编辑效率，证实了非常高的效率。基于使用尿嘧啶特异性切除试剂(USER)的基因切割证实了效率，所述试剂是尿嘧啶DNA糖基化酶和DNA糖基化酶-裂解酶核酸内切酶VIII的混合物(图7)。将TRAC-NC ZFD蛋白以两种方式(电穿孔或直接递送而非电穿孔)递送到难以转染的人白血病K562细胞。ZFD蛋白是非常有效的。C至T碱基编辑效率为26.5％(电穿孔)和17％(直接递送)(图4g)。因此，这些结果表明，编码ZFD的质粒或纯化的重组ZFD蛋白可用于人细胞中核DNA的碱基编辑。
1-5.用ZFD进行线粒体DNA碱基编辑
与基于CRISPR的系统不同，与定制设计的DNA结合蛋白融合的分裂DddAtox系统可用于编辑包括线粒体DNA的细胞器DNA，这是DddA系统优于CRISPR系统的主要优点。为了将ZFD递送到线粒体，通过将线粒体靶向信号(MTS)和核输出信号(NES)连接至设计用于靶向线粒体基因的九个ZFD的N末端部分来构建mitoZFD(图8)。使用公开可用的锌指资源组装ZFD的ZFP片段。ZFD被配置为使得间隔子的长度是7-15bp，并且左和右DNA结合位点都是12bp长。
[构建体]
*C型：MTS-FLAG标签-NES-锌指蛋白-24aa接头-DddAtox半部分-4aa接头-UGI
*N型：MTS-HA标签-NES-DddAtox半部分-24aa接头-锌指蛋白-4aa接头-UGI
-MTS(人线粒体ATP合酶F1β亚基的线粒体靶向序列)
MLGFVGRVAAAPASGALRRLTPSASLPPAQLLLRAAPTAVHPVRDYAAQ(SEQ IDNo:274)
-FLAG标签(C型)
DYKDDDDK(SEQ ID No:275)
-HA标签(N型)
YPYDVPDYA(SEQ ID No:276)
-NES(核输出信号)
VDEMTKKF(小鼠微小病毒；MVM NES)
-分裂DddAtox G1397-N
GSYALGPYQISAPQLPAYNGQTVGTFYYVNDAGGLESKVFSSGGPTPYPNYANAGH VEGQSALFMRDNGISEGLVFHNNPEGTCGFCVNMTETLLPENAKMTVVPPEG
-分裂DddAtox G1397-C
AIPVKRGATGETKVFTGNSNSPKSPTKGGC
-4aa接头
SGGS
-UGI
TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTAYDESTDENVMLLTS DAPEYKPWALVIQDSNGENKIKML
-ZFP
*锌指通过TGEKP接头连接。(ZF1-接头-ZF2-接头-ZF3-接头-ZF4)。
*靶向ND2的mitoZFD具有QQ变体。该变体是Q取代R或K氨基酸。用红色字母标记。
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在HEK293T细胞中mitoZFD的线粒体DNA碱基编辑效率范围为2.6％至30％(平均14％±3％)(图9a)。此外，具有NC构型的mitoZFD(16％±4.5％，n＝6)与CC构型(8.3％±2.7％，n＝3)同样有效。在间隔子区中具有TC背景的大多数胞嘧啶以各种效率被碱基编辑(图9b-9g)。此外，在ND2位点具有ACC(C8和C9)背景的胞嘧啶也分别显示出约7.4％和19.9％的效率(图9b)。这表明ZFD介导的C至T碱基编辑不限于TC基序。
此外，从线粒体DNA(mtDNA)突变细胞中分离单细胞衍生的克隆群体以证明mitoZFD没有细胞毒性，并且mtDNA突变也在克隆群体中维持。在从用ND1特异性mitoZFD处理的HEK293T细胞分离的30个单细胞衍生的克隆群体中，五个克隆群体显示出35％至98％的ND1基因碱基编辑效率(图10a)。同样，在从用ND2特异性mitoZFD处理的HEK293T细胞分离的36个单细胞衍生的克隆群体中，七个克隆群体显示出26％至76％的ND2基因碱基编辑效率(图10b)。对于所有其他克隆群体，观察到0.4％-1.0％的低效率，最可能是由于测序错误。在未用ZFD处理的细胞中获得类似的效率(图10c)。这些结果表明，mitoZFD在细胞群体中不均匀地诱导异质性突变。大多数ZFD处理的细胞是野生型的，而具有异质性突变的细胞具有高达98％的突变率。这些突变甚至在克隆扩增后仍维持(图11和图12)。
1-6.mitoZFD和基于TALE的DdCBE
发现构建的ND1特异性mitoZFD的突变模式不同于靶向相同基因的基于TALE的DdCBE(图9f-图9h)。两种mitoZFD在C5或C8催化C至T碱基编辑(图9f)，而DdCBE在C8、C9和C11位置诱导碱基编辑(图9g)。因此，由mitoZFD引起的氨基酸变化与由DdCBE引起的完全不同(图9h)。与mitoZFD连接的左和右位点被8bp间隔子隔开，但是在DdCBE的情况下，这些位点被16bp间隔子隔开，其可能负责差异突变模式。这些结果表明，mizoZFD和DdCBE可以在线粒体DNA中以互补的方式产生各种突变。
为了产生更多的突变模式，测试是否将ZFD单体和DdCBE单体混合以形成杂交对。靶向ND1基因的10个杂交对在HEK293T细胞中显示出良好的活性，其中平均碱基编辑效率为17％±3.4％(图13)。事实上，其中一个杂交对(TALE-L/ZFD-R1)显示出比靶向相同位点的两个DdCBE对和十个ZFD对更好的效率，其中最高碱基编辑效率为41％(图13b)。此外，杂交对产生与用DdCBE和mitoZFD获得的突变模式不同的突变模式(图13c)。少数杂交对(例如ZFD-L1/TALE-R和ZFD-L2/TALE-R)在单个位置诱导C至T转化而没有旁观者编辑。相比之下，大多数mitoZFD对和DdCBE对在间隔子区的多个位置诱导C至T转化。这些结果表明，ZFD/DdCBE杂交对可以产生独特的突变模式并且产生使用ZFD对或DdCBE对不能获得的某些突变。
1-7.mitoZFD的线粒体全基因组靶特异性
为了证实mitoZFD是否引起脱靶编辑，从用每对靶向ND1或ND2基因的mitoZFD处理的细胞中提取线粒体DNA，然后进行全线粒体基因组测序。将不同量(5-500ng)的编码mitoZFD对的mRNA或质粒转染到HEK293T细胞中。如所预期的，中靶编辑效率是剂量依赖性的。高浓度(100、200和500ng)的mRNA或质粒产生>30％的中靶效率，但也导致>1％的数百个脱靶编辑(图14-17)。低浓度(5和10ng)在很大程度上避免了这些脱靶编辑，但显著降低了中靶效率。中等浓度(50ng)的mRNA是最合适的。在不引起数百个脱靶编辑的情况下保持高中靶效率。为了进一步消除剩余的脱靶编辑，将R(-5)Q突变引入每个锌指以消除非特异性DNA接触。与未用ZFD处理的细胞的mtDNA相比，所得的ZFD变体(在图18中显示为QQ)保持了高中靶活性，并显示了精确的特异性，且几乎没有脱靶编辑(图18a和图18b)。
碱基编辑是一种相对新的方法，其能够编辑靶向碱基而不引起DNA双链断裂或DNA修复模板。碱基编辑使得能够在细胞、动物和植物中进行C至T或A至G转化，从而允许研究单核苷酸多态性(SNP)的功能效应并纠正致病点突变，用于治疗应用。已经开发了两种类型的碱基编辑技术，即基于CRISPR的腺嘌呤和胞嘧啶碱基编辑和基于DddA的碱基编辑技术。基于CRISPR的碱基编辑器由作为DNA结合单元的催化受损的Cas9或Cas12a和源自大鼠或大肠杆菌的单链DNA特异性脱氨酶构成。另一方面，DdCBE由TALE DNA结合阵列和双链DNA特异性DddAtox构成。
与DdCBE相比，ZFD的尺寸更小。这是因为ZFD中的锌指蛋白是紧凑的，而DdCBE中的TALE阵列是庞大的。因此，ZFD对编码基因而不是DdCBE对编码基因可以容易地包装在具有小货物空间的AAV载体中。另外，紧凑的ZFP在工程化上是友好的，使得可以将分裂DddAtox半部分融合到ZFP的N末端或C末端，产生在ZFP结合位点的上游或下游操作的ZFD。此外，重组ZFD蛋白可以在没有电穿孔或脂质转染的情况下自发地穿透人细胞，使得能够进行无基因的基因治疗。ZFD对或ZFD/DdCBE杂交对可以产生独特的突变模式，这不能单独使用DdCBE获得。这些性质使得ZFD成为用于建模和治疗线粒体疾病的强大的新平台。
实施例2.通过TALE-DdCBE进行植物叶绿体和线粒体基因编辑
植物细胞器(包括线粒体和叶绿体)各自具有它们自己的基因组，其编码呼吸和光合作用所必需的许多基因。植物细胞器基因编辑(植物遗传学和生物技术尚未满足的需求)由于缺乏靶向这些细胞器中的DNA的适当工具而受到限制。为了组装DddA衍生的胞嘧啶碱基编辑质粒(DdCBE)，开发了由16个表达质粒(8个用于将所得蛋白质递送到线粒体，另8个用于递送到叶绿体)和424个TALE子阵列质粒构成的Golden Gate克隆系统，并将完成的DdCBE质粒用于在线粒体和叶绿体中高效地诱导点突变。DdCBE碱基编辑在莴苣或油菜籽愈伤组织中以高达25％(线粒体)和38％(叶绿体)的效率诱导突变。为了避免DdCBE编码质粒引起的脱靶突变，将DdCBE信使RNA转染到莴苣原生质体中，证明在叶绿体中进行无DNA碱基编辑。此外，通过在叶绿体16S rRNA基因中引入点突变产生具有高达99％编辑效率的链霉素或大观霉素耐药性莴苣愈伤组织和芽。
DdCBE是包含衍生自细菌胞嘧啶脱氨酶毒素DddAtox的分离的无毒结构域、为特定位置设计的TALE阵列和UGI在内的异二聚体，并在靶DNA中TALE蛋白结合位点之间的间隔子中诱导胞嘧啶至胸腺嘧啶的取代。我们用DdCBE的结果证明了植物中高效的细胞器碱基编辑。
2-1.方法
用于植物原生质体实验的表达质粒的构建
使用Gibson组装方法构建DdCBE Golden Gate目的载体。编码TAL N末端结构域、HA标签、FLAG标签、TAL C末端结构域、分裂DddAtox和UGI的序列经密码子优化用于在双子叶植物(拟南芥(Arabidopsis thaliana))中表达并通过整合DNA技术合成。从拟南芥cDNA扩增编码来自AtinfA和AtRbcS的CTP和来自ATP酶δ亚基和ATP酶γ亚基的MTS的序列。对于植物表达，使用PcUbi启动子和pea3A终止子代替骨架质粒中的哺乳动物CMV启动子。为了构建用于体外DdCBE mRNA转录的载体，将T7启动子盒克隆到PcUbi启动子和DdCBE编码区之间的DdCBE Golden Gate目的载体中。
通过单因素Golden Gate组装构建TALE阵列基因。使用424个TALE阵列质粒和目的载体，通过BsaI消化和Golden Gate组装的T4连接构建DdCBE表达质粒。使用以下步骤进行单因素Golden Gate克隆：37℃和50℃各5分钟，循环20次，随后在50℃下最终反应15分钟，然后在80℃下5分钟。使用Plasmid Plus迷你制备试剂盒(Qiagen)纯化用于植物原生质体转染的所有载体。载体构建中使用的DNA和氨基酸序列如下。
[表2]
DdCBE的特定氨基酸序列和TALE重复序列的特定氨基酸序列如下。
[表3]
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mRNA体外转录
使用Phusion DNA聚合酶(Thermo Fisher)通过PCR制备DdCBE DNA模板。使用体外mRNA合成试剂盒(Enzynomics)合成和纯化DdCBE mRNA。
原生质体分离和转导
将莴苣种子在70％乙醇中表面灭菌30秒，在0.4％次氯酸盐溶液中表面灭菌15分钟，然后用无菌蒸馏水洗涤三次。将莴苣种子在补充有2％蔗糖的0.5xMS培养基上在25℃下在16小时光照和8小时黑暗条件下发芽。将油菜籽种子在70％乙醇中表面灭菌3分钟，在1.0％次氯酸盐溶液中表面灭菌30分钟，然后用无菌蒸馏水洗涤三次。将油菜籽种子在补充有3％蔗糖的1xMS培养基上在25℃下在16小时光照和8小时黑暗条件下发芽。
如前所述进行原生质体分离和转导。将来自7天龄莴苣和14天龄油菜籽植物的子叶用酶溶液在黑暗条件下振荡(40rpm)消化3小时。用等体积的W5溶液洗涤原生质体-酶混合物，然后通过80g离心7分钟从蔗糖溶液获得完整的原生质体。将原生质体在4℃下用W5溶液处理1小时，然后使用聚乙二醇转染。
将重悬于MMG溶液中的莴苣原生质体和油菜籽原生质体用质粒或mRNA使用PEG转染，然后在室温下培养20分钟。将PEG-原生质体混合物用等体积的W5溶液在轻轻倒置下洗涤三次，然后培养10分钟。然后通过100g离心5分钟使原生质体沉淀。
原生质体培养
将用编码DdCBE的质粒转染的莴苣原生质体重悬于莴苣原生质体培养基(LPCM)中。将培养基中的原生质体与含有2.4％低熔点琼脂糖的培养基以1:1混合，然后立即置于6孔板中。混合物固化后，用1ml液体培养基覆盖包埋的原生质体，然后在25℃下，在黑暗条件下培养1周。初始培养后，每周用新鲜培养基替换覆盖的液体培养基，并将包埋的原生质体在16小时光照和8小时黑暗条件下培养1周，然后在16小时光照和8小时黑暗条件下培养2周。将原生质体诱导的微愈伤组织在再生培养基中在25℃下在16小时光照和8小时黑暗条件下培养4周。为了准备分析碱基编辑效率，将原生质体在25℃下，在黑暗条件下在液体培养基中在不进行包埋的情况下培养1周。为了测试抗生素耐药性，将包埋一个月的微愈伤组织在含有50mg/L链霉素或50mg/L壮观霉素的再生培养基中在25℃下在16小时光照和8小时黑暗条件下培养4周。4周后，将抗生素耐药性绿色愈伤组织或不定芽转移到含有200mg/L链霉素或50mg/L壮观霉素的新鲜再生培养基中。
将用编码DdCBE的质粒转染的油菜籽原生质体重悬于油菜籽培养基中。将原生质体-培养基混合物转移到6孔板中并在25℃下，在黑暗条件下培养2周。2周后，将原生质体在16小时光照和8小时黑暗条件下培养3周。用非常新鲜的培养基替换培养基。
DNA和RNA提取
使用DNeasy植物微型试剂盒或RNeasy植物微型试剂盒从液体培养基中培养的细胞或转基因愈伤组织中提取总DNA或RNA。通过以10,000rpm离心1分钟收获培养的细胞或愈伤组织。然后，使用RNA至cDNA EcoDry Premix(TaKaRa)从总RNA逆转录cDNA。
深度测序
使用融合酶和合适的引物扩增靶区域(补充表1)。为了创建DNA测序文库，进行三轮PCR(第一轮，巢式PCR；第二轮，PCR；第三轮，索引PCR)。汇集等量的DNA，然后使用MiniSeq系统(Illumina)测序。通过Cas分析仪和计算机程序的源代码分析配对末端测序文件。
2-2.结果
开发了Golden Gate组装系统以构建靶向叶绿体的DdCBE(cp-DdCBE)和靶向线粒体的DdCBE(mt-DdCBE)(图19)。表达质粒编码由叶绿体转运肽或线粒体靶向序列、TALE的N或C末端结构域、分裂DddAtox半部分(G1333N、G1333C、G1397N和G1397C)和UGI构成的融合蛋白，对所述表达质粒优化以用于在欧芹泛素(PcUbi)启动子和pea3A终止子的控制下在双子叶植物中表达。通过在E管中混合表达载体和六个TALE子阵列质粒，在单个亚克隆步骤中构建具有定制设计的TALE DNA结合序列的DdCBE质粒。共有424个(6×64个三部分+2×16个二部分+2×4个单部分)模块化TALE子阵列质粒可用于制备识别16-20bp长度序列的cp-DdCBE和mt-DdCBE，包括5'端的保守T。因此，DdCBE异二聚体在功能上识别32-40bp DNA序列。
为了确定DdCBE是否能够促进叶绿体中的碱基编辑，构建四对适合于编码30S核糖体亚基的RNA组分的叶绿体16S rRNA基因的cp-DdCBE质粒，并将每对共转染到莴苣和油菜籽原生质体中。7天后，通过深度测序测量碱基编辑效率(图20a和图20b)。具有最高效率的cp-DdCBE对(左-G1397-N+右-G1397-C)在两个TALE结合位点之间的15bp间隔子中诱导C*G至T*A转化，其中在莴苣原生质体中效率为30％，并且在油菜籽原生质体中效率为15％(图20b)。与先前在哺乳动物细胞和小鼠中的结果一样，5’-TC基序中的胞嘧啶(C9和C13)优选通过cp-DdCBE转化为胸腺嘧啶。有趣的是，在莴苣原生质体中，另一种cp-DdCBE(左-G1333-N+右-G1333-C)以4.2％的效率将5'-AC背景中的胞嘧啶(C7)变为胸腺嘧啶。此外，观察到在14天培养期间莴苣原生质体中cp-DdCBE的碱基编辑的持久性(图24)。编辑效率持续增加长达10天，并在整个培养期间维持。
在分别编码光系统II的D1和CP-47光合蛋白的两个另外的叶绿体基因psbA和psbB中测试碱基编辑(图20c，20d和25)。在靶向Psb基因的cp-DdCBE中，最有活性的一个(左-G1397-C+右-G1397-N)能够以高达25％的效率诱导莴苣原生质体中的C*G至T*A转化(图20d)。碱基编辑器仅将5’-TCC背景中的两个胞嘧啶(C11和C12)有效地转化为胸腺嘧啶。可以首先将5’-TCC转化为5’-TTC，然后转化为5’-TTT。在油菜籽原生质体中，其他组合(左-G1333-N+右-G1333-C)在四个胞嘧啶位置(C3，C4，C11和C12)显示出最高的效率，效率高达3.5％(C3)。C3和C4在油菜籽基因中处于5’-TCC背景中，而由于单核苷酸多态性，它们在莴苣对应物中处于5’-ACC背景中，这导致DdCBE在油菜籽基因中而不是在莴苣基因中对两个胞嘧啶(C3和C4)的有效编辑。同样，靶向psbB基因的cp-DdCBE组合在油菜籽原生质体中以0.36％至4.1％的效率在TCC背景中催化两个胞嘧啶的转化(图25)。总之，这些结果表明，编辑效率由包括DddAtox分裂物位置(G1333相对于G1397)和方向(左-G1333-N相对于左-G1333-C)在内的胞嘧啶位置和序列决定，并且cp-DdCBE能够在植物叶绿体基因组中进行有效的碱基编辑。
此外，尝试使用定制设计的mt-DdCBE在植物线粒体DNA中实现碱基编辑。为此，构建靶向莴苣和油菜籽中atp6基因和油菜籽中rps14基因的编码mt-DdCBE的质粒(使用Golden Gate克隆系统)，并将质粒导入莴苣和油菜籽原生质体中。导入后7天，通过深度测序测量碱基编辑效率(图20e，20f和26)。最有效的mt-DdCBE组合(莴苣中的左-G1397-N+右-G1397-C和油菜籽中的左-G1397-C+右-G1397-N)在atp6基因靶位点在莴苣原生质体中以23％的效率和在油菜籽原生质体中以23％的效率催化C*G至T*A转化(图20)。此外，mt-DdCBE组合在rps14靶位点在油菜籽原生质体中以11％效率诱导C*G至T*A转化。这些结果表明，植物中的线粒体DNA易于用mt-DdCBE进行碱基编辑。
为了研究DdCBE对cpDNA和mtDNA的编辑是否在再生过程中维持，在导入后4周从DdCBE处理的原生质体中收集再生的莴苣和油菜籽愈伤组织(图21a)，并使用深度测序和Sanger测序测量每个愈伤组织的碱基编辑效率(图21b和图27)。叶绿体或线粒体基因，其碱基编辑由DdCBE诱导，在26个莴苣愈伤组织中的22个和14个油菜籽愈伤组织中的7个中分别显示高达38％和25％的效率(图21c)。此外，叶绿体psbA基因的碱基编辑在莴苣愈伤组织中显示高达3.9％的效率(图27)。同样，测量油菜籽愈伤组织中线粒体碱基编辑，在atp6和rps14的效率分别高达25％和1.9％(图27)。这些结果表明，DdCBE在植物原生质体中的表达是可以耐受的，并且在原生质体再生过程中DdCBE诱导细胞器碱基编辑。
此外，尝试使用体外转录的cp-DdCBE mRNA而不是质粒来证明细胞器中的无DNA碱基编辑。将编码靶向16S rRNA基因的cp-DdCBE的体外转录物导入莴苣原生质体后，分析靶位点处的碱基编辑效率(图21a)。测量原生质体中的C至T突变，效率高达25％(图21d和28)。如所预期的，在导入原生质体后7天，不存在DdCBE mRNA和DNA序列(图29)。该方法可以避免质粒DNA片段潜在整合到宿主基因组中。
由于从原生质体再生的愈伤组织中细胞器编辑的稳定维持，测量了对链霉素和壮观霉素抗生素的耐药性，所述抗生素通过叶绿体DNA中的16S rRNA基因编辑与16S rRNA基因不可逆结合来抑制蛋白质合成。16S rRNA基因中的多个单核苷酸多态性通常在链霉素耐药性原核生物和真核生物中观察到，特别地，16S rRNA C860T(大肠杆菌坐标C912)突变导致烟草中的链霉素耐药性。烟草中的C860T点突变等同于莴苣中的C9位置(图20a，20b，21b和21d)。将从DdCBE处理的原生质体再生的莴苣愈伤组织转移到补充有链霉素和壮观霉素的培养基中。当暴露于抗生素时，模拟处理组变成白色，表明愈伤组织的原生质体功能障碍。相比之下，DdCBE处理的愈伤组织保持绿色，显示出对这些抗生素的耐药性。在具有耐药性的莴苣愈伤组织和小植株中分析DdCBE的编辑效率。与C860T突变一样，在药物处理后获得的愈伤组织和芽中观察到位置C9的C至T转化，效率高达98.6％(图21e和21f)。有趣的是，在不存在壮观霉素的情况下，在位置C13附近的C至T编辑显示高达20％的效率，但在存在抗生素的情况下根本没有，表明在药物处理时对该突变选择。总之，这些结果表明，原生质体中由DdCBE诱导的植物细胞器突变甚至可以在细胞分裂和植物发育后也可以维持，并且叶绿体编辑的同型异源性可以通过药物选择来实现。
另外，在原生质体、愈伤组织和芽中分析靶向16S rRNA位点的TALE脱氨酶的脱靶活性。在衍生自单细胞的抗生素耐药性愈伤组织或芽中，在靶位点附近(两侧50个碱基对)(图31)或在叶绿体基因组中基于序列同源性选择的前五个候选脱靶位点(图32)没有检测到脱靶突变。相比之下，当将编码DdCBE的质粒导入原生质体时，在五个候选脱靶位点中的三个以1.2％至4.1％的低效率诱导脱靶TC至TT突变。当使用体外转录物(mRNA)代替编码TALE脱氨酶的质粒时，原生质体中的脱靶效率大大降低(图22)。这些结果表明，DdCBE的过量表达或延长的基于质粒的表达增加了脱靶突变，并且使用mRNA的瞬时基于mRNA的表达对于避免脱靶碱基编辑是优选的。
总之，开发了使用424个TALE子阵列质粒和16个表达质粒的Golden Gate克隆系统，以组装编码DdCBE的质粒，用于植物中细胞器碱基编辑。定制设计的靶向叶绿体DNA中的三个基因和线粒体DNA中的两个基因的DdCBE在莴苣和油菜籽原生质体中实现了高效的C至T转化。特别地，在细胞分裂和植物发育期间维持植物细胞器中的编辑。此外，通过叶绿体16S rRNA基因突变获得具有近同型异源性(99％)的抗生素耐药性莴苣愈伤组织和小植株。在没有抗生素选择的情况下，线粒体中的编辑效率为25％，叶绿体为38％。预期GoldenGate克隆系统将是植物中细胞器DNA编辑的有价值的资源。
实施例3.TALE-DdCBE的动物DNA编辑
由分裂细菌间毒素DddAtox、设计为结合DNA的转录激活因子样效应物(TALE)和尿嘧啶糖基化酶抑制剂(UGI)构成的DddA衍生的胞嘧啶碱基编辑器(DdCBE)使得能够在线粒体DNA中进行期望的胞嘧啶至胸腺嘧啶碱基编辑。同样，高效的线粒体DNA编辑在小鼠胚胎中是可能的。在线粒体基因中，靶向编码催化NADH脱水和电子转移到泛醌的NADH脱氢酶的亚基的MT-ND5(ND5)，包括与人线粒体疾病相关的突变，例如m.G12918A，和产生早期终止密码子的突变，例如m.C12336T。因此，有可能在小鼠中产生线粒体疾病模型，表明治疗线粒体疾病的可能性。
3-1.方法
质粒组装。使用TALEN(转录激活因子样效应物核酸酶)系统构建含有DddA半部分和最终TALE-DddAtox构建体的表达质粒。在TALEN系统的表达质粒中，FokI二聚体的核定位信号和单体被线粒体靶向信号(MTS)、DddA脱氨酶半部分和尿嘧啶糖基化酶抑制剂(UGI)取代。通过IDT合成编码MTS、DddA和UGI的序列。为了构建表达载体，使用Q5 DNA聚合酶(NEB)扩增Gibson组装所需的DNA片段，然后纯化。使用HiFi DNA组装试剂盒(NEB)组装纯化的基因片段，化学转化到大肠杆菌DH5ɑ(Enzynomics)中，然后通过Sanger测序进行鉴定。因此，获得了八种不同的表达质粒，其中用于Golden Gate克隆的BsaI限制性位点位于编码N末端和C末端结构域的序列之间。为了组装DdCBE质粒，将表达质粒和模块载体(每个编码TALE序列)、BsaI-HFv2(10U)、T4 DNA连接酶(200U)和反应缓冲液装载在一个试管中。其后，限制酶和连接酶反应在热循环仪中进行在37℃下5分钟和在50℃下5分钟的20个循环，随后进一步反应，在50℃下15分钟和在80℃下5分钟。将缀合的质粒通过化学转化导入大肠杆菌DH5ɑ中，并通过Sanger测序鉴定最终的构建体。对于细胞系导入，将质粒中量制备。
哺乳动物细胞系培养和转染。将NIH3T3(CRL-1658，美国典型培养物保藏中心(ATCC))细胞系在37℃下在5％CO2环境中培养。细胞系在补充有10％(v/v)胎牛血清的DMEM(Gibco)中在没有抗生素的情况下生长，并且不测试支原体。对于脂质转染，在转染前18-24小时将细胞以1.5×104的细胞密度接种在12孔细胞培养板(SPL，韩国首尔)中。使用Lipofectamine 3000(Invitrogen)，使用500ng每个DdCBE分裂物引入总共1,000ng质粒DNA。转染后4天收获细胞。
mRNA制备。使用Q5 DNA聚合酶(NEB)通过PCR扩增mRNA模板，并使用以下引物(F：5’-CATCAA TGGGCGTGGATAG-3’SEQ ID No:268，R：5’-GACACCTACTCAGACAATGC-3SEQ ID No:269)。使用体外RNA转录试剂盒(mMESSAGE mMACHINE T7 Ultra试剂盒，Ambion)合成DdCBEmRNA，然后使用MEGAclear试剂盒(Ambion)纯化。
动物。所有涉及小鼠的实验在基础科学研究所动物护理和使用委员会的批准下进行。将超数排卵的C57BL/6J雌性与C57BL/6J雄性交配，并将ICR品系雌性用作雌性老鼠。将小鼠在12小时昼夜循环条件和恒温恒湿条件(20-26℃，40％-60％)下圈养在特定的无病原体设施中。
微注射到小鼠受精卵中。超数排卵、胚胎收集和显微注射是在临近显微注射之前进行的过程，如前所述。对于显微注射，将左DdCBE mRNA(300ng/μl)和右DdCBE mRNA(300ng/μl)的混合物用DEPC处理的注射缓冲液(0.25mM EDTA，10mM Tris，pH7.4)稀释，并使用Nikon ECLIPSE Ti显微操作器和FemtoJet 4i显微注射器(Eppendorf)注射到受精卵细胞质中。显微注射后，将胚胎置于KSOM+AA(Millipore)微滴中，然后在37℃，5％CO2条件下培养4天。将2-细胞期胚胎转移到0.5-d.p.c.假孕雌性老鼠的输卵管中。
基因分型。将处于胚泡期的胚胎和组织置于消化缓冲液(25mM NaOH，0.2mM EDTA，pH 10)中，接着在95℃下孵育20分钟，之后使用HEPES(游离酸，未经pH调节)将pH调节至7.4，使得最终浓度为50mM。使用DNeasy血液和组织试剂盒(Qiagen)从小鼠后代分离基因组DNA，并通过Sanger测序和靶向深度测序对其进行分析。
用于高通量测序的线粒体DNA分离。为了在12孔板中从培养的NIH3T3细胞中分离线粒体，去除细胞培养基，然后向培养板中添加200μl线粒体分离缓冲液A(ScienCell)。使用细胞刮铲刮取细胞，然后置于微管中，使用一次性杵研磨细胞。研磨15次后，将充分研磨的匀浆在1,000x g和4℃下离心5分钟。将上清液置于新的微管中并在10,000x g，4℃下离心20分钟。将沉淀重悬于20μl裂解缓冲液(25mM NaOH，0.2mM EDTA，pH 10)中，然后在95℃煮沸20分钟。为了降低pH，向线粒体裂解物中添加2μl 1M HEPES(游离酸，未经pH调节)。在PCR模板链中使用1μl溶液用于高通量测序。
高通量测序。为了制备深度测序文库，使用Q5 DNA聚合酶进行巢式初级PCR和次级PCR，然后添加最终的索引序列。使用MiniSeq(Illumina)将文库用于配对末端读取测序。对于全线粒体基因组分析，使用标记DNA制备试剂盒(Illumina)根据制造商的方案制备分离的线粒体DNA。将来自所有分析的配对末端测序结果合并到一个fastq-join文件中，并使用CRISPR RGEN工具(http://www.rgenome.net/)进行分析。
数据分析和显示。使用Microsoft Excel(2019)和PowerPoint(2019)创建图形、图表和表格。使用Geneious(版本2021.0.1)和Snapgene 5.2.3进行基因组序列比对、引物构建和克隆设计，并使用NC_005089作为参考序列。
3-2.结果
DdCBE质粒组装。为了促进定制设计的TALE序列在DdCBE中的组装，构建了编码分裂DddAtox半部分的表达质粒，并使用了使用总共424个(6×64个三部分+2×16个二部分+2×4个单部分)质粒的Golden Gate克隆系统(图33a)。如下表4所示，将六个TALE模块质粒和表达质粒在同一试管中混合以构建即用型DdCBE质粒，其具有15.5-18.5个重复可变双残基序列(图38)。
[表4]
DdCBE构建体的序列如下表5所示。因此，DdCBE识别17-20个DNA序列，包括5'端的保守胸腺嘧啶序列。因此，功能性DdCBE对识别总共32-40个DNA序列。
[表5]
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体外线粒体碱基编辑。为了尝试使用Golden Gate克隆系统在体内编辑线粒体DNA，选择编码小家鼠线粒体NADH泛醌氧化还原酶链5蛋白的ND5基因。ND5蛋白是NADH脱氢酶(泛醌)的关键亚基并催化电子从NADH转移到呼吸链。在人类中，已知ND5基因突变与MELAS(线粒体脑肌病、乳酸性酸中毒和中风样发作)有关，也与Leigh综合征或LHON(Leber遗传性视神经病变)的一些症状有关。尝试建立线粒体基因具有遗传改变的小鼠模型以模拟人的功能障碍。
首先，组装几个DdCBE质粒，这些质粒被设计为产生两个沉默突变即m.C12539T和m.G12542A。将这些质粒转染到NIH3T3小鼠细胞系中，3天后测量碱基编辑频率。如所预期的，靶范围内的胞嘧啶碱基以高达19％的效率被编辑为胸腺嘧啶(图34a)。先前报道了DddAtox仅使“TC”序列中的胞嘧啶脱氨基，但基于实验结果，仅编辑了TC背景中的两个胞嘧啶。在编辑靶范围内没有显著产生插入缺失或其他类型的点突变。
体内线粒体碱基编辑。最有效的DdCBE对(左-G1397-N和右-G1397-C)用于体内实验。将编码该DdCBE对的体外转录物显微注射到1-细胞期的C57BL6/J胚胎中后四天，32个胚胎中有9个被成功编辑(28％，表6)。
[表6]
TALE-DddAtox脱氨酶有效地产生C*G至T*A碱基转化，在m.C12539处效率为2.2％-25％，在m.G12542处效率为0.63％-5.8％。接着，将注射了DdCBE的胚胎转移到雌性老鼠以获得具有m.C12539T和m.G12542T的后代(图39)。四只幼崽(F0)中的三只显示C*G至T*A编辑，效率为1％至27％(图34c)。两只幼崽在脚趾和尾部显示相似的突变水平，并在出生后14天维持效率。此外，在出生后50天，在成年F0小鼠的各种组织中检测到这些线粒体DNA突变(图34d)。这些结果表明，DdCBE诱导的1-细胞期受精卵中线粒体DNA的异质性在发育和分化过程中得以维持。
为了确定DdCBE诱导的突变是否传递到下一代，通过使雌性F0小鼠与野生型C57BL6/J雄性交配产生F1后代。在两只幼崽中观察到m.C12539T和m.G12542T突变，其中效率为6％-26％。此外，在11种不同组织中观察到类似的线粒体编辑(图35b)。
DdCBE介导的MT-ND5 G12918A突变。尝试产生m.G12918A突变，其也引起人的线粒体疾病。这种突变引起各种线粒体疾病，如Leigh综合征、MELAS综合征和LHON综合征。由于在该位置的胞嘧啶碱基具有相邻的胸腺嘧啶，因此使用DdCBE进行碱基编辑是可能的(图36a)。组装四对DdCBE，证实在NIH3T3中编辑是可能的，效率高达6.4％(图36b)。然后，将最有效的DdCBE组合显微注射到小鼠受精卵中，并在胚泡中观察其效率。44个胚胎中有11个(25％)携带m.G12918A突变，效率为0.25％至23％(图36c)。此外，将DdCBE显微注射的胚胎转移到雌性老鼠以获得具有G12918A突变的后代(图39b)。已证实，11只新生小鼠中的4只在约3.9％-31.6％的范围内具有该突变(图36d)。尽管出生后没有立即出现表型，推测是因为后代非常年轻并且野生型线粒体DNA和突变体DNA以杂合状态共存，所以这些结果表明DdCBE可能产生患有线粒体疾病的动物模型。
MT-ND5无义突变。最后，通过在基因中产生无义突变证实了ND5功能丧失突变是否可以在小鼠中维持。使用m.C12336作为靶胞嘧啶，在ND5蛋白的位置199引入早期终止密码子(Q199*；图37a)。具体地，将四种DdCBE组合转染到NIH3T3细胞系中以证实碱基编辑效率，表明最有效的DdCBE对引起无义突变，效率为约5.7％(图37b)。已证实，该DdCBE导致胞嘧啶到胸腺嘧啶编辑，并且产生沉默突变m.G12341A的突变(Q200Q)尽管效率稍低，但在靶范围内被编辑。在37个小鼠胚胎中的19个(＝51％)中，证实了m.C12336T和m.G12341A突变，效率分别为32％和23％(图37c)。
基于这些结果，将小鼠胚胎转移到雌性老鼠以获得具有m.C12336T和m.G12341A突变的后代(图39c)。27只F0小鼠中有9只(23％)显示C*G至T*A编辑，效率为0.22％至57％(图37d和37e)，其显示ND5的无义突变不引起胚胎剔除。
实施例4.动物中线粒体DNA编辑
4-1.构建具有核输出信号的用于动物线粒体的碱基编辑的表达载体
构建在动物细胞中表达具有与TALE-DdCBE融合的核输出信号的蛋白质的载体(图40)。载体使用巨细胞病毒启动子(CMV启动子)。载体包括线粒体靶向信号、蛋白质纯化/检测标签、TALE阵列N末端结构域、重复序列区、C末端结构域、DddA胞嘧啶脱氨酶分裂物半部分、尿嘧啶糖基化酶抑制剂和核输出信号(图40a)。对于核输出信号，例如，可以使用衍生自MVM(小鼠微小病毒)的NS2蛋白，但也可以使用其他序列。由此表达的蛋白质被释放到细胞核外，然后转移到线粒体，并因此进行碱基编辑。这里，靶DNA位点选自线粒体ND5基因-染色体4ND5样基因、线粒体TrnA-染色体5和线粒体Rnr2-染色体6(图40b)。
4-2.动物细胞系中的DdCBE-NES
在转染前一天下午，将NIH3T3细胞系(ATCC CRL-1658)以1.5×104/孔分配到含有1ml细胞生长培养基(DMEM+10％小牛血清)的12孔板中。第二天早上，根据制造商的方案，使用Lipofectamine 3000，用添加了DNA未处理的模拟、DdCBE和DdCBE-MVM NES质粒的实验组转染细胞。在培养箱(37℃，5％CO2)中培养三天后，收获细胞并使用Qiagen血液和组织试剂盒纯化总DNA，随后使用线粒体基因特异性PCR引物扩增，然后使用Illumina MiniSeq系统进行下一代测序，之后使用Cas分析仪(www.rgenome.net)测定碱基编辑效率。
图40c、40d和40e显示用DdCBE-NES转染NIH3T3小鼠细胞系产生的小鼠线粒体基因ND5、TrnA和Rnr2的突变，表明效率根据DdCBE组合而变化。
4-3.动物细胞系中的mitoTALEN
构建识别图40f所示序列的TALEN，并将MTS与该构建体连接以将其与DdCBE一起引入线粒体。实验方法与实施例4-2相同。有意对TALE识别位点进行错配，使得发生与野生型mtDNA的1个错配和与突变体mtDNA的2个错配。这是因为TALE不能区分1个核苷酸错配。结果证实，与单独用DdCBE处理相比，用DdCBE和TALEN处理的+2错配实验组的效率进一步提高。
4-4.动物胚胎中的DdCBE-NES
使用DdCBE表达载体或DdCBE-NES表达载体作为模板，获得含有T7启动子和DdCBE或DdCBE-NES表达位点的PCR扩增子。使用该PCR扩增子作为模板，使用T7聚合酶合成mRNA。
将显微注射溶液中的DdCBE mRNA对或DdCBE-NES mRNA对显微注射到小鼠受精卵中。
受精卵培养四天变成胚泡后，裂解胚泡。使用相同的模板，PCR扩增线粒体DNA中不同于核DNA的一部分靶位点，然后通过另外的PCR扩增索引和测序衔接子。使用IlluminaMiniSeq系统进行高通量测序，然后使用Cas分析仪(www.rgenome.net)分析碱基编辑效率。此外，使用PCR扩增细胞核中具有类似于线粒体靶区域的序列的DNA并测序。
因此，DdCBE不仅在线粒体DNA中而且在细胞核中的类似DNA序列中诱导突变(线粒体：13.1％，细胞核：3.2％)。使用DdCBE-NES，线粒体靶突变效率增加到18.2％，而细胞核DNA突变效率降低到0.2％(图41a)。对于其他靶标TrnA或Rnr2，没有发生核DNA突变，而在线粒体DNA中，碱基编辑效率在统计学上显著增加(*p<0.05，**p<0.01，n.s.：不显著)。
4-5：动物胚胎中的DdCBE和mitoTALEN
除了ND5基因特异性DdCBE之外，还注射切割未编辑的线粒体DNA序列的TALEN，以增加C至T转化后细胞中编辑的线粒体DNA的比例。显微注射方法和测序鉴定方法与实施例4-2和4-4相同。单独用DdCBE显微注射的组显示出11％的编辑效率，并且当用DdCBE和mitoTALEN处理时，效率增加至33.3％，导致编辑效率的统计学显著增加。另外，单独用DdCBE-NES显微注射的组显示编辑效率为20.5％，当用DdCBE-NES和mitoTALEN处理时，观察到效率为36.8％，这也是统计学显著的(图41b)。
同样，通过将显微注射的受精卵转移到雌性老鼠中，DdCBE在新生小鼠幼崽中显示出10.9％的编辑效率，但是当使用DdCBE-NES和mitoTALEN时获得23.4％的效率(图41c)。
当在动物线粒体基因编辑过程中核输出信号与碱基编辑蛋白结合时，碱基编辑以更高的效率实现，并且在动物胚胎中，细胞核中类似序列的非特异性碱基编辑也被抑制。此外，当共注射线粒体序列切割蛋白时，可以预期更高效率的线粒体碱基编辑。
实施例5.分裂DddAtox脱氨酶变体
提供了能够降低DdCBE脱靶效应的高精度DddA衍生的胞嘧啶碱基编辑器。这种脱靶碱基编辑效应是由DddAtox脱氨酶分裂物的自发组装引起的现象，与TALE和DNA之间的相互作用无关。因此，通过用丙氨酸取代位于DddAtox分裂物之间表面上的氨基酸残基来构建HF-DdCBE。HF-DdCBE防止了一对与TALE连接的两个脱氨酶在不与DNA结合时正常发挥功能。通过全线粒体基因组分析，证实了HF-DdCBE是非常有效和精确的，这与在人线粒体DNA中引起许多不希望的脱靶C至T转化的常规DdCBE不同。
5-1.方法
质粒构建。将点突变引入DdCBE表达质粒中。使用Q5定点诱变(NEB)的诱变引物扩增质粒(表7)，并通过Sanger测序证实其结果。
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为了组装界面突变体，将小量制备的突变体表达质粒和模块载体(每个编码TALE序列)、BsaI-HFv2(10U)、T4 DNA连接酶(200U)和反应缓冲液在一个试管中混合。之后，限制酶和连接酶反应在热循环仪中进行在37℃下5分钟和在50℃下20分钟的20个循环，随后是在50℃下15分钟和在80℃下5分钟的进一步反应。将连接的质粒通过化学转化导入大肠杆菌DH5ɑ中，并通过Sanger测序鉴定最终的构建体。为了导入细胞系，将质粒中量制备。
哺乳动物细胞系培养和转染。将HEK 293T/17(CRL-11268，美国典型培养物保藏中心(ATCC))细胞系在37℃下在5％CO2环境中培养。细胞系在补充有10％(v/v)胎牛血清的DMEM(Gibco)中在没有抗生素的情况下生长，并且不测试支原体。对于脂质转染，在转染前18-24小时，以1×105的细胞密度在24孔细胞培养板(SPL，韩国首尔)中开始细胞生长。使用Lipofectamine 2000(Invitrogen)，使用500ng每个DdCBE分裂物引入总共1,000ng质粒DNA。转染后4天收获细胞。
用于高通量测序的基因组和线粒体DNA分离。去除细胞培养基以分离基因组DNA后，将包含DNeasy血液和组织试剂盒(Qiagen)的蛋白酶K的裂解缓冲液添加至细胞培养板，以从板的底部分离细胞。然后根据制造商的方案分离基因组DNA。对于全线粒体基因组测序，将200μl线粒体分离缓冲液A(ScienCell)添加至去除细胞培养基的培养板中。使用细胞刮铲刮取细胞，然后置于微管中，随后使用一次性杵研磨细胞。研磨20次后，将充分研磨的匀浆在1,000x g和4℃下离心5分钟。将上清液置于新的微管中并在10,000x g和4℃下离心20分钟。将沉淀重悬于10μl裂解缓冲液(25mM NaOH，0.2mM EDTA，pH 10)中，然后在95℃煮沸20分钟。为了降低pH，向线粒体裂解物中添加1μl 1M HEPES(游离酸，未经pH调节)。在PCR模板链中使用1μl由此制备的溶液用于高通量测序。
高通量测序。为了构建深度测序文库，使用Q5 DNA聚合酶进行巢式初级PCR和次级PCR，并添加最终的索引序列。使用MiniSeq(Illumina)将文库用于配对末端读取测序。对于全线粒体基因组分析，使用标记DNA制备试剂盒(Illumina)根据制造商的方案制备分离的线粒体DNA。使用一个fastq-join文件合并来自所有分析的配对末端测序结果，并使用CRISPR RGEN工具(http://www.rgenome.net/)进行分析。
5-2.结果
当试图在植物中进行叶绿体编辑时，在叶绿体基因组上出现脱靶碱基突变，产生关于DdCBE准确性的问题。DdCBE的脱靶碱基编辑有两个原因。第一个是TALE蛋白与DNA之间的非特异性结合，第二个是DddAtox半部分之间无意的自发相互作用(图42a)。该研究集中在分裂DddAtox半部分，并设计两个蛋白质分裂物的界面以防止DddAtox半部分的不希望的组装。
具体地，我们检查了靶向线粒体ND1(mtND1)基因的每个亚基(左-TALE或右-TALE)是否结合DNA并与无TALE的DddAtox的另一半相互作用以引起胞嘧啶到胸腺嘧啶碱基编辑。靶向人肾胚胎细胞系(HEK293T)中的人线粒体ND1(mtND1)基因的DdCBE对(左-TALE：G1397N(与识别左半位点的左-TALE阵列的C末端融合的N末端G1397DddAtox半部分)+右-TALE：G1397C(与识别右半位点的右-TALE阵列的C末端融合的C末端G1397 DddAtox半部分))有效地编辑靶序列的C11，以60.7％的效率将胞嘧啶转化为胸腺嘧啶(图43a)。此外，与另一个无TALE的DddAtox半部分配对的每个单独的亚基(左-TALE：G1397N或右-TALE-G1397C)也诱导碱基编辑，虽然效率不如原始对。因此，与另一个无TALE的DddAtox半部分配对的每个与靶ND1序列结合的左-TALE和右-TALE诱导碱基编辑，其中效率为31％或8.1％(图43)。换句话说，具有两个TALE融合物的DdCBE对比仅具有一个TALE融合物的不匹配对仅有效2.0倍(60.7％/31％)或7.5倍(60.7％/8.1％)。显然，与结合半位点的TALE阵列融合的DddAtox N末端部分可募集无TALE阵列的DddAtox C末端部分，反之亦然，以重建功能性脱氨酶。
由于DddAtox可在两个位置(G1333和G1397)分裂，因此还构建靶向位置G1333处的mtND1基因的DdCBE对(左-TALE：G1333-N和右-TALE：G1333-C)，以测试左-TALE：G1333-N和右-TALE：G1333-C构建体是否能够募集无TALE的DddAtox半部分并诱导C至T编辑。如所预期的，与另一个无TALE的DddAtox半部分配对的每个TALE融合物在位置C8处显示出32.7％(左TALE缀合物)或18.1％(右TALE缀合物)的碱基编辑效率，而原始DdCBE对为56.1％。因此，具有两个TALE融合物的原始对比具有一个TALE融合物的不匹配对仅有效1.7倍(56.1％/32.7％)或3.1倍(56.1％/18.1％)。总之，这些结果表明，DdCBE可以在仅一个TALE阵列可以结合的位点引起不希望的脱靶突变。因为TALE蛋白可在发生一些错配下与位点结合，所以DdCBE对可能在细胞器或核基因组中诱导许多脱靶突变。
我们试图开发高保真度DdCBE，其不会表现出由分裂DddAtox半部分的自发组装引起的这种脱靶编辑。我们推测，分裂物二聚体界面可被工程化以抑制或防止自组装。为此，我们在PyMOL软件中使用Python脚本(InterfaceResidues.py)来鉴定在1平方埃范围内的两个分裂DddAtox(在G1333和G1397处分裂)的界面中的氨基酸残基。结果，我们发现G1397-N(位置G1397处的N末端DddAtox半分裂物)中的9个氨基酸残基、G1397-C(位置G1397处的C末端DddAtox半分裂物)中的4个残基、G1333-N(位置G1333处的N末端DddAtox半分裂物)中的14个氨基酸残基、以及G1333-C(位置G1333处的C末端DddAtox半分裂物)中的15个氨基酸残基(图42b和42c)。
随后，我们通过用丙氨酸取代这些氨基酸残基中的每一个产生了各种突变型DddAtox半部分。然后我们在HEK293T细胞中测量这些界面突变型DdCBE与野生型DdCBE配偶体或无TALE的DddAtox半部分组合的编辑频率。许多含有界面突变如C1376A、M1390A和F1412A的G1397-分裂物的DddAtox变体不能在两个TALE结合位点之间的间隔子区中诱导C至T转化，即使当与野生型配偶体组合时，表明这些突变体不能在靶位点附近与其他野生型DddAtox半部分相互作用。其他DddAtox变体(例如含有V1377A和E1381A的那些)与无TALE的半部分一起以高频率诱导C至T编辑，这与野生型DdCBE对是可比较的，从而表明这些突变是中性的并且不阻止分裂物二聚体相互作用。
重要的是，一些突变，例如K1389A、K1410A和T1413A，当与野生型DdCBE配偶体配对时显示高活性，但当与无TALE的半部分配对时显示低活性。例如，K1410A突变显示53.2％的效率，其类似于与野生型DdCBE配偶体配对时的效率(60.7％)，但当与无TALE的半部分配对时显示0.9％的效率，导致59.1倍的差异(＝53.2％/0.9％)。如上所述，野生型对显示出7.5倍(＝60.7％/8.1％)差异。此外，这些变体比野生型DdCBE对更有选择性地编辑碱基。因此，这些变体在编辑窗中优先于C8、C9和C13编辑C11，而野生型DdCBE对区别性小得多，以>6.7％的高频率编辑所有四个胞嘧啶(图43b)。
此外，筛选G1333的29个突变(G1333N的14个突变和G1333C的15个突变)产生了多个所需的界面突变(图44)。含有大多数这些突变(例如I1299A、Y1316A、Y1317A和F1329A)的变体即使与野生型配偶体组合也具有不良活性，或者在其他突变(例如S1300A和T1314A)的情况下，与无TALE的配偶体组合具有不期望的活性。值得注意的是，含有若干个突变(包括K1389A、T1391A和V1393A)的变体，当与野生型配偶体配对时具有高活性，但当与无TALE的配偶体配对时是低效的。例如，K1389A显示38倍差异(＝45.4％/1.2％)，而野生型DdCBE对仅显示3.1倍差异(＝56.1％/18.1％)。此外，含有K1389A变体的对比野生型对更具选择性。因此，该变体优先于C9、C11和C13编辑C8，而野生型对是混杂的，以>19％的高频率编辑所有四个胞嘧啶。还值得注意的是，G1397-C中的K1410A优先编辑C8，而G1333-C中的K1389A以更高的效率选择性编辑C11。相比之下，野生型DdCBE对(G1333或G1397 DddAtox分裂物)显示出选择性不佳。这些结果表明，上述界面突变体具有减少靶位点内多个碱基的不希望的编辑的潜力，通常用DdCBE观察到。
实施例6.全长脱氨酶
DddA衍生的胞嘧啶碱基编辑器(DdCBE)由分裂细菌间毒素DddAtox、TALE阵列和尿嘧啶糖基化酶抑制剂(UGI)构成，使得真核生物核DNA、线粒体DNA(mtDNA)和植物叶绿体DNA中的靶胞嘧啶能够被转化为胸腺嘧啶。DddAtox对细菌具有毒性，是一种衍生自洋葱伯克霍尔德菌的酶并使双链DNA中的胞嘧啶脱氨基。为了避免宿主细胞毒性，将DddAtox分裂成无活性的两个半部分，每个半部分与TALE DNA结合蛋白融合以形成DdCBE对。只有当两个无活性的半部分通过两个相邻结合的TALE蛋白一起结合到靶DNA上时，功能性脱氨酶才能重构。在两个TALE结合位点之间的14-18个碱基对(bp)的间隔子区中诱导C至T碱基转化。
与不能编辑细胞器DNA的CRISPR衍生的碱基编辑器不同，DdCBE使得能够在细胞核和细胞器DNA中进行靶向碱基编辑，但其缺点是需要两个TALE构建体而不是一个构建体来诱导这种编辑。第一个缺点是TALE必须结合在5'和3'端具有胸腺嘧啶的靶DNA位点，因此使用两个TALE阵列限制了可靶向位点。第二，两个TALE构建体代替一个TALE构建体的递送通常效率低且具有挑战性。具有有限容量的病毒载体，例如广泛用于基因治疗的腺相关病毒(AAV)载体(容量：约4.7kbp)，因为二聚体DdCBE组合太大(2×4.1kbp，包括启动子和多聚A信号)，所以不能容纳分裂DdCBE编码序列。此外，由于两个TALE阵列序列的高度相似性，因此将两个编码TALE阵列的DNA片段克隆到单个大容量载体中可能变得困难。最后，使用两个TALE阵列代替一个TALE阵列可能加剧脱靶效应。为了克服具有DddAtox分裂物的二聚体DdCBE的这些限制，我们提供了无毒的全长DddAtox融合的DdCBE，称为mDdCBE(单体DdCBE)，用于细胞核和细胞器DNA中的靶向C至T转化。
6-1.方法
质粒构建。使用合成的全长DddAtox(gBlock,IDT)作为模板并使用下表8中的引物和Q5 DNA聚合酶(NEB)通过PCR扩增DddA变体。使用Gibson组装(NEB)在p3s-BE3位点克隆这些PCR产物，在该位点Apobec1用BamHI和Sma I(NEB)消化。TALE-DddAtox(Addgene#158093、#158095、#157842、#157841)用BamHI和Sma I消化质粒，并使用表8中的引物通过PCR扩增DddA变体，然后使用Gibson组装克隆。通过热休克法将由此获得的质粒转化到化学制备的大肠杆菌DH5ɑ中，并通过Sanger测序法分析存活菌落的质粒序列。将最终的质粒进行中量制备(Macherey-Nagel)以用于细胞转染。
[表8]
随机诱变。使用合成的全长DddAtox(gBlock，IDT)作为模板，通过使用GeneMorphII随机诱变试剂盒(Agilent)根据制造商的方案进行易错PCR。总之，使用1ng、100ng和700ng DddAtox DNA作为模板引入0-16个突变/kb的随机突变。使用表8中的引物通过PCR预先扩增全长DddAtox gBlock。将所有PCR产物合并并使用Gibson组装(NEB)克隆到用Sma1和Xho1消化的p3s-UGI-Cas9(H840A)中。通过热休克法将化学制备的大肠杆菌DH5α用质粒转化，并通过Sanger测序分析存活菌落的质粒序列。在分析的质粒中，将具有编码框架的p3s-UGI-nCas9(H840A)-DddAtox质粒与sgRNA一起转染到HEK293T细胞中，然后通过靶向深度测序测定编辑活性。
哺乳动物细胞培养和转染。将HEK293T(ATCC，CRL-11268)细胞和HeLa(ATCC，CCL-2)细胞在37℃下在5％CO2中培养。将细胞在补充有10％(v/v)胎牛血清(Welgene)和1％青霉素/链霉素(Welgene)的DMEM中培养。在转染前24小时将细胞以3×105细胞(HEK293T)和4×104细胞(HeLa)的密度接种到48孔板(Corning)中，然后使用Lipofectamine 2000(Invitrogen)用Cas9融合的DddA质粒(750ng)和sgRNA(250ng)转染。使用200ng质粒和Lipofectamine 2000将TALE-DddA转染到HEK293T细胞中。在此，sgRNA序列示于下表9中。
[表9]
名称 sgRNA序列5至3 No.
HEK3 GGCCCAGACTGAGCACGTGA 397
TYRO3 GGCCACACTAGCGTTGCTGC 398
ROR1位点1 GCCATAGATGGTGGACCGAA 399
ROR1位点2 CCATCTATGGCTCTCGGCTG 400
ROR1位点3 CCGCAGCCGAGAGCCATAGA 401
FANCF GGAATCCCTTCTGCAGCACC 402
HBB CTTGCCCCACAGGGCAGTAA 403
EMX1-1 TGCCCCTCCCTCCCTGGCCC 404
EMX1-2 CCCTGGCCCAGGTGAAGGTG 405
EMX1-3 GTGAAGGTGTGGTTCCAGAAC 406
EMX1-4 AAAGTACAAACGGCAGAAGC 407
TRAC5位点1 GTGGTAGCGGAACTCACTAAG 408
TRAC5位点2 CACCCAGCCTGCTCTGCCTT 409
基因组和线粒体DNA制备。转染后2天收获用Cas9融合的DddA变体转染的细胞，以及转染后3天收获用TALE-DddA转染的细胞。使用DNeasy血液和组织试剂盒(Qiagen)分离基因组和线粒体DNA。为了大规模分析，使用含有5μl蛋白酶K(Qiagen)的100μl细胞裂解缓冲液(50mM Tris-HCl(pH 8.0)(Sigma-Aldrich)、1mM EDTA(Sigma-Aldrich)、0.005％十二烷基硫酸钠(Sigma-Aldrich))提取DNA。使裂解物在55℃下反应1小时，然后在95℃下反应10分钟。
6-2.结果
比较野生型和新的全长DddA变体的氨基酸序列。改变的氨基酸在图45中用灰色框表示。
如图46所示，使用由16个氨基酸构成的接头将DddA连接到Cas9的N末端的上游，而使用由4个氨基酸构成的接头将UGI(尿嘧啶糖基化酶抑制剂)和NLS(核定位信号)连接到C末端。相反地，使用由16个氨基酸构成的接头将DddA连接到Cas9的C末端的下游，而使用由4个氨基酸构成的接头将UGI和NLS连接到N末端。
在本发明中，我们构建并使用DddA-Cas9(D10A、D10A和H840A)-UGI。与锌指蛋白或TALE模块融合的全长单个DddA模块能够进行胞嘧啶到胸腺嘧啶编辑。当前的分裂物系统需要两个模块，但是全长DddA只需要一个模块。这两个DNA结合蛋白可与NLS(核定位信号)、MTS(线粒体靶向序列)或CTP(叶绿体转运肽)连接，使得不仅在核基因组中，而且在不能使用Cas9编辑的线粒体和植物叶绿体基因组中可以用胸腺嘧啶取代胞嘧啶。如图47所示，在人细胞基因组背景ROR1位点(a)、HEK3位点(b)和TYRO3位点(c)中，证实了用胸腺嘧啶取代TC基序中的胞嘧啶的活性。证实了用胸腺嘧啶取代TC基序中距离靶位置25bp的胞嘧啶的活性(a)。对于A1341D KRKKA，证实了用胸腺嘧啶取代CC基序中的第二个胞嘧啶的活性(a，b)。对于催化突变体E1347A，也证实了用胸腺嘧啶取代TC基序中的胞嘧啶的活性(a，b，c)。红色下划线表示Cas9的结合位点。效率表示为总测序读段中无插入缺失读段中胞嘧啶至胸腺嘧啶转化的百分比。此外，总测序读段中的插入缺失比率以百分比表示。
如图48所示，红色方形框表示证实分裂DddAtox活性的部分，并且通过使用全长DddA将相同部分分成三个靶位点来测量活性。对于分裂物，使用具有不同PAM的正交Cas9将两个Cas9之间的胞嘧啶转化为胸腺嘧啶。因此，用胸腺嘧啶精确取代所需的胞嘧啶是困难的。然而，对于全长DddA，可将其中Cas9结合同一靶位点的部分分为三个并靶向，使得能够用胸腺嘧啶精确取代所需的胞嘧啶。效率表示为总测序读段中无插入缺失读段中胞嘧啶至胸腺嘧啶转化的百分比。此外，总测序读段中的插入缺失比率以百分比表示。
如图49所示，在人细胞基因组背景TRAC位点1(a)、TRAC位点2(b)、FANCF(c)和HBB(d)中测量全长DddA的活性。红色下划线表示Cas9的结合位点。效率表示为总测序读段中无插入缺失读段中胞嘧啶至胸腺嘧啶转化的百分比。此外，总测序读段中的插入缺失比率以百分比表示。
如图50所示，使用DddA-dCas9(D10A,H840A)-UGI在人细胞基因组背景TYRO3(a)、ROR1(b)、HEK3(c)、EMX1位点2(d)、TRAC位点1(e)和HBB(f)中测量DddA的活性。效率表示为总测序读段中胞嘧啶至胸腺嘧啶转化的百分比。没有观察到插入缺失。
为了获得用于碱基编辑的无毒全长DddAtox变体，使用两种方法：基于结构的位点特异性诱变和随机诱变。在第一种方法中，将具有降低的DNA结合或降低的催化活性的DddAtox变体与无活性CRISPR-Cas9(dCas9)或切口酶(nCas9)变体融合，以开发新的碱基编辑器，其中在培养的人细胞中用胸腺嘧啶取代靶胞嘧啶。为此，用丙氨酸取代DddAtox的带正电荷的氨基酸，并将其亚克隆到表达载体中(图51a)。假定这些变体可能通过减弱与带负电荷的dsDNA的结合而避免毒性。大多数丙氨酸取代的变体不能形成大肠杆菌转化体(图51b)。基于从所得转化体分离的质粒DNA的测序分析，在蛋白编码区诱导了各种移码突变。该全长DddAtox变体尽管在哺乳动物启动子的控制下，但在大肠杆菌中表达较弱，从而导致细胞死亡。幸运的是，可以获得多种没有移码突变的三重、四重或五重(称为“AAAAA”)丙氨酸取代的变体。还成功克隆了活性位点突变E1347A。
此外，我们研究了与D10A nCas9或dCas9和UGI融合的AAAAA变体是否可以在人胚肾293T(HEK293T)细胞中诱导碱基编辑(图51c和51d)。由大鼠APBEC1脱氨酶、尿嘧啶糖基化酶抑制剂(UGI)和dCas9(或D10A nCas9)构成的碱基编辑器2(或3)在原型间隔子内的窄区域中具有活性，而AAAAA变体在原型间隔子的5'上游以高达43％的效率诱导胞嘧啶向胸腺嘧啶的转化。出乎意料地，E1347A突变在相同C-3位置以37％(nCas9融合)或16％(dCas9融合)的频率诱导碱基编辑(图51c)，这表明E1347A突变没有完全灭活DddAtox的脱氨酶活性，并且E1347A突变体的残余脱氨酶活性足够高以在人细胞中实现碱基编辑。然而，与五重AAAAAA突变结合的E1347A变体不能诱导碱基编辑。此外，证实了E1347A、AAAAA和其他丙氨酸取代的变体(图53)，在没有与dCas9或nCas9和UGI融合的移码突变的情况下，在距原型间隔子上游最多25个碱基的位置诱导编辑，并且在各种不同位点也表现出高达26％的编辑效率(图54)。此外，编辑在HeLa细胞中是非常有效的，效率高达60％(图55)。融合蛋白诱导的碱基编辑在细胞中维持长达21天，表明这种碱基编辑没有细胞毒性(图56)。
为了改变胞嘧啶碱基编辑器的编辑窗，尝试将丙氨酸取代的变体融合到H840AnCas9的C末端。出乎意料的是，没有获得无移码突变的完整构建体。因此，进行易错PCR以将随机突变引入DddAtox编码序列中，并获得具有四个点突变S1326G、G1348S、A1398V和S1418G的无毒全长DddAtox变体(称为“GSVG”)(对于SEQ ID NO:269的氨基酸序列中的S1326G、G1348S、A1398V和S1418G，位置37处的S被G取代；位置59处的G被S取代；位置109处的A被V取代；位置129位的S被G取代，包括SEQ ID NO:276的序列，图52a)。此外，将这些变体融合到dCas9、D10A nCas9和Cas9的C末端以及dCas9、nCas9和Cas9的N末端。在人细胞中，除了野生型Cas9之外，这些融合蛋白在不同位点以高达38％的效率诱导胞嘧啶到胸腺嘧啶的转化(图52b，57和58)。有趣的是，含有与dCas9、D10AnCas9和H840A nCas9的C末端融合的GSVG变体的融合蛋白引起原型间隔子相邻基序(PAM)3'下游的胞嘧啶碱基编辑，而含有与其N末端融合的相同变体的融合蛋白dCas9和nCas9诱导原型间隔子5'上游的碱基编辑(图52c)。如所预期的，含有Cas9的融合蛋白导致插入缺失而不是碱基取代。
为了找出GSVG变体中哪些突变是重要的，尝试通过定点诱变构建SSVG、GGVG、GSAG和GSVS的四种回复体。获得SSVG、GSAG和GSVS回复体，但未获得与nCas9的C末端融合的GGVG变体。G1348紧靠E1347，E1347是催化位点的关键。G1348S突变降低了催化活性，避免了对大肠杆菌的细胞毒性。测定三种回复体和GSVG变体在转染细胞中两个靶位点处长达21天的编辑频率。由GSAG和GSVS诱导的胞嘧啶到胸腺嘧啶编辑的频率从转染后第3天到第21天逐渐降低到约一半，因此这两种回复体在一定程度上具有细胞毒性，而GSVG和SSVG被保留(图59)。这些结果表明，在GSVG变体中，G1348S是必需的，S1326G是中性的，而A1398V和S1418G降低细胞毒性。
总之，我们的结果表明，对dsDNA具有降低的亲和力(AAAAA)、减弱的脱氨酶活性(E1347A和可能的GSVG)或降低的细胞毒性(GSVG)的无毒全长DddAtox变体可以与dCas9或nCas9融合以产生具有改变的编辑窗的新的碱基编辑器。这些碱基编辑器，称为dCas9-mDdBE(由与dCas9的C末端融合的全长单体DddAtox变体构成的DddA衍生的碱基编辑器)、nCas9-mDdBE、mDdCE-dCas9和mDdCE-nCas9，可用于在超出BE2或BE3范围的原型间隔子区上游或下游位置进行碱基编辑。
我们还研究了无毒全长DddAtox变体是否可用于线粒体DNA编辑。在各种变体中，只有两种变体即GSVG和E1347A成功地融合到设计用于结合线粒体基因ND4和ND6的TALE阵列的C末端。包括GSVG变体的单体DdCBE(mDdCBE)在靶核苷酸位置实现了碱基编辑，其中效率高达31％(ND4)(图60a)和27％(ND6)(图60b)，等同于原始分裂DdCBE对。含有E1347A的mDdCBE也将靶胞嘧啶转化为胸腺嘧啶，但效率降低，其中编辑速率至多7.2％(ND4)和8.9％(ND6)。有趣的是，ND4基因特异性原始DdCBE对(G1333分裂物)在位置C4具有0.8％的编辑效率，而含有GSVG的两个mDdCBE显示26％和31％的高编辑效率。这些结果表明，分裂物二聚体DdCBE和mDdCBE具有不同的突变模式，表明mDdCBE可能与二聚体DdCBE互补，在给定靶位点诱导各种突变。
mDdCBE相对于分裂物二聚体DdCBE的一个潜在优点是由于非特异性TALE-DNA相互作用引起的脱靶效应与二聚体DdCBE相比减半。具有分裂DddAtox的二聚体DdCBE能够在仅一个亚基可结合的半位点上操作，导致不需要的脱靶突变。DdCBE对的失活的DddAtox半部分可募集另一失活的半部分以形成功能性脱氨酶。为了证实该假说，用编码二聚体DdCBE的一个亚基的质粒和编码无TALE的DddAtox半部分的质粒共转染HEK293T细胞，并在两个线粒体靶位点测量编辑频率。如所预期的，在靶位点观察到胞嘧啶到胸腺嘧啶的编辑，频率为0.7％至3.6％(图60c-图60f)。这些结果表明，不需要的脱靶突变可能是由DdCBE对中分裂DddAtox半部分在半位点处彼此相互作用引起的，并且mDdCBE可以避免由二聚体DdCBE引起的一半脱靶突变。
实施例7.使用DdABE在人细胞中进行高效的A至G碱基编辑
通过DddA衍生的胞嘧啶碱基编辑器(DdCBE)进行的线粒体DNA碱基编辑使得能够在各种细胞系和动物中建立疾病模型，开辟了治疗线粒体遗传疾病的新途径。然而，由于DdCBE几乎完全导致TC到TT碱基编辑，因此它能够仅覆盖所有情况的约1/8。因此，TALE连接脱氨酶(TALED)是通过将两种类型的脱氨酶与TALE(转录激活因子样效应物)连接而开发的。在此，TALE被定制设计为结合所需的DNA部分，并融合到没有催化活性的DddAtox胞嘧啶脱氨酶变体和TadA蛋白，所述TadA蛋白是衍生自大肠杆菌的DNA腺嘌呤脱氨酶。TALED能够对A到G转化进行碱基编辑，这与传统的碱基编辑技术不同，在传统的碱基编辑技术中，胞嘧啶碱基编辑仅适用于人线粒体中的TC背景。事实上，定制的TALED能够在人细胞中的各种靶标上高效地(高达约50％)诱导腺嘌呤碱基编辑。
为了开发新的碱基编辑技术，ABE8e的TadA变体(TadA*)选自各种TadA变体。这是因为这样的变体能够以高效率诱导腺嘌呤编辑，并且被改进为与各种DNA结合蛋白相容，因此在实际应用中与TALE或ZFP(锌指蛋白)有效相容。
将TadA*和MTS(线粒体靶向序列)融合至针对ND1或ND4靶位点定制的TALE，并且测试在线粒体DNA中是否可以实际上发生碱基编辑。基于靶向深度测序的结果，发现融合蛋白的腺苷碱基编辑效率非常低但可检测。诱导腺嘌呤碱基编辑，其中在ND1位点的效率高达1.2％(图67a)，在ND4位点的效率高达0.6％(图67b)。虽然已知TadA*仅特异性作用于单链DNA，但发现当TadA*与TALE融合时，双链靶DNA中的碱基编辑也被诱导，尽管效率非常低。
由于腺嘌呤碱基编辑可能发生在线粒体DNA中，我们试图通过融合DddAtox蛋白来提高效率。DddAtox蛋白是衍生自洋葱伯克霍尔德菌的细菌间毒素，其使胞嘧啶脱氨基。该蛋白质作用于双链DNA，因此可以帮助TadA*腺嘌呤脱氨酶更好地接近靶DNA。对于使用DddAtox的现有DdCBE，将DddAtox蛋白分裂成两个半部分，然后分别将这两个半部分融合到识别左-半DNA位点的左-TALE(L-TALE)和识别右-半DNA位点的右-TALE(R-TALE)以及融合到增加胞嘧啶碱基编辑效率的尿嘧啶糖基化酶抑制剂(UGI)(TALE-分裂DddAtox-UGI)。DddAtox以分裂形式使用的原因是使用全长蛋白质引起细胞毒性。具体地，将TadA*代替UGI连接到靶向ND1位点的DdCBE的任一侧，并且制备和测试L-TALE-分裂DddAtox-TadA*和R-TALE-分裂DddAtox-UGI，或L-TALE-分裂DddAtox-UGI和R-TALE-1397C-TadA*形式。奇怪的是，证实了当一侧的TadA*和另一侧的1397N和UGI配对并转移到人细胞时，发生A至G和C至T转化(图62c)。在传统的DdCBE中，胞嘧啶碱基编辑以约20％的效率发生，并且根本没有发生腺嘌呤碱基编辑，而当UGI在任一侧被TadA*替换时，胞嘧啶碱基编辑减少至约一半，并且腺嘌呤碱基编辑以约10％的效率发生(图62c)。简单地说，通过将TadA变体与DdCBE中分裂DddAtox半部分融合产生的TALE脱氨酶可在人mtDNA中同时诱导A至G和C至T编辑(图62c)。在传统的DdCBE中，胞嘧啶碱基编辑以约20％的效率发生，并且根本没有发生腺嘌呤碱基编辑，而当TadA*提供在任一侧时，胞嘧啶碱基编辑减少至约一半，并且腺嘌呤碱基编辑以约10％的效率发生(图62c)。简言之，腺嘌呤碱基编辑和胞嘧啶碱基编辑效率相似(图62c)。
同时进行胞嘧啶碱基编辑和腺嘌呤碱基编辑可用于随机诱变，但在治疗疾病，尤其是线粒体遗传疾病如由C至T突变引起的LOHN和MEALS中，希望仅诱导腺嘌呤碱基编辑。因此，为了消除这种同时发生的胞嘧啶碱基编辑，去除UGI。在DdCBE中，当胞嘧啶脱氨酶DddAtox将C脱氨基为U时，为了防止U被尿嘧啶糖基化酶再次修复，尿嘧啶糖基化酶是DNA修复过程中细胞中的修复蛋白，UGI作为尿嘧啶糖基化酶抑制剂被融合。因此，认为如果去除这种UGI，将维持腺嘌呤碱基编辑效率，并抑制胞嘧啶碱基编辑。令人惊讶的是，证实了不含UGI的靶向ND1的TALE脱氨酶对几乎不引起胞嘧啶碱基编辑(<0.5％)，并且以高效率(约50％)单独诱导腺嘌呤碱基编辑(图63a和图63c)，这比具有UGI高得多。这也在靶向ND4的TALE脱氨酶对中得到证实。与靶向ND1一样，以高效率(约35％)仅检测到腺嘌呤编辑(图63b和图63d)。以这种方式，开发了TALED，它是一种作用于双链DNA的新的腺嘌呤脱氨酶，其中融合了DddAtox系统和TadA*，并且腺嘌呤碱基编辑在人线粒体中首次成为可能。此外，与单独与TadA*融合的TALE相比，腺嘌呤碱基编辑最终以高出约50倍的效率诱导。
此外，尝试使用消除了催化活性的全长E1347A DddAtox变体或维持了催化活性但消除了细胞毒性的变体(AAAAA和GSVG)诱导腺嘌呤碱基编辑。由于腺嘌呤碱基编辑而不是胞嘧啶碱基编辑是在单链DNA中发生，因此缺乏胞嘧啶碱基编辑活性的全长E1347ADddAtox变体仍可用于通过促进双链DNA对TadA的可接近性来增强A至G编辑效率。此外，基于胞嘧啶碱基编辑在不存在UGI时无效的结果，可使用仅消除细胞毒性的变体。制备了两种类型的含有全长变体的TALED(图64a)。第一种类型被配置为使得TadA*(AD)和全长DddAtox变体都包含在一个TALE(mTALED)中，第二种类型被配置为使得TadA*(AD)和全长DddAtox变体分别融合到各自的TALE(dTALD)，并且测试这两种类型(图64a)。令人惊讶的是，证实了两种类型的靶向ND1的TALED以高效率诱导腺嘌呤碱基编辑(图64b)。这里，mTALED表现出高达约45％的效率，dTALED也显示约50％的腺嘌呤碱基编辑效率(图64b)。除了ND1位点外，在ND4位点也进行类似的实验，并以类似的高效率诱导腺嘌呤碱基编辑(图64c)。此外，当使用没有胞嘧啶碱基编辑活性的全长E1347A DddAtox变体时，以高效率诱导腺嘌呤碱基编辑(图64b和图64c)。这被认为是因为即使在不存在胞嘧啶脱氨基活性的情况下也保留了帮助TadA*充分接近双链DNA的作用。当在单核苷酸水平上详细综述上述结果时(图65和图66)，在TALE与DNA结合的紧邻区域诱导腺嘌呤碱基编辑。此外，当使用两个TALE时，仅在它们之间(间隔子)诱导碱基编辑，并且奇怪的是，发现即使在使用一个TALE的mTALED中，靶长度也是相似的(图65和66)。
此外，我们研究了该系统是否在核DNA中与锌指蛋白(ZFP)系统共同起作用。因此，产生靶向核DNA的NC型ZFP，并将分裂DddAtox和TadA*与其融合(图61a)。这里，将TadA*融合到ZFP的不同位置(图61b)。在各种构建体中，产生了能够在核DNA中以高达10％的效率诱导腺嘌呤碱基编辑的构建体(图61d)。由于UGI存在于任一侧，因此胞嘧啶碱基编辑效率也很高(图61c)。ZFP-DddAtox-TadA*系统在人细胞核DNA中起作用，此外，还测试了它是否也在线粒体中起作用。因此，参考在核DNA中以最高效率起作用的构建体。实验以这样的方式进行，其中代替针对核DNA的核定位信号(NLS)，将针对线粒体的线粒体靶向序列(MTS)连接到其上，并且将1397N融合到靶向ND1位点的ZFP的右ZFP，并且将TadA*和1397C融合到左ZFP。结果，以约3％的效率诱导腺嘌呤碱基编辑(图61f)。尽管腺嘌呤碱基编辑效率低于TALED，但是如果优化各种条件如连接蛋白质的接头等，该ZFP系统也能够以良好的效率诱导腺嘌呤碱基编辑。
迄今为止，基因编辑技术已经取得了显著的进步。基于CRISPR的基因剪刀(CRISPRCas9、碱基编辑器、引导编辑器等)已经通过改进脱靶编辑和提高效率以各种方式得到了开发。然而，尽管有许多进展，但线粒体遗传疾病的治疗仍有局限性。这是因为，与蛋白质不同，在包括催化蛋白质和作为靶的指导的gRNA的基于CRISPR的技术中，不存在将gRNA转移到线粒体的方法。因此，除了通过切割DNA消除线粒体DNA之外，还没有处理线粒体基因的技术。David R.Liu在美国的团队首次引入了能够在线粒体中诱导碱基编辑的DdCBE。由于DdCBE含有作用于双链DNA的胞嘧啶脱氨酶DddAtox，因此将其与DNA结合蛋白TALE融合以诱导碱基编辑。然而，由于DdCBE仅在TC背景中引起有限的碱基编辑，因此在创建疾病模型时或在真实世界应用的情况下治疗遗传疾病时存在许多限制。因此，首次产生了能够在线粒体中诱导腺嘌呤碱基编辑的TALED。TALED具有高达50％的高效率，并在靶位点诱导各种类型腺嘌呤的碱基编辑。此外，TALED可在存在UGI的情况下诱导胞嘧啶碱基编辑和腺嘌呤碱基编辑，因此可用于随机诱变，而它可用作特定腺嘌呤碱基编辑技术，因为在不存在UGI的情况下不发生胞嘧啶碱基编辑而仅诱导腺嘌呤碱基编辑。它也适用于ZFP系统，并且腺嘌呤碱基编辑可用于核DNA。TALED的开发将为许多线粒体遗传疾病提供解决方案，使得有可能创建与其对应的疾病模型，并且TALED将可用于尚未开创的许多线粒体基因相关研究。
尽管已在上文详细地公开了本发明的具体实施方案，但是对于本领域的普通技术人员将明显的是描述仅仅是优选的示例性实施方案，而不应被解释为限制本发明的范围。因此，本发明的实质范围将由所附权利要求及其等同物定义。
【工业实用性】
根据本发明，通过在DdCBE中取代胞嘧啶脱氨酶分裂物的界面中的特定氨基酸残基，可以降低不需要的胞嘧啶脱氨酶的非选择性。
关于全长胞嘧啶脱氨酶，可以编辑用传统胞嘧啶碱基编辑器难以编辑的部分。在目前的胞嘧啶碱基编辑器中用作脱氨酶的Apobec1已知是癌基因，并且其用于治疗目的的用途是有限的，但是本文开发的全长脱氨酶可能没有这样的问题。
本发明小至约2.5kb，包括DNA结合蛋白，因此可用于使用AAV载体的基因疗法，促进mRNA和RNP的递送，并能够使用原核生物生产有用的材料。
【序列表自由文本】
附加电子文件。
<110> 基础科学研究院
<120> 靶向脱氨酶及使用其进行的碱基编辑
<130> PP-B2678
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Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln Ser Ala Leu Phe
50 55 60
Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe His Asn Asn Pro
65 70 75 80
Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu Thr Leu Leu Pro
85 90 95
Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly Ala Ile Pro Val
100 105 110
Ala Ala Gly Ala Thr Gly Glu Thr Ala Val Phe Thr Gly Asn Ser Asn
115 120 125
Ser Pro Ala Ser Pro Thr Ala Gly Gly Cys
130 135
<210> 14
<211> 138
<212> PRT
<213> Artificial Sequence
<220>
<223> AAAAK
<400> 14
Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro
1 5 10 15
Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala
20 25 30
Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly Pro Thr Pro Tyr
35 40 45
Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln Ser Ala Leu Phe
50 55 60
Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe His Asn Asn Pro
65 70 75 80
Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu Thr Leu Leu Pro
85 90 95
Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly Ala Ile Pro Val
100 105 110
Ala Ala Gly Ala Thr Gly Glu Thr Ala Val Phe Thr Gly Asn Ser Asn
115 120 125
Ser Pro Ala Ser Pro Thr Lys Gly Gly Cys
130 135
<210> 15
<211> 138
<212> PRT
<213> Artificial Sequence
<220>
<223> AAKAA
<400> 15
Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro
1 5 10 15
Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala
20 25 30
Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly Pro Thr Pro Tyr
35 40 45
Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln Ser Ala Leu Phe
50 55 60
Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe His Asn Asn Pro
65 70 75 80
Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu Thr Leu Leu Pro
85 90 95
Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly Ala Ile Pro Val
100 105 110
Ala Ala Gly Ala Thr Gly Glu Thr Lys Val Phe Thr Gly Asn Ser Asn
115 120 125
Ser Pro Ala Ser Pro Thr Ala Gly Gly Cys
130 135
<210> 16
<211> 138
<212> PRT
<213> Artificial Sequence
<220>
<223> AAKAK
<400> 16
Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro
1 5 10 15
Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala
20 25 30
Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly Pro Thr Pro Tyr
35 40 45
Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln Ser Ala Leu Phe
50 55 60
Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe His Asn Asn Pro
65 70 75 80
Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu Thr Leu Leu Pro
85 90 95
Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly Ala Ile Pro Val
100 105 110
Ala Ala Gly Ala Thr Gly Glu Thr Lys Val Phe Thr Gly Asn Ser Asn
115 120 125
Ser Pro Ala Ser Pro Thr Lys Gly Gly Cys
130 135
<210> 17
<211> 138
<212> PRT
<213> Artificial Sequence
<220>
<223> KAAAA
<400> 17
Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro
1 5 10 15
Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala
20 25 30
Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly Pro Thr Pro Tyr
35 40 45
Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln Ser Ala Leu Phe
50 55 60
Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe His Asn Asn Pro
65 70 75 80
Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu Thr Leu Leu Pro
85 90 95
Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly Ala Ile Pro Val
100 105 110
Lys Ala Gly Ala Thr Gly Glu Thr Ala Val Phe Thr Gly Asn Ser Asn
115 120 125
Ser Pro Ala Ser Pro Thr Ala Gly Gly Cys
130 135
<210> 18
<211> 138
<212> PRT
<213> Artificial Sequence
<220>
<223> E1347A
<400> 18
Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro
1 5 10 15
Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala
20 25 30
Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly Pro Thr Pro Tyr
35 40 45
Pro Asn Tyr Ala Asn Ala Gly His Val Ala Gly Gln Ser Ala Leu Phe
50 55 60
Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe His Asn Asn Pro
65 70 75 80
Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu Thr Leu Leu Pro
85 90 95
Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly Ala Ile Pro Val
100 105 110
Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr Gly Asn Ser Asn
115 120 125
Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys
130 135
<210> 19
<211> 138
<212> PRT
<213> Artificial Sequence
<220>
<223> GSVG
<400> 19
Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro
1 5 10 15
Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala
20 25 30
Gly Gly Leu Glu Gly Lys Val Phe Ser Ser Gly Gly Pro Thr Pro Tyr
35 40 45
Pro Asn Tyr Ala Asn Ala Gly His Val Glu Ser Gln Ser Ala Leu Phe
50 55 60
Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe His Asn Asn Pro
65 70 75 80
Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu Thr Leu Leu Pro
85 90 95
Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly Val Ile Pro Val
100 105 110
Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr Gly Asn Ser Asn
115 120 125
Gly Pro Lys Ser Pro Thr Lys Gly Gly Cys
130 135
<210> 20
<211> 138
<212> PRT
<213> Artificial Sequence
<220>
<223> SSVG
<400> 20
Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro
1 5 10 15
Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala
20 25 30
Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly Pro Thr Pro Tyr
35 40 45
Pro Asn Tyr Ala Asn Ala Gly His Val Glu Ser Gln Ser Ala Leu Phe
50 55 60
Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe His Asn Asn Pro
65 70 75 80
Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu Thr Leu Leu Pro
85 90 95
Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly Val Ile Pro Val
100 105 110
Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr Gly Asn Ser Asn
115 120 125
Gly Pro Lys Ser Pro Thr Lys Gly Gly Cys
130 135
<210> 21
<211> 138
<212> PRT
<213> Artificial Sequence
<220>
<223> GSAG
<400> 21
Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro
1 5 10 15
Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala
20 25 30
Gly Gly Leu Glu Gly Lys Val Phe Ser Ser Gly Gly Pro Thr Pro Tyr
35 40 45
Pro Asn Tyr Ala Asn Ala Gly His Val Glu Ser Gln Ser Ala Leu Phe
50 55 60
Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe His Asn Asn Pro
65 70 75 80
Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu Thr Leu Leu Pro
85 90 95
Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly Ala Ile Pro Val
100 105 110
Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr Gly Asn Ser Asn
115 120 125
Gly Pro Lys Ser Pro Thr Lys Gly Gly Cys
130 135
<210> 22
<211> 138
<212> PRT
<213> Artificial Sequence
<220>
<223> GSVS
<400> 22
Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro
1 5 10 15
Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala
20 25 30
Gly Gly Leu Glu Gly Lys Val Phe Ser Ser Gly Gly Pro Thr Pro Tyr
35 40 45
Pro Asn Tyr Ala Asn Ala Gly His Val Glu Ser Gln Ser Ala Leu Phe
50 55 60
Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe His Asn Asn Pro
65 70 75 80
Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu Thr Leu Leu Pro
85 90 95
Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly Val Ile Pro Val
100 105 110
Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr Gly Asn Ser Asn
115 120 125
Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys
130 135
<210> 23
<211> 44
<212> PRT
<213> Artificial Sequence
<220>
<223> Wild-type DddAtox G1333-N
<400> 23
Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro
1 5 10 15
Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala
20 25 30
Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly
35 40
<210> 24
<211> 94
<212> PRT
<213> Artificial Sequence
<220>
<223> Wild-type G1333-C
<400> 24
Pro Thr Pro Tyr Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln
1 5 10 15
Ser Ala Leu Phe Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe
20 25 30
His Asn Asn Pro Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu
35 40 45
Thr Leu Leu Pro Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly
50 55 60
Ala Ile Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr
65 70 75 80
Gly Asn Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys
85 90
<210> 25
<211> 108
<212> PRT
<213> Artificial Sequence
<220>
<223> Wild-type DddAtox G1397-N
<400> 25
Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro
1 5 10 15
Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala
20 25 30
Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly Pro Thr Pro Tyr
35 40 45
Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln Ser Ala Leu Phe
50 55 60
Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe His Asn Asn Pro
65 70 75 80
Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu Thr Leu Leu Pro
85 90 95
Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly
100 105
<210> 26
<211> 30
<212> PRT
<213> Artificial Sequence
<220>
<223> Wild-type DddAtox G1397-C
<400> 26
Ala Ile Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr
1 5 10 15
Gly Asn Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys
20 25 30
<210> 27
<211> 138
<212> DNA
<213> Artificial Sequence
<220>
<223> Wild-type DddAtox G1333-N
<400> 27
ggctctggtt cctacgccct gggtccatat cagattagtg ctccccaact ccccgcctac 60
aacggtcaga cagtggggac cttttactat gtcaacgacg ccgggggatt ggaatccaag 120
gttttctcta gcggtggg 138
<210> 28
<211> 282
<212> DNA
<213> Artificial Sequence
<220>
<223> Wild-type G1333-C
<400> 28
ccaacacctt atcctaacta cgctaacgcc gggcacgtcg aggggcagtc agctcttttt 60
atgagagata acggcattag cgaagggctt gtgttccata ataatcctga gggcacctgt 120
ggcttctgtg taaatatgac cgaaacactt ctgcctgaga acgctaaaat gactgtcgta 180
ccacccgaag gcgcaatccc agttaaacgg ggcgcaaccg gcgaaaccaa agtattcacc 240
ggaaacagca atagtccaaa gtcccccacc aagggaggtt gc 282
<210> 29
<211> 324
<212> DNA
<213> Artificial Sequence
<220>
<223> Wild-type DddAtox G1397-N
<400> 29
ggtagctacg cacttggtcc ttaccagatt agcgcacccc aactccccgc ctataatggt 60
caaaccgtcg ggacctttta ctacgtaaac gatgctggtg ggctggaatc caaagtattc 120
tcctcagggg gccctacacc ctaccccaac tacgccaatg ctggtcatgt agaagggcag 180
tcagcactgt ttatgcgcga taatggtata agcgaggggt tggtcttcca taacaaccca 240
gagggtactt gtggcttctg tgtgaatatg actgaaaccc ttctgcccga aaatgccaag 300
atgactgtcg tcccacctga aggc 324
<210> 30
<211> 90
<212> DNA
<213> Artificial Sequence
<220>
<223> Wild-type DddAtox G1397-C
<400> 30
gccatacctg tgaagcgggg agcaacaggg gagacaaagg tgttcacagg caactctaac 60
agtccaaaga gccccaccaa aggcgggtgt 90
<210> 31
<211> 44
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 31
Gly Ser Ala Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro
1 5 10 15
Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala
20 25 30
Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly
35 40
<210> 32
<211> 138
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 32
ggctctggtt ccgcagccct gggtccatat cagattagtg ctccccaact ccccgcctac 60
aacggtcaga cagtggggac cttttactat gtcaacgacg ccgggggatt ggaatccaag 120
gttttctcta gcggtggg 138
<210> 33
<211> 44
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 33
Gly Ser Tyr Ala Ala Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro
1 5 10 15
Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala
20 25 30
Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly
35 40
<210> 34
<211> 138
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 34
ggctctggtt cctacgccgc aggtccatat cagattagtg ctccccaact ccccgcctac 60
aacggtcaga cagtggggac cttttactat gtcaacgacg ccgggggatt ggaatccaag 120
gttttctcta gcggtggg 138
<210> 35
<211> 44
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 35
Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ala Ser Ala Pro Gln Leu Pro
1 5 10 15
Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala
20 25 30
Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly
35 40
<210> 36
<211> 138
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 36
ggctctggtt cctacgccct gggtccatat caggcaagtg ctccccaact ccccgcctac 60
aacggtcaga cagtggggac cttttactat gtcaacgacg ccgggggatt ggaatccaag 120
gttttctcta gcggtggg 138
<210> 37
<211> 44
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 37
Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ala Ala Pro Gln Leu Pro
1 5 10 15
Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala
20 25 30
Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly
35 40
<210> 38
<211> 138
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 38
ggctctggtt cctacgccct gggtccatat cagattgcag ctccccaact ccccgcctac 60
aacggtcaga cagtggggac cttttactat gtcaacgacg ccgggggatt ggaatccaag 120
gttttctcta gcggtggg 138
<210> 39
<211> 44
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 39
Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro
1 5 10 15
Ala Tyr Asn Gly Gln Thr Ala Gly Thr Phe Tyr Tyr Val Asn Asp Ala
20 25 30
Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly
35 40
<210> 40
<211> 138
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 40
ggctctggtt cctacgccct gggtccatat cagattagtg ctccccaact ccccgcctac 60
aacggtcaga cagcagggac cttttactat gtcaacgacg ccgggggatt ggaatccaag 120
gttttctcta gcggtggg 138
<210> 41
<211> 44
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 41
Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro
1 5 10 15
Ala Tyr Asn Gly Gln Thr Val Ala Thr Phe Tyr Tyr Val Asn Asp Ala
20 25 30
Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly
35 40
<210> 42
<211> 138
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 42
ggctctggtt cctacgccct gggtccatat cagattagtg ctccccaact ccccgcctac 60
aacggtcaga cagtggcaac cttttactat gtcaacgacg ccgggggatt ggaatccaag 120
gttttctcta gcggtggg 138
<210> 43
<211> 44
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 43
Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro
1 5 10 15
Ala Tyr Asn Gly Gln Thr Val Gly Ala Phe Tyr Tyr Val Asn Asp Ala
20 25 30
Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly
35 40
<210> 44
<211> 138
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 44
ggctctggtt cctacgccct gggtccatat cagattagtg ctccccaact ccccgcctac 60
aacggtcaga cagtgggggc attttactat gtcaacgacg ccgggggatt ggaatccaag 120
gttttctcta gcggtggg 138
<210> 45
<211> 44
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 45
Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro
1 5 10 15
Ala Tyr Asn Gly Gln Thr Val Gly Thr Ala Tyr Tyr Val Asn Asp Ala
20 25 30
Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly
35 40
<210> 46
<211> 138
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 46
ggctctggtt cctacgccct gggtccatat cagattagtg ctccccaact ccccgcctac 60
aacggtcaga cagtggggac cgcatactat gtcaacgacg ccgggggatt ggaatccaag 120
gttttctcta gcggtggg 138
<210> 47
<211> 44
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 47
Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro
1 5 10 15
Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Ala Tyr Val Asn Asp Ala
20 25 30
Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly
35 40
<210> 48
<211> 138
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 48
ggctctggtt cctacgccct gggtccatat cagattagtg ctccccaact ccccgcctac 60
aacggtcaga cagtggggac ctttgcatat gtcaacgacg ccgggggatt ggaatccaag 120
gttttctcta gcggtggg 138
<210> 49
<211> 44
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 49
Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro
1 5 10 15
Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Ala Val Asn Asp Ala
20 25 30
Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly
35 40
<210> 50
<211> 138
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 50
ggctctggtt cctacgccct gggtccatat cagattagtg ctccccaact ccccgcctac 60
aacggtcaga cagtggggac cttttacgca gtcaacgacg ccgggggatt ggaatccaag 120
gttttctcta gcggtggg 138
<210> 51
<211> 44
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 51
Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro
1 5 10 15
Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Ala Asn Asp Ala
20 25 30
Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly
35 40
<210> 52
<211> 138
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 52
ggctctggtt cctacgccct gggtccatat cagattagtg ctccccaact ccccgcctac 60
aacggtcaga cagtggggac cttttactat gcaaacgacg ccgggggatt ggaatccaag 120
gttttctcta gcggtggg 138
<210> 53
<211> 44
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 53
Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro
1 5 10 15
Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala
20 25 30
Gly Gly Leu Glu Ser Ala Val Phe Ser Ser Gly Gly
35 40
<210> 54
<211> 138
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 54
ggctctggtt cctacgccct gggtccatat cagattagtg ctccccaact ccccgcctac 60
aacggtcaga cagtggggac cttttactat gtcaacgacg ccgggggatt ggaatccgca 120
gttttctcta gcggtggg 138
<210> 55
<211> 44
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 55
Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro
1 5 10 15
Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala
20 25 30
Gly Gly Leu Glu Ser Lys Val Ala Ser Ser Gly Gly
35 40
<210> 56
<211> 138
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 56
ggctctggtt cctacgccct gggtccatat cagattagtg ctccccaact ccccgcctac 60
aacggtcaga cagtggggac cttttactat gtcaacgacg ccgggggatt ggaatccaag 120
gttgcatcta gcggtggg 138
<210> 57
<211> 44
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 57
Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro
1 5 10 15
Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala
20 25 30
Gly Gly Leu Glu Ser Lys Val Phe Ala Ser Gly Gly
35 40
<210> 58
<211> 138
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 58
ggctctggtt cctacgccct gggtccatat cagattagtg ctccccaact ccccgcctac 60
aacggtcaga cagtggggac cttttactat gtcaacgacg ccgggggatt ggaatccaag 120
gttttcgcaa gcggtggg 138
<210> 59
<211> 94
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 59
Pro Thr Pro Tyr Pro Asn Tyr Ala Asn Ala Gly His Ala Glu Gly Gln
1 5 10 15
Ser Ala Leu Phe Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe
20 25 30
His Asn Asn Pro Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu
35 40 45
Thr Leu Leu Pro Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly
50 55 60
Ala Ile Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr
65 70 75 80
Gly Asn Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys
85 90
<210> 60
<211> 282
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 60
ccaacacctt atcctaacta cgctaacgcc gggcacgcag aggggcagtc agctcttttt 60
atgagagata acggcattag cgaagggctt gtgttccata ataatcctga gggcacctgt 120
ggcttctgtg taaatatgac cgaaacactt ctgcctgaga acgctaaaat gactgtcgta 180
ccacccgaag gcgcaatccc agttaaacgg ggcgcaaccg gcgaaaccaa agtattcacc 240
ggaaacagca atagtccaaa gtcccccacc aagggaggtt gc 282
<210> 61
<211> 94
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 61
Pro Thr Pro Tyr Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Ala
1 5 10 15
Ser Ala Leu Phe Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe
20 25 30
His Asn Asn Pro Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu
35 40 45
Thr Leu Leu Pro Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly
50 55 60
Ala Ile Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr
65 70 75 80
Gly Asn Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys
85 90
<210> 62
<211> 282
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 62
ccaacacctt atcctaacta cgctaacgcc gggcacgtcg agggggcatc agctcttttt 60
atgagagata acggcattag cgaagggctt gtgttccata ataatcctga gggcacctgt 120
ggcttctgtg taaatatgac cgaaacactt ctgcctgaga acgctaaaat gactgtcgta 180
ccacccgaag gcgcaatccc agttaaacgg ggcgcaaccg gcgaaaccaa agtattcacc 240
ggaaacagca atagtccaaa gtcccccacc aagggaggtt gc 282
<210> 63
<211> 94
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 63
Pro Thr Pro Tyr Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln
1 5 10 15
Ala Ala Leu Phe Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe
20 25 30
His Asn Asn Pro Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu
35 40 45
Thr Leu Leu Pro Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly
50 55 60
Ala Ile Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr
65 70 75 80
Gly Asn Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys
85 90
<210> 64
<211> 282
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 64
ccaacacctt atcctaacta cgctaacgcc gggcacgtcg aggggcaggc agctcttttt 60
atgagagata acggcattag cgaagggctt gtgttccata ataatcctga gggcacctgt 120
ggcttctgtg taaatatgac cgaaacactt ctgcctgaga acgctaaaat gactgtcgta 180
ccacccgaag gcgcaatccc agttaaacgg ggcgcaaccg gcgaaaccaa agtattcacc 240
ggaaacagca atagtccaaa gtcccccacc aagggaggtt gc 282
<210> 65
<211> 94
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 65
Pro Thr Pro Tyr Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln
1 5 10 15
Ser Ala Leu Ala Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe
20 25 30
His Asn Asn Pro Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu
35 40 45
Thr Leu Leu Pro Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly
50 55 60
Ala Ile Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr
65 70 75 80
Gly Asn Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys
85 90
<210> 66
<211> 282
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 66
ccaacacctt atcctaacta cgctaacgcc gggcacgtcg aggggcagtc agctcttgca 60
atgagagata acggcattag cgaagggctt gtgttccata ataatcctga gggcacctgt 120
ggcttctgtg taaatatgac cgaaacactt ctgcctgaga acgctaaaat gactgtcgta 180
ccacccgaag gcgcaatccc agttaaacgg ggcgcaaccg gcgaaaccaa agtattcacc 240
ggaaacagca atagtccaaa gtcccccacc aagggaggtt gc 282
<210> 67
<211> 94
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 67
Pro Thr Pro Tyr Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln
1 5 10 15
Ser Ala Leu Phe Ala Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe
20 25 30
His Asn Asn Pro Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu
35 40 45
Thr Leu Leu Pro Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly
50 55 60
Ala Ile Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr
65 70 75 80
Gly Asn Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys
85 90
<210> 68
<211> 282
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 68
ccaacacctt atcctaacta cgctaacgcc gggcacgtcg aggggcagtc agctcttttt 60
gcaagagata acggcattag cgaagggctt gtgttccata ataatcctga gggcacctgt 120
ggcttctgtg taaatatgac cgaaacactt ctgcctgaga acgctaaaat gactgtcgta 180
ccacccgaag gcgcaatccc agttaaacgg ggcgcaaccg gcgaaaccaa agtattcacc 240
ggaaacagca atagtccaaa gtcccccacc aagggaggtt gc 282
<210> 69
<211> 94
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 69
Pro Thr Pro Tyr Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln
1 5 10 15
Ser Ala Leu Phe Met Arg Asp Asn Gly Ile Ser Ala Gly Leu Val Phe
20 25 30
His Asn Asn Pro Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu
35 40 45
Thr Leu Leu Pro Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly
50 55 60
Ala Ile Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr
65 70 75 80
Gly Asn Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys
85 90
<210> 70
<211> 282
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 70
ccaacacctt atcctaacta cgctaacgcc gggcacgtcg aggggcagtc agctcttttt 60
atgagagata acggcattag cgcagggctt gtgttccata ataatcctga gggcacctgt 120
ggcttctgtg taaatatgac cgaaacactt ctgcctgaga acgctaaaat gactgtcgta 180
ccacccgaag gcgcaatccc agttaaacgg ggcgcaaccg gcgaaaccaa agtattcacc 240
ggaaacagca atagtccaaa gtcccccacc aagggaggtt gc 282
<210> 71
<211> 94
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 71
Pro Thr Pro Tyr Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln
1 5 10 15
Ser Ala Leu Phe Met Arg Asp Asn Gly Ile Ser Glu Ala Leu Val Phe
20 25 30
His Asn Asn Pro Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu
35 40 45
Thr Leu Leu Pro Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly
50 55 60
Ala Ile Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr
65 70 75 80
Gly Asn Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys
85 90
<210> 72
<211> 282
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 72
ccaacacctt atcctaacta cgctaacgcc gggcacgtcg aggggcagtc agctcttttt 60
atgagagata acggcattag cgaagcactt gtgttccata ataatcctga gggcacctgt 120
ggcttctgtg taaatatgac cgaaacactt ctgcctgaga acgctaaaat gactgtcgta 180
ccacccgaag gcgcaatccc agttaaacgg ggcgcaaccg gcgaaaccaa agtattcacc 240
ggaaacagca atagtccaaa gtcccccacc aagggaggtt gc 282
<210> 73
<211> 94
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 73
Pro Thr Pro Tyr Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln
1 5 10 15
Ser Ala Leu Phe Met Arg Asp Asn Gly Ile Ser Glu Gly Ala Val Phe
20 25 30
His Asn Asn Pro Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu
35 40 45
Thr Leu Leu Pro Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly
50 55 60
Ala Ile Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr
65 70 75 80
Gly Asn Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys
85 90
<210> 74
<211> 282
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 74
ccaacacctt atcctaacta cgctaacgcc gggcacgtcg aggggcagtc agctcttttt 60
atgagagata acggcattag cgaaggggca gtgttccata ataatcctga gggcacctgt 120
ggcttctgtg taaatatgac cgaaacactt ctgcctgaga acgctaaaat gactgtcgta 180
ccacccgaag gcgcaatccc agttaaacgg ggcgcaaccg gcgaaaccaa agtattcacc 240
ggaaacagca atagtccaaa gtcccccacc aagggaggtt gc 282
<210> 75
<211> 94
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 75
Pro Thr Pro Tyr Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln
1 5 10 15
Ser Ala Leu Phe Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Ala Phe
20 25 30
His Asn Asn Pro Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu
35 40 45
Thr Leu Leu Pro Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly
50 55 60
Ala Ile Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr
65 70 75 80
Gly Asn Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys
85 90
<210> 76
<211> 282
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 76
ccaacacctt atcctaacta cgctaacgcc gggcacgtcg aggggcagtc agctcttttt 60
atgagagata acggcattag cgaagggctt gcattccata ataatcctga gggcacctgt 120
ggcttctgtg taaatatgac cgaaacactt ctgcctgaga acgctaaaat gactgtcgta 180
ccacccgaag gcgcaatccc agttaaacgg ggcgcaaccg gcgaaaccaa agtattcacc 240
ggaaacagca atagtccaaa gtcccccacc aagggaggtt gc 282
<210> 77
<211> 94
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 77
Pro Thr Pro Tyr Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln
1 5 10 15
Ser Ala Leu Phe Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Ala
20 25 30
His Asn Asn Pro Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu
35 40 45
Thr Leu Leu Pro Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly
50 55 60
Ala Ile Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr
65 70 75 80
Gly Asn Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys
85 90
<210> 78
<211> 282
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 78
ccaacacctt atcctaacta cgctaacgcc gggcacgtcg aggggcagtc agctcttttt 60
atgagagata acggcattag cgaagggctt gtggcacata ataatcctga gggcacctgt 120
ggcttctgtg taaatatgac cgaaacactt ctgcctgaga acgctaaaat gactgtcgta 180
ccacccgaag gcgcaatccc agttaaacgg ggcgcaaccg gcgaaaccaa agtattcacc 240
ggaaacagca atagtccaaa gtcccccacc aagggaggtt gc 282
<210> 79
<211> 94
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 79
Pro Thr Pro Tyr Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln
1 5 10 15
Ser Ala Leu Phe Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe
20 25 30
Ala Asn Asn Pro Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu
35 40 45
Thr Leu Leu Pro Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly
50 55 60
Ala Ile Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr
65 70 75 80
Gly Asn Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys
85 90
<210> 80
<211> 282
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 80
ccaacacctt atcctaacta cgctaacgcc gggcacgtcg aggggcagtc agctcttttt 60
atgagagata acggcattag cgaagggctt gtgttcgcaa ataatcctga gggcacctgt 120
ggcttctgtg taaatatgac cgaaacactt ctgcctgaga acgctaaaat gactgtcgta 180
ccacccgaag gcgcaatccc agttaaacgg ggcgcaaccg gcgaaaccaa agtattcacc 240
ggaaacagca atagtccaaa gtcccccacc aagggaggtt gc 282
<210> 81
<211> 94
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 81
Pro Thr Pro Tyr Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln
1 5 10 15
Ser Ala Leu Phe Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe
20 25 30
His Asn Asn Pro Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu
35 40 45
Thr Leu Leu Pro Glu Asn Ala Ala Met Thr Val Val Pro Pro Glu Gly
50 55 60
Ala Ile Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr
65 70 75 80
Gly Asn Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys
85 90
<210> 82
<211> 282
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 82
ccaacacctt atcctaacta cgctaacgcc gggcacgtcg aggggcagtc agctcttttt 60
atgagagata acggcattag cgaagggctt gtgttccata ataatcctga gggcacctgt 120
ggcttctgtg taaatatgac cgaaacactt ctgcctgaga acgctgcaat gactgtcgta 180
ccacccgaag gcgcaatccc agttaaacgg ggcgcaaccg gcgaaaccaa agtattcacc 240
ggaaacagca atagtccaaa gtcccccacc aagggaggtt gc 282
<210> 83
<211> 94
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 83
Pro Thr Pro Tyr Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln
1 5 10 15
Ser Ala Leu Phe Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe
20 25 30
His Asn Asn Pro Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu
35 40 45
Thr Leu Leu Pro Glu Asn Ala Lys Ala Thr Val Val Pro Pro Glu Gly
50 55 60
Ala Ile Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr
65 70 75 80
Gly Asn Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys
85 90
<210> 84
<211> 282
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 84
ccaacacctt atcctaacta cgctaacgcc gggcacgtcg aggggcagtc agctcttttt 60
atgagagata acggcattag cgaagggctt gtgttccata ataatcctga gggcacctgt 120
ggcttctgtg taaatatgac cgaaacactt ctgcctgaga acgctaaagc aactgtcgta 180
ccacccgaag gcgcaatccc agttaaacgg ggcgcaaccg gcgaaaccaa agtattcacc 240
ggaaacagca atagtccaaa gtcccccacc aagggaggtt gc 282
<210> 85
<211> 94
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 85
Pro Thr Pro Tyr Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln
1 5 10 15
Ser Ala Leu Phe Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe
20 25 30
His Asn Asn Pro Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu
35 40 45
Thr Leu Leu Pro Glu Asn Ala Lys Met Ala Val Val Pro Pro Glu Gly
50 55 60
Ala Ile Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr
65 70 75 80
Gly Asn Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys
85 90
<210> 86
<211> 282
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 86
ccaacacctt atcctaacta cgctaacgcc gggcacgtcg aggggcagtc agctcttttt 60
atgagagata acggcattag cgaagggctt gtgttccata ataatcctga gggcacctgt 120
ggcttctgtg taaatatgac cgaaacactt ctgcctgaga acgctaaaat ggcagtcgta 180
ccacccgaag gcgcaatccc agttaaacgg ggcgcaaccg gcgaaaccaa agtattcacc 240
ggaaacagca atagtccaaa gtcccccacc aagggaggtt gc 282
<210> 87
<211> 94
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 87
Pro Thr Pro Tyr Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln
1 5 10 15
Ser Ala Leu Phe Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe
20 25 30
His Asn Asn Pro Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu
35 40 45
Thr Leu Leu Pro Glu Asn Ala Lys Met Thr Val Ala Pro Pro Glu Gly
50 55 60
Ala Ile Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr
65 70 75 80
Gly Asn Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys
85 90
<210> 88
<211> 282
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 88
ccaacacctt atcctaacta cgctaacgcc gggcacgtcg aggggcagtc agctcttttt 60
atgagagata acggcattag cgaagggctt gtgttccata ataatcctga gggcacctgt 120
ggcttctgtg taaatatgac cgaaacactt ctgcctgaga acgctaaaat gactgtcgca 180
ccacccgaag gcgcaatccc agttaaacgg ggcgcaaccg gcgaaaccaa agtattcacc 240
ggaaacagca atagtccaaa gtcccccacc aagggaggtt gc 282
<210> 89
<211> 108
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 89
Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro
1 5 10 15
Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala
20 25 30
Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly Pro Thr Pro Tyr
35 40 45
Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln Ser Ala Leu Phe
50 55 60
Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe His Asn Asn Pro
65 70 75 80
Glu Gly Thr Cys Gly Phe Ala Val Asn Met Thr Glu Thr Leu Leu Pro
85 90 95
Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly
100 105
<210> 90
<211> 324
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 90
ggtagctacg cacttggtcc ttaccagatt agcgcacccc aactccccgc ctataatggt 60
caaaccgtcg ggacctttta ctacgtaaac gatgctggtg ggctggaatc caaagtattc 120
tcctcagggg gccctacacc ctaccccaac tacgccaatg ctggtcatgt agaagggcag 180
tcagcactgt ttatgcgcga taatggtata agcgaggggt tggtcttcca taacaaccca 240
gagggtactt gtggcttcgc agtgaatatg actgaaaccc ttctgcccga aaatgccaag 300
atgactgtcg tcccacctga aggc 324
<210> 91
<211> 108
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 91
Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro
1 5 10 15
Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala
20 25 30
Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly Pro Thr Pro Tyr
35 40 45
Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln Ser Ala Leu Phe
50 55 60
Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe His Asn Asn Pro
65 70 75 80
Glu Gly Thr Cys Gly Phe Cys Ala Asn Met Thr Glu Thr Leu Leu Pro
85 90 95
Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly
100 105
<210> 92
<211> 324
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 92
ggtagctacg cacttggtcc ttaccagatt agcgcacccc aactccccgc ctataatggt 60
caaaccgtcg ggacctttta ctacgtaaac gatgctggtg ggctggaatc caaagtattc 120
tcctcagggg gccctacacc ctaccccaac tacgccaatg ctggtcatgt agaagggcag 180
tcagcactgt ttatgcgcga taatggtata agcgaggggt tggtcttcca taacaaccca 240
gagggtactt gtggcttctg tgcaaatatg actgaaaccc ttctgcccga aaatgccaag 300
atgactgtcg tcccacctga aggc 324
<210> 93
<211> 108
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 93
Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro
1 5 10 15
Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala
20 25 30
Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly Pro Thr Pro Tyr
35 40 45
Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln Ser Ala Leu Phe
50 55 60
Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe His Asn Asn Pro
65 70 75 80
Glu Gly Thr Cys Gly Phe Cys Val Asn Met Ala Glu Thr Leu Leu Pro
85 90 95
Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly
100 105
<210> 94
<211> 324
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 94
ggtagctacg cacttggtcc ttaccagatt agcgcacccc aactccccgc ctataatggt 60
caaaccgtcg ggacctttta ctacgtaaac gatgctggtg ggctggaatc caaagtattc 120
tcctcagggg gccctacacc ctaccccaac tacgccaatg ctggtcatgt agaagggcag 180
tcagcactgt ttatgcgcga taatggtata agcgaggggt tggtcttcca taacaaccca 240
gagggtactt gtggcttctg tgtgaatatg gcagaaaccc ttctgcccga aaatgccaag 300
atgactgtcg tcccacctga aggc 324
<210> 95
<211> 108
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 95
Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro
1 5 10 15
Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala
20 25 30
Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly Pro Thr Pro Tyr
35 40 45
Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln Ser Ala Leu Phe
50 55 60
Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe His Asn Asn Pro
65 70 75 80
Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Ala Thr Leu Leu Pro
85 90 95
Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly
100 105
<210> 96
<211> 324
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 96
ggtagctacg cacttggtcc ttaccagatt agcgcacccc aactccccgc ctataatggt 60
caaaccgtcg ggacctttta ctacgtaaac gatgctggtg ggctggaatc caaagtattc 120
tcctcagggg gccctacacc ctaccccaac tacgccaatg ctggtcatgt agaagggcag 180
tcagcactgt ttatgcgcga taatggtata agcgaggggt tggtcttcca taacaaccca 240
gagggtactt gtggcttctg tgtgaatatg actgcaaccc ttctgcccga aaatgccaag 300
atgactgtcg tcccacctga aggc 324
<210> 97
<211> 108
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 97
Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro
1 5 10 15
Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala
20 25 30
Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly Pro Thr Pro Tyr
35 40 45
Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln Ser Ala Leu Phe
50 55 60
Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe His Asn Asn Pro
65 70 75 80
Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu Thr Leu Ala Pro
85 90 95
Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly
100 105
<210> 98
<211> 324
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 98
ggtagctacg cacttggtcc ttaccagatt agcgcacccc aactccccgc ctataatggt 60
caaaccgtcg ggacctttta ctacgtaaac gatgctggtg ggctggaatc caaagtattc 120
tcctcagggg gccctacacc ctaccccaac tacgccaatg ctggtcatgt agaagggcag 180
tcagcactgt ttatgcgcga taatggtata agcgaggggt tggtcttcca taacaaccca 240
gagggtactt gtggcttctg tgtgaatatg actgaaaccc ttgcacccga aaatgccaag 300
atgactgtcg tcccacctga aggc 324
<210> 99
<211> 108
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 99
Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro
1 5 10 15
Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala
20 25 30
Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly Pro Thr Pro Tyr
35 40 45
Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln Ser Ala Leu Phe
50 55 60
Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe His Asn Asn Pro
65 70 75 80
Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu Thr Leu Leu Pro
85 90 95
Glu Asn Ala Ala Met Thr Val Val Pro Pro Glu Gly
100 105
<210> 100
<211> 324
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 100
ggtagctacg cacttggtcc ttaccagatt agcgcacccc aactccccgc ctataatggt 60
caaaccgtcg ggacctttta ctacgtaaac gatgctggtg ggctggaatc caaagtattc 120
tcctcagggg gccctacacc ctaccccaac tacgccaatg ctggtcatgt agaagggcag 180
tcagcactgt ttatgcgcga taatggtata agcgaggggt tggtcttcca taacaaccca 240
gagggtactt gtggcttctg tgtgaatatg actgaaaccc ttctgcccga aaatgccgca 300
atgactgtcg tcccacctga aggc 324
<210> 101
<211> 108
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 101
Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro
1 5 10 15
Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala
20 25 30
Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly Pro Thr Pro Tyr
35 40 45
Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln Ser Ala Leu Phe
50 55 60
Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe His Asn Asn Pro
65 70 75 80
Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu Thr Leu Leu Pro
85 90 95
Glu Asn Ala Lys Ala Thr Val Val Pro Pro Glu Gly
100 105
<210> 102
<211> 324
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 102
ggtagctacg cacttggtcc ttaccagatt agcgcacccc aactccccgc ctataatggt 60
caaaccgtcg ggacctttta ctacgtaaac gatgctggtg ggctggaatc caaagtattc 120
tcctcagggg gccctacacc ctaccccaac tacgccaatg ctggtcatgt agaagggcag 180
tcagcactgt ttatgcgcga taatggtata agcgaggggt tggtcttcca taacaaccca 240
gagggtactt gtggcttctg tgtgaatatg actgaaaccc ttctgcccga aaatgccaag 300
gcaactgtcg tcccacctga aggc 324
<210> 103
<211> 108
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 103
Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro
1 5 10 15
Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala
20 25 30
Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly Pro Thr Pro Tyr
35 40 45
Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln Ser Ala Leu Phe
50 55 60
Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe His Asn Asn Pro
65 70 75 80
Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu Thr Leu Leu Pro
85 90 95
Glu Asn Ala Lys Met Ala Val Val Pro Pro Glu Gly
100 105
<210> 104
<211> 324
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 104
ggtagctacg cacttggtcc ttaccagatt agcgcacccc aactccccgc ctataatggt 60
caaaccgtcg ggacctttta ctacgtaaac gatgctggtg ggctggaatc caaagtattc 120
tcctcagggg gccctacacc ctaccccaac tacgccaatg ctggtcatgt agaagggcag 180
tcagcactgt ttatgcgcga taatggtata agcgaggggt tggtcttcca taacaaccca 240
gagggtactt gtggcttctg tgtgaatatg actgaaaccc ttctgcccga aaatgccaag 300
atggcagtcg tcccacctga aggc 324
<210> 105
<211> 108
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 105
Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro
1 5 10 15
Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala
20 25 30
Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly Pro Thr Pro Tyr
35 40 45
Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln Ser Ala Leu Phe
50 55 60
Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe His Asn Asn Pro
65 70 75 80
Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu Thr Leu Leu Pro
85 90 95
Glu Asn Ala Lys Met Thr Ala Val Pro Pro Glu Gly
100 105
<210> 106
<211> 324
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 106
ggtagctacg cacttggtcc ttaccagatt agcgcacccc aactccccgc ctataatggt 60
caaaccgtcg ggacctttta ctacgtaaac gatgctggtg ggctggaatc caaagtattc 120
tcctcagggg gccctacacc ctaccccaac tacgccaatg ctggtcatgt agaagggcag 180
tcagcactgt ttatgcgcga taatggtata agcgaggggt tggtcttcca taacaaccca 240
gagggtactt gtggcttctg tgtgaatatg actgaaaccc ttctgcccga aaatgccaag 300
atgactgcag tcccacctga aggc 324
<210> 107
<211> 30
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 107
Ala Ile Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Ala Val Phe Thr
1 5 10 15
Gly Asn Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys
20 25 30
<210> 108
<211> 90
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 108
gccatacctg tgaagcgggg agcaacaggg gagacagcag tgttcacagg caactctaac 60
agtccaaaga gccccaccaa aggcgggtgt 90
<210> 109
<211> 30
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 109
Ala Ile Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Ala Phe Thr
1 5 10 15
Gly Asn Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys
20 25 30
<210> 110
<211> 90
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 110
gccatacctg tgaagcgggg agcaacaggg gagacaaagg cattcacagg caactctaac 60
agtccaaaga gccccaccaa aggcgggtgt 90
<210> 111
<211> 30
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 111
Ala Ile Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Ala Thr
1 5 10 15
Gly Asn Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys
20 25 30
<210> 112
<211> 90
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 112
gccatacctg tgaagcgggg agcaacaggg gagacaaagg tggcaacagg caactctaac 60
agtccaaaga gccccaccaa aggcgggtgt 90
<210> 113
<211> 30
<212> PRT
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 113
Ala Ile Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Ala
1 5 10 15
Gly Asn Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys
20 25 30
<210> 114
<211> 90
<212> DNA
<213> Artificial Sequence
<220>
<223> Deaminase Variant
<400> 114
gccatacctg tgaagcgggg agcaacaggg gagacaaagg tgttcgcagg caactctaac 60
agtccaaaga gccccaccaa aggcgggtgt 90
<210> 115
<211> 24
<212> PRT
<213> Artificial Sequence
<220>
<223> Linker
<400> 115
Ser Gly Thr Pro His Glu Val Gly Val Tyr Thr Leu Ser Gly Thr Pro
1 5 10 15
His Glu Val Gly Val Tyr Thr Leu
20
<210> 116
<211> 19
<212> DNA
<213> Artificial Sequence
<220>
<223> Primer
<400> 116
catcaatggg cgtggatag 19
<210> 117
<211> 19
<212> DNA
<213> Artificial Sequence
<220>
<223> Primer
<400> 117
catcaatggg cgtggatag 19
<210> 118
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> Primer
<400> 118
gacacctact cagacaatgc 20
<210> 119
<211> 12
<212> DNA
<213> Artificial Sequence
<220>
<223> Left-ZFD DNA binding region
<400> 119
gtcatcctca tc 12
<210> 120
<211> 12
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 120
aaactgcaaa ag 12
<210> 121
<211> 3
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 121
tcg 3
<210> 122
<211> 4
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 122
ctga 4
<210> 123
<211> 5
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 123
ctgat 5
<210> 124
<211> 6
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 124
ctgatc 6
<210> 125
<211> 7
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 125
ctgatcg 7
<210> 126
<211> 8
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 126
ctgatcga 8
<210> 127
<211> 9
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 127
ctgatcgat 9
<210> 128
<211> 10
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 128
ctgatcgatc 10
<210> 129
<211> 11
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 129
ctgatcgatc g 11
<210> 130
<211> 12
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 130
ctgatcgatc ga 12
<210> 131
<211> 13
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 131
ctgatcgatc gat 13
<210> 132
<211> 14
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 132
ctgatcgatc gatc 14
<210> 133
<211> 15
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 133
ctgatcgatc gatcg 15
<210> 134
<211> 16
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 134
ctgatcgatc gatcga 16
<210> 135
<211> 17
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 135
ctgatcgatc gatcgat 17
<210> 136
<211> 18
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 136
ctgatcgatc gatcgatc 18
<210> 137
<211> 19
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 137
ctgatcgatc gatcgatcg 19
<210> 138
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 138
ctgatcgatc gatcgatcga 20
<210> 139
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 139
ctgatcgatc gatcgatcga t 21
<210> 140
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 140
ctgatcgatc gatcgatcga tc 22
<210> 141
<211> 23
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 141
ctgatcgatc gatcgatcga tcg 23
<210> 142
<211> 24
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 142
ctgatcgatc gatcgatcga tcga 24
<210> 143
<211> 33
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 143
gtttactcaa tcctctgatc agggtgagca tca 33
<210> 144
<211> 11
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 144
Val Tyr Ser Ile Leu Trp Ser Gly Trp Ala Ser
1 5 10
<210> 145
<211> 33
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 145
gtttactcaa tcctctgatt agggtgagca tca 33
<210> 146
<211> 11
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 146
Val Tyr Ser Ile Leu Trp Leu Gly Trp Ala Ser
1 5 10
<210> 147
<211> 33
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 147
gtttactcaa tcctctaatc agggtgagca tca 33
<210> 148
<211> 10
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 148
Val Tyr Ser Ile Leu Ser Gly Trp Ala Ser
1 5 10
<210> 149
<211> 39
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 149
ctacgcctaa tctactccac ctcaatcaca ctactcccc 39
<210> 150
<211> 39
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 150
ctacgcctaa tctactttac ctcaatcaca ctactcccc 39
<210> 151
<211> 13
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 151
Leu Arg Leu Ile Tyr Phe Thr Ser Ile Thr Leu Leu Pro
1 5 10
<210> 152
<211> 39
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 152
ctacgcctaa tctactccac ctcaatcaca ctactcccc 39
<210> 153
<211> 13
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 153
Leu Arg Leu Ile Tyr Ser Thr Ser Ile Thr Leu Leu Pro
1 5 10
<210> 154
<211> 39
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 154
ctacgcctaa tctactttat ttcaatcaca ctactcccc 39
<210> 155
<211> 13
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 155
Leu Arg Leu Ile Tyr Phe Ile Ser Ile Thr Leu Leu Pro
1 5 10
<210> 156
<211> 39
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 156
ctacgcctaa tctacttcac ctcaatcaca ctactcccc 39
<210> 157
<211> 13
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 157
Leu Arg Leu Ile Tyr Phe Thr Ser Ile Thr Leu Leu Pro
1 5 10
<210> 158
<211> 39
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 158
ctacgcctaa tctactttat ctcaatcaca ctactcccc 39
<210> 159
<211> 13
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 159
Leu Arg Leu Ile Tyr Phe Ile Ser Ile Thr Leu Leu Pro
1 5 10
<210> 160
<211> 39
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 160
ctacgcctaa tctactttac cttaatcaca ctactcccc 39
<210> 161
<211> 13
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 161
Leu Arg Leu Ile Tyr Phe Thr Leu Ile Thr Leu Leu Pro
1 5 10
<210> 162
<211> 917
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 162
ctagcaacga ttgtacaatt gcttctttaa aaaaggaaga aagaaagaaa gaaaagaatc 60
aacatcagcg ttaacaaacg gccccgttac ggcccaaacg gtcatataga gtaacggcgt 120
taagcgttga aagactccta tcgaaatacg taaccgcaaa cgtgtcatag tcagatcccc 180
tcttccttca ccgcctcaaa cacaaaaata atcttctaca gcctatatat acaacccccc 240
cttctatctc tcctttctca caattcatca tctttctttc tctaccccca attttaagaa 300
atcctctctt ctcctcttca ttttcaaggt aaatctctct ctctctctct ctctctgtta 360
ttccttgttt taattaggta tgtattattg ctagtttgtt aatctgctta tcttatgtat 420
gccttatgtg aatatcttta tcttgttcat ctcatccgtt tagaagctat aaatttgttg 480
atttgactgt gtatctacac gtggttatgt ttatatctaa tcagatatga atttcttcat 540
attgttgcgt ttgtgtgtac caatccgaaa tcgttgattt ttttcattta atcgtgtagc 600
taattgtacg tatacatatg gatctacgta tcaattgttc atctgtttgt gtttgtatgt 660
atacagatct gaaaacatca cttctctcat ctgattgtgt tgttacatac atagatatag 720
atctgttata tcattttttt tattaattgt gtatatatat atgtgcatag atctggatta 780
catgattgtg attatttaca tgattttgtt atttacgtat gtatatatgt agatctggac 840
tttttggagt tgttgacttg attgtatttg tgtgtgtata tgtgtgttct gatcttgata 900
tgttatgtat gtgcagc 917
<210> 163
<211> 469
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 163
caggcctccc agctttcgtc cgtatcatcg gtttcgacaa cgttcgtcaa gttcaatgca 60
tcagtttcat tgcccacaca ccagaatcct actaagtttg agtattatgg cattggaaaa 120
gctgttttct tctatcattt gttctgcttg taatttactg tgttctttca gtttttgttt 180
tcggacatca aaatgcaaat ggatggataa gagttaataa atgatatggt ccttttgttc 240
attctcaaat tattattatc tgttgttttt actttaatgg gttgaattta agtaagaaag 300
gaactaacag tgtgatatta aggtgcaatg ttagacatat aaaacagtct ttcacctctc 360
tttggttatg tcttgaattg gtttgtttct tcacttatct gtgtaatcaa gtttactatg 420
agtctatgat caagtaatta tgcaatcaag ttaagtacag tataggctt 469
<210> 164
<211> 19
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 164
taatacgact cactatagg 19
<210> 165
<211> 308
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 165
Met Leu Gln Leu Cys Ser Thr Phe Arg Pro Gln Leu Leu Leu Pro Cys
1 5 10 15
Gln Phe Arg Phe Thr Asn Gly Val Leu Ile Pro Gln Ile Asn Tyr Val
20 25 30
Ala Ser Asn Ser Val Val Asn Ile Arg Pro Met Ile Arg Cys Gln Arg
35 40 45
Ala Ser Gly Gly Arg Gly Gly Ala Asn Arg Ser Lys Pro Ala Lys Pro
50 55 60
Gln Val Lys Glu Gly Ser Asn Lys Thr Val Ile Glu Gly Leu Val Thr
65 70 75 80
Glu Ser Leu Pro Asn Gly Met Phe Arg Val Asp Leu Glu Asn Gly Asp
85 90 95
Asn Ile Leu Gly Tyr Ile Cys Gly Lys Ile Arg Lys Asn Phe Ile Arg
100 105 110
Ile Leu Pro Gly Asp Lys Val Lys Val Glu Met Ser Val Tyr Asp Ser
115 120 125
Thr Lys Gly Arg Ile Ile Phe Arg Met Ser Ser Arg Asp Tyr Pro Tyr
130 135 140
Asp Val Pro Asp Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp Tyr Ala
145 150 155 160
Gly Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Met Asp Ile Ala Asp Leu
165 170 175
Arg Thr Leu Gly Tyr Ser Gln Gln Gln Gln Glu Lys Ile Lys Pro Lys
180 185 190
Val Arg Ser Thr Val Ala Gln His His Glu Ala Leu Val Gly His Gly
195 200 205
Phe Thr His Ala His Ile Val Ala Leu Ser Gln His Pro Ala Ala Leu
210 215 220
Gly Thr Val Ala Val Lys Tyr Gln Asp Met Ile Ala Ala Leu Pro Glu
225 230 235 240
Ala Thr His Glu Ala Ile Val Gly Val Gly Lys Gln Trp Ser Gly Ala
245 250 255
Arg Ala Leu Glu Ala Leu Leu Thr Val Ala Gly Glu Leu Arg Gly Pro
260 265 270
Pro Leu Gln Leu Asp Thr Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly
275 280 285
Gly Val Thr Ala Val Glu Ala Val His Ala Trp Arg Asn Ala Leu Thr
290 295 300
Gly Ala Pro Leu
305
<210> 166
<211> 240
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 166
Met Ala Ser Ser Met Leu Ser Ser Ala Thr Met Val Ala Ser Pro Ala
1 5 10 15
Gln Ala Thr Met Val Ala Pro Phe Asn Gly Leu Lys Ser Ser Ala Ala
20 25 30
Phe Pro Ala Thr Arg Lys Ala Asn Asn Asp Ile Thr Ser Ile Thr Ser
35 40 45
Asn Gly Gly Arg Val Asn Cys Met Gln Val Trp Pro Pro Ile Gly Lys
50 55 60
Lys Lys Phe Glu Thr Leu Ser Tyr Leu Pro Asp Leu Thr Asp Ser Glu
65 70 75 80
Asp Tyr Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp Ile Asp Tyr
85 90 95
Lys Asp Asp Asp Asp Lys Met Asp Ile Ala Asp Leu Arg Thr Leu Gly
100 105 110
Tyr Ser Gln Gln Gln Gln Glu Lys Ile Lys Pro Lys Val Arg Ser Thr
115 120 125
Val Ala Gln His His Glu Ala Leu Val Gly His Gly Phe Thr His Ala
130 135 140
His Ile Val Ala Leu Ser Gln His Pro Ala Ala Leu Gly Thr Val Ala
145 150 155 160
Val Lys Tyr Gln Asp Met Ile Ala Ala Leu Pro Glu Ala Thr His Glu
165 170 175
Ala Ile Val Gly Val Gly Lys Arg Gly Ala Gly Ala Arg Ala Leu Glu
180 185 190
Ala Leu Leu Thr Val Ala Gly Glu Leu Arg Gly Pro Pro Leu Gln Leu
195 200 205
Asp Thr Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly Gly Val Thr Ala
210 215 220
Val Glu Ala Val His Ala Trp Arg Asn Ala Leu Thr Gly Ala Pro Leu
225 230 235 240
<210> 167
<211> 244
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 167
Met Ala Met Ala Val Phe Arg Arg Glu Gly Arg Arg Leu Leu Pro Ser
1 5 10 15
Ile Ala Ala Arg Pro Ile Ala Ala Ile Arg Ser Pro Leu Ser Ser Asp
20 25 30
Gln Glu Glu Gly Leu Leu Gly Val Arg Ser Ile Ser Thr Gln Val Val
35 40 45
Arg Asn Arg Met Lys Ser Val Lys Asn Ile Gln Lys Ile Thr Lys Ala
50 55 60
Met Lys Met Val Ala Ala Ser Lys Leu Arg Ala Val Gln Tyr Pro Tyr
65 70 75 80
Asp Val Pro Asp Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp Tyr Ala
85 90 95
Gly Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Met Asp Ile Ala Asp Leu
100 105 110
Arg Thr Leu Gly Tyr Ser Gln Gln Gln Gln Glu Lys Ile Lys Pro Lys
115 120 125
Val Arg Ser Thr Val Ala Gln His His Glu Ala Leu Val Gly His Gly
130 135 140
Phe Thr His Ala His Ile Val Ala Leu Ser Gln His Pro Ala Ala Leu
145 150 155 160
Gly Thr Val Ala Val Lys Tyr Gln Asp Met Ile Ala Ala Leu Pro Glu
165 170 175
Ala Thr His Glu Ala Ile Val Gly Val Gly Lys Gln Trp Ser Gly Ala
180 185 190
Arg Ala Leu Glu Ala Leu Leu Thr Val Ala Gly Glu Leu Arg Gly Pro
195 200 205
Pro Leu Gln Leu Asp Thr Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly
210 215 220
Gly Val Thr Ala Val Glu Ala Val His Ala Trp Arg Asn Ala Leu Thr
225 230 235 240
Gly Ala Pro Leu
<210> 168
<211> 196
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 168
Met Phe Lys Gln Ala Ser Arg Leu Leu Ser Arg Ser Val Ala Ala Ala
1 5 10 15
Ser Ser Lys Ser Val Thr Thr Arg Ala Phe Ser Thr Glu Leu Pro Ser
20 25 30
Thr Leu Asp Ser Asp Tyr Lys Asp His Asp Gly Asp Tyr Lys Asp His
35 40 45
Asp Ile Asp Tyr Lys Asp Asp Asp Asp Lys Met Asp Ile Ala Asp Leu
50 55 60
Arg Thr Leu Gly Tyr Ser Gln Gln Gln Gln Glu Lys Ile Lys Pro Lys
65 70 75 80
Val Arg Ser Thr Val Ala Gln His His Glu Ala Leu Val Gly His Gly
85 90 95
Phe Thr His Ala His Ile Val Ala Leu Ser Gln His Pro Ala Ala Leu
100 105 110
Gly Thr Val Ala Val Lys Tyr Gln Asp Met Ile Ala Ala Leu Pro Glu
115 120 125
Ala Thr His Glu Ala Ile Val Gly Val Gly Lys Arg Gly Ala Gly Ala
130 135 140
Arg Ala Leu Glu Ala Leu Leu Thr Val Ala Gly Glu Leu Arg Gly Pro
145 150 155 160
Pro Leu Gln Leu Asp Thr Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly
165 170 175
Gly Val Thr Ala Val Glu Ala Val His Ala Trp Arg Asn Ala Leu Thr
180 185 190
Gly Ala Pro Leu
195
<210> 169
<211> 195
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 169
Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp Pro
20 25 30
Ala Leu Ala Ala Leu Thr Asn Asp His Leu Val Ala Leu Ala Cys Leu
35 40 45
Gly Gly Arg Pro Ala Leu Asp Ala Val Lys Lys Gly Leu Gly Gly Ser
50 55 60
Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro
65 70 75 80
Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala
85 90 95
Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly Ser Gly Gly Ser
100 105 110
Thr Asn Leu Ser Asp Ile Ile Glu Lys Glu Thr Gly Lys Gln Leu Val
115 120 125
Ile Gln Glu Ser Ile Leu Met Leu Pro Glu Glu Val Glu Glu Val Ile
130 135 140
Gly Asn Lys Pro Glu Ser Asp Ile Leu Val His Thr Ala Tyr Asp Glu
145 150 155 160
Ser Thr Asp Glu Asn Val Met Leu Leu Thr Ser Asp Ala Pro Glu Tyr
165 170 175
Lys Pro Trp Ala Leu Val Ile Gln Asp Ser Asn Gly Glu Asn Lys Ile
180 185 190
Lys Met Leu
195
<210> 170
<211> 245
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 170
Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp Pro
20 25 30
Ala Leu Ala Ala Leu Thr Asn Asp His Leu Val Ala Leu Ala Cys Leu
35 40 45
Gly Gly Arg Pro Ala Leu Asp Ala Val Lys Lys Gly Leu Gly Gly Ser
50 55 60
Pro Thr Pro Tyr Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln
65 70 75 80
Ser Ala Leu Phe Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe
85 90 95
His Asn Asn Pro Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu
100 105 110
Thr Leu Leu Pro Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly
115 120 125
Ala Ile Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr
130 135 140
Gly Asn Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys Ser Gly
145 150 155 160
Gly Ser Thr Asn Leu Ser Asp Ile Ile Glu Lys Glu Thr Gly Lys Gln
165 170 175
Leu Val Ile Gln Glu Ser Ile Leu Met Leu Pro Glu Glu Val Glu Glu
180 185 190
Val Ile Gly Asn Lys Pro Glu Ser Asp Ile Leu Val His Thr Ala Tyr
195 200 205
Asp Glu Ser Thr Asp Glu Asn Val Met Leu Leu Thr Ser Asp Ala Pro
210 215 220
Glu Tyr Lys Pro Trp Ala Leu Val Ile Gln Asp Ser Asn Gly Glu Asn
225 230 235 240
Lys Ile Lys Met Leu
245
<210> 171
<211> 259
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 171
Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp Pro
20 25 30
Ala Leu Ala Ala Leu Thr Asn Asp His Leu Val Ala Leu Ala Cys Leu
35 40 45
Gly Gly Arg Pro Ala Leu Asp Ala Val Lys Lys Gly Leu Gly Gly Ser
50 55 60
Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro
65 70 75 80
Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala
85 90 95
Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly Pro Thr Pro Tyr
100 105 110
Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln Ser Ala Leu Phe
115 120 125
Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe His Asn Asn Pro
130 135 140
Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu Thr Leu Leu Pro
145 150 155 160
Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly Ser Gly Gly Ser
165 170 175
Thr Asn Leu Ser Asp Ile Ile Glu Lys Glu Thr Gly Lys Gln Leu Val
180 185 190
Ile Gln Glu Ser Ile Leu Met Leu Pro Glu Glu Val Glu Glu Val Ile
195 200 205
Gly Asn Lys Pro Glu Ser Asp Ile Leu Val His Thr Ala Tyr Asp Glu
210 215 220
Ser Thr Asp Glu Asn Val Met Leu Leu Thr Ser Asp Ala Pro Glu Tyr
225 230 235 240
Lys Pro Trp Ala Leu Val Ile Gln Asp Ser Asn Gly Glu Asn Lys Ile
245 250 255
Lys Met Leu
<210> 172
<211> 181
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 172
Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp Pro
20 25 30
Ala Leu Ala Ala Leu Thr Asn Asp His Leu Val Ala Leu Ala Cys Leu
35 40 45
Gly Gly Arg Pro Ala Leu Asp Ala Val Lys Lys Gly Leu Gly Gly Ser
50 55 60
Ala Ile Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr
65 70 75 80
Gly Asn Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys Ser Gly
85 90 95
Gly Ser Thr Asn Leu Ser Asp Ile Ile Glu Lys Glu Thr Gly Lys Gln
100 105 110
Leu Val Ile Gln Glu Ser Ile Leu Met Leu Pro Glu Glu Val Glu Glu
115 120 125
Val Ile Gly Asn Lys Pro Glu Ser Asp Ile Leu Val His Thr Ala Tyr
130 135 140
Asp Glu Ser Thr Asp Glu Asn Val Met Leu Leu Thr Ser Asp Ala Pro
145 150 155 160
Glu Tyr Lys Pro Trp Ala Leu Val Ile Gln Asp Ser Asn Gly Glu Asn
165 170 175
Lys Ile Lys Met Leu
180
<210> 173
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 173
Asn Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 174
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 174
Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 175
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 175
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 176
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 176
Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 177
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 177
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser His Asp Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Asn
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 178
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 178
Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 179
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 179
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser His Asp Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 180
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 180
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Asp His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 181
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 181
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Asp His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 182
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 182
Asn Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 183
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 183
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Asp His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Asn
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 184
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 184
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Asp His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Asn
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 185
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 185
Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Asn
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 186
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 186
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser His Asp
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 187
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 187
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Asp His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 188
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 188
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Asp His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 189
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 189
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser His Asp
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 190
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 190
Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 191
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 191
Asn Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 192
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 192
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 193
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 193
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Asp His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 194
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 194
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 195
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 195
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 196
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 196
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser His Asp Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Asp His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 197
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 197
Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 198
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 198
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 199
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 199
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Asp His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 200
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 200
Asn Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 201
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 201
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Asp His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 202
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 202
Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 203
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 203
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Asp His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 204
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 204
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser His Asp
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 205
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 205
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Asp His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 206
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 206
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Asp His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Asn
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 207
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 207
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 208
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 208
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Asn
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 209
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 209
Asn Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 210
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 210
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 211
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 211
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Asp His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 212
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 212
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 213
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 213
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser His Asp
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 214
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 214
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 215
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 215
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 216
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 216
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 217
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 217
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser His Asp
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 218
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 218
Asn Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 219
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 219
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 220
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 220
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Asp His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 221
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 221
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Asp His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 222
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 222
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Asn
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 223
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 223
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Asp His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 224
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 224
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 225
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 225
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser His Asp
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 226
<211> 34
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 226
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser His Asp Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His
<210> 227
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 227
Asn Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 228
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 228
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 229
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 229
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Asp His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 230
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 230
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 231
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 231
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser His Asp
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 232
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 232
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 233
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 233
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 234
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 234
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Asp His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Asn
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 235
<211> 34
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 235
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Asp His
<210> 236
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 236
Asn Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 237
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 237
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 238
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 238
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Asp His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 239
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 239
Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 240
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 240
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 241
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 241
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Asp His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 242
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 242
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser His Asp Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 243
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 243
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Asp His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 244
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 244
Asn Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 245
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 245
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 246
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 246
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Asp His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 247
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 247
Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 248
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 248
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 249
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 249
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Asp His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 250
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 250
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser His Asp Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Asp His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser His Asp
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 251
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 251
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser His Asp Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Asp His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 252
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 252
Asn Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Asp His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 253
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 253
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 254
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 254
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Asp His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 255
<211> 69
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 255
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His Gly
65
<210> 256
<211> 67
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 256
Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys
1 5 10 15
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
20 25 30
His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly
35 40 45
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
50 55 60
Gln Ala His
65
<210> 257
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 257
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Asp His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 258
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 258
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 259
<211> 34
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 259
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His
<210> 260
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 260
Asn Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 261
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 261
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser His Asp
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His
65
<210> 262
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 262
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 263
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 263
Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser His Asp Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Asn
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 264
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 264
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 265
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 265
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Asp His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 266
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 266
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Asp His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 267
<211> 68
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 267
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Asp His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His
65
<210> 268
<211> 19
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 268
catcaatggg cgtggatag 19
<210> 269
<211> 136
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 269
Asp Ile Ala Asp Leu Arg Thr Leu Gly Tyr Ser Gln Gln Gln Gln Glu
1 5 10 15
Lys Ile Lys Pro Lys Val Arg Ser Thr Val Ala Gln His His Glu Ala
20 25 30
Leu Val Gly His Gly Phe Thr His Ala His Ile Val Ala Leu Ser Gln
35 40 45
His Pro Ala Ala Leu Gly Thr Val Ala Val Lys Tyr Gln Asp Met Ile
50 55 60
Ala Ala Leu Pro Glu Ala Thr His Glu Ala Ile Val Gly Val Gly Lys
65 70 75 80
Gln Trp Ser Gly Ala Arg Ala Leu Glu Ala Leu Leu Thr Val Ala Gly
85 90 95
Glu Leu Arg Gly Pro Pro Leu Gln Leu Asp Thr Gly Gln Leu Leu Lys
100 105 110
Ile Ala Lys Arg Gly Gly Val Thr Ala Val Glu Ala Val His Ala Trp
115 120 125
Arg Asn Ala Leu Thr Gly Ala Pro
130 135
<210> 270
<211> 137
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 270
Asp Ile Ala Asp Leu Arg Thr Leu Gly Tyr Ser Gln Gln Gln Gln Glu
1 5 10 15
Lys Ile Lys Pro Lys Val Arg Ser Thr Val Ala Gln His His Glu Ala
20 25 30
Leu Val Gly His Gly Phe Thr His Ala His Ile Val Ala Leu Ser Gln
35 40 45
His Pro Ala Ala Leu Gly Thr Val Ala Val Lys Tyr Gln Asp Met Ile
50 55 60
Ala Ala Leu Pro Glu Ala Thr His Glu Ala Ile Val Gly Val Gly Lys
65 70 75 80
Arg Gly Ala Gly Ala Arg Ala Leu Glu Ala Leu Leu Thr Val Ala Gly
85 90 95
Glu Leu Arg Gly Pro Pro Leu Gln Leu Asp Thr Gly Gln Leu Leu Lys
100 105 110
Ile Ala Lys Arg Gly Gly Val Thr Ala Val Glu Ala Val His Ala Trp
115 120 125
Arg Asn Ala Leu Thr Gly Ala Pro Leu
130 135
<210> 271
<211> 41
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 271
Ser Ile Val Ala Gln Leu Ser Arg Pro Asp Pro Ala Leu Ala Ala Leu
1 5 10 15
Thr Asn Asp His Leu Val Ala Leu Ala Cys Leu Gly Gly Arg Pro Ala
20 25 30
Leu Asp Ala Val Lys Lys Gly Leu Gly
35 40
<210> 272
<211> 94
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 272
Pro Thr Pro Tyr Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln
1 5 10 15
Ser Ala Leu Phe Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe
20 25 30
His Asn Asn Pro Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu
35 40 45
Thr Leu Leu Pro Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly
50 55 60
Ala Ile Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr
65 70 75 80
Gly Asn Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys
85 90
<210> 273
<211> 108
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 273
Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro
1 5 10 15
Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala
20 25 30
Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly Pro Thr Pro Tyr
35 40 45
Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln Ser Ala Leu Phe
50 55 60
Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe His Asn Asn Pro
65 70 75 80
Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu Thr Leu Leu Pro
85 90 95
Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly
100 105
<210> 274
<211> 49
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 274
Met Leu Gly Phe Val Gly Arg Val Ala Ala Ala Pro Ala Ser Gly Ala
1 5 10 15
Leu Arg Arg Leu Thr Pro Ser Ala Ser Leu Pro Pro Ala Gln Leu Leu
20 25 30
Leu Arg Ala Ala Pro Thr Ala Val His Pro Val Arg Asp Tyr Ala Ala
35 40 45
Gln
<210> 275
<211> 8
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 275
Asp Tyr Lys Asp Asp Asp Asp Lys
1 5
<210> 276
<211> 9
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 276
Tyr Pro Tyr Asp Val Pro Asp Tyr Ala
1 5
<210> 277
<211> 19
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 277
ttttcctact ggtccgatt 19
<210> 278
<211> 19
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 278
ttgcaggtat taattgctt 19
<210> 279
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 279
ttccctaaac ataaactcat 20
<210> 280
<211> 19
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 280
ttaaaataaa gttattatt 19
<210> 281
<211> 19
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 281
ttcctaacag ggttctact 19
<210> 282
<211> 17
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 282
ttgttgttgg agaatat 17
<210> 283
<211> 199
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 283
Met Ala Leu Ser Arg Ala Val Cys Gly Thr Ser Arg Gln Leu Ala Pro
1 5 10 15
Val Leu Gly Tyr Leu Gly Ser Arg Gln Lys His Ser Leu Pro Asp Tyr
20 25 30
Pro Tyr Asp Val Pro Asp Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp
35 40 45
Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Gly Ile Arg Ile
50 55 60
Gln Asp Leu Arg Thr Leu Gly Tyr Ser Gln Gln Gln Gln Glu Lys Ile
65 70 75 80
Lys Pro Lys Val Arg Ser Thr Val Ala Gln His His Glu Ala Leu Val
85 90 95
Gly His Gly Phe Thr His Ala His Ile Val Ala Leu Ser Gln His Pro
100 105 110
Ala Ala Leu Gly Thr Val Ala Val Lys Tyr Gln Asp Met Ile Ala Ala
115 120 125
Leu Pro Glu Ala Thr His Glu Ala Ile Val Gly Val Gly Lys Gln Trp
130 135 140
Ser Gly Ala Arg Ala Leu Glu Ala Leu Leu Thr Val Ala Gly Glu Leu
145 150 155 160
Arg Gly Pro Pro Leu Gln Leu Asp Thr Gly Gln Leu Leu Lys Ile Ala
165 170 175
Lys Arg Gly Gly Val Thr Ala Val Glu Ala Val His Ala Trp Arg Asn
180 185 190
Ala Leu Thr Gly Ala Pro Leu
195
<210> 284
<211> 191
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 284
Met Ala Ser Val Leu Thr Pro Leu Leu Leu Arg Gly Leu Thr Gly Ser
1 5 10 15
Ala Arg Arg Leu Pro Val Pro Arg Ala Lys Ile His Ser Leu Asp Tyr
20 25 30
Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp Ile Asp Tyr Lys Asp
35 40 45
Asp Asp Asp Lys Gly Ile Arg Ile Gln Asp Leu Arg Thr Leu Gly Tyr
50 55 60
Ser Gln Gln Gln Gln Glu Lys Ile Lys Pro Lys Val Arg Ser Thr Val
65 70 75 80
Ala Gln His His Glu Ala Leu Val Gly His Gly Phe Thr His Ala His
85 90 95
Ile Val Ala Leu Ser Gln His Pro Ala Ala Leu Gly Thr Val Ala Val
100 105 110
Lys Tyr Gln Asp Met Ile Ala Ala Leu Pro Glu Ala Thr His Glu Ala
115 120 125
Ile Val Gly Val Gly Lys Gln Trp Ser Gly Ala Arg Ala Leu Glu Ala
130 135 140
Leu Leu Thr Val Ala Gly Glu Leu Arg Gly Pro Pro Leu Gln Leu Asp
145 150 155 160
Thr Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly Gly Val Thr Ala Val
165 170 175
Glu Ala Val His Ala Trp Arg Asn Ala Leu Thr Gly Ala Pro Leu
180 185 190
<210> 285
<211> 197
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 285
Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp Pro
20 25 30
Ala Leu Ala Ala Leu Thr Asn Asp His Leu Val Ala Leu Ala Cys Leu
35 40 45
Gly Gly Arg Pro Ala Leu Asp Ala Val Lys Lys Gly Leu Gly Gly Ser
50 55 60
Gly Ser Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln
65 70 75 80
Leu Pro Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn
85 90 95
Asp Ala Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly Ser Gly
100 105 110
Gly Ser Thr Asn Leu Ser Asp Ile Ile Glu Lys Glu Thr Gly Lys Gln
115 120 125
Leu Val Ile Gln Glu Ser Ile Leu Met Leu Pro Glu Glu Val Glu Glu
130 135 140
Val Ile Gly Asn Lys Pro Glu Ser Asp Ile Leu Val His Thr Ala Tyr
145 150 155 160
Asp Glu Ser Thr Asp Glu Asn Val Met Leu Leu Thr Ser Asp Ala Pro
165 170 175
Glu Tyr Lys Pro Trp Ala Leu Val Ile Gln Asp Ser Asn Gly Glu Asn
180 185 190
Lys Ile Lys Met Leu
195
<210> 286
<211> 247
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 286
Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp Pro
20 25 30
Ala Leu Ala Ala Leu Thr Asn Asp His Leu Val Ala Leu Ala Cys Leu
35 40 45
Gly Gly Arg Pro Ala Leu Asp Ala Val Lys Lys Gly Leu Gly Gly Ser
50 55 60
Gly Ser Pro Thr Pro Tyr Pro Asn Tyr Ala Asn Ala Gly His Val Glu
65 70 75 80
Gly Gln Ser Ala Leu Phe Met Arg Asp Asn Gly Ile Ser Glu Gly Leu
85 90 95
Val Phe His Asn Asn Pro Glu Gly Thr Cys Gly Phe Cys Val Asn Met
100 105 110
Thr Glu Thr Leu Leu Pro Glu Asn Ala Lys Met Thr Val Val Pro Pro
115 120 125
Glu Gly Ala Ile Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val
130 135 140
Phe Thr Gly Asn Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys
145 150 155 160
Ser Gly Gly Ser Thr Asn Leu Ser Asp Ile Ile Glu Lys Glu Thr Gly
165 170 175
Lys Gln Leu Val Ile Gln Glu Ser Ile Leu Met Leu Pro Glu Glu Val
180 185 190
Glu Glu Val Ile Gly Asn Lys Pro Glu Ser Asp Ile Leu Val His Thr
195 200 205
Ala Tyr Asp Glu Ser Thr Asp Glu Asn Val Met Leu Leu Thr Ser Asp
210 215 220
Ala Pro Glu Tyr Lys Pro Trp Ala Leu Val Ile Gln Asp Ser Asn Gly
225 230 235 240
Glu Asn Lys Ile Lys Met Leu
245
<210> 287
<211> 261
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 287
Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp Pro
20 25 30
Ala Leu Ala Ala Leu Thr Asn Asp His Leu Val Ala Leu Ala Cys Leu
35 40 45
Gly Gly Arg Pro Ala Leu Asp Ala Val Lys Lys Gly Leu Gly Gly Ser
50 55 60
Gly Ser Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln
65 70 75 80
Leu Pro Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn
85 90 95
Asp Ala Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly Pro Thr
100 105 110
Pro Tyr Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln Ser Ala
115 120 125
Leu Phe Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe His Asn
130 135 140
Asn Pro Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu Thr Leu
145 150 155 160
Leu Pro Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly Ser Gly
165 170 175
Gly Ser Thr Asn Leu Ser Asp Ile Ile Glu Lys Glu Thr Gly Lys Gln
180 185 190
Leu Val Ile Gln Glu Ser Ile Leu Met Leu Pro Glu Glu Val Glu Glu
195 200 205
Val Ile Gly Asn Lys Pro Glu Ser Asp Ile Leu Val His Thr Ala Tyr
210 215 220
Asp Glu Ser Thr Asp Glu Asn Val Met Leu Leu Thr Ser Asp Ala Pro
225 230 235 240
Glu Tyr Lys Pro Trp Ala Leu Val Ile Gln Asp Ser Asn Gly Glu Asn
245 250 255
Lys Ile Lys Met Leu
260
<210> 288
<211> 183
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 288
Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp Pro
20 25 30
Ala Leu Ala Ala Leu Thr Asn Asp His Leu Val Ala Leu Ala Cys Leu
35 40 45
Gly Gly Arg Pro Ala Leu Asp Ala Val Lys Lys Gly Leu Gly Gly Ser
50 55 60
Gly Ser Ala Ile Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val
65 70 75 80
Phe Thr Gly Asn Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys
85 90 95
Ser Gly Gly Ser Thr Asn Leu Ser Asp Ile Ile Glu Lys Glu Thr Gly
100 105 110
Lys Gln Leu Val Ile Gln Glu Ser Ile Leu Met Leu Pro Glu Glu Val
115 120 125
Glu Glu Val Ile Gly Asn Lys Pro Glu Ser Asp Ile Leu Val His Thr
130 135 140
Ala Tyr Asp Glu Ser Thr Asp Glu Asn Val Met Leu Leu Thr Ser Asp
145 150 155 160
Ala Pro Glu Tyr Lys Pro Trp Ala Leu Val Ile Gln Asp Ser Asn Gly
165 170 175
Glu Asn Lys Ile Lys Met Leu
180
<210> 289
<211> 577
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 289
Asn Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser
65 70 75 80
Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro
85 90 95
Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile
100 105 110
Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu
115 120 125
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val Val
130 135 140
Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
145 150 155 160
Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Ala Gln
165 170 175
Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr
180 185 190
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro
195 200 205
Glu Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu
210 215 220
Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
225 230 235 240
Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln
245 250 255
Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His
260 265 270
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
275 280 285
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
290 295 300
Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Asn
305 310 315 320
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
325 330 335
Cys Gln Asp His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser
340 345 350
Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro
355 360 365
Val Leu Cys Gln Asp His Gly Leu Thr Pro Asp Gln Val Val Ala Ile
370 375 380
Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu
385 390 395 400
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val Val
405 410 415
Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
420 425 430
Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Asp Gln
435 440 445
Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr
450 455 460
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
465 470 475 480
Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu
485 490 495
Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
500 505 510
Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln
515 520 525
Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His
530 535 540
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
545 550 555 560
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
565 570 575
Asp
<210> 290
<211> 577
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 290
Asn Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Asp His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser
65 70 75 80
Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro
85 90 95
Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile
100 105 110
Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu
115 120 125
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val
130 135 140
Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
145 150 155 160
Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Asp Gln
165 170 175
Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr
180 185 190
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
195 200 205
Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu
210 215 220
Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
225 230 235 240
Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln
245 250 255
Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His
260 265 270
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
275 280 285
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
290 295 300
Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Asn
305 310 315 320
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
325 330 335
Cys Gln Asp His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser
340 345 350
Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro
355 360 365
Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile
370 375 380
Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu
385 390 395 400
Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Ala Gln Val Val
405 410 415
Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
420 425 430
Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln
435 440 445
Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr
450 455 460
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
465 470 475 480
Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu
485 490 495
Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
500 505 510
Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln
515 520 525
Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His
530 535 540
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
545 550 555 560
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
565 570 575
Asp
<210> 291
<211> 577
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 291
Asn Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser
65 70 75 80
His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro
85 90 95
Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile
100 105 110
Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu
115 120 125
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val Val
130 135 140
Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
145 150 155 160
Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Ala Gln
165 170 175
Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr
180 185 190
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
195 200 205
Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu
210 215 220
Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
225 230 235 240
Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln
245 250 255
Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His
260 265 270
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
275 280 285
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
290 295 300
Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly
305 310 315 320
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
325 330 335
Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser
340 345 350
Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro
355 360 365
Val Leu Cys Gln Asp His Gly Leu Thr Pro Ala Gln Val Val Ala Ile
370 375 380
Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu
385 390 395 400
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val
405 410 415
Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
420 425 430
Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln
435 440 445
Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr
450 455 460
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
465 470 475 480
Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu
485 490 495
Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu
500 505 510
Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln
515 520 525
Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His
530 535 540
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
545 550 555 560
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
565 570 575
Asp
<210> 292
<211> 577
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 292
Asn Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Asp His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser His Asp
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser
65 70 75 80
His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro
85 90 95
Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile
100 105 110
Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu
115 120 125
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val Val
130 135 140
Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
145 150 155 160
Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Ala Gln
165 170 175
Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr
180 185 190
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
195 200 205
Glu Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu
210 215 220
Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
225 230 235 240
Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln
245 250 255
Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His
260 265 270
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly
275 280 285
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
290 295 300
Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Asn
305 310 315 320
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
325 330 335
Cys Gln Asp His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser
340 345 350
Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro
355 360 365
Val Leu Cys Gln Asp His Gly Leu Thr Pro Asp Gln Val Val Ala Ile
370 375 380
Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu
385 390 395 400
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val
405 410 415
Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
420 425 430
Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln
435 440 445
Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr
450 455 460
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro
465 470 475 480
Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu
485 490 495
Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
500 505 510
Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln
515 520 525
Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His
530 535 540
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser His Asp Gly Gly
545 550 555 560
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
565 570 575
Asp
<210> 293
<211> 611
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 293
Asn Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Asp His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser His Asp
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser
65 70 75 80
His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro
85 90 95
Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile
100 105 110
Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu
115 120 125
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val
130 135 140
Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
145 150 155 160
Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Ala Gln
165 170 175
Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr
180 185 190
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro
195 200 205
Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu
210 215 220
Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
225 230 235 240
Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln
245 250 255
Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His
260 265 270
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser His Asp Gly Gly
275 280 285
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
290 295 300
Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile
305 310 315 320
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
325 330 335
Cys Gln Asp His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser
340 345 350
Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro
355 360 365
Val Leu Cys Gln Asp His Gly Leu Thr Pro Asp Gln Val Val Ala Ile
370 375 380
Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu
385 390 395 400
Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Asp Gln Val Val
405 410 415
Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
420 425 430
Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln
435 440 445
Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr
450 455 460
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
465 470 475 480
Ala Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu
485 490 495
Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
500 505 510
Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln
515 520 525
Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His
530 535 540
Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser His Asp Gly Gly
545 550 555 560
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
565 570 575
Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile
580 585 590
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
595 600 605
Cys Gln Asp
610
<210> 294
<211> 509
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 294
Asn Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
1 5 10 15
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
20 25 30
Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn
35 40 45
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
50 55 60
Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser
65 70 75 80
Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro
85 90 95
Val Leu Cys Gln Asp His Gly Leu Thr Pro Ala Gln Val Val Ala Ile
100 105 110
Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu
115 120 125
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val
130 135 140
Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
145 150 155 160
Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln
165 170 175
Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr
180 185 190
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro
195 200 205
Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu
210 215 220
Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
225 230 235 240
Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln
245 250 255
Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His
260 265 270
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly
275 280 285
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
290 295 300
Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile
305 310 315 320
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
325 330 335
Cys Gln Asp His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser
340 345 350
Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro
355 360 365
Val Leu Cys Gln Asp His Gly Leu Thr Pro Ala Gln Val Val Ala Ile
370 375 380
Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu
385 390 395 400
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val
405 410 415
Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
420 425 430
Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln
435 440 445
Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr
450 455 460
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro
465 470 475 480
Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu
485 490 495
Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
500 505
<210> 295
<211> 28
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 295
agtggggacc gcatactatg tcaacgac 28
<210> 296
<211> 18
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 296
gtctgaccgt tgtaggcg 18
<210> 297
<211> 30
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 297
atccaaggtt gcatctagcg gtgggtccgg 30
<210> 298
<211> 18
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 298
tccaatcccc cggcgtcg 18
<210> 299
<211> 33
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 299
cagacagtgg caacctttta ctatgtcaac gac 33
<210> 300
<211> 18
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 300
accgttgtag gcggggag 18
<210> 301
<211> 30
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 301
tccatatcag gcaagtgctc cccaactccc 30
<210> 302
<211> 18
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 302
cccagggcgt aggaacca 18
<210> 303
<211> 36
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 303
attggaatcc gcagttttct ctagcggtgg gtccgg 36
<210> 304
<211> 18
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 304
cccccggcgt cgttgaca 18
<210> 305
<211> 36
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 305
ctctggttcc tacgccgcag gtccatatca gattag 36
<210> 306
<211> 36
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 306
ctaatctgat atggacctgc ggcgtaggaa ccagag 36
<210> 307
<211> 30
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 307
atatcagatt gcagctcccc aactccccgc 30
<210> 308
<211> 18
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 308
ggacccaggg cgtaggaa 18
<210> 309
<211> 24
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 309
caaggttttc gcaagcggtg ggtc 24
<210> 310
<211> 18
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 310
gattccaatc ccccggcg 18
<210> 311
<211> 30
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 311
ggtcagacag tgggggcatt ttactatgtc 30
<210> 312
<211> 30
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 312
gacatagtaa aatgccccca ctgtctgacc 30
<210> 313
<211> 28
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 313
ggtcagacag cagggacctt ttactatg 28
<210> 314
<211> 18
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 314
gttgtaggcg gggagttg 18
<210> 315
<211> 25
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 315
ttttactatg caaacgacgc cgggg 25
<210> 316
<211> 18
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 316
ggtccccact gtctgacc 18
<210> 317
<211> 42
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 317
ctctggttcc gcagccctgg gtccatatca gattagtgct cc 42
<210> 318
<211> 18
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 318
ccagagcccc ccaacccc 18
<210> 319
<211> 31
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 319
cagtggggac ctttgcatat gtcaacgacg c 31
<210> 320
<211> 31
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 320
gcgtcgttga catatgcaaa ggtccccact g 31
<210> 321
<211> 28
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 321
gaccttttac gcagtcaacg acgccggg 28
<210> 322
<211> 18
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 322
cccactgtct gaccgttg 18
<210> 323
<211> 36
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 323
tgtggcttcg cagtgaatat gactgaaacc cttctg 36
<210> 324
<211> 32
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 324
catattcact gcgaagccac aagtaccctc tg 32
<210> 325
<211> 36
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 325
gtgtgaatat gactgcaacc cttctgcccg aaaatg 36
<210> 326
<211> 36
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 326
cattttcggg cagaagggtt gcagtcatat tcacac 36
<210> 327
<211> 35
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 327
gaaaatgccg caatgactgt cgtcccacct gaagg 35
<210> 328
<211> 31
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 328
gacagtcatt gcggcatttt cgggcagaag g 31
<210> 329
<211> 32
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 329
gaaacccttg cacccgaaaa tgccaagatg ac 32
<210> 330
<211> 40
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 330
attttcgggt gcaagggttt cagtcatatt cacacagaag 40
<210> 331
<211> 32
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 331
aatgccaagg caactgtcgt cccacctgaa gg 32
<210> 332
<211> 34
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 332
gacgacagtt gccttggcat tttcgggcag aagg 34
<210> 333
<211> 34
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 333
gtgaatatgg cagaaaccct tctgcccgaa aatg 34
<210> 334
<211> 35
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 334
aagggtttct gccatattca cacagaagcc acaag 35
<210> 335
<211> 35
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 335
cgaaaatgcc aagatggcag tcgtcccacc tgaag 35
<210> 336
<211> 35
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 336
cttcaggtgg gacgactgcc atcttggcat tttcg 35
<210> 337
<211> 36
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 337
ggtacttgtg gcttctgtgc aaatatgact gaaacc 36
<210> 338
<211> 36
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 338
ggtttcagtc atatttgcac agaagccaca agtacc 36
<210> 339
<211> 35
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 339
gaaaatgcca agatgactgc agtcccacct gaagg 35
<210> 340
<211> 35
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 340
ccttcaggtg ggactgcagt catcttggca ttttc 35
<210> 341
<211> 35
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 341
ggcattagcg cagggcttgt gttccataat aatcc 35
<210> 342
<211> 42
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 342
cacaagccct gcgctaatgc cgttatctct cataaaaaga gc 42
<210> 343
<211> 40
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 343
gtcagctctt gcaatgagag ataacggcat tagcgaaggg 40
<210> 344
<211> 18
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 344
tgcccctcga cgtgcccg 18
<210> 345
<211> 35
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 345
gggcttgtgg cacataataa tcctgagggc acctg 35
<210> 346
<211> 32
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 346
gattattatg tgccacaagc ccttcgctaa tg 32
<210> 347
<211> 23
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 347
cattagcgaa gcacttgtgt tcc 23
<210> 348
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 348
ccgttatctc tcataaaaag 20
<210> 349
<211> 30
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 349
gcttgtgttc gcaaataatc ctgagggcac 30
<210> 350
<211> 18
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 350
ccttcgctaa tgccgtta 18
<210> 351
<211> 23
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 351
tgagaacgct gcaatgactg tcg 23
<210> 352
<211> 18
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 352
ggcagaagtg tttcggtc 18
<210> 353
<211> 37
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 353
tagcgaaggg gcagtgttcc ataataatcc tgagggc 37
<210> 354
<211> 35
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 354
atggaacact gccccttcgc taatgccgtt atctc 35
<210> 355
<211> 36
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 355
agctcttttt gcaagagata acggcattag cgaagg 36
<210> 356
<211> 18
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 356
gactgcccct cgacgtgc 18
<210> 357
<211> 26
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 357
gaacgctaaa gcaactgtcg taccac 26
<210> 358
<211> 18
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 358
tcaggcagaa gtgtttcg 18
<210> 359
<211> 40
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 359
cgtcgagggg gcatcagctc tttttatgag agataacggc 40
<210> 360
<211> 18
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 360
tgcccggcgt tagcgtag 18
<210> 361
<211> 25
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 361
cgaggggcag gcagctcttt ttatg 25
<210> 362
<211> 18
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 362
acgtgcccgg cgttagcg 18
<210> 363
<211> 23
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 363
cgctaaaatg gcagtcgtac cac 23
<210> 364
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 364
ttctcaggca gaagtgtttc 20
<210> 365
<211> 22
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 365
gccgggcacg cagaggggca gt 22
<210> 366
<211> 26
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 366
gttagcgtag ttaggataag gtgttg 26
<210> 367
<211> 27
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 367
cgaagggctt gcattccata ataatcc 27
<210> 368
<211> 18
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 368
ctaatgccgt tatctctc 18
<210> 369
<211> 23
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 369
aatgactgtc gcaccacccg aag 23
<210> 370
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 370
ttagcgttct caggcagaag 20
<210> 371
<211> 36
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 371
acaaaggtgg caacaggcaa ctctaacagt ccaaag 36
<210> 372
<211> 36
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 372
gttgcctgtt gccacctttg tctcccctgt tgctcc 36
<210> 373
<211> 31
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 373
caacagggga gacagcagtg ttcacaggca a 31
<210> 374
<211> 31
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 374
ttgcctgtga acactgctgt ctcccctgtt g 31
<210> 375
<211> 36
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 375
ggagacaaag gtgttcgcag gcaactctaa cagtcc 36
<210> 376
<211> 36
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 376
ggactgttag agttgcctgc gaacaccttt gtctcc 36
<210> 377
<211> 34
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 377
gagacaaagg cattcacagg caactctaac agtc 34
<210> 378
<211> 33
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 378
gcctgtgaat gcctttgtct cccctgttgc tcc 33
<210> 379
<211> 138
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 379
Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro
1 5 10 15
Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala
20 25 30
Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly Pro Thr Pro Tyr
35 40 45
Pro Asn Tyr Ala Asn Ala Gly His Val Ala Gly Gln Ser Ala Leu Phe
50 55 60
Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe His Asn Asn Pro
65 70 75 80
Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu Thr Leu Leu Pro
85 90 95
Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly Ala Ile Pro Val
100 105 110
Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr Gly Asn Ser Asn
115 120 125
Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys
130 135
<210> 380
<211> 51
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 380
cccaagcttg ccaccatggg atccggcagc tacgccctgg gtccgtatca g 51
<210> 381
<211> 49
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 381
cccgggagtc tcgctgccgc tgcaaccgcc tttggtcggg ctcttcggg 49
<210> 382
<211> 110
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 382
cccgggagtc tcgctgccgc tgcaaccgcc tttggtcggg ctcttcgggc tgttgctgtt 60
accggtaaac actttggttt caccggtagc gccgcgcgcc actggaatgg 110
<210> 383
<211> 108
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 383
cccgggagtc tcgctgccgc tgcaaccgcc tttggtcggg ctcttcgggc tgttgctgtt 60
accggtaaac actttggttt caccggtagc gccggccttc ttcactgg 108
<210> 384
<211> 110
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 384
cccgggagtc tcgctgccgc tgcaaccgcc tttggtcggg ctcttcgggc tgttgctgtt 60
accggtaaac actttggttt caccggtagc gccggccgcc actggaatgg 110
<210> 385
<211> 84
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 385
cccgggagtc tcgctgccgc tgcaaccgcc tttggtcggg ctcttcgggc tgttgctgtt 60
accggtaaac acggcggttt cacc 84
<210> 386
<211> 58
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 386
cccgggagtc tcgctgccgc tgcaaccgcc tttggtcggg ctggccgggc tgttgctg 58
<210> 387
<211> 49
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 387
cccgggagtc tcgctgccgc tgcaaccgcc ggcggtcggg ctcttcggg 49
<210> 388
<211> 46
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 388
ctatgccaat gccggtcatg tggccggtca gagcgccctg ttcatg 46
<210> 389
<211> 46
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 389
catgaacagg gcgctctgac cggccacatg accggcattg gcatag 46
<210> 390
<211> 88
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 390
tgagccagct gggcggcgac agcggcagcg agactcccgg gacctcagag tccgccacac 60
ccgaaagtgg cagctacgcc ctgggtcc 88
<210> 391
<211> 50
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 391
aatagggccc tctagatgca tgctcgagtt agcaaccgcc tttggtcggg 50
<210> 392
<211> 50
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 392
tgaagaaagg tctgggcgga tccggcagct acgccctggg tccgtatcag 50
<210> 393
<211> 50
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 393
tgaagaaagg cctgggtgga tccggcagct acgccctggg tccgtatcag 50
<210> 394
<211> 50
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 394
tcagattagt tgagccgcca gagcaaccgc ctttggtcgg gctcttcggg 50
<210> 395
<211> 34
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 395
tctggcggct caactaatct gagcgacatc attg 34
<210> 396
<211> 29
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 396
cagatccgaa aatggatata caagctccc 29
<210> 397
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 397
ggcccagact gagcacgtga 20
<210> 398
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 398
ggccacacta gcgttgctgc 20
<210> 399
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 399
gccatagatg gtggaccgaa 20
<210> 400
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 400
ccatctatgg ctctcggctg 20
<210> 401
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 401
ccgcagccga gagccataga 20
<210> 402
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 402
ggaatccctt ctgcagcacc 20
<210> 403
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 403
cttgccccac agggcagtaa 20
<210> 404
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 404
tgcccctccc tccctggccc 20
<210> 405
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 405
ccctggccca ggtgaaggtg 20
<210> 406
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 406
gtgaaggtgt ggttccagaa c 21
<210> 407
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 407
aaagtacaaa cggcagaagc 20
<210> 408
<211> 21
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 408
gtggtagcgg aactcactaa g 21
<210> 409
<211> 20
<212> DNA
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 409
cacccagcct gctctgcctt 20
<210> 410
<211> 434
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 410
Met Leu Gly Phe Val Gly Arg Val Ala Ala Ala Pro Ala Ser Gly Ala
1 5 10 15
Leu Arg Arg Leu Thr Pro Ser Ala Ser Leu Pro Pro Ala Gln Leu Leu
20 25 30
Leu Arg Ala Ala Pro Thr Ala Val His Pro Val Arg Asp Tyr Ala Ala
35 40 45
Gln Asp Tyr Lys Asp Asp Asp Asp Lys Val Asp Glu Met Thr Lys Lys
50 55 60
Phe Gly Thr Leu Thr Ile His Asp Thr Glu Lys Ala Ala Glu Phe Gly
65 70 75 80
Ile Arg Ile Pro Gly Glu Lys Pro Phe Gln Cys Arg Ile Cys Met Arg
85 90 95
Asn Phe Ser Asp Ser Gly Asn Leu Arg Val His Ile Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Asp Cys Gly Lys Ser Phe Ser Gln
115 120 125
Ser Ser Ser Leu Ile Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Glu Cys Asp His Cys Gly Lys Ser Phe Ser Gln Ser Ser His Leu
145 150 155 160
Asn Val His Lys Arg Thr His Thr Gly Glu Lys Pro Tyr Arg Cys Lys
165 170 175
Tyr Cys Asp Arg Ser Phe Ser Ile Ser Ser Asn Leu Gln Arg His Val
180 185 190
Arg Asn Ile His Leu Arg Ser Gly Thr Pro His Glu Val Gly Val Tyr
195 200 205
Thr Leu Ser Gly Thr Pro His Glu Val Gly Val Tyr Thr Leu Ala Ile
210 215 220
Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr Gly Asn
225 230 235 240
Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys Ser Gly Ser Glu
245 250 255
Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser Ser Glu Val Glu
260 265 270
Phe Ser His Glu Tyr Trp Met Arg His Ala Leu Thr Leu Ala Lys Arg
275 280 285
Ala Arg Asp Glu Arg Glu Val Pro Val Gly Ala Val Leu Val Leu Asn
290 295 300
Asn Arg Val Ile Gly Glu Gly Trp Asn Arg Ala Ile Gly Leu His Asp
305 310 315 320
Pro Thr Ala His Ala Glu Ile Met Ala Leu Arg Gln Gly Gly Leu Val
325 330 335
Met Gln Asn Tyr Arg Leu Ile Asp Ala Thr Leu Tyr Val Thr Phe Glu
340 345 350
Pro Cys Val Met Cys Ala Gly Ala Met Ile His Ser Arg Ile Gly Arg
355 360 365
Val Val Phe Gly Val Arg Asn Ser Lys Arg Gly Ala Ala Gly Ser Leu
370 375 380
Met Asn Val Leu Asn Tyr Pro Gly Met Asn His Arg Val Glu Ile Thr
385 390 395 400
Glu Gly Ile Leu Ala Asp Glu Cys Ala Ala Leu Leu Cys Asp Phe Tyr
405 410 415
Arg Met Pro Arg Gln Val Phe Asn Ala Gln Lys Lys Ala Gln Ser Ser
420 425 430
Ile Asn
<210> 411
<211> 339
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 411
Met Leu Gly Phe Val Gly Arg Val Ala Ala Ala Pro Ala Ser Gly Ala
1 5 10 15
Leu Arg Arg Leu Thr Pro Ser Ala Ser Leu Pro Pro Ala Gln Leu Leu
20 25 30
Leu Arg Ala Ala Pro Thr Ala Val His Pro Val Arg Asp Tyr Ala Ala
35 40 45
Gln Asp Tyr Lys Asp Asp Asp Asp Lys Val Asp Glu Met Thr Lys Lys
50 55 60
Phe Gly Thr Leu Thr Ile His Asp Thr Glu Lys Ala Ala Glu Phe Gly
65 70 75 80
Ile Arg Ile Pro Gly Glu Lys Pro Phe Gln Cys Arg Ile Cys Met Arg
85 90 95
Asn Phe Ser Asp Ser Gly Asn Leu Arg Val His Ile Arg Thr His Thr
100 105 110
Gly Glu Lys Pro Tyr Lys Cys Pro Asp Cys Gly Lys Ser Phe Ser Gln
115 120 125
Ser Ser Ser Leu Ile Arg His Gln Arg Thr His Thr Gly Glu Lys Pro
130 135 140
Tyr Glu Cys Asp His Cys Gly Lys Ser Phe Ser Gln Ser Ser His Leu
145 150 155 160
Asn Val His Lys Arg Thr His Thr Gly Glu Lys Pro Tyr Arg Cys Lys
165 170 175
Tyr Cys Asp Arg Ser Phe Ser Ile Ser Ser Asn Leu Gln Arg His Val
180 185 190
Arg Asn Ile His Leu Arg Ser Gly Thr Pro His Glu Val Gly Val Tyr
195 200 205
Thr Leu Ser Gly Thr Pro His Glu Val Gly Val Tyr Thr Leu Ala Ile
210 215 220
Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr Gly Asn
225 230 235 240
Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys Ser Gly Gly Ser
245 250 255
Thr Asn Leu Ser Asp Ile Ile Glu Lys Glu Thr Gly Lys Gln Leu Val
260 265 270
Ile Gln Glu Ser Ile Leu Met Leu Pro Glu Glu Val Glu Glu Val Ile
275 280 285
Gly Asn Lys Pro Glu Ser Asp Ile Leu Val His Thr Ala Tyr Asp Glu
290 295 300
Ser Thr Asp Glu Asn Val Met Leu Leu Thr Ser Asp Ala Pro Glu Tyr
305 310 315 320
Lys Pro Trp Ala Leu Val Ile Gln Asp Ser Asn Gly Glu Asn Lys Ile
325 330 335
Lys Met Leu
<210> 412
<211> 419
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 412
Met Leu Gly Phe Val Gly Arg Val Ala Ala Ala Pro Ala Ser Gly Ala
1 5 10 15
Leu Arg Arg Leu Thr Pro Ser Ala Ser Leu Pro Pro Ala Gln Leu Leu
20 25 30
Leu Arg Ala Ala Pro Thr Ala Val His Pro Val Arg Asp Tyr Ala Ala
35 40 45
Gln Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Val Asp Glu Met Thr Lys
50 55 60
Lys Phe Gly Thr Leu Thr Ile His Asp Thr Glu Lys Ala Ala Glu Phe
65 70 75 80
Gly Ile His Gly Val Pro Ala Ala Met Gly Gly Ser Tyr Ala Leu Gly
85 90 95
Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro Ala Tyr Asn Gly Gln Thr
100 105 110
Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala Gly Gly Leu Glu Ser Lys
115 120 125
Val Phe Ser Ser Gly Gly Pro Thr Pro Tyr Pro Asn Tyr Ala Asn Ala
130 135 140
Gly His Val Glu Gly Gln Ser Ala Leu Phe Met Arg Asp Asn Gly Ile
145 150 155 160
Ser Glu Gly Leu Val Phe His Asn Asn Pro Glu Gly Thr Cys Gly Phe
165 170 175
Cys Val Asn Met Thr Glu Thr Leu Leu Pro Glu Asn Ala Lys Met Thr
180 185 190
Val Val Pro Pro Glu Gly Ser Gly Thr Pro His Glu Val Gly Val Tyr
195 200 205
Thr Leu Ser Gly Thr Pro His Glu Val Gly Val Tyr Thr Leu Tyr Lys
210 215 220
Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Lys Asn Ser Leu Thr Glu
225 230 235 240
His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro Glu Cys
245 250 255
Gly Lys Ser Phe Ser Ser Lys Lys Ala Leu Thr Glu His Gln Arg Thr
260 265 270
His Thr Gly Glu Lys Pro Tyr Glu Cys Asn Tyr Cys Gly Lys Thr Phe
275 280 285
Ser Val Ser Ser Thr Leu Ile Arg His Gln Arg Ile His Thr Gly Glu
290 295 300
Lys Pro Tyr Arg Cys Lys Tyr Cys Asp Arg Ser Phe Ser Ile Ser Ser
305 310 315 320
Asn Leu Gln Arg His Val Arg Asn Ile His Leu Arg Ser Gly Gly Ser
325 330 335
Thr Asn Leu Ser Asp Ile Ile Glu Lys Glu Thr Gly Lys Gln Leu Val
340 345 350
Ile Gln Glu Ser Ile Leu Met Leu Pro Glu Glu Val Glu Glu Val Ile
355 360 365
Gly Asn Lys Pro Glu Ser Asp Ile Leu Val His Thr Ala Tyr Asp Glu
370 375 380
Ser Thr Asp Glu Asn Val Met Leu Leu Thr Ser Asp Ala Pro Glu Tyr
385 390 395 400
Lys Pro Trp Ala Leu Val Ile Gln Asp Ser Asn Gly Glu Asn Lys Ile
405 410 415
Lys Met Leu
<210> 413
<211> 380
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 413
Met Ala Pro Lys Lys Lys Arg Lys Val Gly Ile His Gly Val Pro Ala
1 5 10 15
Ala Met Gly Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro
20 25 30
Gln Leu Pro Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val
35 40 45
Asn Asp Ala Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly Pro
50 55 60
Thr Pro Tyr Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln Ser
65 70 75 80
Ala Leu Phe Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe His
85 90 95
Asn Asn Pro Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu Thr
100 105 110
Leu Leu Pro Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly Ser
115 120 125
Gly Thr Pro His Glu Val Gly Val Tyr Thr Leu Ser Gly Thr Pro His
130 135 140
Glu Val Gly Val Tyr Thr Leu Phe Gln Cys Arg Ile Cys Met Arg Lys
145 150 155 160
Phe Ala Thr Ser Gly Ser Leu Thr Arg His Thr Lys Ile His Thr Gly
165 170 175
Glu Lys Pro Phe Gln Cys Arg Ile Cys Met Arg Asn Phe Ser Arg Ser
180 185 190
Asp His Leu Ser Thr His Ile Arg Thr His Thr Gly Glu Lys Pro Phe
195 200 205
Ala Cys Asp Ile Cys Gly Arg Lys Phe Ala Thr Ser Ser Asn Arg Thr
210 215 220
Lys His Thr Lys Ile His Thr His Pro Arg Ala Pro Ile Pro Lys Pro
225 230 235 240
Phe Gln Cys Arg Ile Cys Met Arg Asn Phe Ser Arg Ser Asp Asn Leu
245 250 255
Ser Glu His Ile Arg Thr His Thr Gly Glu Lys Pro Phe Ala Cys Asp
260 265 270
Ile Cys Gly Arg Lys Phe Ala Trp His Ser Ser Leu Arg Val His Thr
275 280 285
Lys Ile His Leu Arg Ser Gly Gly Ser Thr Asn Leu Ser Asp Ile Ile
290 295 300
Glu Lys Glu Thr Gly Lys Gln Leu Val Ile Gln Glu Ser Ile Leu Met
305 310 315 320
Leu Pro Glu Glu Val Glu Glu Val Ile Gly Asn Lys Pro Glu Ser Asp
325 330 335
Ile Leu Val His Thr Ala Tyr Asp Glu Ser Thr Asp Glu Asn Val Met
340 345 350
Leu Leu Thr Ser Asp Ala Pro Glu Tyr Lys Pro Trp Ala Leu Val Ile
355 360 365
Gln Asp Ser Asn Gly Glu Asn Lys Ile Lys Met Leu
370 375 380
<210> 414
<211> 396
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 414
Met Ala Pro Lys Lys Lys Arg Lys Val Gly Ile His Gly Val Pro Ala
1 5 10 15
Ala Met Ala Glu Arg Pro Phe Gln Cys Arg Ile Cys Met Arg Asn Phe
20 25 30
Ser Arg Ser Asp His Leu Ser Thr His Ile Arg Thr His Thr Gly Glu
35 40 45
Lys Pro Phe Ala Cys Asp Ile Cys Gly Arg Lys Phe Ala Asp Arg Ser
50 55 60
His Leu Ala Arg His Thr Lys Ile His Thr Gly Ser Gln Lys Pro Phe
65 70 75 80
Gln Cys Arg Ile Cys Met Arg Lys Phe Ala Leu Lys Gln His Leu Asn
85 90 95
Glu His Thr Lys Ile His Thr Gly Glu Lys Pro Phe Gln Cys Arg Ile
100 105 110
Cys Met Arg Asn Phe Ser Gln Ser Gly Asn Leu Ala Arg His Ile Arg
115 120 125
Thr His Thr Gly Glu Lys Pro Phe Ala Cys Asp Ile Cys Gly Arg Lys
130 135 140
Phe Ala His Asn Ser Ser Leu Lys Asp His Thr Lys Ile His Leu Arg
145 150 155 160
Ser Gly Thr Pro His Glu Val Gly Val Tyr Thr Leu Ser Gly Thr Pro
165 170 175
His Glu Val Gly Val Tyr Thr Leu Ala Ile Pro Val Lys Arg Gly Ala
180 185 190
Thr Gly Glu Thr Lys Val Phe Thr Gly Asn Ser Asn Ser Pro Lys Ser
195 200 205
Pro Thr Lys Gly Gly Cys Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu
210 215 220
Ser Ala Thr Pro Glu Ser Ser Glu Val Glu Phe Ser His Glu Tyr Trp
225 230 235 240
Met Arg His Ala Leu Thr Leu Ala Lys Arg Ala Arg Asp Glu Arg Glu
245 250 255
Val Pro Val Gly Ala Val Leu Val Leu Asn Asn Arg Val Ile Gly Glu
260 265 270
Gly Trp Asn Arg Ala Ile Gly Leu His Asp Pro Thr Ala His Ala Glu
275 280 285
Ile Met Ala Leu Arg Gln Gly Gly Leu Val Met Gln Asn Tyr Arg Leu
290 295 300
Ile Asp Ala Thr Leu Tyr Val Thr Phe Glu Pro Cys Val Met Cys Ala
305 310 315 320
Gly Ala Met Ile His Ser Arg Ile Gly Arg Val Val Phe Gly Val Arg
325 330 335
Asn Ser Lys Arg Gly Ala Ala Gly Ser Leu Met Asn Val Leu Asn Tyr
340 345 350
Pro Gly Met Asn His Arg Val Glu Ile Thr Glu Gly Ile Leu Ala Asp
355 360 365
Glu Cys Ala Ala Leu Leu Cys Asp Phe Tyr Arg Met Pro Arg Gln Val
370 375 380
Phe Asn Ala Gln Lys Lys Ala Gln Ser Ser Ile Asn
385 390 395
<210> 415
<211> 911
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 415
Met Ala Leu Ser Arg Ala Val Cys Gly Thr Ser Arg Gln Leu Ala Pro
1 5 10 15
Val Leu Gly Tyr Leu Gly Ser Arg Gln Lys His Ser Leu Pro Asp Tyr
20 25 30
Pro Tyr Asp Val Pro Asp Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp
35 40 45
Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Gly Ile Arg Ile
50 55 60
Gln Asp Leu Arg Thr Leu Gly Tyr Ser Gln Gln Gln Gln Glu Lys Ile
65 70 75 80
Lys Pro Lys Val Arg Ser Thr Val Ala Gln His His Glu Ala Leu Val
85 90 95
Gly His Gly Phe Thr His Ala His Ile Val Ala Leu Ser Gln His Pro
100 105 110
Ala Ala Leu Gly Thr Val Ala Val Lys Tyr Gln Asp Met Ile Ala Ala
115 120 125
Leu Pro Glu Ala Thr His Glu Ala Ile Val Gly Val Gly Lys Gln Trp
130 135 140
Ser Gly Ala Arg Ala Leu Glu Ala Leu Leu Thr Val Ala Gly Glu Leu
145 150 155 160
Arg Gly Pro Pro Leu Gln Leu Asp Thr Gly Gln Leu Leu Lys Ile Ala
165 170 175
Lys Arg Gly Gly Val Thr Ala Val Glu Ala Val His Ala Trp Arg Asn
180 185 190
Ala Leu Thr Gly Ala Pro Leu Asn Leu Thr Pro Asp Gln Val Val Ala
195 200 205
Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
210 215 220
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val
225 230 235 240
Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val
245 250 255
Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Ala
260 265 270
Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu
275 280 285
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr
290 295 300
Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala
305 310 315 320
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
325 330 335
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys
340 345 350
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp
355 360 365
His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly
370 375 380
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
385 390 395 400
Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn
405 410 415
Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
420 425 430
Leu Cys Gln Asp His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala
435 440 445
Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
450 455 460
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala
465 470 475 480
Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
485 490 495
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val
500 505 510
Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val
515 520 525
Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Asp
530 535 540
Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu
545 550 555 560
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr
565 570 575
Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala
580 585 590
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
595 600 605
Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys
610 615 620
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
625 630 635 640
His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly
645 650 655
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
660 665 670
Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn
675 680 685
Gly Gly Gly Lys Gln Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg
690 695 700
Pro Asp Pro Ala Leu Ala Ala Leu Leu Val Gly Ser Gly Ser Tyr Ala
705 710 715 720
Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro Ala Tyr Asn Gly
725 730 735
Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala Gly Gly Leu Glu
740 745 750
Ser Lys Val Phe Ser Ser Gly Gly Pro Thr Pro Tyr Pro Asn Tyr Ala
755 760 765
Asn Ala Gly His Val Glu Gly Gln Ser Ala Leu Phe Met Arg Asp Asn
770 775 780
Gly Ile Ser Glu Gly Leu Val Phe His Asn Asn Pro Glu Gly Thr Cys
785 790 795 800
Gly Phe Cys Val Asn Met Thr Glu Thr Leu Leu Pro Glu Asn Ala Lys
805 810 815
Met Thr Val Val Pro Pro Glu Gly Ser Gly Gly Ser Thr Asn Leu Ser
820 825 830
Asp Ile Ile Glu Lys Glu Thr Gly Lys Gln Leu Val Ile Gln Glu Ser
835 840 845
Ile Leu Met Leu Pro Glu Glu Val Glu Glu Val Ile Gly Asn Lys Pro
850 855 860
Glu Ser Asp Ile Leu Val His Thr Ala Tyr Asp Glu Ser Thr Asp Glu
865 870 875 880
Asn Val Met Leu Leu Thr Ser Asp Ala Pro Glu Tyr Lys Pro Trp Ala
885 890 895
Leu Val Ile Gln Asp Ser Asn Gly Glu Asn Lys Ile Lys Met Leu
900 905 910
<210> 416
<211> 833
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 416
Met Ala Leu Ser Arg Ala Val Cys Gly Thr Ser Arg Gln Leu Ala Pro
1 5 10 15
Val Leu Gly Tyr Leu Gly Ser Arg Gln Lys His Ser Leu Pro Asp Tyr
20 25 30
Pro Tyr Asp Val Pro Asp Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp
35 40 45
Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Gly Ile Arg Ile
50 55 60
Gln Asp Leu Arg Thr Leu Gly Tyr Ser Gln Gln Gln Gln Glu Lys Ile
65 70 75 80
Lys Pro Lys Val Arg Ser Thr Val Ala Gln His His Glu Ala Leu Val
85 90 95
Gly His Gly Phe Thr His Ala His Ile Val Ala Leu Ser Gln His Pro
100 105 110
Ala Ala Leu Gly Thr Val Ala Val Lys Tyr Gln Asp Met Ile Ala Ala
115 120 125
Leu Pro Glu Ala Thr His Glu Ala Ile Val Gly Val Gly Lys Gln Trp
130 135 140
Ser Gly Ala Arg Ala Leu Glu Ala Leu Leu Thr Val Ala Gly Glu Leu
145 150 155 160
Arg Gly Pro Pro Leu Gln Leu Asp Thr Gly Gln Leu Leu Lys Ile Ala
165 170 175
Lys Arg Gly Gly Val Thr Ala Val Glu Ala Val His Ala Trp Arg Asn
180 185 190
Ala Leu Thr Gly Ala Pro Leu Asn Leu Thr Pro Asp Gln Val Val Ala
195 200 205
Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
210 215 220
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val
225 230 235 240
Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val
245 250 255
Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Ala
260 265 270
Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu
275 280 285
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr
290 295 300
Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala
305 310 315 320
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
325 330 335
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys
340 345 350
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp
355 360 365
His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly
370 375 380
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
385 390 395 400
Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn
405 410 415
Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
420 425 430
Leu Cys Gln Asp His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala
435 440 445
Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
450 455 460
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala
465 470 475 480
Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
485 490 495
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val
500 505 510
Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val
515 520 525
Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Asp
530 535 540
Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu
545 550 555 560
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr
565 570 575
Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala
580 585 590
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
595 600 605
Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys
610 615 620
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
625 630 635 640
His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly
645 650 655
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
660 665 670
Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn
675 680 685
Gly Gly Gly Lys Gln Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg
690 695 700
Pro Asp Pro Ala Leu Ala Ala Leu Leu Val Gly Ser Ala Ile Pro Val
705 710 715 720
Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr Gly Asn Ser Asn
725 730 735
Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys Ser Gly Gly Ser Thr Asn
740 745 750
Leu Ser Asp Ile Ile Glu Lys Glu Thr Gly Lys Gln Leu Val Ile Gln
755 760 765
Glu Ser Ile Leu Met Leu Pro Glu Glu Val Glu Glu Val Ile Gly Asn
770 775 780
Lys Pro Glu Ser Asp Ile Leu Val His Thr Ala Tyr Asp Glu Ser Thr
785 790 795 800
Asp Glu Asn Val Met Leu Leu Thr Ser Asp Ala Pro Glu Tyr Lys Pro
805 810 815
Trp Ala Leu Val Ile Gln Asp Ser Asn Gly Glu Asn Lys Ile Lys Met
820 825 830
Leu
<210> 417
<211> 1005
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 417
Met Ala Ser Val Leu Thr Pro Leu Leu Leu Arg Gly Leu Thr Gly Ser
1 5 10 15
Ala Arg Arg Leu Pro Val Pro Arg Ala Lys Ile His Ser Leu Asp Tyr
20 25 30
Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp Ile Asp Tyr Lys Asp
35 40 45
Asp Asp Asp Lys Gly Ile Arg Ile Gln Asp Leu Arg Thr Leu Gly Tyr
50 55 60
Ser Gln Gln Gln Gln Glu Lys Ile Lys Pro Lys Val Arg Ser Thr Val
65 70 75 80
Ala Gln His His Glu Ala Leu Val Gly His Gly Phe Thr His Ala His
85 90 95
Ile Val Ala Leu Ser Gln His Pro Ala Ala Leu Gly Thr Val Ala Val
100 105 110
Lys Tyr Gln Asp Met Ile Ala Ala Leu Pro Glu Ala Thr His Glu Ala
115 120 125
Ile Val Gly Val Gly Lys Gln Trp Ser Gly Ala Arg Ala Leu Glu Ala
130 135 140
Leu Leu Thr Val Ala Gly Glu Leu Arg Gly Pro Pro Leu Gln Leu Asp
145 150 155 160
Thr Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly Gly Val Thr Ala Val
165 170 175
Glu Ala Val His Ala Trp Arg Asn Ala Leu Thr Gly Ala Pro Leu Asn
180 185 190
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys
195 200 205
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp
210 215 220
His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile Gly
225 230 235 240
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
245 250 255
Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn
260 265 270
Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
275 280 285
Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala
290 295 300
Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
305 310 315 320
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala
325 330 335
Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
340 345 350
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val
355 360 365
Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val
370 375 380
Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala
385 390 395 400
Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu
405 410 415
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr
420 425 430
Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala
435 440 445
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
450 455 460
Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys
465 470 475 480
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
485 490 495
His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly
500 505 510
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
515 520 525
Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser His
530 535 540
Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
545 550 555 560
Leu Cys Gln Asp His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala
565 570 575
Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
580 585 590
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala
595 600 605
Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
610 615 620
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val
625 630 635 640
Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val
645 650 655
Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala
660 665 670
Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu
675 680 685
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr
690 695 700
Pro Ala Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala
705 710 715 720
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly
725 730 735
Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys
740 745 750
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
755 760 765
His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly
770 775 780
Gly Lys Gln Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp
785 790 795 800
Pro Ala Leu Ala Ala Leu Leu Val Gly Ser Gly Ser Tyr Ala Leu Gly
805 810 815
Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro Ala Tyr Asn Gly Gln Thr
820 825 830
Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala Gly Gly Leu Glu Ser Lys
835 840 845
Val Phe Ser Ser Gly Gly Pro Thr Pro Tyr Pro Asn Tyr Ala Asn Ala
850 855 860
Gly His Val Glu Gly Gln Ser Ala Leu Phe Met Arg Asp Asn Gly Ile
865 870 875 880
Ser Glu Gly Leu Val Phe His Asn Asn Pro Glu Gly Thr Cys Gly Phe
885 890 895
Cys Val Asn Met Thr Glu Thr Leu Leu Pro Glu Asn Ala Lys Met Thr
900 905 910
Val Val Pro Pro Glu Gly Ser Gly Gly Ser Thr Asn Leu Ser Asp Ile
915 920 925
Ile Glu Lys Glu Thr Gly Lys Gln Leu Val Ile Gln Glu Ser Ile Leu
930 935 940
Met Leu Pro Glu Glu Val Glu Glu Val Ile Gly Asn Lys Pro Glu Ser
945 950 955 960
Asp Ile Leu Val His Thr Ala Tyr Asp Glu Ser Thr Asp Glu Asn Val
965 970 975
Met Leu Leu Thr Ser Asp Ala Pro Glu Tyr Lys Pro Trp Ala Leu Val
980 985 990
Ile Gln Asp Ser Asn Gly Glu Asn Lys Ile Lys Met Leu
995 1000 1005
<210> 418
<211> 927
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 418
Met Ala Ser Val Leu Thr Pro Leu Leu Leu Arg Gly Leu Thr Gly Ser
1 5 10 15
Ala Arg Arg Leu Pro Val Pro Arg Ala Lys Ile His Ser Leu Asp Tyr
20 25 30
Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp Ile Asp Tyr Lys Asp
35 40 45
Asp Asp Asp Lys Gly Ile Arg Ile Gln Asp Leu Arg Thr Leu Gly Tyr
50 55 60
Ser Gln Gln Gln Gln Glu Lys Ile Lys Pro Lys Val Arg Ser Thr Val
65 70 75 80
Ala Gln His His Glu Ala Leu Val Gly His Gly Phe Thr His Ala His
85 90 95
Ile Val Ala Leu Ser Gln His Pro Ala Ala Leu Gly Thr Val Ala Val
100 105 110
Lys Tyr Gln Asp Met Ile Ala Ala Leu Pro Glu Ala Thr His Glu Ala
115 120 125
Ile Val Gly Val Gly Lys Gln Trp Ser Gly Ala Arg Ala Leu Glu Ala
130 135 140
Leu Leu Thr Val Ala Gly Glu Leu Arg Gly Pro Pro Leu Gln Leu Asp
145 150 155 160
Thr Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly Gly Val Thr Ala Val
165 170 175
Glu Ala Val His Ala Trp Arg Asn Ala Leu Thr Gly Ala Pro Leu Asn
180 185 190
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys
195 200 205
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp
210 215 220
His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile Gly
225 230 235 240
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
245 250 255
Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn
260 265 270
Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
275 280 285
Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala
290 295 300
Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
305 310 315 320
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala
325 330 335
Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
340 345 350
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val
355 360 365
Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val
370 375 380
Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala
385 390 395 400
Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu
405 410 415
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr
420 425 430
Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala
435 440 445
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
450 455 460
Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys
465 470 475 480
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
485 490 495
His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly
500 505 510
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
515 520 525
Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser His
530 535 540
Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
545 550 555 560
Leu Cys Gln Asp His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala
565 570 575
Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
580 585 590
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala
595 600 605
Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
610 615 620
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val
625 630 635 640
Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val
645 650 655
Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala
660 665 670
Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu
675 680 685
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr
690 695 700
Pro Ala Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala
705 710 715 720
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly
725 730 735
Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys
740 745 750
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
755 760 765
His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly
770 775 780
Gly Lys Gln Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp
785 790 795 800
Pro Ala Leu Ala Ala Leu Leu Val Gly Ser Ala Ile Pro Val Lys Arg
805 810 815
Gly Ala Thr Gly Glu Thr Lys Val Phe Thr Gly Asn Ser Asn Ser Pro
820 825 830
Lys Ser Pro Thr Lys Gly Gly Cys Ser Gly Gly Ser Thr Asn Leu Ser
835 840 845
Asp Ile Ile Glu Lys Glu Thr Gly Lys Gln Leu Val Ile Gln Glu Ser
850 855 860
Ile Leu Met Leu Pro Glu Glu Val Glu Glu Val Ile Gly Asn Lys Pro
865 870 875 880
Glu Ser Asp Ile Leu Val His Thr Ala Tyr Asp Glu Ser Thr Asp Glu
885 890 895
Asn Val Met Leu Leu Thr Ser Asp Ala Pro Glu Tyr Lys Pro Trp Ala
900 905 910
Leu Val Ile Gln Asp Ser Asn Gly Glu Asn Lys Ile Lys Met Leu
915 920 925
<210> 419
<211> 988
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 419
Met Ala Leu Ser Arg Ala Val Cys Gly Thr Ser Arg Gln Leu Ala Pro
1 5 10 15
Val Leu Gly Tyr Leu Gly Ser Arg Gln Lys His Ser Leu Pro Asp Tyr
20 25 30
Pro Tyr Asp Val Pro Asp Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp
35 40 45
Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Met Asp Ile Ala
50 55 60
Asp Leu Arg Thr Leu Gly Tyr Ser Gln Gln Gln Gln Glu Lys Ile Lys
65 70 75 80
Pro Lys Val Arg Ser Thr Val Ala Gln His His Glu Ala Leu Val Gly
85 90 95
His Gly Phe Thr His Ala His Ile Val Ala Leu Ser Gln His Pro Ala
100 105 110
Ala Leu Gly Thr Val Ala Val Lys Tyr Gln Asp Met Ile Ala Ala Leu
115 120 125
Pro Glu Ala Thr His Glu Ala Ile Val Gly Val Gly Lys Gln Trp Ser
130 135 140
Gly Ala Arg Ala Leu Glu Ala Leu Leu Thr Val Ala Gly Glu Leu Arg
145 150 155 160
Gly Pro Pro Leu Gln Leu Asp Thr Gly Gln Leu Leu Lys Ile Ala Lys
165 170 175
Arg Gly Gly Val Thr Ala Val Glu Ala Val His Ala Trp Arg Asn Ala
180 185 190
Leu Thr Gly Ala Pro Leu Asn Leu Thr Pro Glu Gln Val Val Ala Ile
195 200 205
Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu
210 215 220
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Gln Gln Val Val
225 230 235 240
Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
245 250 255
Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Gln Gln
260 265 270
Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr
275 280 285
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
290 295 300
Glu Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu
305 310 315 320
Glu Thr Val Gln Ala Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
325 330 335
Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln
340 345 350
Ala Leu Glu Thr Val Gln Ala Leu Leu Pro Val Leu Cys Gln Ala His
355 360 365
Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
370 375 380
Lys Gln Ala Leu Glu Thr Val Gln Ala Leu Leu Pro Val Leu Cys Gln
385 390 395 400
Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile
405 410 415
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
420 425 430
Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser
435 440 445
Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu Leu Pro
450 455 460
Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile
465 470 475 480
Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu
485 490 495
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val
500 505 510
Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
515 520 525
Ala Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln
530 535 540
Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr
545 550 555 560
Val Gln Ala Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
565 570 575
Glu Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu
580 585 590
Glu Thr Val Gln Ala Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
595 600 605
Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln
610 615 620
Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His
625 630 635 640
Gly Leu Thr Pro Gln Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
645 650 655
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
660 665 670
Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly
675 680 685
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu Leu Pro Val Leu
690 695 700
Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser
705 710 715 720
His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu Leu Pro
725 730 735
Val Leu Cys Gln Ala His Gly Leu Thr Pro Gln Gln Val Val Ala Ile
740 745 750
Ala Ser Asn Gly Gly Gly Arg Pro Ala Leu Glu Ser Ile Val Ala Gln
755 760 765
Leu Ser Arg Pro Asp Pro Ala Leu Ala Ala Leu Thr Asn Asp His Leu
770 775 780
Val Ala Leu Ala Cys Leu Gly Gly Arg Pro Ala Leu Asp Ala Val Lys
785 790 795 800
Lys Gly Leu Gly Gly Ser Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu
805 810 815
Ser Ala Thr Pro Glu Ser Ser Glu Val Glu Phe Ser His Glu Tyr Trp
820 825 830
Met Arg His Ala Leu Thr Leu Ala Lys Arg Ala Arg Asp Glu Arg Glu
835 840 845
Val Pro Val Gly Ala Val Leu Val Leu Asn Asn Arg Val Ile Gly Glu
850 855 860
Gly Trp Asn Arg Ala Ile Gly Leu His Asp Pro Thr Ala His Ala Glu
865 870 875 880
Ile Met Ala Leu Arg Gln Gly Gly Leu Val Met Gln Asn Tyr Arg Leu
885 890 895
Ile Asp Ala Thr Leu Tyr Val Thr Phe Glu Pro Cys Val Met Cys Ala
900 905 910
Gly Ala Met Ile His Ser Arg Ile Gly Arg Val Val Phe Gly Val Arg
915 920 925
Asn Ser Lys Arg Gly Ala Ala Gly Ser Leu Met Asn Val Leu Asn Tyr
930 935 940
Pro Gly Met Asn His Arg Val Glu Ile Thr Glu Gly Ile Leu Ala Asp
945 950 955 960
Glu Cys Ala Ala Leu Leu Cys Asp Phe Tyr Arg Met Pro Arg Gln Val
965 970 975
Phe Asn Ala Gln Lys Lys Ala Gln Ser Ser Ile Asn
980 985
<210> 420
<211> 980
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 420
Met Ala Ser Val Leu Thr Pro Leu Leu Leu Arg Gly Leu Thr Gly Ser
1 5 10 15
Ala Arg Arg Leu Pro Val Pro Arg Ala Lys Ile His Ser Leu Asp Tyr
20 25 30
Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp Ile Asp Tyr Lys Asp
35 40 45
Asp Asp Asp Lys Met Asp Ile Ala Asp Leu Arg Thr Leu Gly Tyr Ser
50 55 60
Gln Gln Gln Gln Glu Lys Ile Lys Pro Lys Val Arg Ser Thr Val Ala
65 70 75 80
Gln His His Glu Ala Leu Val Gly His Gly Phe Thr His Ala His Ile
85 90 95
Val Ala Leu Ser Gln His Pro Ala Ala Leu Gly Thr Val Ala Val Lys
100 105 110
Tyr Gln Asp Met Ile Ala Ala Leu Pro Glu Ala Thr His Glu Ala Ile
115 120 125
Val Gly Val Gly Lys Gln Trp Ser Gly Ala Arg Ala Leu Glu Ala Leu
130 135 140
Leu Thr Val Ala Gly Glu Leu Arg Gly Pro Pro Leu Gln Leu Asp Thr
145 150 155 160
Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly Gly Val Thr Ala Val Glu
165 170 175
Ala Val His Ala Trp Arg Asn Ala Leu Thr Gly Ala Pro Leu Asn Leu
180 185 190
Thr Pro Glu Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln
195 200 205
Ala Leu Glu Thr Val Gln Ala Leu Leu Pro Val Leu Cys Gln Ala His
210 215 220
Gly Leu Thr Pro Gln Gln Val Val Ala Ile Ala Ser His Asp Gly Gly
225 230 235 240
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
245 250 255
Ala His Gly Leu Thr Pro Gln Gln Val Val Ala Ile Ala Ser Asn Gly
260 265 270
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
275 280 285
Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser
290 295 300
Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu Leu Pro
305 310 315 320
Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile
325 330 335
Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu
340 345 350
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val
355 360 365
Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
370 375 380
Ala Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln
385 390 395 400
Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr
405 410 415
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
420 425 430
Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu
435 440 445
Glu Thr Val Gln Ala Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
450 455 460
Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln
465 470 475 480
Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His
485 490 495
Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
500 505 510
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
515 520 525
Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn
530 535 540
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu Leu Pro Val Leu
545 550 555 560
Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser
565 570 575
Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu Leu Pro
580 585 590
Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile
595 600 605
Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu
610 615 620
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val
625 630 635 640
Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
645 650 655
Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Gln Gln
660 665 670
Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr
675 680 685
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
690 695 700
Gln Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu
705 710 715 720
Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
725 730 735
Thr Pro Gln Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Arg Pro
740 745 750
Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp Pro Ala Leu
755 760 765
Ala Ala Leu Thr Asn Asp His Leu Val Ala Leu Ala Cys Leu Gly Gly
770 775 780
Arg Pro Ala Leu Asp Ala Val Lys Lys Gly Leu Gly Gly Ser Ser Gly
785 790 795 800
Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser Ser Glu
805 810 815
Val Glu Phe Ser His Glu Tyr Trp Met Arg His Ala Leu Thr Leu Ala
820 825 830
Lys Arg Ala Arg Asp Glu Arg Glu Val Pro Val Gly Ala Val Leu Val
835 840 845
Leu Asn Asn Arg Val Ile Gly Glu Gly Trp Asn Arg Ala Ile Gly Leu
850 855 860
His Asp Pro Thr Ala His Ala Glu Ile Met Ala Leu Arg Gln Gly Gly
865 870 875 880
Leu Val Met Gln Asn Tyr Arg Leu Ile Asp Ala Thr Leu Tyr Val Thr
885 890 895
Phe Glu Pro Cys Val Met Cys Ala Gly Ala Met Ile His Ser Arg Ile
900 905 910
Gly Arg Val Val Phe Gly Val Arg Asn Ser Lys Arg Gly Ala Ala Gly
915 920 925
Ser Leu Met Asn Val Leu Asn Tyr Pro Gly Met Asn His Arg Val Glu
930 935 940
Ile Thr Glu Gly Ile Leu Ala Asp Glu Cys Ala Ala Leu Leu Cys Asp
945 950 955 960
Phe Tyr Arg Met Pro Arg Gln Val Phe Asn Ala Gln Lys Lys Ala Gln
965 970 975
Ser Ser Ile Asn
980
<210> 421
<211> 942
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 421
Met Ala Leu Ser Arg Ala Val Cys Gly Thr Ser Arg Gln Leu Ala Pro
1 5 10 15
Val Leu Gly Tyr Leu Gly Ser Arg Gln Lys His Ser Leu Pro Asp Tyr
20 25 30
Pro Tyr Asp Val Pro Asp Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp
35 40 45
Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Gly Ile Arg Ile
50 55 60
Gln Asp Leu Arg Thr Leu Gly Tyr Ser Gln Gln Gln Gln Glu Lys Ile
65 70 75 80
Lys Pro Lys Val Arg Ser Thr Val Ala Gln His His Glu Ala Leu Val
85 90 95
Gly His Gly Phe Thr His Ala His Ile Val Ala Leu Ser Gln His Pro
100 105 110
Ala Ala Leu Gly Thr Val Ala Val Lys Tyr Gln Asp Met Ile Ala Ala
115 120 125
Leu Pro Glu Ala Thr His Glu Ala Ile Val Gly Val Gly Lys Gln Trp
130 135 140
Ser Gly Ala Arg Ala Leu Glu Ala Leu Leu Thr Val Ala Gly Glu Leu
145 150 155 160
Arg Gly Pro Pro Leu Gln Leu Asp Thr Gly Gln Leu Leu Lys Ile Ala
165 170 175
Lys Arg Gly Gly Val Thr Ala Val Glu Ala Val His Ala Trp Arg Asn
180 185 190
Ala Leu Thr Gly Ala Pro Leu Asn Leu Thr Pro Asp Gln Val Val Ala
195 200 205
Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
210 215 220
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val
225 230 235 240
Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val
245 250 255
Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Ala
260 265 270
Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu
275 280 285
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr
290 295 300
Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala
305 310 315 320
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
325 330 335
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys
340 345 350
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp
355 360 365
His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly
370 375 380
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
385 390 395 400
Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn
405 410 415
Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
420 425 430
Leu Cys Gln Asp His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala
435 440 445
Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
450 455 460
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala
465 470 475 480
Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
485 490 495
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val
500 505 510
Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val
515 520 525
Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Asp
530 535 540
Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu
545 550 555 560
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr
565 570 575
Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala
580 585 590
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
595 600 605
Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys
610 615 620
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
625 630 635 640
His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly
645 650 655
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
660 665 670
Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn
675 680 685
Gly Gly Gly Lys Gln Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg
690 695 700
Pro Asp Pro Ala Leu Ala Ala Leu Leu Val Gly Ser Gly Ser Tyr Ala
705 710 715 720
Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro Ala Tyr Asn Gly
725 730 735
Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala Gly Gly Leu Glu
740 745 750
Ser Lys Val Phe Ser Ser Gly Gly Ser Gly Ser Glu Thr Pro Gly Thr
755 760 765
Ser Glu Ser Ala Thr Pro Glu Ser Ser Glu Val Glu Phe Ser His Glu
770 775 780
Tyr Trp Met Arg His Ala Leu Thr Leu Ala Lys Arg Ala Arg Asp Glu
785 790 795 800
Arg Glu Val Pro Val Gly Ala Val Leu Val Leu Asn Asn Arg Val Ile
805 810 815
Gly Glu Gly Trp Asn Arg Ala Ile Gly Leu His Asp Pro Thr Ala His
820 825 830
Ala Glu Ile Met Ala Leu Arg Gln Gly Gly Leu Val Met Gln Asn Tyr
835 840 845
Arg Leu Ile Asp Ala Thr Leu Tyr Val Thr Phe Glu Pro Cys Val Met
850 855 860
Cys Ala Gly Ala Met Ile His Ser Arg Ile Gly Arg Val Val Phe Gly
865 870 875 880
Val Arg Asn Ser Lys Arg Gly Ala Ala Gly Ser Leu Met Asn Val Leu
885 890 895
Asn Tyr Pro Gly Met Asn His Arg Val Glu Ile Thr Glu Gly Ile Leu
900 905 910
Ala Asp Glu Cys Ala Ala Leu Leu Cys Asp Phe Tyr Arg Met Pro Arg
915 920 925
Gln Val Phe Asn Ala Gln Lys Lys Ala Gln Ser Ser Ile Asn
930 935 940
<210> 422
<211> 992
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 422
Met Ala Leu Ser Arg Ala Val Cys Gly Thr Ser Arg Gln Leu Ala Pro
1 5 10 15
Val Leu Gly Tyr Leu Gly Ser Arg Gln Lys His Ser Leu Pro Asp Tyr
20 25 30
Pro Tyr Asp Val Pro Asp Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp
35 40 45
Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Gly Ile Arg Ile
50 55 60
Gln Asp Leu Arg Thr Leu Gly Tyr Ser Gln Gln Gln Gln Glu Lys Ile
65 70 75 80
Lys Pro Lys Val Arg Ser Thr Val Ala Gln His His Glu Ala Leu Val
85 90 95
Gly His Gly Phe Thr His Ala His Ile Val Ala Leu Ser Gln His Pro
100 105 110
Ala Ala Leu Gly Thr Val Ala Val Lys Tyr Gln Asp Met Ile Ala Ala
115 120 125
Leu Pro Glu Ala Thr His Glu Ala Ile Val Gly Val Gly Lys Gln Trp
130 135 140
Ser Gly Ala Arg Ala Leu Glu Ala Leu Leu Thr Val Ala Gly Glu Leu
145 150 155 160
Arg Gly Pro Pro Leu Gln Leu Asp Thr Gly Gln Leu Leu Lys Ile Ala
165 170 175
Lys Arg Gly Gly Val Thr Ala Val Glu Ala Val His Ala Trp Arg Asn
180 185 190
Ala Leu Thr Gly Ala Pro Leu Asn Leu Thr Pro Asp Gln Val Val Ala
195 200 205
Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
210 215 220
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val
225 230 235 240
Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val
245 250 255
Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Ala
260 265 270
Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu
275 280 285
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr
290 295 300
Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala
305 310 315 320
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
325 330 335
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys
340 345 350
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp
355 360 365
His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly
370 375 380
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
385 390 395 400
Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn
405 410 415
Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
420 425 430
Leu Cys Gln Asp His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala
435 440 445
Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
450 455 460
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala
465 470 475 480
Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
485 490 495
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val
500 505 510
Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val
515 520 525
Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Asp
530 535 540
Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu
545 550 555 560
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr
565 570 575
Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala
580 585 590
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
595 600 605
Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys
610 615 620
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
625 630 635 640
His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly
645 650 655
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
660 665 670
Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn
675 680 685
Gly Gly Gly Lys Gln Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg
690 695 700
Pro Asp Pro Ala Leu Ala Ala Leu Leu Val Gly Ser Pro Thr Pro Tyr
705 710 715 720
Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln Ser Ala Leu Phe
725 730 735
Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe His Asn Asn Pro
740 745 750
Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu Thr Leu Leu Pro
755 760 765
Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly Ala Ile Pro Val
770 775 780
Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr Gly Asn Ser Asn
785 790 795 800
Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys Ser Gly Ser Glu Thr Pro
805 810 815
Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser Ser Glu Val Glu Phe Ser
820 825 830
His Glu Tyr Trp Met Arg His Ala Leu Thr Leu Ala Lys Arg Ala Arg
835 840 845
Asp Glu Arg Glu Val Pro Val Gly Ala Val Leu Val Leu Asn Asn Arg
850 855 860
Val Ile Gly Glu Gly Trp Asn Arg Ala Ile Gly Leu His Asp Pro Thr
865 870 875 880
Ala His Ala Glu Ile Met Ala Leu Arg Gln Gly Gly Leu Val Met Gln
885 890 895
Asn Tyr Arg Leu Ile Asp Ala Thr Leu Tyr Val Thr Phe Glu Pro Cys
900 905 910
Val Met Cys Ala Gly Ala Met Ile His Ser Arg Ile Gly Arg Val Val
915 920 925
Phe Gly Val Arg Asn Ser Lys Arg Gly Ala Ala Gly Ser Leu Met Asn
930 935 940
Val Leu Asn Tyr Pro Gly Met Asn His Arg Val Glu Ile Thr Glu Gly
945 950 955 960
Ile Leu Ala Asp Glu Cys Ala Ala Leu Leu Cys Asp Phe Tyr Arg Met
965 970 975
Pro Arg Gln Val Phe Asn Ala Gln Lys Lys Ala Gln Ser Ser Ile Asn
980 985 990
<210> 423
<211> 1036
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 423
Met Ala Ser Val Leu Thr Pro Leu Leu Leu Arg Gly Leu Thr Gly Ser
1 5 10 15
Ala Arg Arg Leu Pro Val Pro Arg Ala Lys Ile His Ser Leu Asp Tyr
20 25 30
Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp Ile Asp Tyr Lys Asp
35 40 45
Asp Asp Asp Lys Gly Ile Arg Ile Gln Asp Leu Arg Thr Leu Gly Tyr
50 55 60
Ser Gln Gln Gln Gln Glu Lys Ile Lys Pro Lys Val Arg Ser Thr Val
65 70 75 80
Ala Gln His His Glu Ala Leu Val Gly His Gly Phe Thr His Ala His
85 90 95
Ile Val Ala Leu Ser Gln His Pro Ala Ala Leu Gly Thr Val Ala Val
100 105 110
Lys Tyr Gln Asp Met Ile Ala Ala Leu Pro Glu Ala Thr His Glu Ala
115 120 125
Ile Val Gly Val Gly Lys Gln Trp Ser Gly Ala Arg Ala Leu Glu Ala
130 135 140
Leu Leu Thr Val Ala Gly Glu Leu Arg Gly Pro Pro Leu Gln Leu Asp
145 150 155 160
Thr Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly Gly Val Thr Ala Val
165 170 175
Glu Ala Val His Ala Trp Arg Asn Ala Leu Thr Gly Ala Pro Leu Asn
180 185 190
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys
195 200 205
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp
210 215 220
His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile Gly
225 230 235 240
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
245 250 255
Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn
260 265 270
Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
275 280 285
Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala
290 295 300
Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
305 310 315 320
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala
325 330 335
Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
340 345 350
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val
355 360 365
Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val
370 375 380
Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala
385 390 395 400
Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu
405 410 415
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr
420 425 430
Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala
435 440 445
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
450 455 460
Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys
465 470 475 480
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
485 490 495
His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly
500 505 510
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
515 520 525
Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser His
530 535 540
Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
545 550 555 560
Leu Cys Gln Asp His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala
565 570 575
Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
580 585 590
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala
595 600 605
Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
610 615 620
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val
625 630 635 640
Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val
645 650 655
Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala
660 665 670
Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu
675 680 685
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr
690 695 700
Pro Ala Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala
705 710 715 720
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly
725 730 735
Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys
740 745 750
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
755 760 765
His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly
770 775 780
Gly Lys Gln Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp
785 790 795 800
Pro Ala Leu Ala Ala Leu Leu Val Gly Ser Gly Ser Tyr Ala Leu Gly
805 810 815
Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro Ala Tyr Asn Gly Gln Thr
820 825 830
Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala Gly Gly Leu Glu Ser Lys
835 840 845
Val Phe Ser Ser Gly Gly Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu
850 855 860
Ser Ala Thr Pro Glu Ser Ser Glu Val Glu Phe Ser His Glu Tyr Trp
865 870 875 880
Met Arg His Ala Leu Thr Leu Ala Lys Arg Ala Arg Asp Glu Arg Glu
885 890 895
Val Pro Val Gly Ala Val Leu Val Leu Asn Asn Arg Val Ile Gly Glu
900 905 910
Gly Trp Asn Arg Ala Ile Gly Leu His Asp Pro Thr Ala His Ala Glu
915 920 925
Ile Met Ala Leu Arg Gln Gly Gly Leu Val Met Gln Asn Tyr Arg Leu
930 935 940
Ile Asp Ala Thr Leu Tyr Val Thr Phe Glu Pro Cys Val Met Cys Ala
945 950 955 960
Gly Ala Met Ile His Ser Arg Ile Gly Arg Val Val Phe Gly Val Arg
965 970 975
Asn Ser Lys Arg Gly Ala Ala Gly Ser Leu Met Asn Val Leu Asn Tyr
980 985 990
Pro Gly Met Asn His Arg Val Glu Ile Thr Glu Gly Ile Leu Ala Asp
995 1000 1005
Glu Cys Ala Ala Leu Leu Cys Asp Phe Tyr Arg Met Pro Arg Gln Val
1010 1015 1020
Phe Asn Ala Gln Lys Lys Ala Gln Ser Ser Ile Asn
1025 1030 1035
<210> 424
<211> 1086
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 424
Met Ala Ser Val Leu Thr Pro Leu Leu Leu Arg Gly Leu Thr Gly Ser
1 5 10 15
Ala Arg Arg Leu Pro Val Pro Arg Ala Lys Ile His Ser Leu Asp Tyr
20 25 30
Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp Ile Asp Tyr Lys Asp
35 40 45
Asp Asp Asp Lys Gly Ile Arg Ile Gln Asp Leu Arg Thr Leu Gly Tyr
50 55 60
Ser Gln Gln Gln Gln Glu Lys Ile Lys Pro Lys Val Arg Ser Thr Val
65 70 75 80
Ala Gln His His Glu Ala Leu Val Gly His Gly Phe Thr His Ala His
85 90 95
Ile Val Ala Leu Ser Gln His Pro Ala Ala Leu Gly Thr Val Ala Val
100 105 110
Lys Tyr Gln Asp Met Ile Ala Ala Leu Pro Glu Ala Thr His Glu Ala
115 120 125
Ile Val Gly Val Gly Lys Gln Trp Ser Gly Ala Arg Ala Leu Glu Ala
130 135 140
Leu Leu Thr Val Ala Gly Glu Leu Arg Gly Pro Pro Leu Gln Leu Asp
145 150 155 160
Thr Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly Gly Val Thr Ala Val
165 170 175
Glu Ala Val His Ala Trp Arg Asn Ala Leu Thr Gly Ala Pro Leu Asn
180 185 190
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys
195 200 205
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp
210 215 220
His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile Gly
225 230 235 240
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
245 250 255
Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn
260 265 270
Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
275 280 285
Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala
290 295 300
Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
305 310 315 320
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala
325 330 335
Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
340 345 350
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val
355 360 365
Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val
370 375 380
Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala
385 390 395 400
Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu
405 410 415
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr
420 425 430
Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala
435 440 445
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
450 455 460
Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys
465 470 475 480
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
485 490 495
His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly
500 505 510
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
515 520 525
Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser His
530 535 540
Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
545 550 555 560
Leu Cys Gln Asp His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala
565 570 575
Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
580 585 590
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala
595 600 605
Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
610 615 620
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val
625 630 635 640
Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val
645 650 655
Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala
660 665 670
Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu
675 680 685
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr
690 695 700
Pro Ala Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala
705 710 715 720
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly
725 730 735
Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys
740 745 750
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
755 760 765
His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly
770 775 780
Gly Lys Gln Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp
785 790 795 800
Pro Ala Leu Ala Ala Leu Leu Val Gly Ser Pro Thr Pro Tyr Pro Asn
805 810 815
Tyr Ala Asn Ala Gly His Val Glu Gly Gln Ser Ala Leu Phe Met Arg
820 825 830
Asp Asn Gly Ile Ser Glu Gly Leu Val Phe His Asn Asn Pro Glu Gly
835 840 845
Thr Cys Gly Phe Cys Val Asn Met Thr Glu Thr Leu Leu Pro Glu Asn
850 855 860
Ala Lys Met Thr Val Val Pro Pro Glu Gly Ala Ile Pro Val Lys Arg
865 870 875 880
Gly Ala Thr Gly Glu Thr Lys Val Phe Thr Gly Asn Ser Asn Ser Pro
885 890 895
Lys Ser Pro Thr Lys Gly Gly Cys Ser Gly Ser Glu Thr Pro Gly Thr
900 905 910
Ser Glu Ser Ala Thr Pro Glu Ser Ser Glu Val Glu Phe Ser His Glu
915 920 925
Tyr Trp Met Arg His Ala Leu Thr Leu Ala Lys Arg Ala Arg Asp Glu
930 935 940
Arg Glu Val Pro Val Gly Ala Val Leu Val Leu Asn Asn Arg Val Ile
945 950 955 960
Gly Glu Gly Trp Asn Arg Ala Ile Gly Leu His Asp Pro Thr Ala His
965 970 975
Ala Glu Ile Met Ala Leu Arg Gln Gly Gly Leu Val Met Gln Asn Tyr
980 985 990
Arg Leu Ile Asp Ala Thr Leu Tyr Val Thr Phe Glu Pro Cys Val Met
995 1000 1005
Cys Ala Gly Ala Met Ile His Ser Arg Ile Gly Arg Val Val Phe Gly
1010 1015 1020
Val Arg Asn Ser Lys Arg Gly Ala Ala Gly Ser Leu Met Asn Val Leu
1025 1030 1035 1040
Asn Tyr Pro Gly Met Asn His Arg Val Glu Ile Thr Glu Gly Ile Leu
1045 1050 1055
Ala Asp Glu Cys Ala Ala Leu Leu Cys Asp Phe Tyr Arg Met Pro Arg
1060 1065 1070
Gln Val Phe Asn Ala Gln Lys Lys Ala Gln Ser Ser Ile Asn
1075 1080 1085
<210> 425
<211> 952
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 425
Met Ala Leu Ser Arg Ala Val Cys Gly Thr Ser Arg Gln Leu Ala Pro
1 5 10 15
Val Leu Gly Tyr Leu Gly Ser Arg Gln Lys His Ser Leu Pro Asp Tyr
20 25 30
Pro Tyr Asp Val Pro Asp Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp
35 40 45
Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Gly Ile Arg Ile
50 55 60
Gln Asp Leu Arg Thr Leu Gly Tyr Ser Gln Gln Gln Gln Glu Lys Ile
65 70 75 80
Lys Pro Lys Val Arg Ser Thr Val Ala Gln His His Glu Ala Leu Val
85 90 95
Gly His Gly Phe Thr His Ala His Ile Val Ala Leu Ser Gln His Pro
100 105 110
Ala Ala Leu Gly Thr Val Ala Val Lys Tyr Gln Asp Met Ile Ala Ala
115 120 125
Leu Pro Glu Ala Thr His Glu Ala Ile Val Gly Val Gly Lys Gln Trp
130 135 140
Ser Gly Ala Arg Ala Leu Glu Ala Leu Leu Thr Val Ala Gly Glu Leu
145 150 155 160
Arg Gly Pro Pro Leu Gln Leu Asp Thr Gly Gln Leu Leu Lys Ile Ala
165 170 175
Lys Arg Gly Gly Val Thr Ala Val Glu Ala Val His Ala Trp Arg Asn
180 185 190
Ala Leu Thr Gly Ala Pro Leu Asn Leu Thr Pro Asp Gln Val Val Ala
195 200 205
Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
210 215 220
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val
225 230 235 240
Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val
245 250 255
Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Ala
260 265 270
Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu
275 280 285
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr
290 295 300
Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala
305 310 315 320
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
325 330 335
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys
340 345 350
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp
355 360 365
His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly
370 375 380
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
385 390 395 400
Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn
405 410 415
Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
420 425 430
Leu Cys Gln Asp His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala
435 440 445
Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
450 455 460
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala
465 470 475 480
Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
485 490 495
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val
500 505 510
Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val
515 520 525
Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Asp
530 535 540
Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu
545 550 555 560
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr
565 570 575
Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala
580 585 590
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
595 600 605
Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys
610 615 620
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
625 630 635 640
His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly
645 650 655
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
660 665 670
Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn
675 680 685
Gly Gly Gly Lys Gln Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg
690 695 700
Pro Asp Pro Ala Leu Ala Ala Leu Thr Asn Asp His Leu Val Ala Leu
705 710 715 720
Ala Cys Leu Gly Gly Arg Pro Ala Leu Asp Ala Val Lys Lys Gly Leu
725 730 735
Leu Val Gly Ser Ala Ile Pro Val Lys Arg Gly Ala Thr Gly Glu Thr
740 745 750
Lys Val Phe Thr Gly Asn Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly
755 760 765
Gly Cys Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro
770 775 780
Glu Ser Ser Glu Val Glu Phe Ser His Glu Tyr Trp Met Arg His Ala
785 790 795 800
Leu Thr Leu Ala Lys Arg Ala Arg Asp Glu Arg Glu Val Pro Val Gly
805 810 815
Ala Val Leu Val Leu Asn Asn Arg Val Ile Gly Glu Gly Trp Asn Arg
820 825 830
Ala Ile Gly Leu His Asp Pro Thr Ala His Ala Glu Ile Met Ala Leu
835 840 845
Arg Gln Gly Gly Leu Val Met Gln Asn Tyr Arg Leu Ile Asp Ala Thr
850 855 860
Leu Tyr Val Thr Phe Glu Pro Cys Val Met Cys Ala Gly Ala Met Ile
865 870 875 880
His Ser Arg Ile Gly Arg Val Val Phe Gly Val Arg Asn Ser Lys Arg
885 890 895
Gly Ala Ala Gly Ser Leu Met Asn Val Leu Asn Tyr Pro Gly Met Asn
900 905 910
His Arg Val Glu Ile Thr Glu Gly Ile Leu Ala Asp Glu Cys Ala Ala
915 920 925
Leu Leu Cys Asp Phe Tyr Arg Met Pro Arg Gln Val Phe Asn Ala Gln
930 935 940
Lys Lys Ala Gln Ser Ser Ile Asn
945 950
<210> 426
<211> 1047
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 426
Met Ala Ser Val Leu Thr Pro Leu Leu Leu Arg Gly Leu Thr Gly Ser
1 5 10 15
Ala Arg Arg Leu Pro Val Pro Arg Ala Lys Ile His Ser Leu Asp Tyr
20 25 30
Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp Ile Asp Tyr Lys Asp
35 40 45
Asp Asp Asp Lys Gly Ile Arg Ile Gln Asp Leu Arg Thr Leu Gly Tyr
50 55 60
Ser Gln Gln Gln Gln Glu Lys Ile Lys Pro Lys Val Arg Ser Thr Val
65 70 75 80
Ala Gln His His Glu Ala Leu Val Gly His Gly Phe Thr His Ala His
85 90 95
Ile Val Ala Leu Ser Gln His Pro Ala Ala Leu Gly Thr Val Ala Val
100 105 110
Lys Tyr Gln Asp Met Ile Ala Ala Leu Pro Glu Ala Thr His Glu Ala
115 120 125
Ile Val Gly Val Gly Lys Gln Trp Ser Gly Ala Arg Ala Leu Glu Ala
130 135 140
Leu Leu Thr Val Ala Gly Glu Leu Arg Gly Pro Pro Leu Gln Leu Asp
145 150 155 160
Thr Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly Gly Val Thr Ala Val
165 170 175
Glu Ala Val His Ala Trp Arg Asn Ala Leu Thr Gly Ala Pro Leu Asn
180 185 190
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys
195 200 205
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp
210 215 220
His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile Gly
225 230 235 240
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
245 250 255
Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn
260 265 270
Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
275 280 285
Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala
290 295 300
Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
305 310 315 320
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala
325 330 335
Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
340 345 350
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val
355 360 365
Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val
370 375 380
Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala
385 390 395 400
Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu
405 410 415
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr
420 425 430
Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala
435 440 445
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
450 455 460
Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys
465 470 475 480
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
485 490 495
His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly
500 505 510
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
515 520 525
Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser His
530 535 540
Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
545 550 555 560
Leu Cys Gln Asp His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala
565 570 575
Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
580 585 590
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala
595 600 605
Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
610 615 620
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val
625 630 635 640
Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val
645 650 655
Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala
660 665 670
Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu
675 680 685
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr
690 695 700
Pro Ala Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala
705 710 715 720
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly
725 730 735
Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys
740 745 750
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
755 760 765
His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly
770 775 780
Gly Lys Gln Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp
785 790 795 800
Pro Ala Leu Ala Ala Leu Thr Asn Asp His Leu Val Ala Leu Ala Cys
805 810 815
Leu Gly Gly Arg Pro Ala Leu Asp Ala Val Lys Lys Gly Leu Gly Gly
820 825 830
Ser Gly Ser Ala Ile Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys
835 840 845
Val Phe Thr Gly Asn Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly
850 855 860
Cys Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro Glu
865 870 875 880
Ser Ser Glu Val Glu Phe Ser His Glu Tyr Trp Met Arg His Ala Leu
885 890 895
Thr Leu Ala Lys Arg Ala Arg Asp Glu Arg Glu Val Pro Val Gly Ala
900 905 910
Val Leu Val Leu Asn Asn Arg Val Ile Gly Glu Gly Trp Asn Arg Ala
915 920 925
Ile Gly Leu His Asp Pro Thr Ala His Ala Glu Ile Met Ala Leu Arg
930 935 940
Gln Gly Gly Leu Val Met Gln Asn Tyr Arg Leu Ile Asp Ala Thr Leu
945 950 955 960
Tyr Val Thr Phe Glu Pro Cys Val Met Cys Ala Gly Ala Met Ile His
965 970 975
Ser Arg Ile Gly Arg Val Val Phe Gly Val Arg Asn Ser Lys Arg Gly
980 985 990
Ala Ala Gly Ser Leu Met Asn Val Leu Asn Tyr Pro Gly Met Asn His
995 1000 1005
Arg Val Glu Ile Thr Glu Gly Ile Leu Ala Asp Glu Cys Ala Ala Leu
1010 1015 1020
Leu Cys Asp Phe Tyr Arg Met Pro Arg Gln Val Phe Asn Ala Gln Lys
1025 1030 1035 1040
Lys Ala Gln Ser Ser Ile Asn
1045
<210> 427
<211> 785
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 427
Met Ala Leu Ser Arg Ala Val Cys Gly Thr Ser Arg Gln Leu Ala Pro
1 5 10 15
Val Leu Gly Tyr Leu Gly Ser Arg Gln Lys His Ser Leu Pro Asp Tyr
20 25 30
Pro Tyr Asp Val Pro Asp Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp
35 40 45
Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Gly Ile Arg Ile
50 55 60
Gln Asp Leu Arg Thr Leu Gly Tyr Ser Gln Gln Gln Gln Glu Lys Ile
65 70 75 80
Lys Pro Lys Val Arg Ser Thr Val Ala Gln His His Glu Ala Leu Val
85 90 95
Gly His Gly Phe Thr His Ala His Ile Val Ala Leu Ser Gln His Pro
100 105 110
Ala Ala Leu Gly Thr Val Ala Val Lys Tyr Gln Asp Met Ile Ala Ala
115 120 125
Leu Pro Glu Ala Thr His Glu Ala Ile Val Gly Val Gly Lys Gln Trp
130 135 140
Ser Gly Ala Arg Ala Leu Glu Ala Leu Leu Thr Val Ala Gly Glu Leu
145 150 155 160
Arg Gly Pro Pro Leu Gln Leu Asp Thr Gly Gln Leu Leu Lys Ile Ala
165 170 175
Lys Arg Gly Gly Val Thr Ala Val Glu Ala Val His Ala Trp Arg Asn
180 185 190
Ala Leu Thr Gly Ala Pro Leu Asn Leu Thr Pro Asp Gln Val Val Ala
195 200 205
Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
210 215 220
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val
225 230 235 240
Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val
245 250 255
Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Ala
260 265 270
Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu
275 280 285
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr
290 295 300
Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala
305 310 315 320
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
325 330 335
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys
340 345 350
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp
355 360 365
His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly
370 375 380
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
385 390 395 400
Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn
405 410 415
Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
420 425 430
Leu Cys Gln Asp His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala
435 440 445
Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
450 455 460
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala
465 470 475 480
Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
485 490 495
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val
500 505 510
Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val
515 520 525
Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Asp
530 535 540
Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu
545 550 555 560
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr
565 570 575
Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala
580 585 590
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
595 600 605
Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys
610 615 620
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
625 630 635 640
His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly
645 650 655
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
660 665 670
Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn
675 680 685
Gly Gly Gly Lys Gln Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg
690 695 700
Pro Asp Pro Ala Leu Ala Ala Leu Thr Asn Asp His Leu Val Ala Leu
705 710 715 720
Ala Cys Leu Gly Gly Arg Pro Ala Leu Asp Ala Val Lys Lys Gly Leu
725 730 735
Gly Gly Ser Gly Ser Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser
740 745 750
Ala Pro Gln Leu Pro Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr
755 760 765
Tyr Val Asn Asp Ala Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly
770 775 780
Gly
785
<210> 428
<211> 835
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 428
Met Ala Leu Ser Arg Ala Val Cys Gly Thr Ser Arg Gln Leu Ala Pro
1 5 10 15
Val Leu Gly Tyr Leu Gly Ser Arg Gln Lys His Ser Leu Pro Asp Tyr
20 25 30
Pro Tyr Asp Val Pro Asp Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp
35 40 45
Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Gly Ile Arg Ile
50 55 60
Gln Asp Leu Arg Thr Leu Gly Tyr Ser Gln Gln Gln Gln Glu Lys Ile
65 70 75 80
Lys Pro Lys Val Arg Ser Thr Val Ala Gln His His Glu Ala Leu Val
85 90 95
Gly His Gly Phe Thr His Ala His Ile Val Ala Leu Ser Gln His Pro
100 105 110
Ala Ala Leu Gly Thr Val Ala Val Lys Tyr Gln Asp Met Ile Ala Ala
115 120 125
Leu Pro Glu Ala Thr His Glu Ala Ile Val Gly Val Gly Lys Gln Trp
130 135 140
Ser Gly Ala Arg Ala Leu Glu Ala Leu Leu Thr Val Ala Gly Glu Leu
145 150 155 160
Arg Gly Pro Pro Leu Gln Leu Asp Thr Gly Gln Leu Leu Lys Ile Ala
165 170 175
Lys Arg Gly Gly Val Thr Ala Val Glu Ala Val His Ala Trp Arg Asn
180 185 190
Ala Leu Thr Gly Ala Pro Leu Asn Leu Thr Pro Asp Gln Val Val Ala
195 200 205
Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
210 215 220
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val
225 230 235 240
Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val
245 250 255
Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Ala
260 265 270
Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu
275 280 285
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr
290 295 300
Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala
305 310 315 320
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
325 330 335
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys
340 345 350
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp
355 360 365
His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly
370 375 380
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
385 390 395 400
Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn
405 410 415
Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
420 425 430
Leu Cys Gln Asp His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala
435 440 445
Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
450 455 460
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala
465 470 475 480
Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
485 490 495
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val
500 505 510
Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val
515 520 525
Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Asp
530 535 540
Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu
545 550 555 560
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr
565 570 575
Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala
580 585 590
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
595 600 605
Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys
610 615 620
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
625 630 635 640
His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly
645 650 655
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
660 665 670
Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn
675 680 685
Gly Gly Gly Lys Gln Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg
690 695 700
Pro Asp Pro Ala Leu Ala Ala Leu Thr Asn Asp His Leu Val Ala Leu
705 710 715 720
Ala Cys Leu Gly Gly Arg Pro Ala Leu Asp Ala Val Lys Lys Gly Leu
725 730 735
Gly Gly Ser Gly Ser Pro Thr Pro Tyr Pro Asn Tyr Ala Asn Ala Gly
740 745 750
His Val Glu Gly Gln Ser Ala Leu Phe Met Arg Asp Asn Gly Ile Ser
755 760 765
Glu Gly Leu Val Phe His Asn Asn Pro Glu Gly Thr Cys Gly Phe Cys
770 775 780
Val Asn Met Thr Glu Thr Leu Leu Pro Glu Asn Ala Lys Met Thr Val
785 790 795 800
Val Pro Pro Glu Gly Ala Ile Pro Val Lys Arg Gly Ala Thr Gly Glu
805 810 815
Thr Lys Val Phe Thr Gly Asn Ser Asn Ser Pro Lys Ser Pro Thr Lys
820 825 830
Gly Gly Cys
835
<210> 429
<211> 849
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 429
Met Ala Leu Ser Arg Ala Val Cys Gly Thr Ser Arg Gln Leu Ala Pro
1 5 10 15
Val Leu Gly Tyr Leu Gly Ser Arg Gln Lys His Ser Leu Pro Asp Tyr
20 25 30
Pro Tyr Asp Val Pro Asp Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp
35 40 45
Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Gly Ile Arg Ile
50 55 60
Gln Asp Leu Arg Thr Leu Gly Tyr Ser Gln Gln Gln Gln Glu Lys Ile
65 70 75 80
Lys Pro Lys Val Arg Ser Thr Val Ala Gln His His Glu Ala Leu Val
85 90 95
Gly His Gly Phe Thr His Ala His Ile Val Ala Leu Ser Gln His Pro
100 105 110
Ala Ala Leu Gly Thr Val Ala Val Lys Tyr Gln Asp Met Ile Ala Ala
115 120 125
Leu Pro Glu Ala Thr His Glu Ala Ile Val Gly Val Gly Lys Gln Trp
130 135 140
Ser Gly Ala Arg Ala Leu Glu Ala Leu Leu Thr Val Ala Gly Glu Leu
145 150 155 160
Arg Gly Pro Pro Leu Gln Leu Asp Thr Gly Gln Leu Leu Lys Ile Ala
165 170 175
Lys Arg Gly Gly Val Thr Ala Val Glu Ala Val His Ala Trp Arg Asn
180 185 190
Ala Leu Thr Gly Ala Pro Leu Asn Leu Thr Pro Asp Gln Val Val Ala
195 200 205
Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
210 215 220
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val
225 230 235 240
Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val
245 250 255
Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Ala
260 265 270
Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu
275 280 285
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr
290 295 300
Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala
305 310 315 320
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
325 330 335
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys
340 345 350
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp
355 360 365
His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly
370 375 380
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
385 390 395 400
Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn
405 410 415
Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
420 425 430
Leu Cys Gln Asp His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala
435 440 445
Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
450 455 460
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala
465 470 475 480
Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
485 490 495
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val
500 505 510
Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val
515 520 525
Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Asp
530 535 540
Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu
545 550 555 560
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr
565 570 575
Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala
580 585 590
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
595 600 605
Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys
610 615 620
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
625 630 635 640
His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly
645 650 655
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
660 665 670
Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn
675 680 685
Gly Gly Gly Lys Gln Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg
690 695 700
Pro Asp Pro Ala Leu Ala Ala Leu Thr Asn Asp His Leu Val Ala Leu
705 710 715 720
Ala Cys Leu Gly Gly Arg Pro Ala Leu Asp Ala Val Lys Lys Gly Leu
725 730 735
Gly Gly Ser Gly Ser Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser
740 745 750
Ala Pro Gln Leu Pro Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr
755 760 765
Tyr Val Asn Asp Ala Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly
770 775 780
Gly Pro Thr Pro Tyr Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly
785 790 795 800
Gln Ser Ala Leu Phe Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val
805 810 815
Phe His Asn Asn Pro Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr
820 825 830
Glu Thr Leu Leu Pro Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu
835 840 845
Gly
<210> 430
<211> 854
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 430
Met Ala Ser Val Leu Thr Pro Leu Leu Leu Arg Gly Leu Thr Gly Ser
1 5 10 15
Ala Arg Arg Leu Pro Val Pro Arg Ala Lys Ile His Ser Leu Asp Tyr
20 25 30
Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp Ile Asp Tyr Lys Asp
35 40 45
Asp Asp Asp Lys Gly Ile Arg Ile Gln Asp Leu Arg Thr Leu Gly Tyr
50 55 60
Ser Gln Gln Gln Gln Glu Lys Ile Lys Pro Lys Val Arg Ser Thr Val
65 70 75 80
Ala Gln His His Glu Ala Leu Val Gly His Gly Phe Thr His Ala His
85 90 95
Ile Val Ala Leu Ser Gln His Pro Ala Ala Leu Gly Thr Val Ala Val
100 105 110
Lys Tyr Gln Asp Met Ile Ala Ala Leu Pro Glu Ala Thr His Glu Ala
115 120 125
Ile Val Gly Val Gly Lys Gln Trp Ser Gly Ala Arg Ala Leu Glu Ala
130 135 140
Leu Leu Thr Val Ala Gly Glu Leu Arg Gly Pro Pro Leu Gln Leu Asp
145 150 155 160
Thr Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly Gly Val Thr Ala Val
165 170 175
Glu Ala Val His Ala Trp Arg Asn Ala Leu Thr Gly Ala Pro Leu Asn
180 185 190
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys
195 200 205
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp
210 215 220
His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile Gly
225 230 235 240
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
245 250 255
Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn
260 265 270
Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
275 280 285
Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala
290 295 300
Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
305 310 315 320
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala
325 330 335
Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
340 345 350
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val
355 360 365
Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val
370 375 380
Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala
385 390 395 400
Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu
405 410 415
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr
420 425 430
Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala
435 440 445
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
450 455 460
Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys
465 470 475 480
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
485 490 495
His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly
500 505 510
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
515 520 525
Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser His
530 535 540
Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
545 550 555 560
Leu Cys Gln Asp His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala
565 570 575
Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
580 585 590
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala
595 600 605
Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
610 615 620
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val
625 630 635 640
Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val
645 650 655
Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala
660 665 670
Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu
675 680 685
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr
690 695 700
Pro Ala Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala
705 710 715 720
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly
725 730 735
Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys
740 745 750
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
755 760 765
His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly
770 775 780
Gly Lys Gln Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp
785 790 795 800
Pro Ala Leu Ala Ala Leu Leu Val Gly Ser Gly Ser Tyr Ala Leu Gly
805 810 815
Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro Ala Tyr Asn Gly Gln Thr
820 825 830
Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala Gly Gly Leu Glu Ser Lys
835 840 845
Val Phe Ser Ser Gly Gly
850
<210> 431
<211> 904
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 431
Met Ala Ser Val Leu Thr Pro Leu Leu Leu Arg Gly Leu Thr Gly Ser
1 5 10 15
Ala Arg Arg Leu Pro Val Pro Arg Ala Lys Ile His Ser Leu Asp Tyr
20 25 30
Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp Ile Asp Tyr Lys Asp
35 40 45
Asp Asp Asp Lys Gly Ile Arg Ile Gln Asp Leu Arg Thr Leu Gly Tyr
50 55 60
Ser Gln Gln Gln Gln Glu Lys Ile Lys Pro Lys Val Arg Ser Thr Val
65 70 75 80
Ala Gln His His Glu Ala Leu Val Gly His Gly Phe Thr His Ala His
85 90 95
Ile Val Ala Leu Ser Gln His Pro Ala Ala Leu Gly Thr Val Ala Val
100 105 110
Lys Tyr Gln Asp Met Ile Ala Ala Leu Pro Glu Ala Thr His Glu Ala
115 120 125
Ile Val Gly Val Gly Lys Gln Trp Ser Gly Ala Arg Ala Leu Glu Ala
130 135 140
Leu Leu Thr Val Ala Gly Glu Leu Arg Gly Pro Pro Leu Gln Leu Asp
145 150 155 160
Thr Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly Gly Val Thr Ala Val
165 170 175
Glu Ala Val His Ala Trp Arg Asn Ala Leu Thr Gly Ala Pro Leu Asn
180 185 190
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys
195 200 205
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp
210 215 220
His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile Gly
225 230 235 240
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
245 250 255
Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn
260 265 270
Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
275 280 285
Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala
290 295 300
Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
305 310 315 320
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala
325 330 335
Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
340 345 350
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val
355 360 365
Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val
370 375 380
Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala
385 390 395 400
Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu
405 410 415
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr
420 425 430
Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala
435 440 445
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
450 455 460
Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys
465 470 475 480
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
485 490 495
His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly
500 505 510
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
515 520 525
Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser His
530 535 540
Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
545 550 555 560
Leu Cys Gln Asp His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala
565 570 575
Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
580 585 590
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala
595 600 605
Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
610 615 620
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val
625 630 635 640
Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val
645 650 655
Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala
660 665 670
Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu
675 680 685
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr
690 695 700
Pro Ala Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala
705 710 715 720
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly
725 730 735
Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys
740 745 750
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
755 760 765
His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly
770 775 780
Gly Lys Gln Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp
785 790 795 800
Pro Ala Leu Ala Ala Leu Leu Val Gly Ser Pro Thr Pro Tyr Pro Asn
805 810 815
Tyr Ala Asn Ala Gly His Val Glu Gly Gln Ser Ala Leu Phe Met Arg
820 825 830
Asp Asn Gly Ile Ser Glu Gly Leu Val Phe His Asn Asn Pro Glu Gly
835 840 845
Thr Cys Gly Phe Cys Val Asn Met Thr Glu Thr Leu Leu Pro Glu Asn
850 855 860
Ala Lys Met Thr Val Val Pro Pro Glu Gly Ala Ile Pro Val Lys Arg
865 870 875 880
Gly Ala Thr Gly Glu Thr Lys Val Phe Thr Gly Asn Ser Asn Ser Pro
885 890 895
Lys Ser Pro Thr Lys Gly Gly Cys
900
<210> 432
<211> 918
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 432
Met Ala Ser Val Leu Thr Pro Leu Leu Leu Arg Gly Leu Thr Gly Ser
1 5 10 15
Ala Arg Arg Leu Pro Val Pro Arg Ala Lys Ile His Ser Leu Asp Tyr
20 25 30
Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp Ile Asp Tyr Lys Asp
35 40 45
Asp Asp Asp Lys Gly Ile Arg Ile Gln Asp Leu Arg Thr Leu Gly Tyr
50 55 60
Ser Gln Gln Gln Gln Glu Lys Ile Lys Pro Lys Val Arg Ser Thr Val
65 70 75 80
Ala Gln His His Glu Ala Leu Val Gly His Gly Phe Thr His Ala His
85 90 95
Ile Val Ala Leu Ser Gln His Pro Ala Ala Leu Gly Thr Val Ala Val
100 105 110
Lys Tyr Gln Asp Met Ile Ala Ala Leu Pro Glu Ala Thr His Glu Ala
115 120 125
Ile Val Gly Val Gly Lys Gln Trp Ser Gly Ala Arg Ala Leu Glu Ala
130 135 140
Leu Leu Thr Val Ala Gly Glu Leu Arg Gly Pro Pro Leu Gln Leu Asp
145 150 155 160
Thr Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly Gly Val Thr Ala Val
165 170 175
Glu Ala Val His Ala Trp Arg Asn Ala Leu Thr Gly Ala Pro Leu Asn
180 185 190
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys
195 200 205
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp
210 215 220
His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile Gly
225 230 235 240
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
245 250 255
Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn
260 265 270
Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
275 280 285
Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala
290 295 300
Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
305 310 315 320
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala
325 330 335
Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
340 345 350
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val
355 360 365
Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val
370 375 380
Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala
385 390 395 400
Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu
405 410 415
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr
420 425 430
Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala
435 440 445
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
450 455 460
Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys
465 470 475 480
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
485 490 495
His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly
500 505 510
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
515 520 525
Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser His
530 535 540
Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
545 550 555 560
Leu Cys Gln Asp His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala
565 570 575
Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
580 585 590
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala
595 600 605
Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
610 615 620
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val
625 630 635 640
Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val
645 650 655
Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala
660 665 670
Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu
675 680 685
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr
690 695 700
Pro Ala Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala
705 710 715 720
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly
725 730 735
Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys
740 745 750
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
755 760 765
His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly
770 775 780
Gly Lys Gln Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp
785 790 795 800
Pro Ala Leu Ala Ala Leu Leu Val Gly Ser Gly Ser Tyr Ala Leu Gly
805 810 815
Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro Ala Tyr Asn Gly Gln Thr
820 825 830
Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala Gly Gly Leu Glu Ser Lys
835 840 845
Val Phe Ser Ser Gly Gly Pro Thr Pro Tyr Pro Asn Tyr Ala Asn Ala
850 855 860
Gly His Val Glu Gly Gln Ser Ala Leu Phe Met Arg Asp Asn Gly Ile
865 870 875 880
Ser Glu Gly Leu Val Phe His Asn Asn Pro Glu Gly Thr Cys Gly Phe
885 890 895
Cys Val Asn Met Thr Glu Thr Leu Leu Pro Glu Asn Ala Lys Met Thr
900 905 910
Val Val Pro Pro Glu Gly
915
<210> 433
<211> 1032
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 433
Met Ala Leu Ser Arg Ala Val Cys Gly Thr Ser Arg Gln Leu Ala Pro
1 5 10 15
Val Leu Gly Tyr Leu Gly Ser Arg Gln Lys His Ser Leu Pro Asp Tyr
20 25 30
Pro Tyr Asp Val Pro Asp Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp
35 40 45
Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Met Asp Ile Ala
50 55 60
Asp Leu Arg Thr Leu Gly Tyr Ser Gln Gln Gln Gln Glu Lys Ile Lys
65 70 75 80
Pro Lys Val Arg Ser Thr Val Ala Gln His His Glu Ala Leu Val Gly
85 90 95
His Gly Phe Thr His Ala His Ile Val Ala Leu Ser Gln His Pro Ala
100 105 110
Ala Leu Gly Thr Val Ala Val Lys Tyr Gln Asp Met Ile Ala Ala Leu
115 120 125
Pro Glu Ala Thr His Glu Ala Ile Val Gly Val Gly Lys Gln Trp Ser
130 135 140
Gly Ala Arg Ala Leu Glu Ala Leu Leu Thr Val Ala Gly Glu Leu Arg
145 150 155 160
Gly Pro Pro Leu Gln Leu Asp Thr Gly Gln Leu Leu Lys Ile Ala Lys
165 170 175
Arg Gly Gly Val Thr Ala Val Glu Ala Val His Ala Trp Arg Asn Ala
180 185 190
Leu Thr Gly Ala Pro Leu Asn Leu Thr Pro Glu Gln Val Val Ala Ile
195 200 205
Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu
210 215 220
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Gln Gln Val Val
225 230 235 240
Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
245 250 255
Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Gln Gln
260 265 270
Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr
275 280 285
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
290 295 300
Glu Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu
305 310 315 320
Glu Thr Val Gln Ala Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
325 330 335
Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln
340 345 350
Ala Leu Glu Thr Val Gln Ala Leu Leu Pro Val Leu Cys Gln Ala His
355 360 365
Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
370 375 380
Lys Gln Ala Leu Glu Thr Val Gln Ala Leu Leu Pro Val Leu Cys Gln
385 390 395 400
Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile
405 410 415
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
420 425 430
Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser
435 440 445
Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu Leu Pro
450 455 460
Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile
465 470 475 480
Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu
485 490 495
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val
500 505 510
Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
515 520 525
Ala Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln
530 535 540
Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr
545 550 555 560
Val Gln Ala Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
565 570 575
Glu Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu
580 585 590
Glu Thr Val Gln Ala Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
595 600 605
Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln
610 615 620
Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His
625 630 635 640
Gly Leu Thr Pro Gln Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
645 650 655
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
660 665 670
Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly
675 680 685
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu Leu Pro Val Leu
690 695 700
Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser
705 710 715 720
His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu Leu Pro
725 730 735
Val Leu Cys Gln Ala His Gly Leu Thr Pro Gln Gln Val Val Ala Ile
740 745 750
Ala Ser Asn Gly Gly Gly Arg Pro Ala Leu Glu Ser Ile Val Ala Gln
755 760 765
Leu Ser Arg Pro Asp Pro Ala Leu Ala Ala Leu Thr Asn Asp His Leu
770 775 780
Val Ala Leu Ala Cys Leu Gly Gly Arg Pro Ala Leu Asp Ala Val Lys
785 790 795 800
Lys Gly Leu Gly Gly Ser Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile
805 810 815
Ser Ala Pro Gln Leu Pro Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe
820 825 830
Tyr Tyr Val Asn Asp Ala Gly Gly Leu Glu Ser Lys Val Phe Ser Ser
835 840 845
Gly Gly Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro
850 855 860
Glu Ser Ser Glu Val Glu Phe Ser His Glu Tyr Trp Met Arg His Ala
865 870 875 880
Leu Thr Leu Ala Lys Arg Ala Arg Asp Glu Arg Glu Val Pro Val Gly
885 890 895
Ala Val Leu Val Leu Asn Asn Arg Val Ile Gly Glu Gly Trp Asn Arg
900 905 910
Ala Ile Gly Leu His Asp Pro Thr Ala His Ala Glu Ile Met Ala Leu
915 920 925
Arg Gln Gly Gly Leu Val Met Gln Asn Tyr Arg Leu Ile Asp Ala Thr
930 935 940
Leu Tyr Val Thr Phe Glu Pro Cys Val Met Cys Ala Gly Ala Met Ile
945 950 955 960
His Ser Arg Ile Gly Arg Val Val Phe Gly Val Arg Asn Ser Lys Arg
965 970 975
Gly Ala Ala Gly Ser Leu Met Asn Val Leu Asn Tyr Pro Gly Met Asn
980 985 990
His Arg Val Glu Ile Thr Glu Gly Ile Leu Ala Asp Glu Cys Ala Ala
995 1000 1005
Leu Leu Cys Asp Phe Tyr Arg Met Pro Arg Gln Val Phe Asn Ala Gln
1010 1015 1020
Lys Lys Ala Gln Ser Ser Ile Asn
1025 1030
<210> 434
<211> 1082
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 434
Met Ala Leu Ser Arg Ala Val Cys Gly Thr Ser Arg Gln Leu Ala Pro
1 5 10 15
Val Leu Gly Tyr Leu Gly Ser Arg Gln Lys His Ser Leu Pro Asp Tyr
20 25 30
Pro Tyr Asp Val Pro Asp Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp
35 40 45
Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Met Asp Ile Ala
50 55 60
Asp Leu Arg Thr Leu Gly Tyr Ser Gln Gln Gln Gln Glu Lys Ile Lys
65 70 75 80
Pro Lys Val Arg Ser Thr Val Ala Gln His His Glu Ala Leu Val Gly
85 90 95
His Gly Phe Thr His Ala His Ile Val Ala Leu Ser Gln His Pro Ala
100 105 110
Ala Leu Gly Thr Val Ala Val Lys Tyr Gln Asp Met Ile Ala Ala Leu
115 120 125
Pro Glu Ala Thr His Glu Ala Ile Val Gly Val Gly Lys Gln Trp Ser
130 135 140
Gly Ala Arg Ala Leu Glu Ala Leu Leu Thr Val Ala Gly Glu Leu Arg
145 150 155 160
Gly Pro Pro Leu Gln Leu Asp Thr Gly Gln Leu Leu Lys Ile Ala Lys
165 170 175
Arg Gly Gly Val Thr Ala Val Glu Ala Val His Ala Trp Arg Asn Ala
180 185 190
Leu Thr Gly Ala Pro Leu Asn Leu Thr Pro Glu Gln Val Val Ala Ile
195 200 205
Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu
210 215 220
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Gln Gln Val Val
225 230 235 240
Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
245 250 255
Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Gln Gln
260 265 270
Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr
275 280 285
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
290 295 300
Glu Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu
305 310 315 320
Glu Thr Val Gln Ala Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
325 330 335
Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln
340 345 350
Ala Leu Glu Thr Val Gln Ala Leu Leu Pro Val Leu Cys Gln Ala His
355 360 365
Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
370 375 380
Lys Gln Ala Leu Glu Thr Val Gln Ala Leu Leu Pro Val Leu Cys Gln
385 390 395 400
Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile
405 410 415
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
420 425 430
Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser
435 440 445
Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu Leu Pro
450 455 460
Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile
465 470 475 480
Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu
485 490 495
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val
500 505 510
Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
515 520 525
Ala Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln
530 535 540
Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr
545 550 555 560
Val Gln Ala Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
565 570 575
Glu Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu
580 585 590
Glu Thr Val Gln Ala Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
595 600 605
Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln
610 615 620
Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His
625 630 635 640
Gly Leu Thr Pro Gln Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
645 650 655
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
660 665 670
Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly
675 680 685
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu Leu Pro Val Leu
690 695 700
Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser
705 710 715 720
His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu Leu Pro
725 730 735
Val Leu Cys Gln Ala His Gly Leu Thr Pro Gln Gln Val Val Ala Ile
740 745 750
Ala Ser Asn Gly Gly Gly Arg Pro Ala Leu Glu Ser Ile Val Ala Gln
755 760 765
Leu Ser Arg Pro Asp Pro Ala Leu Ala Ala Leu Thr Asn Asp His Leu
770 775 780
Val Ala Leu Ala Cys Leu Gly Gly Arg Pro Ala Leu Asp Ala Val Lys
785 790 795 800
Lys Gly Leu Gly Gly Ser Pro Thr Pro Tyr Pro Asn Tyr Ala Asn Ala
805 810 815
Gly His Val Glu Gly Gln Ser Ala Leu Phe Met Arg Asp Asn Gly Ile
820 825 830
Ser Glu Gly Leu Val Phe His Asn Asn Pro Glu Gly Thr Cys Gly Phe
835 840 845
Cys Val Asn Met Thr Glu Thr Leu Leu Pro Glu Asn Ala Lys Met Thr
850 855 860
Val Val Pro Pro Glu Gly Ala Ile Pro Val Lys Arg Gly Ala Thr Gly
865 870 875 880
Glu Thr Lys Val Phe Thr Gly Asn Ser Asn Ser Pro Lys Ser Pro Thr
885 890 895
Lys Gly Gly Cys Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala
900 905 910
Thr Pro Glu Ser Ser Glu Val Glu Phe Ser His Glu Tyr Trp Met Arg
915 920 925
His Ala Leu Thr Leu Ala Lys Arg Ala Arg Asp Glu Arg Glu Val Pro
930 935 940
Val Gly Ala Val Leu Val Leu Asn Asn Arg Val Ile Gly Glu Gly Trp
945 950 955 960
Asn Arg Ala Ile Gly Leu His Asp Pro Thr Ala His Ala Glu Ile Met
965 970 975
Ala Leu Arg Gln Gly Gly Leu Val Met Gln Asn Tyr Arg Leu Ile Asp
980 985 990
Ala Thr Leu Tyr Val Thr Phe Glu Pro Cys Val Met Cys Ala Gly Ala
995 1000 1005
Met Ile His Ser Arg Ile Gly Arg Val Val Phe Gly Val Arg Asn Ser
1010 1015 1020
Lys Arg Gly Ala Ala Gly Ser Leu Met Asn Val Leu Asn Tyr Pro Gly
1025 1030 1035 1040
Met Asn His Arg Val Glu Ile Thr Glu Gly Ile Leu Ala Asp Glu Cys
1045 1050 1055
Ala Ala Leu Leu Cys Asp Phe Tyr Arg Met Pro Arg Gln Val Phe Asn
1060 1065 1070
Ala Gln Lys Lys Ala Gln Ser Ser Ile Asn
1075 1080
<210> 435
<211> 1020
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 435
Met Ala Leu Ser Arg Ala Val Cys Gly Thr Ser Arg Gln Leu Ala Pro
1 5 10 15
Val Leu Gly Tyr Leu Gly Ser Arg Gln Lys His Ser Leu Pro Asp Tyr
20 25 30
Pro Tyr Asp Val Pro Asp Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp
35 40 45
Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Met Asp Ile Ala
50 55 60
Asp Leu Arg Thr Leu Gly Tyr Ser Gln Gln Gln Gln Glu Lys Ile Lys
65 70 75 80
Pro Lys Val Arg Ser Thr Val Ala Gln His His Glu Ala Leu Val Gly
85 90 95
His Gly Phe Thr His Ala His Ile Val Ala Leu Ser Gln His Pro Ala
100 105 110
Ala Leu Gly Thr Val Ala Val Lys Tyr Gln Asp Met Ile Ala Ala Leu
115 120 125
Pro Glu Ala Thr His Glu Ala Ile Val Gly Val Gly Lys Gln Trp Ser
130 135 140
Gly Ala Arg Ala Leu Glu Ala Leu Leu Thr Val Ala Gly Glu Leu Arg
145 150 155 160
Gly Pro Pro Leu Gln Leu Asp Thr Gly Gln Leu Leu Lys Ile Ala Lys
165 170 175
Arg Gly Gly Val Thr Ala Val Glu Ala Val His Ala Trp Arg Asn Ala
180 185 190
Leu Thr Gly Ala Pro Leu Asn Leu Thr Pro Glu Gln Val Val Ala Ile
195 200 205
Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu
210 215 220
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Gln Gln Val Val
225 230 235 240
Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
245 250 255
Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Gln Gln
260 265 270
Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr
275 280 285
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
290 295 300
Glu Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu
305 310 315 320
Glu Thr Val Gln Ala Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
325 330 335
Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln
340 345 350
Ala Leu Glu Thr Val Gln Ala Leu Leu Pro Val Leu Cys Gln Ala His
355 360 365
Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
370 375 380
Lys Gln Ala Leu Glu Thr Val Gln Ala Leu Leu Pro Val Leu Cys Gln
385 390 395 400
Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile
405 410 415
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
420 425 430
Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser
435 440 445
Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu Leu Pro
450 455 460
Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile
465 470 475 480
Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu
485 490 495
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val
500 505 510
Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
515 520 525
Ala Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln
530 535 540
Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr
545 550 555 560
Val Gln Ala Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
565 570 575
Glu Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu
580 585 590
Glu Thr Val Gln Ala Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
595 600 605
Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln
610 615 620
Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His
625 630 635 640
Gly Leu Thr Pro Gln Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
645 650 655
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
660 665 670
Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly
675 680 685
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu Leu Pro Val Leu
690 695 700
Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser
705 710 715 720
His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu Leu Pro
725 730 735
Val Leu Cys Gln Ala His Gly Leu Thr Pro Gln Gln Val Val Ala Ile
740 745 750
Ala Ser Asn Gly Gly Gly Arg Pro Ala Leu Glu Ser Ile Val Ala Gln
755 760 765
Leu Ser Arg Pro Asp Pro Ala Leu Ala Ala Leu Thr Asn Asp His Leu
770 775 780
Val Ala Leu Ala Cys Leu Gly Gly Arg Pro Ala Leu Asp Ala Val Lys
785 790 795 800
Lys Gly Leu Gly Gly Ser Gly Ser Ala Ile Pro Val Lys Arg Gly Ala
805 810 815
Thr Gly Glu Thr Lys Val Phe Thr Gly Asn Ser Asn Ser Pro Lys Ser
820 825 830
Pro Thr Lys Gly Gly Cys Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu
835 840 845
Ser Ala Thr Pro Glu Ser Ser Glu Val Glu Phe Ser His Glu Tyr Trp
850 855 860
Met Arg His Ala Leu Thr Leu Ala Lys Arg Ala Arg Asp Glu Arg Glu
865 870 875 880
Val Pro Val Gly Ala Val Leu Val Leu Asn Asn Arg Val Ile Gly Glu
885 890 895
Gly Trp Asn Arg Ala Ile Gly Leu His Asp Pro Thr Ala His Ala Glu
900 905 910
Ile Met Ala Leu Arg Gln Gly Gly Leu Val Met Gln Asn Tyr Arg Leu
915 920 925
Ile Asp Ala Thr Leu Tyr Val Thr Phe Glu Pro Cys Val Met Cys Ala
930 935 940
Gly Ala Met Ile His Ser Arg Ile Gly Arg Val Val Phe Gly Val Arg
945 950 955 960
Asn Ser Lys Arg Gly Ala Ala Gly Ser Leu Met Asn Val Leu Asn Tyr
965 970 975
Pro Gly Met Asn His Arg Val Glu Ile Thr Glu Gly Ile Leu Ala Asp
980 985 990
Glu Cys Ala Ala Leu Leu Cys Asp Phe Tyr Arg Met Pro Arg Gln Val
995 1000 1005
Phe Asn Ala Gln Lys Lys Ala Gln Ser Ser Ile Asn
1010 1015 1020
<210> 436
<211> 1024
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 436
Met Ala Ser Val Leu Thr Pro Leu Leu Leu Arg Gly Leu Thr Gly Ser
1 5 10 15
Ala Arg Arg Leu Pro Val Pro Arg Ala Lys Ile His Ser Leu Asp Tyr
20 25 30
Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp Ile Asp Tyr Lys Asp
35 40 45
Asp Asp Asp Lys Met Asp Ile Ala Asp Leu Arg Thr Leu Gly Tyr Ser
50 55 60
Gln Gln Gln Gln Glu Lys Ile Lys Pro Lys Val Arg Ser Thr Val Ala
65 70 75 80
Gln His His Glu Ala Leu Val Gly His Gly Phe Thr His Ala His Ile
85 90 95
Val Ala Leu Ser Gln His Pro Ala Ala Leu Gly Thr Val Ala Val Lys
100 105 110
Tyr Gln Asp Met Ile Ala Ala Leu Pro Glu Ala Thr His Glu Ala Ile
115 120 125
Val Gly Val Gly Lys Gln Trp Ser Gly Ala Arg Ala Leu Glu Ala Leu
130 135 140
Leu Thr Val Ala Gly Glu Leu Arg Gly Pro Pro Leu Gln Leu Asp Thr
145 150 155 160
Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly Gly Val Thr Ala Val Glu
165 170 175
Ala Val His Ala Trp Arg Asn Ala Leu Thr Gly Ala Pro Leu Asn Leu
180 185 190
Thr Pro Glu Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln
195 200 205
Ala Leu Glu Thr Val Gln Ala Leu Leu Pro Val Leu Cys Gln Ala His
210 215 220
Gly Leu Thr Pro Gln Gln Val Val Ala Ile Ala Ser His Asp Gly Gly
225 230 235 240
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
245 250 255
Ala His Gly Leu Thr Pro Gln Gln Val Val Ala Ile Ala Ser Asn Gly
260 265 270
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
275 280 285
Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser
290 295 300
Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu Leu Pro
305 310 315 320
Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile
325 330 335
Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu
340 345 350
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val
355 360 365
Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
370 375 380
Ala Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln
385 390 395 400
Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr
405 410 415
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
420 425 430
Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu
435 440 445
Glu Thr Val Gln Ala Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
450 455 460
Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln
465 470 475 480
Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His
485 490 495
Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
500 505 510
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
515 520 525
Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn
530 535 540
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu Leu Pro Val Leu
545 550 555 560
Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser
565 570 575
Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu Leu Pro
580 585 590
Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile
595 600 605
Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu
610 615 620
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val
625 630 635 640
Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
645 650 655
Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Gln Gln
660 665 670
Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr
675 680 685
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
690 695 700
Gln Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu
705 710 715 720
Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
725 730 735
Thr Pro Gln Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Arg Pro
740 745 750
Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp Pro Ala Leu
755 760 765
Ala Ala Leu Thr Asn Asp His Leu Val Ala Leu Ala Cys Leu Gly Gly
770 775 780
Arg Pro Ala Leu Asp Ala Val Lys Lys Gly Leu Gly Gly Ser Gly Ser
785 790 795 800
Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro Ala Tyr
805 810 815
Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala Gly Gly
820 825 830
Leu Glu Ser Lys Val Phe Ser Ser Gly Gly Ser Gly Ser Glu Thr Pro
835 840 845
Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser Ser Glu Val Glu Phe Ser
850 855 860
His Glu Tyr Trp Met Arg His Ala Leu Thr Leu Ala Lys Arg Ala Arg
865 870 875 880
Asp Glu Arg Glu Val Pro Val Gly Ala Val Leu Val Leu Asn Asn Arg
885 890 895
Val Ile Gly Glu Gly Trp Asn Arg Ala Ile Gly Leu His Asp Pro Thr
900 905 910
Ala His Ala Glu Ile Met Ala Leu Arg Gln Gly Gly Leu Val Met Gln
915 920 925
Asn Tyr Arg Leu Ile Asp Ala Thr Leu Tyr Val Thr Phe Glu Pro Cys
930 935 940
Val Met Cys Ala Gly Ala Met Ile His Ser Arg Ile Gly Arg Val Val
945 950 955 960
Phe Gly Val Arg Asn Ser Lys Arg Gly Ala Ala Gly Ser Leu Met Asn
965 970 975
Val Leu Asn Tyr Pro Gly Met Asn His Arg Val Glu Ile Thr Glu Gly
980 985 990
Ile Leu Ala Asp Glu Cys Ala Ala Leu Leu Cys Asp Phe Tyr Arg Met
995 1000 1005
Pro Arg Gln Val Phe Asn Ala Gln Lys Lys Ala Gln Ser Ser Ile Asn
1010 1015 1020
<210> 437
<211> 1074
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 437
Met Ala Ser Val Leu Thr Pro Leu Leu Leu Arg Gly Leu Thr Gly Ser
1 5 10 15
Ala Arg Arg Leu Pro Val Pro Arg Ala Lys Ile His Ser Leu Asp Tyr
20 25 30
Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp Ile Asp Tyr Lys Asp
35 40 45
Asp Asp Asp Lys Met Asp Ile Ala Asp Leu Arg Thr Leu Gly Tyr Ser
50 55 60
Gln Gln Gln Gln Glu Lys Ile Lys Pro Lys Val Arg Ser Thr Val Ala
65 70 75 80
Gln His His Glu Ala Leu Val Gly His Gly Phe Thr His Ala His Ile
85 90 95
Val Ala Leu Ser Gln His Pro Ala Ala Leu Gly Thr Val Ala Val Lys
100 105 110
Tyr Gln Asp Met Ile Ala Ala Leu Pro Glu Ala Thr His Glu Ala Ile
115 120 125
Val Gly Val Gly Lys Gln Trp Ser Gly Ala Arg Ala Leu Glu Ala Leu
130 135 140
Leu Thr Val Ala Gly Glu Leu Arg Gly Pro Pro Leu Gln Leu Asp Thr
145 150 155 160
Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly Gly Val Thr Ala Val Glu
165 170 175
Ala Val His Ala Trp Arg Asn Ala Leu Thr Gly Ala Pro Leu Asn Leu
180 185 190
Thr Pro Glu Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln
195 200 205
Ala Leu Glu Thr Val Gln Ala Leu Leu Pro Val Leu Cys Gln Ala His
210 215 220
Gly Leu Thr Pro Gln Gln Val Val Ala Ile Ala Ser His Asp Gly Gly
225 230 235 240
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
245 250 255
Ala His Gly Leu Thr Pro Gln Gln Val Val Ala Ile Ala Ser Asn Gly
260 265 270
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
275 280 285
Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser
290 295 300
Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu Leu Pro
305 310 315 320
Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile
325 330 335
Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu
340 345 350
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val
355 360 365
Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
370 375 380
Ala Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln
385 390 395 400
Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr
405 410 415
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
420 425 430
Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu
435 440 445
Glu Thr Val Gln Ala Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
450 455 460
Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln
465 470 475 480
Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His
485 490 495
Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
500 505 510
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
515 520 525
Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn
530 535 540
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu Leu Pro Val Leu
545 550 555 560
Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser
565 570 575
Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu Leu Pro
580 585 590
Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile
595 600 605
Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu
610 615 620
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val
625 630 635 640
Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
645 650 655
Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Gln Gln
660 665 670
Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr
675 680 685
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
690 695 700
Gln Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu
705 710 715 720
Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
725 730 735
Thr Pro Gln Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Arg Pro
740 745 750
Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp Pro Ala Leu
755 760 765
Ala Ala Leu Thr Asn Asp His Leu Val Ala Leu Ala Cys Leu Gly Gly
770 775 780
Arg Pro Ala Leu Asp Ala Val Lys Lys Gly Leu Gly Gly Ser Pro Thr
785 790 795 800
Pro Tyr Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln Ser Ala
805 810 815
Leu Phe Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe His Asn
820 825 830
Asn Pro Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu Thr Leu
835 840 845
Leu Pro Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly Ala Ile
850 855 860
Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr Gly Asn
865 870 875 880
Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys Ser Gly Ser Glu
885 890 895
Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser Ser Glu Val Glu
900 905 910
Phe Ser His Glu Tyr Trp Met Arg His Ala Leu Thr Leu Ala Lys Arg
915 920 925
Ala Arg Asp Glu Arg Glu Val Pro Val Gly Ala Val Leu Val Leu Asn
930 935 940
Asn Arg Val Ile Gly Glu Gly Trp Asn Arg Ala Ile Gly Leu His Asp
945 950 955 960
Pro Thr Ala His Ala Glu Ile Met Ala Leu Arg Gln Gly Gly Leu Val
965 970 975
Met Gln Asn Tyr Arg Leu Ile Asp Ala Thr Leu Tyr Val Thr Phe Glu
980 985 990
Pro Cys Val Met Cys Ala Gly Ala Met Ile His Ser Arg Ile Gly Arg
995 1000 1005
Val Val Phe Gly Val Arg Asn Ser Lys Arg Gly Ala Ala Gly Ser Leu
1010 1015 1020
Met Asn Val Leu Asn Tyr Pro Gly Met Asn His Arg Val Glu Ile Thr
1025 1030 1035 1040
Glu Gly Ile Leu Ala Asp Glu Cys Ala Ala Leu Leu Cys Asp Phe Tyr
1045 1050 1055
Arg Met Pro Arg Gln Val Phe Asn Ala Gln Lys Lys Ala Gln Ser Ser
1060 1065 1070
Ile Asn
<210> 438
<211> 1012
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 438
Met Ala Ser Val Leu Thr Pro Leu Leu Leu Arg Gly Leu Thr Gly Ser
1 5 10 15
Ala Arg Arg Leu Pro Val Pro Arg Ala Lys Ile His Ser Leu Asp Tyr
20 25 30
Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp Ile Asp Tyr Lys Asp
35 40 45
Asp Asp Asp Lys Met Asp Ile Ala Asp Leu Arg Thr Leu Gly Tyr Ser
50 55 60
Gln Gln Gln Gln Glu Lys Ile Lys Pro Lys Val Arg Ser Thr Val Ala
65 70 75 80
Gln His His Glu Ala Leu Val Gly His Gly Phe Thr His Ala His Ile
85 90 95
Val Ala Leu Ser Gln His Pro Ala Ala Leu Gly Thr Val Ala Val Lys
100 105 110
Tyr Gln Asp Met Ile Ala Ala Leu Pro Glu Ala Thr His Glu Ala Ile
115 120 125
Val Gly Val Gly Lys Gln Trp Ser Gly Ala Arg Ala Leu Glu Ala Leu
130 135 140
Leu Thr Val Ala Gly Glu Leu Arg Gly Pro Pro Leu Gln Leu Asp Thr
145 150 155 160
Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly Gly Val Thr Ala Val Glu
165 170 175
Ala Val His Ala Trp Arg Asn Ala Leu Thr Gly Ala Pro Leu Asn Leu
180 185 190
Thr Pro Glu Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln
195 200 205
Ala Leu Glu Thr Val Gln Ala Leu Leu Pro Val Leu Cys Gln Ala His
210 215 220
Gly Leu Thr Pro Gln Gln Val Val Ala Ile Ala Ser His Asp Gly Gly
225 230 235 240
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
245 250 255
Ala His Gly Leu Thr Pro Gln Gln Val Val Ala Ile Ala Ser Asn Gly
260 265 270
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
275 280 285
Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser
290 295 300
Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu Leu Pro
305 310 315 320
Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile
325 330 335
Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu
340 345 350
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val
355 360 365
Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
370 375 380
Ala Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln
385 390 395 400
Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr
405 410 415
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
420 425 430
Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu
435 440 445
Glu Thr Val Gln Ala Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
450 455 460
Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln
465 470 475 480
Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His
485 490 495
Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
500 505 510
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
515 520 525
Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn
530 535 540
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu Leu Pro Val Leu
545 550 555 560
Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser
565 570 575
Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu Leu Pro
580 585 590
Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile
595 600 605
Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu
610 615 620
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val
625 630 635 640
Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
645 650 655
Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Gln Gln
660 665 670
Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr
675 680 685
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
690 695 700
Gln Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu
705 710 715 720
Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
725 730 735
Thr Pro Gln Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Arg Pro
740 745 750
Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp Pro Ala Leu
755 760 765
Ala Ala Leu Thr Asn Asp His Leu Val Ala Leu Ala Cys Leu Gly Gly
770 775 780
Arg Pro Ala Leu Asp Ala Val Lys Lys Gly Leu Gly Gly Ser Gly Ser
785 790 795 800
Ala Ile Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr
805 810 815
Gly Asn Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys Ser Gly
820 825 830
Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser Ser Glu
835 840 845
Val Glu Phe Ser His Glu Tyr Trp Met Arg His Ala Leu Thr Leu Ala
850 855 860
Lys Arg Ala Arg Asp Glu Arg Glu Val Pro Val Gly Ala Val Leu Val
865 870 875 880
Leu Asn Asn Arg Val Ile Gly Glu Gly Trp Asn Arg Ala Ile Gly Leu
885 890 895
His Asp Pro Thr Ala His Ala Glu Ile Met Ala Leu Arg Gln Gly Gly
900 905 910
Leu Val Met Gln Asn Tyr Arg Leu Ile Asp Ala Thr Leu Tyr Val Thr
915 920 925
Phe Glu Pro Cys Val Met Cys Ala Gly Ala Met Ile His Ser Arg Ile
930 935 940
Gly Arg Val Val Phe Gly Val Arg Asn Ser Lys Arg Gly Ala Ala Gly
945 950 955 960
Ser Leu Met Asn Val Leu Asn Tyr Pro Gly Met Asn His Arg Val Glu
965 970 975
Ile Thr Glu Gly Ile Leu Ala Asp Glu Cys Ala Ala Leu Leu Cys Asp
980 985 990
Phe Tyr Arg Met Pro Arg Gln Val Phe Asn Ala Gln Lys Lys Ala Gln
995 1000 1005
Ser Ser Ile Asn
1010
<210> 439
<211> 1038
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 439
Met Ala Leu Ser Arg Ala Val Cys Gly Thr Ser Arg Gln Leu Ala Pro
1 5 10 15
Val Leu Gly Tyr Leu Gly Ser Arg Gln Lys His Ser Leu Pro Asp Tyr
20 25 30
Pro Tyr Asp Val Pro Asp Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp
35 40 45
Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Gly Ile Arg Ile
50 55 60
Gln Asp Leu Arg Thr Leu Gly Tyr Ser Gln Gln Gln Gln Glu Lys Ile
65 70 75 80
Lys Pro Lys Val Arg Ser Thr Val Ala Gln His His Glu Ala Leu Val
85 90 95
Gly His Gly Phe Thr His Ala His Ile Val Ala Leu Ser Gln His Pro
100 105 110
Ala Ala Leu Gly Thr Val Ala Val Lys Tyr Gln Asp Met Ile Ala Ala
115 120 125
Leu Pro Glu Ala Thr His Glu Ala Ile Val Gly Val Gly Lys Gln Trp
130 135 140
Ser Gly Ala Arg Ala Leu Glu Ala Leu Leu Thr Val Ala Gly Glu Leu
145 150 155 160
Arg Gly Pro Pro Leu Gln Leu Asp Thr Gly Gln Leu Leu Lys Ile Ala
165 170 175
Lys Arg Gly Gly Val Thr Ala Val Glu Ala Val His Ala Trp Arg Asn
180 185 190
Ala Leu Thr Gly Ala Pro Leu Asn Leu Thr Pro Asp Gln Val Val Ala
195 200 205
Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
210 215 220
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val
225 230 235 240
Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val
245 250 255
Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Ala
260 265 270
Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu
275 280 285
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr
290 295 300
Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala
305 310 315 320
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
325 330 335
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys
340 345 350
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp
355 360 365
His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly
370 375 380
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
385 390 395 400
Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn
405 410 415
Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
420 425 430
Leu Cys Gln Asp His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala
435 440 445
Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
450 455 460
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala
465 470 475 480
Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
485 490 495
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val
500 505 510
Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val
515 520 525
Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Asp
530 535 540
Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu
545 550 555 560
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr
565 570 575
Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala
580 585 590
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
595 600 605
Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys
610 615 620
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
625 630 635 640
His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly
645 650 655
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
660 665 670
Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn
675 680 685
Gly Gly Gly Lys Gln Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg
690 695 700
Pro Asp Pro Ala Leu Ala Ala Leu Leu Val Ser Gly Ser Glu Thr Pro
705 710 715 720
Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser Ser Glu Val Glu Phe Ser
725 730 735
His Glu Tyr Trp Met Arg His Ala Leu Thr Leu Ala Lys Arg Ala Arg
740 745 750
Asp Glu Arg Glu Val Pro Val Gly Ala Val Leu Val Leu Asn Asn Arg
755 760 765
Val Ile Gly Glu Gly Trp Asn Arg Ala Ile Gly Leu His Asp Pro Thr
770 775 780
Ala His Ala Glu Ile Met Ala Leu Arg Gln Gly Gly Leu Val Met Gln
785 790 795 800
Asn Tyr Arg Leu Ile Asp Ala Thr Leu Tyr Val Thr Phe Glu Pro Cys
805 810 815
Val Met Cys Ala Gly Ala Met Ile His Ser Arg Ile Gly Arg Val Val
820 825 830
Phe Gly Val Arg Asn Ser Lys Arg Gly Ala Ala Gly Ser Leu Met Asn
835 840 845
Val Leu Asn Tyr Pro Gly Met Asn His Arg Val Glu Ile Thr Glu Gly
850 855 860
Ile Leu Ala Asp Glu Cys Ala Ala Leu Leu Cys Asp Phe Tyr Arg Met
865 870 875 880
Pro Arg Gln Val Phe Asn Ala Gln Lys Lys Ala Gln Ser Ser Ile Asn
885 890 895
Leu Val Gly Ser Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala
900 905 910
Pro Gln Leu Pro Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr
915 920 925
Val Asn Asp Ala Gly Gly Leu Glu Gly Lys Val Phe Ser Ser Gly Gly
930 935 940
Pro Thr Pro Tyr Pro Asn Tyr Ala Asn Ala Gly His Val Glu Ser Gln
945 950 955 960
Ser Ala Leu Phe Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe
965 970 975
His Asn Asn Pro Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu
980 985 990
Thr Leu Leu Pro Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly
995 1000 1005
Val Ile Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr
1010 1015 1020
Gly Asn Ser Asn Gly Pro Lys Ser Pro Thr Lys Gly Gly Cys
1025 1030 1035
<210> 440
<211> 1038
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 440
Met Ala Leu Ser Arg Ala Val Cys Gly Thr Ser Arg Gln Leu Ala Pro
1 5 10 15
Val Leu Gly Tyr Leu Gly Ser Arg Gln Lys His Ser Leu Pro Asp Tyr
20 25 30
Pro Tyr Asp Val Pro Asp Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp
35 40 45
Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Gly Ile Arg Ile
50 55 60
Gln Asp Leu Arg Thr Leu Gly Tyr Ser Gln Gln Gln Gln Glu Lys Ile
65 70 75 80
Lys Pro Lys Val Arg Ser Thr Val Ala Gln His His Glu Ala Leu Val
85 90 95
Gly His Gly Phe Thr His Ala His Ile Val Ala Leu Ser Gln His Pro
100 105 110
Ala Ala Leu Gly Thr Val Ala Val Lys Tyr Gln Asp Met Ile Ala Ala
115 120 125
Leu Pro Glu Ala Thr His Glu Ala Ile Val Gly Val Gly Lys Gln Trp
130 135 140
Ser Gly Ala Arg Ala Leu Glu Ala Leu Leu Thr Val Ala Gly Glu Leu
145 150 155 160
Arg Gly Pro Pro Leu Gln Leu Asp Thr Gly Gln Leu Leu Lys Ile Ala
165 170 175
Lys Arg Gly Gly Val Thr Ala Val Glu Ala Val His Ala Trp Arg Asn
180 185 190
Ala Leu Thr Gly Ala Pro Leu Asn Leu Thr Pro Asp Gln Val Val Ala
195 200 205
Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
210 215 220
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val
225 230 235 240
Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val
245 250 255
Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Ala
260 265 270
Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu
275 280 285
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr
290 295 300
Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala
305 310 315 320
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
325 330 335
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys
340 345 350
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp
355 360 365
His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly
370 375 380
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
385 390 395 400
Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn
405 410 415
Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
420 425 430
Leu Cys Gln Asp His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala
435 440 445
Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
450 455 460
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala
465 470 475 480
Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
485 490 495
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val
500 505 510
Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val
515 520 525
Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Asp
530 535 540
Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu
545 550 555 560
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr
565 570 575
Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala
580 585 590
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
595 600 605
Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys
610 615 620
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
625 630 635 640
His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly
645 650 655
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
660 665 670
Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn
675 680 685
Gly Gly Gly Lys Gln Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg
690 695 700
Pro Asp Pro Ala Leu Ala Ala Leu Leu Val Ser Gly Ser Glu Thr Pro
705 710 715 720
Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser Ser Glu Val Glu Phe Ser
725 730 735
His Glu Tyr Trp Met Arg His Ala Leu Thr Leu Ala Lys Arg Ala Arg
740 745 750
Asp Glu Arg Glu Val Pro Val Gly Ala Val Leu Val Leu Asn Asn Arg
755 760 765
Val Ile Gly Glu Gly Trp Asn Arg Ala Ile Gly Leu His Asp Pro Thr
770 775 780
Ala His Ala Glu Ile Met Ala Leu Arg Gln Gly Gly Leu Val Met Gln
785 790 795 800
Asn Tyr Arg Leu Ile Asp Ala Thr Leu Tyr Val Thr Phe Glu Pro Cys
805 810 815
Val Met Cys Ala Gly Ala Met Ile His Ser Arg Ile Gly Arg Val Val
820 825 830
Phe Gly Val Arg Asn Ser Lys Arg Gly Ala Ala Gly Ser Leu Met Asn
835 840 845
Val Leu Asn Tyr Pro Gly Met Asn His Arg Val Glu Ile Thr Glu Gly
850 855 860
Ile Leu Ala Asp Glu Cys Ala Ala Leu Leu Cys Asp Phe Tyr Arg Met
865 870 875 880
Pro Arg Gln Val Phe Asn Ala Gln Lys Lys Ala Gln Ser Ser Ile Asn
885 890 895
Leu Val Gly Ser Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala
900 905 910
Pro Gln Leu Pro Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr
915 920 925
Val Asn Asp Ala Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly
930 935 940
Pro Thr Pro Tyr Pro Asn Tyr Ala Asn Ala Gly His Val Ala Gly Gln
945 950 955 960
Ser Ala Leu Phe Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe
965 970 975
His Asn Asn Pro Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu
980 985 990
Thr Leu Leu Pro Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly
995 1000 1005
Ala Ile Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr
1010 1015 1020
Gly Asn Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys
1025 1030 1035
<210> 441
<211> 1038
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 441
Met Ala Leu Ser Arg Ala Val Cys Gly Thr Ser Arg Gln Leu Ala Pro
1 5 10 15
Val Leu Gly Tyr Leu Gly Ser Arg Gln Lys His Ser Leu Pro Asp Tyr
20 25 30
Pro Tyr Asp Val Pro Asp Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp
35 40 45
Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Gly Ile Arg Ile
50 55 60
Gln Asp Leu Arg Thr Leu Gly Tyr Ser Gln Gln Gln Gln Glu Lys Ile
65 70 75 80
Lys Pro Lys Val Arg Ser Thr Val Ala Gln His His Glu Ala Leu Val
85 90 95
Gly His Gly Phe Thr His Ala His Ile Val Ala Leu Ser Gln His Pro
100 105 110
Ala Ala Leu Gly Thr Val Ala Val Lys Tyr Gln Asp Met Ile Ala Ala
115 120 125
Leu Pro Glu Ala Thr His Glu Ala Ile Val Gly Val Gly Lys Gln Trp
130 135 140
Ser Gly Ala Arg Ala Leu Glu Ala Leu Leu Thr Val Ala Gly Glu Leu
145 150 155 160
Arg Gly Pro Pro Leu Gln Leu Asp Thr Gly Gln Leu Leu Lys Ile Ala
165 170 175
Lys Arg Gly Gly Val Thr Ala Val Glu Ala Val His Ala Trp Arg Asn
180 185 190
Ala Leu Thr Gly Ala Pro Leu Asn Leu Thr Pro Asp Gln Val Val Ala
195 200 205
Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
210 215 220
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val
225 230 235 240
Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val
245 250 255
Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Ala
260 265 270
Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu
275 280 285
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr
290 295 300
Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala
305 310 315 320
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
325 330 335
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys
340 345 350
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp
355 360 365
His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly
370 375 380
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
385 390 395 400
Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn
405 410 415
Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
420 425 430
Leu Cys Gln Asp His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala
435 440 445
Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
450 455 460
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala
465 470 475 480
Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
485 490 495
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val
500 505 510
Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val
515 520 525
Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Asp
530 535 540
Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu
545 550 555 560
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr
565 570 575
Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala
580 585 590
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
595 600 605
Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys
610 615 620
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
625 630 635 640
His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly
645 650 655
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
660 665 670
Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn
675 680 685
Gly Gly Gly Lys Gln Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg
690 695 700
Pro Asp Pro Ala Leu Ala Ala Leu Leu Val Ser Gly Ser Glu Thr Pro
705 710 715 720
Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser Ser Glu Val Glu Phe Ser
725 730 735
His Glu Tyr Trp Met Arg His Ala Leu Thr Leu Ala Lys Arg Ala Arg
740 745 750
Asp Glu Arg Glu Val Pro Val Gly Ala Val Leu Val Leu Asn Asn Arg
755 760 765
Val Ile Gly Glu Gly Trp Asn Arg Ala Ile Gly Leu His Asp Pro Thr
770 775 780
Ala His Ala Glu Ile Met Ala Leu Arg Gln Gly Gly Leu Val Met Gln
785 790 795 800
Asn Tyr Arg Leu Ile Asp Ala Thr Leu Tyr Val Thr Phe Glu Pro Cys
805 810 815
Val Met Cys Ala Gly Ala Met Ile His Ser Arg Ile Gly Arg Val Val
820 825 830
Phe Gly Val Arg Asn Ser Lys Arg Gly Ala Ala Gly Ser Leu Met Asn
835 840 845
Val Leu Asn Tyr Pro Gly Met Asn His Arg Val Glu Ile Thr Glu Gly
850 855 860
Ile Leu Ala Asp Glu Cys Ala Ala Leu Leu Cys Asp Phe Tyr Arg Met
865 870 875 880
Pro Arg Gln Val Phe Asn Ala Gln Lys Lys Ala Gln Ser Ser Ile Asn
885 890 895
Leu Val Gly Ser Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala
900 905 910
Pro Gln Leu Pro Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr
915 920 925
Val Asn Asp Ala Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly
930 935 940
Pro Thr Pro Tyr Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln
945 950 955 960
Ser Ala Leu Phe Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe
965 970 975
His Asn Asn Pro Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu
980 985 990
Thr Leu Leu Pro Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly
995 1000 1005
Ala Ile Pro Val Ala Ala Gly Ala Thr Gly Glu Thr Ala Val Phe Thr
1010 1015 1020
Gly Asn Ser Asn Ser Pro Ala Ser Pro Thr Ala Gly Gly Cys
1025 1030 1035
<210> 442
<211> 990
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 442
Met Ala Ser Val Leu Thr Pro Leu Leu Leu Arg Gly Leu Thr Gly Ser
1 5 10 15
Ala Arg Arg Leu Pro Val Pro Arg Ala Lys Ile His Ser Leu Asp Tyr
20 25 30
Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp Ile Asp Tyr Lys Asp
35 40 45
Asp Asp Asp Lys Gly Ile Arg Ile Gln Asp Leu Arg Thr Leu Gly Tyr
50 55 60
Ser Gln Gln Gln Gln Glu Lys Ile Lys Pro Lys Val Arg Ser Thr Val
65 70 75 80
Ala Gln His His Glu Ala Leu Val Gly His Gly Phe Thr His Ala His
85 90 95
Ile Val Ala Leu Ser Gln His Pro Ala Ala Leu Gly Thr Val Ala Val
100 105 110
Lys Tyr Gln Asp Met Ile Ala Ala Leu Pro Glu Ala Thr His Glu Ala
115 120 125
Ile Val Gly Val Gly Lys Gln Trp Ser Gly Ala Arg Ala Leu Glu Ala
130 135 140
Leu Leu Thr Val Ala Gly Glu Leu Arg Gly Pro Pro Leu Gln Leu Asp
145 150 155 160
Thr Gly Gln Leu Leu Lys Ile Ala Lys Arg Gly Gly Val Thr Ala Val
165 170 175
Glu Ala Val His Ala Trp Arg Asn Ala Leu Thr Gly Ala Pro Leu Asn
180 185 190
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys
195 200 205
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp
210 215 220
His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile Gly
225 230 235 240
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
245 250 255
Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn
260 265 270
Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
275 280 285
Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala
290 295 300
Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
305 310 315 320
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala
325 330 335
Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
340 345 350
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val
355 360 365
Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val
370 375 380
Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala
385 390 395 400
Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu
405 410 415
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr
420 425 430
Pro Ala Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala
435 440 445
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
450 455 460
Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys
465 470 475 480
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
485 490 495
His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly
500 505 510
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
515 520 525
Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser His
530 535 540
Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
545 550 555 560
Leu Cys Gln Asp His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala
565 570 575
Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
580 585 590
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala
595 600 605
Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
610 615 620
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val
625 630 635 640
Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val
645 650 655
Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala
660 665 670
Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu
675 680 685
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr
690 695 700
Pro Ala Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala
705 710 715 720
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly
725 730 735
Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys
740 745 750
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
755 760 765
His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly
770 775 780
Gly Lys Gln Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg Pro Asp
785 790 795 800
Pro Ala Leu Ala Ala Leu Leu Val Ser Gly Ser Glu Thr Pro Gly Thr
805 810 815
Ser Glu Ser Ala Thr Pro Glu Ser Ser Glu Val Glu Phe Ser His Glu
820 825 830
Tyr Trp Met Arg His Ala Leu Thr Leu Ala Lys Arg Ala Arg Asp Glu
835 840 845
Arg Glu Val Pro Val Gly Ala Val Leu Val Leu Asn Asn Arg Val Ile
850 855 860
Gly Glu Gly Trp Asn Arg Ala Ile Gly Leu His Asp Pro Thr Ala His
865 870 875 880
Ala Glu Ile Met Ala Leu Arg Gln Gly Gly Leu Val Met Gln Asn Tyr
885 890 895
Arg Leu Ile Asp Ala Thr Leu Tyr Val Thr Phe Glu Pro Cys Val Met
900 905 910
Cys Ala Gly Ala Met Ile His Ser Arg Ile Gly Arg Val Val Phe Gly
915 920 925
Val Arg Asn Ser Lys Arg Gly Ala Ala Gly Ser Leu Met Asn Val Leu
930 935 940
Asn Tyr Pro Gly Met Asn His Arg Val Glu Ile Thr Glu Gly Ile Leu
945 950 955 960
Ala Asp Glu Cys Ala Ala Leu Leu Cys Asp Phe Tyr Arg Met Pro Arg
965 970 975
Gln Val Phe Asn Ala Gln Lys Lys Ala Gln Ser Ser Ile Asn
980 985 990
<210> 443
<211> 854
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 443
Met Ala Leu Ser Arg Ala Val Cys Gly Thr Ser Arg Gln Leu Ala Pro
1 5 10 15
Val Leu Gly Tyr Leu Gly Ser Arg Gln Lys His Ser Leu Pro Asp Tyr
20 25 30
Pro Tyr Asp Val Pro Asp Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp
35 40 45
Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Gly Ile Arg Ile
50 55 60
Gln Asp Leu Arg Thr Leu Gly Tyr Ser Gln Gln Gln Gln Glu Lys Ile
65 70 75 80
Lys Pro Lys Val Arg Ser Thr Val Ala Gln His His Glu Ala Leu Val
85 90 95
Gly His Gly Phe Thr His Ala His Ile Val Ala Leu Ser Gln His Pro
100 105 110
Ala Ala Leu Gly Thr Val Ala Val Lys Tyr Gln Asp Met Ile Ala Ala
115 120 125
Leu Pro Glu Ala Thr His Glu Ala Ile Val Gly Val Gly Lys Gln Trp
130 135 140
Ser Gly Ala Arg Ala Leu Glu Ala Leu Leu Thr Val Ala Gly Glu Leu
145 150 155 160
Arg Gly Pro Pro Leu Gln Leu Asp Thr Gly Gln Leu Leu Lys Ile Ala
165 170 175
Lys Arg Gly Gly Val Thr Ala Val Glu Ala Val His Ala Trp Arg Asn
180 185 190
Ala Leu Thr Gly Ala Pro Leu Asn Leu Thr Pro Asp Gln Val Val Ala
195 200 205
Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
210 215 220
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val
225 230 235 240
Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val
245 250 255
Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Ala
260 265 270
Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu
275 280 285
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr
290 295 300
Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala
305 310 315 320
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
325 330 335
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys
340 345 350
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp
355 360 365
His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly
370 375 380
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
385 390 395 400
Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn
405 410 415
Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
420 425 430
Leu Cys Gln Asp His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala
435 440 445
Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
450 455 460
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala
465 470 475 480
Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
485 490 495
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val
500 505 510
Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val
515 520 525
Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Asp
530 535 540
Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu
545 550 555 560
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr
565 570 575
Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala
580 585 590
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
595 600 605
Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys
610 615 620
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
625 630 635 640
His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly
645 650 655
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
660 665 670
Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn
675 680 685
Gly Gly Gly Lys Gln Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg
690 695 700
Pro Asp Pro Ala Leu Ala Ala Leu Leu Val Gly Ser Gly Ser Tyr Ala
705 710 715 720
Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro Ala Tyr Asn Gly
725 730 735
Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala Gly Gly Leu Glu
740 745 750
Gly Lys Val Phe Ser Ser Gly Gly Pro Thr Pro Tyr Pro Asn Tyr Ala
755 760 765
Asn Ala Gly His Val Glu Ser Gln Ser Ala Leu Phe Met Arg Asp Asn
770 775 780
Gly Ile Ser Glu Gly Leu Val Phe His Asn Asn Pro Glu Gly Thr Cys
785 790 795 800
Gly Phe Cys Val Asn Met Thr Glu Thr Leu Leu Pro Glu Asn Ala Lys
805 810 815
Met Thr Val Val Pro Pro Glu Gly Val Ile Pro Val Lys Arg Gly Ala
820 825 830
Thr Gly Glu Thr Lys Val Phe Thr Gly Asn Ser Asn Gly Pro Lys Ser
835 840 845
Pro Thr Lys Gly Gly Cys
850
<210> 444
<211> 854
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 444
Met Ala Leu Ser Arg Ala Val Cys Gly Thr Ser Arg Gln Leu Ala Pro
1 5 10 15
Val Leu Gly Tyr Leu Gly Ser Arg Gln Lys His Ser Leu Pro Asp Tyr
20 25 30
Pro Tyr Asp Val Pro Asp Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp
35 40 45
Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Gly Ile Arg Ile
50 55 60
Gln Asp Leu Arg Thr Leu Gly Tyr Ser Gln Gln Gln Gln Glu Lys Ile
65 70 75 80
Lys Pro Lys Val Arg Ser Thr Val Ala Gln His His Glu Ala Leu Val
85 90 95
Gly His Gly Phe Thr His Ala His Ile Val Ala Leu Ser Gln His Pro
100 105 110
Ala Ala Leu Gly Thr Val Ala Val Lys Tyr Gln Asp Met Ile Ala Ala
115 120 125
Leu Pro Glu Ala Thr His Glu Ala Ile Val Gly Val Gly Lys Gln Trp
130 135 140
Ser Gly Ala Arg Ala Leu Glu Ala Leu Leu Thr Val Ala Gly Glu Leu
145 150 155 160
Arg Gly Pro Pro Leu Gln Leu Asp Thr Gly Gln Leu Leu Lys Ile Ala
165 170 175
Lys Arg Gly Gly Val Thr Ala Val Glu Ala Val His Ala Trp Arg Asn
180 185 190
Ala Leu Thr Gly Ala Pro Leu Asn Leu Thr Pro Asp Gln Val Val Ala
195 200 205
Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
210 215 220
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val
225 230 235 240
Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val
245 250 255
Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Ala
260 265 270
Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu
275 280 285
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr
290 295 300
Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala
305 310 315 320
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
325 330 335
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys
340 345 350
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp
355 360 365
His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly
370 375 380
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
385 390 395 400
Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn
405 410 415
Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
420 425 430
Leu Cys Gln Asp His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala
435 440 445
Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
450 455 460
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala
465 470 475 480
Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
485 490 495
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val
500 505 510
Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val
515 520 525
Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Asp
530 535 540
Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu
545 550 555 560
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr
565 570 575
Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala
580 585 590
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
595 600 605
Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys
610 615 620
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
625 630 635 640
His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly
645 650 655
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
660 665 670
Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn
675 680 685
Gly Gly Gly Lys Gln Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg
690 695 700
Pro Asp Pro Ala Leu Ala Ala Leu Leu Val Gly Ser Gly Ser Tyr Ala
705 710 715 720
Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro Ala Tyr Asn Gly
725 730 735
Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala Gly Gly Leu Glu
740 745 750
Ser Lys Val Phe Ser Ser Gly Gly Pro Thr Pro Tyr Pro Asn Tyr Ala
755 760 765
Asn Ala Gly His Val Ala Gly Gln Ser Ala Leu Phe Met Arg Asp Asn
770 775 780
Gly Ile Ser Glu Gly Leu Val Phe His Asn Asn Pro Glu Gly Thr Cys
785 790 795 800
Gly Phe Cys Val Asn Met Thr Glu Thr Leu Leu Pro Glu Asn Ala Lys
805 810 815
Met Thr Val Val Pro Pro Glu Gly Ala Ile Pro Val Lys Arg Gly Ala
820 825 830
Thr Gly Glu Thr Lys Val Phe Thr Gly Asn Ser Asn Ser Pro Lys Ser
835 840 845
Pro Thr Lys Gly Gly Cys
850
<210> 445
<211> 854
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 445
Met Ala Leu Ser Arg Ala Val Cys Gly Thr Ser Arg Gln Leu Ala Pro
1 5 10 15
Val Leu Gly Tyr Leu Gly Ser Arg Gln Lys His Ser Leu Pro Asp Tyr
20 25 30
Pro Tyr Asp Val Pro Asp Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp
35 40 45
Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Gly Ile Arg Ile
50 55 60
Gln Asp Leu Arg Thr Leu Gly Tyr Ser Gln Gln Gln Gln Glu Lys Ile
65 70 75 80
Lys Pro Lys Val Arg Ser Thr Val Ala Gln His His Glu Ala Leu Val
85 90 95
Gly His Gly Phe Thr His Ala His Ile Val Ala Leu Ser Gln His Pro
100 105 110
Ala Ala Leu Gly Thr Val Ala Val Lys Tyr Gln Asp Met Ile Ala Ala
115 120 125
Leu Pro Glu Ala Thr His Glu Ala Ile Val Gly Val Gly Lys Gln Trp
130 135 140
Ser Gly Ala Arg Ala Leu Glu Ala Leu Leu Thr Val Ala Gly Glu Leu
145 150 155 160
Arg Gly Pro Pro Leu Gln Leu Asp Thr Gly Gln Leu Leu Lys Ile Ala
165 170 175
Lys Arg Gly Gly Val Thr Ala Val Glu Ala Val His Ala Trp Arg Asn
180 185 190
Ala Leu Thr Gly Ala Pro Leu Asn Leu Thr Pro Asp Gln Val Val Ala
195 200 205
Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
210 215 220
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val
225 230 235 240
Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val
245 250 255
Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Ala
260 265 270
Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu
275 280 285
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr
290 295 300
Pro Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala
305 310 315 320
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
325 330 335
Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys
340 345 350
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp
355 360 365
His Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly
370 375 380
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
385 390 395 400
Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn
405 410 415
Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val
420 425 430
Leu Cys Gln Asp His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala
435 440 445
Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu
450 455 460
Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala
465 470 475 480
Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg
485 490 495
Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val
500 505 510
Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val
515 520 525
Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Asp
530 535 540
Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu
545 550 555 560
Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr
565 570 575
Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala
580 585 590
Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly
595 600 605
Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys
610 615 620
Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala
625 630 635 640
His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly
645 650 655
Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys
660 665 670
Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn
675 680 685
Gly Gly Gly Lys Gln Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg
690 695 700
Pro Asp Pro Ala Leu Ala Ala Leu Leu Val Gly Ser Gly Ser Tyr Ala
705 710 715 720
Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro Ala Tyr Asn Gly
725 730 735
Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala Gly Gly Leu Glu
740 745 750
Ser Lys Val Phe Ser Ser Gly Gly Pro Thr Pro Tyr Pro Asn Tyr Ala
755 760 765
Asn Ala Gly His Val Glu Gly Gln Ser Ala Leu Phe Met Arg Asp Asn
770 775 780
Gly Ile Ser Glu Gly Leu Val Phe His Asn Asn Pro Glu Gly Thr Cys
785 790 795 800
Gly Phe Cys Val Asn Met Thr Glu Thr Leu Leu Pro Glu Asn Ala Lys
805 810 815
Met Thr Val Val Pro Pro Glu Gly Ala Ile Pro Val Ala Ala Gly Ala
820 825 830
Thr Gly Glu Thr Ala Val Phe Thr Gly Asn Ser Asn Ser Pro Ala Ser
835 840 845
Pro Thr Ala Gly Gly Cys
850
<210> 446
<211> 647
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 446
Asp Leu Arg Thr Leu Gly Tyr Ser Gln Gln Gln Gln Glu Lys Ile Lys
1 5 10 15
Pro Lys Val Arg Ser Thr Val Ala Gln His His Glu Ala Leu Val Gly
20 25 30
His Gly Phe Thr His Ala His Ile Val Ala Leu Ser Gln His Pro Ala
35 40 45
Ala Leu Gly Thr Val Ala Val Lys Tyr Gln Asp Met Ile Ala Ala Leu
50 55 60
Pro Glu Ala Thr His Glu Ala Ile Val Gly Val Gly Lys Gln Trp Ser
65 70 75 80
Gly Ala Arg Ala Leu Glu Ala Leu Leu Thr Val Ala Gly Glu Leu Arg
85 90 95
Gly Pro Pro Leu Gln Leu Asp Thr Gly Gln Leu Leu Lys Ile Ala Lys
100 105 110
Arg Gly Gly Val Thr Ala Val Glu Ala Val His Ala Trp Arg Asn Ala
115 120 125
Leu Thr Gly Ala Pro Leu Asn Leu Thr Pro Asp Gln Val Val Ala Ile
130 135 140
Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu
145 150 155 160
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val
165 170 175
Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
180 185 190
Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Ala Gln
195 200 205
Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr
210 215 220
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro
225 230 235 240
Asp Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu
245 250 255
Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
260 265 270
Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln
275 280 285
Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His
290 295 300
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly
305 310 315 320
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
325 330 335
Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly
340 345 350
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
355 360 365
Cys Gln Asp His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser
370 375 380
Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro
385 390 395 400
Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val Ala Ile
405 410 415
Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu
420 425 430
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val Val
435 440 445
Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
450 455 460
Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Asp Gln
465 470 475 480
Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr
485 490 495
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
500 505 510
Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu
515 520 525
Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
530 535 540
Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln
545 550 555 560
Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His
565 570 575
Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
580 585 590
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
595 600 605
Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly
610 615 620
Gly Gly Lys Gln Ala Leu Glu Ser Ile Val Ala Gln Leu Ser Arg Pro
625 630 635 640
Asp Pro Ala Leu Ala Ala Leu
645
<210> 447
<211> 749
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 447
Asp Leu Arg Thr Leu Gly Tyr Ser Gln Gln Gln Gln Glu Lys Ile Lys
1 5 10 15
Pro Lys Val Arg Ser Thr Val Ala Gln His His Glu Ala Leu Val Gly
20 25 30
His Gly Phe Thr His Ala His Ile Val Ala Leu Ser Gln His Pro Ala
35 40 45
Ala Leu Gly Thr Val Ala Val Lys Tyr Gln Asp Met Ile Ala Ala Leu
50 55 60
Pro Glu Ala Thr His Glu Ala Ile Val Gly Val Gly Lys Gln Trp Ser
65 70 75 80
Gly Ala Arg Ala Leu Glu Ala Leu Leu Thr Val Ala Gly Glu Leu Arg
85 90 95
Gly Pro Pro Leu Gln Leu Asp Thr Gly Gln Leu Leu Lys Ile Ala Lys
100 105 110
Arg Gly Gly Val Thr Ala Val Glu Ala Val His Ala Trp Arg Asn Ala
115 120 125
Leu Thr Gly Ala Pro Leu Asn Leu Thr Pro Asp Gln Val Val Ala Ile
130 135 140
Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu
145 150 155 160
Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro Ala Gln Val Val
165 170 175
Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
180 185 190
Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln
195 200 205
Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr
210 215 220
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
225 230 235 240
Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu
245 250 255
Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
260 265 270
Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln
275 280 285
Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His
290 295 300
Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly
305 310 315 320
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
325 330 335
Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser Asn Asn
340 345 350
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
355 360 365
Cys Gln Asp His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser
370 375 380
Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro
385 390 395 400
Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile
405 410 415
Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu
420 425 430
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Asp Gln Val Val
435 440 445
Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
450 455 460
Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Ala Gln
465 470 475 480
Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr
485 490 495
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Asp His Gly Leu Thr Pro
500 505 510
Asp Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu
515 520 525
Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
530 535 540
Thr Pro Asp Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln
545 550 555 560
Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His
565 570 575
Gly Leu Thr Pro Asp Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
580 585 590
Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln
595 600 605
Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser His Asp
610 615 620
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
625 630 635 640
Cys Gln Ala His Gly Leu Thr Pro Ala Gln Val Val Ala Ile Ala Ser
645 650 655
His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro
660 665 670
Val Leu Cys Gln Asp His Gly Leu Thr Pro Ala Gln Val Val Ala Ile
675 680 685
Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu
690 695 700
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val
705 710 715 720
Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Ser Ile Val
725 730 735
Ala Gln Leu Ser Arg Pro Asp Pro Ala Leu Ala Ala Leu
740 745
<210> 448
<211> 744
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 448
Met Asp Ile Ala Asp Leu Arg Thr Leu Gly Tyr Ser Gln Gln Gln Gln
1 5 10 15
Glu Lys Ile Lys Pro Lys Val Arg Ser Thr Val Ala Gln His His Glu
20 25 30
Ala Leu Val Gly His Gly Phe Thr His Ala His Ile Val Ala Leu Ser
35 40 45
Gln His Pro Ala Ala Leu Gly Thr Val Ala Val Lys Tyr Gln Asp Met
50 55 60
Ile Ala Ala Leu Pro Glu Ala Thr His Glu Ala Ile Val Gly Val Gly
65 70 75 80
Lys Gln Trp Ser Gly Ala Arg Ala Leu Glu Ala Leu Leu Thr Val Ala
85 90 95
Gly Glu Leu Arg Gly Pro Pro Leu Gln Leu Asp Thr Gly Gln Leu Leu
100 105 110
Lys Ile Ala Lys Arg Gly Gly Val Thr Ala Val Glu Ala Val His Ala
115 120 125
Trp Arg Asn Ala Leu Thr Gly Ala Pro Leu Asn Leu Thr Pro Glu Gln
130 135 140
Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr
145 150 155 160
Val Gln Ala Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
165 170 175
Gln Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu
180 185 190
Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
195 200 205
Thr Pro Gln Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln
210 215 220
Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His
225 230 235 240
Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly
245 250 255
Lys Gln Ala Leu Glu Thr Val Gln Ala Leu Leu Pro Val Leu Cys Gln
260 265 270
Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn
275 280 285
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu Leu Pro Val Leu
290 295 300
Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser
305 310 315 320
Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu Leu Pro
325 330 335
Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile
340 345 350
Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu
355 360 365
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val
370 375 380
Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
385 390 395 400
Ala Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln
405 410 415
Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr
420 425 430
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
435 440 445
Glu Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu
450 455 460
Glu Thr Val Gln Ala Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
465 470 475 480
Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln
485 490 495
Ala Leu Glu Thr Val Gln Ala Leu Leu Pro Val Leu Cys Gln Ala His
500 505 510
Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser His Asp Gly Gly
515 520 525
Lys Gln Ala Leu Glu Thr Val Gln Ala Leu Leu Pro Val Leu Cys Gln
530 535 540
Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn
545 550 555 560
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu
565 570 575
Cys Gln Ala His Gly Leu Thr Pro Gln Gln Val Val Ala Ile Ala Ser
580 585 590
Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro
595 600 605
Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile
610 615 620
Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu
625 630 635 640
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val
645 650 655
Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
660 665 670
Ala Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Gln Gln
675 680 685
Val Val Ala Ile Ala Ser Asn Gly Gly Gly Arg Pro Ala Leu Glu Ser
690 695 700
Ile Val Ala Gln Leu Ser Arg Pro Asp Pro Ala Leu Ala Ala Leu Thr
705 710 715 720
Asn Asp His Leu Val Ala Leu Ala Cys Leu Gly Gly Arg Pro Ala Leu
725 730 735
Asp Ala Val Lys Lys Gly Leu Gly
740
<210> 449
<211> 744
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 449
Met Asp Ile Ala Asp Leu Arg Thr Leu Gly Tyr Ser Gln Gln Gln Gln
1 5 10 15
Glu Lys Ile Lys Pro Lys Val Arg Ser Thr Val Ala Gln His His Glu
20 25 30
Ala Leu Val Gly His Gly Phe Thr His Ala His Ile Val Ala Leu Ser
35 40 45
Gln His Pro Ala Ala Leu Gly Thr Val Ala Val Lys Tyr Gln Asp Met
50 55 60
Ile Ala Ala Leu Pro Glu Ala Thr His Glu Ala Ile Val Gly Val Gly
65 70 75 80
Lys Gln Trp Ser Gly Ala Arg Ala Leu Glu Ala Leu Leu Thr Val Ala
85 90 95
Gly Glu Leu Arg Gly Pro Pro Leu Gln Leu Asp Thr Gly Gln Leu Leu
100 105 110
Lys Ile Ala Lys Arg Gly Gly Val Thr Ala Val Glu Ala Val His Ala
115 120 125
Trp Arg Asn Ala Leu Thr Gly Ala Pro Leu Asn Leu Thr Pro Glu Gln
130 135 140
Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu Glu Thr
145 150 155 160
Val Gln Ala Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
165 170 175
Gln Gln Val Val Ala Ile Ala Ser His Asp Gly Gly Lys Gln Ala Leu
180 185 190
Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
195 200 205
Thr Pro Gln Gln Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln
210 215 220
Ala Leu Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His
225 230 235 240
Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly
245 250 255
Lys Gln Ala Leu Glu Thr Val Gln Ala Leu Leu Pro Val Leu Cys Gln
260 265 270
Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Gly
275 280 285
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu Leu Pro Val Leu
290 295 300
Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser
305 310 315 320
Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu Leu Pro
325 330 335
Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile
340 345 350
Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu
355 360 365
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val
370 375 380
Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
385 390 395 400
Ala Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Glu Gln
405 410 415
Val Val Ala Ile Ala Ser Asn Gly Gly Gly Lys Gln Ala Leu Glu Thr
420 425 430
Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro
435 440 445
Glu Gln Val Val Ala Ile Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu
450 455 460
Glu Thr Val Gln Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu
465 470 475 480
Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln
485 490 495
Ala Leu Glu Thr Val Gln Ala Leu Leu Pro Val Leu Cys Gln Ala His
500 505 510
Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Asn Gly Gly
515 520 525
Lys Gln Ala Leu Glu Thr Val Gln Ala Leu Leu Pro Val Leu Cys Gln
530 535 540
Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser Asn Ile
545 550 555 560
Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Ala Leu Leu Pro Val Leu
565 570 575
Cys Gln Ala His Gly Leu Thr Pro Glu Gln Val Val Ala Ile Ala Ser
580 585 590
Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu Leu Pro
595 600 605
Val Leu Cys Gln Ala His Gly Leu Thr Pro Gln Gln Val Val Ala Ile
610 615 620
Ala Ser Asn Ile Gly Gly Lys Gln Ala Leu Glu Thr Val Gln Arg Leu
625 630 635 640
Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Gln Gln Val Val
645 650 655
Ala Ile Ala Ser Asn Asn Gly Gly Lys Gln Ala Leu Glu Thr Val Gln
660 665 670
Arg Leu Leu Pro Val Leu Cys Gln Ala His Gly Leu Thr Pro Gln Gln
675 680 685
Val Val Ala Ile Ala Ser Asn Gly Gly Gly Arg Pro Ala Leu Glu Ser
690 695 700
Ile Val Ala Gln Leu Ser Arg Pro Asp Pro Ala Leu Ala Ala Leu Thr
705 710 715 720
Asn Asp His Leu Val Ala Leu Ala Cys Leu Gly Gly Arg Pro Ala Leu
725 730 735
Asp Ala Val Lys Lys Gly Leu Gly
740
<210> 450
<211> 142
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 450
Phe Gln Cys Arg Ile Cys Met Arg Lys Phe Ala Thr Ser Gly Ser Leu
1 5 10 15
Thr Arg His Thr Lys Ile His Thr Gly Glu Lys Pro Phe Gln Cys Arg
20 25 30
Ile Cys Met Arg Asn Phe Ser Arg Ser Asp His Leu Ser Thr His Ile
35 40 45
Arg Thr His Thr Gly Glu Lys Pro Phe Ala Cys Asp Ile Cys Gly Arg
50 55 60
Lys Phe Ala Thr Ser Ser Asn Arg Thr Lys His Thr Lys Ile His Thr
65 70 75 80
His Pro Arg Ala Pro Ile Pro Lys Pro Phe Gln Cys Arg Ile Cys Met
85 90 95
Arg Asn Phe Ser Arg Ser Asp Asn Leu Ser Glu His Ile Arg Thr His
100 105 110
Thr Gly Glu Lys Pro Phe Ala Cys Asp Ile Cys Gly Arg Lys Phe Ala
115 120 125
Trp His Ser Ser Leu Arg Val His Thr Lys Ile His Leu Arg
130 135 140
<210> 451
<211> 138
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 451
Phe Gln Cys Arg Ile Cys Met Arg Asn Phe Ser Arg Ser Asp His Leu
1 5 10 15
Ser Thr His Ile Arg Thr His Thr Gly Glu Lys Pro Phe Ala Cys Asp
20 25 30
Ile Cys Gly Arg Lys Phe Ala Asp Arg Ser His Leu Ala Arg His Thr
35 40 45
Lys Ile His Thr Gly Ser Gln Lys Pro Phe Gln Cys Arg Ile Cys Met
50 55 60
Arg Lys Phe Ala Leu Lys Gln His Leu Asn Glu His Thr Lys Ile His
65 70 75 80
Thr Gly Glu Lys Pro Phe Gln Cys Arg Ile Cys Met Arg Asn Phe Ser
85 90 95
Gln Ser Gly Asn Leu Ala Arg His Ile Arg Thr His Thr Gly Glu Lys
100 105 110
Pro Phe Ala Cys Asp Ile Cys Gly Arg Lys Phe Ala His Asn Ser Ser
115 120 125
Leu Lys Asp His Thr Lys Ile His Leu Arg
130 135
<210> 452
<211> 110
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 452
Phe Gln Cys Arg Ile Cys Met Arg Asn Phe Ser Asp Ser Gly Asn Leu
1 5 10 15
Arg Val His Ile Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
20 25 30
Asp Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu Ile Arg His Gln
35 40 45
Arg Thr His Thr Gly Glu Lys Pro Tyr Glu Cys Asp His Cys Gly Lys
50 55 60
Ser Phe Ser Gln Ser Ser His Leu Asn Val His Lys Arg Thr His Thr
65 70 75 80
Gly Glu Lys Pro Tyr Arg Cys Lys Tyr Cys Asp Arg Ser Phe Ser Ile
85 90 95
Ser Ser Asn Leu Gln Arg His Val Arg Asn Ile His Leu Arg
100 105 110
<210> 453
<211> 110
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 453
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Lys Asn Ser Leu
1 5 10 15
Thr Glu His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr Lys Cys Pro
20 25 30
Glu Cys Gly Lys Ser Phe Ser Ser Lys Lys Ala Leu Thr Glu His Gln
35 40 45
Arg Thr His Thr Gly Glu Lys Pro Tyr Glu Cys Asn Tyr Cys Gly Lys
50 55 60
Thr Phe Ser Val Ser Ser Thr Leu Ile Arg His Gln Arg Ile His Thr
65 70 75 80
Gly Glu Lys Pro Tyr Arg Cys Lys Tyr Cys Asp Arg Ser Phe Ser Ile
85 90 95
Ser Ser Asn Leu Gln Arg His Val Arg Asn Ile His Leu Arg
100 105 110
<210> 454
<211> 46
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 454
Gly Ser Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln
1 5 10 15
Leu Pro Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn
20 25 30
Asp Ala Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly
35 40 45
<210> 455
<211> 96
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 455
Gly Ser Pro Thr Pro Tyr Pro Asn Tyr Ala Asn Ala Gly His Val Glu
1 5 10 15
Gly Gln Ser Ala Leu Phe Met Arg Asp Asn Gly Ile Ser Glu Gly Leu
20 25 30
Val Phe His Asn Asn Pro Glu Gly Thr Cys Gly Phe Cys Val Asn Met
35 40 45
Thr Glu Thr Leu Leu Pro Glu Asn Ala Lys Met Thr Val Val Pro Pro
50 55 60
Glu Gly Ala Ile Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val
65 70 75 80
Phe Thr Gly Asn Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys
85 90 95
<210> 456
<211> 110
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 456
Gly Ser Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln
1 5 10 15
Leu Pro Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn
20 25 30
Asp Ala Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly Pro Thr
35 40 45
Pro Tyr Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln Ser Ala
50 55 60
Leu Phe Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe His Asn
65 70 75 80
Asn Pro Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu Thr Leu
85 90 95
Leu Pro Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly
100 105 110
<210> 457
<211> 32
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 457
Gly Ser Ala Ile Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val
1 5 10 15
Phe Thr Gly Asn Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys
20 25 30
<210> 458
<211> 166
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 458
Ser Glu Val Glu Phe Ser His Glu Tyr Trp Met Arg His Ala Leu Thr
1 5 10 15
Leu Ala Lys Arg Ala Arg Asp Glu Arg Glu Val Pro Val Gly Ala Val
20 25 30
Leu Val Leu Asn Asn Arg Val Ile Gly Glu Gly Trp Asn Arg Ala Ile
35 40 45
Gly Leu His Asp Pro Thr Ala His Ala Glu Ile Met Ala Leu Arg Gln
50 55 60
Gly Gly Leu Val Met Gln Asn Tyr Arg Leu Ile Asp Ala Thr Leu Tyr
65 70 75 80
Val Thr Phe Glu Pro Cys Val Met Cys Ala Gly Ala Met Ile His Ser
85 90 95
Arg Ile Gly Arg Val Val Phe Gly Val Arg Asn Ser Lys Arg Gly Ala
100 105 110
Ala Gly Ser Leu Met Asn Val Leu Asn Tyr Pro Gly Met Asn His Arg
115 120 125
Val Glu Ile Thr Glu Gly Ile Leu Ala Asp Glu Cys Ala Ala Leu Leu
130 135 140
Cys Asp Phe Tyr Arg Met Pro Arg Gln Val Phe Asn Ala Gln Lys Lys
145 150 155 160
Ala Gln Ser Ser Ile Asn
165
<210> 459
<211> 140
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 459
Gly Ser Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln
1 5 10 15
Leu Pro Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn
20 25 30
Asp Ala Gly Gly Leu Glu Gly Lys Val Phe Ser Ser Gly Gly Pro Thr
35 40 45
Pro Tyr Pro Asn Tyr Ala Asn Ala Gly His Val Glu Ser Gln Ser Ala
50 55 60
Leu Phe Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe His Asn
65 70 75 80
Asn Pro Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu Thr Leu
85 90 95
Leu Pro Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly Val Ile
100 105 110
Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr Gly Asn
115 120 125
Ser Asn Gly Pro Lys Ser Pro Thr Lys Gly Gly Cys
130 135 140
<210> 460
<211> 140
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 460
Gly Ser Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln
1 5 10 15
Leu Pro Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn
20 25 30
Asp Ala Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly Pro Thr
35 40 45
Pro Tyr Pro Asn Tyr Ala Asn Ala Gly His Val Ala Gly Gln Ser Ala
50 55 60
Leu Phe Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe His Asn
65 70 75 80
Asn Pro Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu Thr Leu
85 90 95
Leu Pro Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly Ala Ile
100 105 110
Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr Gly Asn
115 120 125
Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys
130 135 140
<210> 461
<211> 140
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 461
Gly Ser Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln
1 5 10 15
Leu Pro Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn
20 25 30
Asp Ala Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly Pro Thr
35 40 45
Pro Tyr Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln Ser Ala
50 55 60
Leu Phe Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe His Asn
65 70 75 80
Asn Pro Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu Thr Leu
85 90 95
Leu Pro Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly Ala Ile
100 105 110
Pro Val Ala Ala Gly Ala Thr Gly Glu Thr Ala Val Phe Thr Gly Asn
115 120 125
Ser Asn Ser Pro Ala Ser Pro Thr Ala Gly Gly Cys
130 135 140
<210> 462
<211> 83
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 462
Thr Asn Leu Ser Asp Ile Ile Glu Lys Glu Thr Gly Lys Gln Leu Val
1 5 10 15
Ile Gln Glu Ser Ile Leu Met Leu Pro Glu Glu Val Glu Glu Val Ile
20 25 30
Gly Asn Lys Pro Glu Ser Asp Ile Leu Val His Thr Ala Tyr Asp Glu
35 40 45
Ser Thr Asp Glu Asn Val Met Leu Leu Thr Ser Asp Ala Pro Glu Tyr
50 55 60
Lys Pro Trp Ala Leu Val Ile Gln Asp Ser Asn Gly Glu Asn Lys Ile
65 70 75 80
Lys Met Leu
<210> 463
<211> 324
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 463
Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser
1 5 10 15
Ser Glu Val Glu Phe Ser His Glu Tyr Trp Met Arg His Ala Leu Thr
20 25 30
Leu Ala Lys Arg Ala Arg Asp Glu Arg Glu Val Pro Val Gly Ala Val
35 40 45
Leu Val Leu Asn Asn Arg Val Ile Gly Glu Gly Trp Asn Arg Ala Ile
50 55 60
Gly Leu His Asp Pro Thr Ala His Ala Glu Ile Met Ala Leu Arg Gln
65 70 75 80
Gly Gly Leu Val Met Gln Asn Tyr Arg Leu Ile Asp Ala Thr Leu Tyr
85 90 95
Val Thr Phe Glu Pro Cys Val Met Cys Ala Gly Ala Met Ile His Ser
100 105 110
Arg Ile Gly Arg Val Val Phe Gly Val Arg Asn Ser Lys Arg Gly Ala
115 120 125
Ala Gly Ser Leu Met Asn Val Leu Asn Tyr Pro Gly Met Asn His Arg
130 135 140
Val Glu Ile Thr Glu Gly Ile Leu Ala Asp Glu Cys Ala Ala Leu Leu
145 150 155 160
Cys Asp Phe Tyr Arg Met Pro Arg Gln Val Phe Asn Ala Gln Lys Lys
165 170 175
Ala Gln Ser Ser Ile Asn Leu Val Gly Ser Gly Ser Tyr Ala Leu Gly
180 185 190
Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro Ala Tyr Asn Gly Gln Thr
195 200 205
Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala Gly Gly Leu Glu Gly Lys
210 215 220
Val Phe Ser Ser Gly Gly Pro Thr Pro Tyr Pro Asn Tyr Ala Asn Ala
225 230 235 240
Gly His Val Glu Ser Gln Ser Ala Leu Phe Met Arg Asp Asn Gly Ile
245 250 255
Ser Glu Gly Leu Val Phe His Asn Asn Pro Glu Gly Thr Cys Gly Phe
260 265 270
Cys Val Asn Met Thr Glu Thr Leu Leu Pro Glu Asn Ala Lys Met Thr
275 280 285
Val Val Pro Pro Glu Gly Val Ile Pro Val Lys Arg Gly Ala Thr Gly
290 295 300
Glu Thr Lys Val Phe Thr Gly Asn Ser Asn Gly Pro Lys Ser Pro Thr
305 310 315 320
Lys Gly Gly Cys
<210> 464
<211> 324
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 464
Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser
1 5 10 15
Ser Glu Val Glu Phe Ser His Glu Tyr Trp Met Arg His Ala Leu Thr
20 25 30
Leu Ala Lys Arg Ala Arg Asp Glu Arg Glu Val Pro Val Gly Ala Val
35 40 45
Leu Val Leu Asn Asn Arg Val Ile Gly Glu Gly Trp Asn Arg Ala Ile
50 55 60
Gly Leu His Asp Pro Thr Ala His Ala Glu Ile Met Ala Leu Arg Gln
65 70 75 80
Gly Gly Leu Val Met Gln Asn Tyr Arg Leu Ile Asp Ala Thr Leu Tyr
85 90 95
Val Thr Phe Glu Pro Cys Val Met Cys Ala Gly Ala Met Ile His Ser
100 105 110
Arg Ile Gly Arg Val Val Phe Gly Val Arg Asn Ser Lys Arg Gly Ala
115 120 125
Ala Gly Ser Leu Met Asn Val Leu Asn Tyr Pro Gly Met Asn His Arg
130 135 140
Val Glu Ile Thr Glu Gly Ile Leu Ala Asp Glu Cys Ala Ala Leu Leu
145 150 155 160
Cys Asp Phe Tyr Arg Met Pro Arg Gln Val Phe Asn Ala Gln Lys Lys
165 170 175
Ala Gln Ser Ser Ile Asn Leu Val Gly Ser Gly Ser Tyr Ala Leu Gly
180 185 190
Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro Ala Tyr Asn Gly Gln Thr
195 200 205
Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala Gly Gly Leu Glu Ser Lys
210 215 220
Val Phe Ser Ser Gly Gly Pro Thr Pro Tyr Pro Asn Tyr Ala Asn Ala
225 230 235 240
Gly His Val Ala Gly Gln Ser Ala Leu Phe Met Arg Asp Asn Gly Ile
245 250 255
Ser Glu Gly Leu Val Phe His Asn Asn Pro Glu Gly Thr Cys Gly Phe
260 265 270
Cys Val Asn Met Thr Glu Thr Leu Leu Pro Glu Asn Ala Lys Met Thr
275 280 285
Val Val Pro Pro Glu Gly Ala Ile Pro Val Lys Arg Gly Ala Thr Gly
290 295 300
Glu Thr Lys Val Phe Thr Gly Asn Ser Asn Ser Pro Lys Ser Pro Thr
305 310 315 320
Lys Gly Gly Cys
<210> 465
<211> 324
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 465
Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser
1 5 10 15
Ser Glu Val Glu Phe Ser His Glu Tyr Trp Met Arg His Ala Leu Thr
20 25 30
Leu Ala Lys Arg Ala Arg Asp Glu Arg Glu Val Pro Val Gly Ala Val
35 40 45
Leu Val Leu Asn Asn Arg Val Ile Gly Glu Gly Trp Asn Arg Ala Ile
50 55 60
Gly Leu His Asp Pro Thr Ala His Ala Glu Ile Met Ala Leu Arg Gln
65 70 75 80
Gly Gly Leu Val Met Gln Asn Tyr Arg Leu Ile Asp Ala Thr Leu Tyr
85 90 95
Val Thr Phe Glu Pro Cys Val Met Cys Ala Gly Ala Met Ile His Ser
100 105 110
Arg Ile Gly Arg Val Val Phe Gly Val Arg Asn Ser Lys Arg Gly Ala
115 120 125
Ala Gly Ser Leu Met Asn Val Leu Asn Tyr Pro Gly Met Asn His Arg
130 135 140
Val Glu Ile Thr Glu Gly Ile Leu Ala Asp Glu Cys Ala Ala Leu Leu
145 150 155 160
Cys Asp Phe Tyr Arg Met Pro Arg Gln Val Phe Asn Ala Gln Lys Lys
165 170 175
Ala Gln Ser Ser Ile Asn Leu Val Gly Ser Gly Ser Tyr Ala Leu Gly
180 185 190
Pro Tyr Gln Ile Ser Ala Pro Gln Leu Pro Ala Tyr Asn Gly Gln Thr
195 200 205
Val Gly Thr Phe Tyr Tyr Val Asn Asp Ala Gly Gly Leu Glu Ser Lys
210 215 220
Val Phe Ser Ser Gly Gly Pro Thr Pro Tyr Pro Asn Tyr Ala Asn Ala
225 230 235 240
Gly His Val Glu Gly Gln Ser Ala Leu Phe Met Arg Asp Asn Gly Ile
245 250 255
Ser Glu Gly Leu Val Phe His Asn Asn Pro Glu Gly Thr Cys Gly Phe
260 265 270
Cys Val Asn Met Thr Glu Thr Leu Leu Pro Glu Asn Ala Lys Met Thr
275 280 285
Val Val Pro Pro Glu Gly Ala Ile Pro Val Ala Ala Gly Ala Thr Gly
290 295 300
Glu Thr Ala Val Phe Thr Gly Asn Ser Asn Ser Pro Ala Ser Pro Thr
305 310 315 320
Ala Gly Gly Cys
<210> 466
<211> 182
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 466
Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser
1 5 10 15
Ser Glu Val Glu Phe Ser His Glu Tyr Trp Met Arg His Ala Leu Thr
20 25 30
Leu Ala Lys Arg Ala Arg Asp Glu Arg Glu Val Pro Val Gly Ala Val
35 40 45
Leu Val Leu Asn Asn Arg Val Ile Gly Glu Gly Trp Asn Arg Ala Ile
50 55 60
Gly Leu His Asp Pro Thr Ala His Ala Glu Ile Met Ala Leu Arg Gln
65 70 75 80
Gly Gly Leu Val Met Gln Asn Tyr Arg Leu Ile Asp Ala Thr Leu Tyr
85 90 95
Val Thr Phe Glu Pro Cys Val Met Cys Ala Gly Ala Met Ile His Ser
100 105 110
Arg Ile Gly Arg Val Val Phe Gly Val Arg Asn Ser Lys Arg Gly Ala
115 120 125
Ala Gly Ser Leu Met Asn Val Leu Asn Tyr Pro Gly Met Asn His Arg
130 135 140
Val Glu Ile Thr Glu Gly Ile Leu Ala Asp Glu Cys Ala Ala Leu Leu
145 150 155 160
Cys Asp Phe Tyr Arg Met Pro Arg Gln Val Phe Asn Ala Gln Lys Lys
165 170 175
Ala Gln Ser Ser Ile Asn
180
<210> 467
<211> 140
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 467
Gly Ser Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln
1 5 10 15
Leu Pro Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn
20 25 30
Asp Ala Gly Gly Leu Glu Gly Lys Val Phe Ser Ser Gly Gly Pro Thr
35 40 45
Pro Tyr Pro Asn Tyr Ala Asn Ala Gly His Val Glu Ser Gln Ser Ala
50 55 60
Leu Phe Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe His Asn
65 70 75 80
Asn Pro Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu Thr Leu
85 90 95
Leu Pro Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly Val Ile
100 105 110
Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr Gly Asn
115 120 125
Ser Asn Gly Pro Lys Ser Pro Thr Lys Gly Gly Cys
130 135 140
<210> 468
<211> 140
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 468
Gly Ser Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln
1 5 10 15
Leu Pro Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn
20 25 30
Asp Ala Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly Pro Thr
35 40 45
Pro Tyr Pro Asn Tyr Ala Asn Ala Gly His Val Ala Gly Gln Ser Ala
50 55 60
Leu Phe Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe His Asn
65 70 75 80
Asn Pro Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu Thr Leu
85 90 95
Leu Pro Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly Ala Ile
100 105 110
Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val Phe Thr Gly Asn
115 120 125
Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys
130 135 140
<210> 469
<211> 140
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 469
Gly Ser Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln
1 5 10 15
Leu Pro Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn
20 25 30
Asp Ala Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly Pro Thr
35 40 45
Pro Tyr Pro Asn Tyr Ala Asn Ala Gly His Val Glu Gly Gln Ser Ala
50 55 60
Leu Phe Met Arg Asp Asn Gly Ile Ser Glu Gly Leu Val Phe His Asn
65 70 75 80
Asn Pro Glu Gly Thr Cys Gly Phe Cys Val Asn Met Thr Glu Thr Leu
85 90 95
Leu Pro Glu Asn Ala Lys Met Thr Val Val Pro Pro Glu Gly Ala Ile
100 105 110
Pro Val Ala Ala Gly Ala Thr Gly Glu Thr Ala Val Phe Thr Gly Asn
115 120 125
Ser Asn Ser Pro Ala Ser Pro Thr Ala Gly Gly Cys
130 135 140
<210> 470
<211> 214
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 470
Gly Ser Ala Ile Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val
1 5 10 15
Phe Thr Gly Asn Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys
20 25 30
Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser
35 40 45
Ser Glu Val Glu Phe Ser His Glu Tyr Trp Met Arg His Ala Leu Thr
50 55 60
Leu Ala Lys Arg Ala Arg Asp Glu Arg Glu Val Pro Val Gly Ala Val
65 70 75 80
Leu Val Leu Asn Asn Arg Val Ile Gly Glu Gly Trp Asn Arg Ala Ile
85 90 95
Gly Leu His Asp Pro Thr Ala His Ala Glu Ile Met Ala Leu Arg Gln
100 105 110
Gly Gly Leu Val Met Gln Asn Tyr Arg Leu Ile Asp Ala Thr Leu Tyr
115 120 125
Val Thr Phe Glu Pro Cys Val Met Cys Ala Gly Ala Met Ile His Ser
130 135 140
Arg Ile Gly Arg Val Val Phe Gly Val Arg Asn Ser Lys Arg Gly Ala
145 150 155 160
Ala Gly Ser Leu Met Asn Val Leu Asn Tyr Pro Gly Met Asn His Arg
165 170 175
Val Glu Ile Thr Glu Gly Ile Leu Ala Asp Glu Cys Ala Ala Leu Leu
180 185 190
Cys Asp Phe Tyr Arg Met Pro Arg Gln Val Phe Asn Ala Gln Lys Lys
195 200 205
Ala Gln Ser Ser Ile Asn
210
<210> 471
<211> 228
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 471
Gly Ser Gly Ser Tyr Ala Leu Gly Pro Tyr Gln Ile Ser Ala Pro Gln
1 5 10 15
Leu Pro Ala Tyr Asn Gly Gln Thr Val Gly Thr Phe Tyr Tyr Val Asn
20 25 30
Asp Ala Gly Gly Leu Glu Ser Lys Val Phe Ser Ser Gly Gly Ser Gly
35 40 45
Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser Ser Glu
50 55 60
Val Glu Phe Ser His Glu Tyr Trp Met Arg His Ala Leu Thr Leu Ala
65 70 75 80
Lys Arg Ala Arg Asp Glu Arg Glu Val Pro Val Gly Ala Val Leu Val
85 90 95
Leu Asn Asn Arg Val Ile Gly Glu Gly Trp Asn Arg Ala Ile Gly Leu
100 105 110
His Asp Pro Thr Ala His Ala Glu Ile Met Ala Leu Arg Gln Gly Gly
115 120 125
Leu Val Met Gln Asn Tyr Arg Leu Ile Asp Ala Thr Leu Tyr Val Thr
130 135 140
Phe Glu Pro Cys Val Met Cys Ala Gly Ala Met Ile His Ser Arg Ile
145 150 155 160
Gly Arg Val Val Phe Gly Val Arg Asn Ser Lys Arg Gly Ala Ala Gly
165 170 175
Ser Leu Met Asn Val Leu Asn Tyr Pro Gly Met Asn His Arg Val Glu
180 185 190
Ile Thr Glu Gly Ile Leu Ala Asp Glu Cys Ala Ala Leu Leu Cys Asp
195 200 205
Phe Tyr Arg Met Pro Arg Gln Val Phe Asn Ala Gln Lys Lys Ala Gln
210 215 220
Ser Ser Ile Asn
225
<210> 472
<211> 278
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 472
Gly Ser Pro Thr Pro Tyr Pro Asn Tyr Ala Asn Ala Gly His Val Glu
1 5 10 15
Gly Gln Ser Ala Leu Phe Met Arg Asp Asn Gly Ile Ser Glu Gly Leu
20 25 30
Val Phe His Asn Asn Pro Glu Gly Thr Cys Gly Phe Cys Val Asn Met
35 40 45
Thr Glu Thr Leu Leu Pro Glu Asn Ala Lys Met Thr Val Val Pro Pro
50 55 60
Glu Gly Ala Ile Pro Val Lys Arg Gly Ala Thr Gly Glu Thr Lys Val
65 70 75 80
Phe Thr Gly Asn Ser Asn Ser Pro Lys Ser Pro Thr Lys Gly Gly Cys
85 90 95
Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser
100 105 110
Ser Glu Val Glu Phe Ser His Glu Tyr Trp Met Arg His Ala Leu Thr
115 120 125
Leu Ala Lys Arg Ala Arg Asp Glu Arg Glu Val Pro Val Gly Ala Val
130 135 140
Leu Val Leu Asn Asn Arg Val Ile Gly Glu Gly Trp Asn Arg Ala Ile
145 150 155 160
Gly Leu His Asp Pro Thr Ala His Ala Glu Ile Met Ala Leu Arg Gln
165 170 175
Gly Gly Leu Val Met Gln Asn Tyr Arg Leu Ile Asp Ala Thr Leu Tyr
180 185 190
Val Thr Phe Glu Pro Cys Val Met Cys Ala Gly Ala Met Ile His Ser
195 200 205
Arg Ile Gly Arg Val Val Phe Gly Val Arg Asn Ser Lys Arg Gly Ala
210 215 220
Ala Gly Ser Leu Met Asn Val Leu Asn Tyr Pro Gly Met Asn His Arg
225 230 235 240
Val Glu Ile Thr Glu Gly Ile Leu Ala Asp Glu Cys Ala Ala Leu Leu
245 250 255
Cys Asp Phe Tyr Arg Met Pro Arg Gln Val Phe Asn Ala Gln Lys Lys
260 265 270
Ala Gln Ser Ser Ile Asn
275
<210> 473
<211> 8
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 473
Ser Gly Gly Gly Leu Gly Ser Thr
1 5
<210> 474
<211> 16
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 474
Ser Gly Ser Glu Thr Pro Gly Thr Ser Glu Ser Ala Thr Pro Glu Ser
1 5 10 15
<210> 475
<211> 32
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 475
Ser Gly Gly Ser Ser Gly Gly Ser Ser Gly Ser Glu Thr Pro Gly Thr
1 5 10 15
Ser Glu Ser Ala Thr Pro Glu Ser Ser Gly Gly Ser Ser Gly Gly Ser
20 25 30
<210> 476
<211> 60
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 476
Met Ala Leu Ser Arg Ala Val Cys Gly Thr Ser Arg Gln Leu Ala Pro
1 5 10 15
Val Leu Gly Tyr Leu Gly Ser Arg Gln Lys His Ser Leu Pro Asp Tyr
20 25 30
Pro Tyr Asp Val Pro Asp Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp
35 40 45
Tyr Ala Gly Tyr Pro Tyr Asp Val Pro Asp Tyr Ala
50 55 60
<210> 477
<211> 52
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 477
Met Ala Ser Val Leu Thr Pro Leu Leu Leu Arg Gly Leu Thr Gly Ser
1 5 10 15
Ala Arg Arg Leu Pro Val Pro Arg Ala Lys Ile His Ser Leu Asp Tyr
20 25 30
Lys Asp His Asp Gly Asp Tyr Lys Asp His Asp Ile Asp Tyr Lys Asp
35 40 45
Asp Asp Asp Lys
50
<210> 478
<211> 7
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 478
Pro Lys Lys Lys Arg Lys Val
1 5
<210> 479
<211> 5
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 479
Thr Gly Glu Lys Pro
1 5
<210> 480
<211> 4
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 480
Ser Gly Gly Ser
1
<210> 481
<211> 83
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 481
Thr Asn Leu Ser Asp Ile Ile Glu Lys Glu Thr Gly Lys Gln Leu Val
1 5 10 15
Ile Gln Glu Ser Ile Leu Met Leu Pro Glu Glu Val Glu Glu Val Ile
20 25 30
Gly Asn Lys Pro Glu Ser Asp Ile Leu Val His Thr Ala Tyr Asp Glu
35 40 45
Ser Thr Asp Glu Asn Val Met Leu Leu Thr Ser Asp Ala Pro Glu Tyr
50 55 60
Lys Pro Trp Ala Leu Val Ile Gln Asp Ser Asn Gly Glu Asn Lys Ile
65 70 75 80
Lys Met Leu
<210> 482
<211> 123
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 482
Gly Ile His Gly Val Pro Ala Ala Met Ala Glu Arg Pro Phe Gln Cys
1 5 10 15
Arg Ile Cys Met Arg Asn Phe Ser Gln Ser Gly Asp Leu Arg Arg His
20 25 30
Ile Arg Thr His Thr Gly Glu Lys Pro Phe Ala Cys Asp Ile Cys Gly
35 40 45
Arg Lys Phe Ala Arg Ser Asp Asn Leu Ser Val His Thr Lys Ile His
50 55 60
Thr Gly Ser Gln Lys Pro Phe Gln Cys Arg Ile Cys Met Arg Asn Phe
65 70 75 80
Ser Gln Lys Ile Asn Leu Gln Val His Ile Arg Thr His Thr Gly Glu
85 90 95
Lys Pro Phe Ala Cys Asp Ile Cys Gly Arg Lys Phe Ala Arg Ser Asp
100 105 110
Val Leu Ser Glu His Thr Lys Ile His Leu Arg
115 120
<210> 483
<211> 123
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 483
Gly Ile His Gly Val Pro Ala Ala Met Ala Glu Arg Pro Phe Gln Cys
1 5 10 15
Arg Ile Cys Met Arg Asn Phe Ser Asp Gln Ser Asn Leu Arg Ala His
20 25 30
Ile Arg Thr His Thr Gly Glu Lys Pro Phe Ala Cys Asp Ile Cys Gly
35 40 45
Arg Lys Phe Ala Thr Ser Ser Asn Arg Lys Thr His Thr Lys Ile His
50 55 60
Thr Gly Ser Gln Lys Pro Phe Gln Cys Arg Ile Cys Met Arg Asn Phe
65 70 75 80
Ser Leu Gln Gln Thr Leu Ala Asp His Ile Arg Thr His Thr Gly Glu
85 90 95
Lys Pro Phe Ala Cys Asp Ile Cys Gly Arg Lys Phe Ala Gln Ser Gly
100 105 110
Asn Leu Ala Arg His Thr Lys Ile His Leu Arg
115 120
<210> 484
<211> 142
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 484
Phe Gln Cys Arg Ile Cys Met Arg Lys Phe Ala Thr Ser Gly Ser Leu
1 5 10 15
Thr Arg His Thr Lys Ile His Thr Gly Glu Lys Pro Phe Gln Cys Arg
20 25 30
Ile Cys Met Arg Asn Phe Ser Arg Ser Asp His Leu Ser Thr His Ile
35 40 45
Arg Thr His Thr Gly Glu Lys Pro Phe Ala Cys Asp Ile Cys Gly Arg
50 55 60
Lys Phe Ala Thr Ser Ser Asn Arg Thr Lys His Thr Lys Ile His Thr
65 70 75 80
His Pro Arg Ala Pro Ile Pro Lys Pro Phe Gln Cys Arg Ile Cys Met
85 90 95
Arg Asn Phe Ser Arg Ser Asp Asn Leu Ser Glu His Ile Arg Thr His
100 105 110
Thr Gly Glu Lys Pro Phe Ala Cys Asp Ile Cys Gly Arg Lys Phe Ala
115 120 125
Trp His Ser Ser Leu Arg Val His Thr Lys Ile His Leu Arg
130 135 140
<210> 485
<211> 151
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 485
Gly Ile His Gly Val Pro Ala Ala Met Ala Glu Arg Pro Phe Gln Cys
1 5 10 15
Arg Ile Cys Met Arg Asn Phe Ser Arg Ser Asp His Leu Ser Thr His
20 25 30
Ile Arg Thr His Thr Gly Glu Lys Pro Phe Ala Cys Asp Ile Cys Gly
35 40 45
Arg Lys Phe Ala Asp Arg Ser His Leu Ala Arg His Thr Lys Ile His
50 55 60
Thr Gly Ser Gln Lys Pro Phe Gln Cys Arg Ile Cys Met Arg Lys Phe
65 70 75 80
Ala Leu Lys Gln His Leu Asn Glu His Thr Lys Ile His Thr Gly Glu
85 90 95
Lys Pro Phe Gln Cys Arg Ile Cys Met Arg Asn Phe Ser Gln Ser Gly
100 105 110
Asn Leu Ala Arg His Ile Arg Thr His Thr Gly Glu Lys Pro Phe Ala
115 120 125
Cys Asp Ile Cys Gly Arg Lys Phe Ala His Asn Ser Ser Leu Lys Asp
130 135 140
His Thr Lys Ile His Leu Arg
145 150
<210> 486
<211> 118
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 486
Gly Ile Arg Ile Pro Gly Glu Lys Pro Tyr Ser Cys Gly Ile Cys Gly
1 5 10 15
Lys Ser Phe Ser Asp Ser Ser Ala Lys Arg Arg His Cys Ile Leu His
20 25 30
Thr Gly Glu Lys Pro Tyr Thr Cys Ser Asp Cys Gly Lys Ala Phe Arg
35 40 45
Asp Lys Ser Cys Leu Asn Arg His Arg Arg Thr His Thr Gly Glu Lys
50 55 60
Pro Tyr Lys Cys Met Glu Cys Gly Lys Ala Phe Asn Arg Arg Ser His
65 70 75 80
Leu Thr Arg His Gln Arg Ile His Thr Gly Glu Lys Pro Tyr Glu Cys
85 90 95
Asn Tyr Cys Gly Lys Thr Phe Ser Val Ser Ser Thr Leu Ile Arg His
100 105 110
Gln Arg Ile His Leu Arg
115
<210> 487
<211> 117
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 487
Gly Ile Arg Glu Arg Pro Tyr Ala Cys Pro Val Glu Ser Cys Asp Arg
1 5 10 15
Arg Phe Ser Thr Ser Gly Ser Leu Val Arg His Ile Arg Ile His Thr
20 25 30
Gly Gln Lys Pro Phe Gln Cys Arg Ile Cys Met Arg Asn Phe Ser Arg
35 40 45
Ser Asp Glu Leu Thr Arg His Ile Arg Thr His Thr Gly Glu Lys Pro
50 55 60
Phe Ala Cys Asp Ile Cys Gly Arg Lys Phe Ala Arg Ser Asp His Leu
65 70 75 80
Thr Thr His Thr Lys Ile His Thr Gly Glu Lys Pro Phe Gln Cys Arg
85 90 95
Ile Cys Met Arg Lys Phe Ala Gln Ser Ser Asn Leu Val Arg His Thr
100 105 110
Lys Ile His Leu Arg
115
<210> 488
<211> 117
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 488
Gly Ile Arg Glu Arg Pro Tyr Ala Cys Pro Val Glu Ser Cys Asp Arg
1 5 10 15
Arg Phe Ser Asp Pro Gly His Leu Val Arg His Ile Arg Ile His Thr
20 25 30
Gly Gln Lys Pro Phe Gln Cys Arg Ile Cys Met Arg Asn Phe Ser Arg
35 40 45
Ser Asp Glu Leu Thr Arg His Ile Arg Thr His Thr Gly Glu Lys Pro
50 55 60
Phe Ala Cys Asp Ile Cys Gly Arg Lys Phe Ala Arg Ser Asp His Leu
65 70 75 80
Thr Thr His Thr Lys Ile His Thr Gly Glu Lys Pro Phe Gln Cys Arg
85 90 95
Ile Cys Met Arg Lys Phe Ala Arg Ser Asp Lys Leu Val Arg His Thr
100 105 110
Lys Ile His Leu Arg
115
<210> 489
<211> 122
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 489
Gly Ile Arg Ile Pro Gly Glu Lys Pro Tyr Glu Cys Asn Tyr Cys Gly
1 5 10 15
Lys Thr Phe Ser Val Ser Ser Thr Leu Ile Arg His Gln Arg Ile His
20 25 30
Thr Gly Glu Lys Pro Tyr His Cys Asp Trp Asp Gly Cys Gly Trp Lys
35 40 45
Phe Ala Arg Ser Asp Glu Leu Thr Arg His Tyr Arg Lys His Thr Gly
50 55 60
Glu Lys Pro Phe Glu Cys Lys Asp Cys Gly Lys Ala Phe Ile Gln Lys
65 70 75 80
Ser Asn Leu Ile Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr
85 90 95
His Cys Asp Trp Asp Gly Cys Gly Trp Lys Phe Ala Arg Ser Asp Glu
100 105 110
Leu Thr Arg His Tyr Arg Lys His Leu Arg
115 120
<210> 490
<211> 117
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 490
Gly Ile Arg Glu Arg Pro Tyr Ala Cys Pro Val Glu Ser Cys Asp Arg
1 5 10 15
Arg Phe Ser Asp Cys Arg Asp Leu Ala Arg His Ile Arg Ile His Thr
20 25 30
Gly Gln Lys Pro Phe Gln Cys Arg Ile Cys Met Arg Asn Phe Ser Arg
35 40 45
Ser Asp Glu Leu Thr Arg His Ile Arg Thr His Thr Gly Glu Lys Pro
50 55 60
Phe Ala Cys Asp Ile Cys Gly Arg Lys Phe Ala Arg Ser Asp His Leu
65 70 75 80
Thr Thr His Thr Lys Ile His Thr Gly Glu Lys Pro Phe Gln Cys Arg
85 90 95
Ile Cys Met Arg Lys Phe Ala Arg Ser Asp Lys Leu Val Arg His Thr
100 105 110
Lys Ile His Leu Arg
115
<210> 491
<211> 118
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 491
Gly Ile Arg Ile Pro Gly Glu Lys Pro Phe Glu Cys Lys Asp Cys Gly
1 5 10 15
Lys Ala Phe Ile Gln Lys Ser Asn Leu Ile Arg His Gln Arg Thr His
20 25 30
Thr Gly Glu Lys Pro Tyr Arg Cys Glu Glu Cys Gly Lys Ala Phe Arg
35 40 45
Trp Pro Ser Asn Leu Thr Arg His Lys Arg Ile His Thr Gly Glu Lys
50 55 60
Pro Tyr Arg Cys Glu Glu Cys Gly Lys Ala Phe Arg Trp Pro Ser Asn
65 70 75 80
Leu Thr Arg His Lys Arg Ile His Thr Gly Glu Lys Pro Tyr Ser Cys
85 90 95
Gly Ile Cys Gly Lys Ser Phe Ser Asp Ser Ser Ala Lys Arg Arg His
100 105 110
Cys Ile Leu His Leu Arg
115
<210> 492
<211> 117
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 492
Gly Ile Arg Glu Arg Pro Tyr Ala Cys Pro Val Glu Ser Cys Asp Arg
1 5 10 15
Arg Phe Ser Asp Cys Arg Asp Leu Ala Arg His Ile Arg Ile His Thr
20 25 30
Gly Gln Lys Pro Phe Gln Cys Arg Ile Cys Met Arg Asn Phe Ser Arg
35 40 45
Ser Asp Glu Leu Thr Arg His Ile Arg Thr His Thr Gly Glu Lys Pro
50 55 60
Phe Ala Cys Asp Ile Cys Gly Arg Lys Phe Ala Arg Ser Asp His Leu
65 70 75 80
Thr Thr His Thr Lys Ile His Thr Gly Glu Lys Pro Phe Gln Cys Arg
85 90 95
Ile Cys Met Arg Lys Phe Ala Arg Ser Asp Lys Leu Val Arg His Thr
100 105 110
Lys Ile His Leu Arg
115
<210> 493
<211> 120
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 493
Gly Ile Arg Ile Pro Gly Glu Lys Pro Tyr Thr Cys Ser Asp Cys Gly
1 5 10 15
Lys Ala Phe Arg Asp Lys Ser Cys Leu Asn Arg His Arg Arg Thr His
20 25 30
Thr Gly Glu Lys Pro Tyr Lys Cys Gly Gln Cys Gly Lys Phe Tyr Ser
35 40 45
Gln Val Ser His Leu Thr Arg His Gln Lys Ile His Thr Gly Glu Lys
50 55 60
Pro Phe Glu Cys Lys Asp Cys Gly Lys Ala Phe Ile Gln Lys Ser Asn
65 70 75 80
Leu Ile Arg His Gln Arg Thr His Thr Gly Glu Lys Pro Tyr His Cys
85 90 95
Asp Trp Asp Gly Cys Gly Trp Lys Phe Ala Arg Ser Asp Glu Leu Thr
100 105 110
Arg His Tyr Arg Lys His Leu Arg
115 120
<210> 494
<211> 117
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 494
Gly Ile Arg Glu Arg Pro Tyr Ala Cys Pro Val Glu Ser Cys Asp Arg
1 5 10 15
Arg Phe Ser Arg Ser Asp Lys Leu Val Arg His Ile Arg Ile His Thr
20 25 30
Gly Gln Lys Pro Phe Gln Cys Arg Ile Cys Met Arg Asn Phe Ser Arg
35 40 45
Ser Asp Glu Leu Thr Arg His Ile Arg Thr His Thr Gly Glu Lys Pro
50 55 60
Phe Ala Cys Asp Ile Cys Gly Arg Lys Phe Ala Arg Ser Asp His Leu
65 70 75 80
Thr Thr His Thr Lys Ile His Thr Gly Glu Lys Pro Phe Gln Cys Arg
85 90 95
Ile Cys Met Arg Lys Phe Ala Arg Ser Asp Lys Leu Val Arg His Thr
100 105 110
Lys Ile His Leu Arg
115
<210> 495
<211> 118
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 495
Gly Ile Arg Ile Pro Gly Glu Lys Pro Tyr Thr Cys Ser Asp Cys Gly
1 5 10 15
Lys Ala Phe Arg Asp Lys Ser Cys Leu Asn Arg His Arg Arg Thr His
20 25 30
Thr Gly Glu Lys Pro Tyr Glu Cys Asn Tyr Cys Gly Lys Thr Phe Ser
35 40 45
Val Ser Ser Thr Leu Ile Arg His Gln Arg Ile His Thr Gly Glu Lys
50 55 60
Pro Tyr Thr Cys Ser Asp Cys Gly Lys Ala Phe Arg Asp Lys Ser Cys
65 70 75 80
Leu Asn Arg His Arg Arg Thr His Thr Gly Glu Lys Pro Tyr Thr Cys
85 90 95
Ser Asp Cys Gly Lys Ala Phe Arg Asp Lys Ser Cys Leu Asn Arg His
100 105 110
Arg Arg Thr His Leu Arg
115
<210> 496
<211> 117
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 496
Gly Ile Arg Glu Arg Pro Tyr Ala Cys Pro Val Glu Ser Cys Asp Arg
1 5 10 15
Arg Phe Ser Arg Ser Asp Lys Leu Val Arg His Ile Arg Ile His Thr
20 25 30
Gly Gln Lys Pro Phe Gln Cys Arg Ile Cys Met Arg Asn Phe Ser Arg
35 40 45
Ser Asp Glu Leu Thr Arg His Ile Arg Thr His Thr Gly Glu Lys Pro
50 55 60
Phe Ala Cys Asp Ile Cys Gly Arg Lys Phe Ala Arg Ser Asp His Leu
65 70 75 80
Thr Thr His Thr Lys Ile His Thr Gly Glu Lys Pro Phe Gln Cys Arg
85 90 95
Ile Cys Met Arg Lys Phe Ala Arg Ser Asp Lys Leu Val Arg His Thr
100 105 110
Lys Ile His Leu Arg
115
<210> 497
<211> 118
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 497
Gly Ile Arg Ile Pro Gly Glu Lys Pro Tyr Glu Cys Asn Tyr Cys Gly
1 5 10 15
Lys Thr Phe Ser Val Ser Ser Thr Leu Ile Arg His Gln Arg Ile His
20 25 30
Thr Gly Glu Lys Pro Tyr Thr Cys Ser Asp Cys Gly Lys Ala Phe Arg
35 40 45
Asp Lys Ser Cys Leu Asn Arg His Arg Arg Thr His Thr Gly Glu Lys
50 55 60
Pro Tyr Thr Cys Ser Asp Cys Gly Lys Ala Phe Arg Asp Lys Ser Cys
65 70 75 80
Leu Asn Arg His Arg Arg Thr His Thr Gly Glu Lys Pro Tyr His Cys
85 90 95
Asp Trp Asp Gly Cys Gly Trp Lys Phe Ala Arg Ser Asp Glu Leu Thr
100 105 110
Arg His Tyr Arg Lys His
115
<210> 498
<211> 122
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 498
Gly Ile His Gly Val Pro Ala Ala Met Ala Glu Arg Pro Phe Gln Cys
1 5 10 15
Arg Ile Cys Met Arg Asn Phe Ser Thr Ser Gly His Leu Val Arg His
20 25 30
Ile Arg Thr His Thr Gly Glu Lys Pro Phe Ala Cys Asp Ile Cys Gly
35 40 45
Arg Lys Phe Ala Thr Ser Gly His Leu Val Arg His Thr Lys Ile His
50 55 60
Thr Gly Glu Lys Pro Phe Gln Cys Arg Ile Cys Met Arg Asn Phe Ser
65 70 75 80
Thr Ser Gly Glu Leu Val Arg His Ile Arg Thr His Thr Gly Glu Lys
85 90 95
Pro Phe Ala Cys Asp Ile Cys Gly Arg Lys Phe Ala Gln Ser Ser Asn
100 105 110
Leu Val Arg His Thr Lys Ile His Leu Arg
115 120
<210> 499
<211> 123
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 499
Gly Ile His Gly Val Pro Ala Ala Met Ala Glu Arg Pro Phe Gln Cys
1 5 10 15
Arg Ile Cys Met Arg Asn Phe Ser Asp Arg Ser Asn Leu Ser Arg His
20 25 30
Ile Arg Thr His Thr Gly Glu Lys Pro Phe Ala Cys Asp Ile Cys Gly
35 40 45
Arg Lys Phe Ala Ile Ser Ser Asn Leu Asn Ser His Thr Lys Ile His
50 55 60
Thr Gly Ser Gln Lys Pro Phe Gln Cys Arg Ile Cys Met Arg Asn Phe
65 70 75 80
Ser Arg Ser Asp Asn Leu Ala Arg His Ile Arg Thr His Thr Gly Glu
85 90 95
Lys Pro Phe Ala Cys Asp Ile Cys Gly Arg Lys Phe Ala Thr Ser Gly
100 105 110
Asn Leu Thr Arg His Thr Lys Ile His Leu Arg
115 120
<210> 500
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 500
Phe Gln Cys Arg Ile Cys Met Arg Asn Phe Ser Asp Ser Gly Asn Leu
1 5 10 15
Arg Val His Ile Arg Thr His
20
<210> 501
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 501
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr Lys Asn Ser Leu
1 5 10 15
Thr Glu His Gln Arg Thr His
20
<210> 502
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 502
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Glu Asp Asn Leu
1 5 10 15
His Thr His Gln Arg Thr His
20
<210> 503
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 503
Tyr Glu Cys Asp His Cys Gly Lys Ser Phe Ser Gln Ser Ser His Leu
1 5 10 15
Asn Val His Lys Arg Thr His
20
<210> 504
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 504
Tyr Glu Cys His Asp Cys Gly Lys Ser Phe Arg Gln Ser Thr His Leu
1 5 10 15
Thr Gln His Arg Arg Ile His
20
<210> 505
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 505
Tyr Lys Cys Pro Asp Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu
1 5 10 15
Ile Arg His Gln Arg Thr His
20
<210> 506
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 506
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu
1 5 10 15
Val Arg His Gln Arg Thr His
20
<210> 507
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 507
Tyr Gly Cys His Leu Cys Gly Lys Ala Phe Ser Lys Ser Ser Asn Leu
1 5 10 15
Arg Arg His Glu Met Ile His
20
<210> 508
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 508
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu
1 5 10 15
Val Arg His Gln Arg Thr His
20
<210> 509
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 509
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu
1 5 10 15
Val Arg His Gln Arg Thr His
20
<210> 510
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 510
Phe His Cys Gly Tyr Cys Glu Lys Ser Phe Ser Val Lys Asp Tyr Leu
1 5 10 15
Thr Lys His Ile Arg Thr His
20
<210> 511
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 511
Phe His Cys Gly Tyr Cys Glu Lys Ser Phe Ser Val Lys Asp Tyr Leu
1 5 10 15
Thr Lys His Ile Arg Thr His
20
<210> 512
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 512
Phe His Cys Gly Tyr Cys Glu Lys Ser Phe Ser Val Lys Asp Tyr Leu
1 5 10 15
Thr Lys His Ile Arg Thr His
20
<210> 513
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 513
Phe Ala Cys Pro Glu Cys Pro Lys Arg Phe Met Arg Ser Asp Asn Leu
1 5 10 15
Thr Gln His Ile Lys Thr His
20
<210> 514
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 514
Tyr Glu Cys His Asp Cys Gly Lys Ser Phe Arg Gln Ser Thr His Leu
1 5 10 15
Thr Gln His Arg Arg Ile His
20
<210> 515
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 515
Tyr Arg Cys Glu Glu Cys Gly Lys Ala Phe Arg Trp Pro Ser Asn Leu
1 5 10 15
Thr Arg His Lys Arg Ile His
20
<210> 516
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 516
Phe Glu Cys Lys Asp Cys Gly Lys Ala Phe Ile Gln Lys Ser Asn Leu
1 5 10 15
Ile Arg His Gln Arg Thr His
20
<210> 517
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 517
Tyr Thr Cys Ser Tyr Cys Gly Lys Ser Phe Thr Gln Ser Asn Thr Leu
1 5 10 15
Lys Gln His Thr Arg Ile His
20
<210> 518
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 518
Tyr Lys Cys Pro Asp Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu
1 5 10 15
Ile Arg His Gln Arg Thr His
20
<210> 519
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 519
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Ser Lys Lys Ala Leu
1 5 10 15
Thr Glu His Gln Arg Thr His
20
<210> 520
<211> 25
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 520
Tyr His Cys Asp Trp Asp Gly Cys Gly Trp Lys Phe Ala Arg Ser Asp
1 5 10 15
Glu Leu Thr Arg His Tyr Arg Lys His
20 25
<210> 521
<211> 25
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 521
Tyr His Cys Asp Trp Asp Gly Cys Gly Trp Lys Phe Ala Arg Ser Asp
1 5 10 15
Glu Leu Thr Arg His Tyr Arg Lys His
20 25
<210> 522
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 522
Tyr Ser Cys Gly Ile Cys Gly Lys Ser Phe Ser Asp Ser Ser Ala Lys
1 5 10 15
Arg Arg His Cys Ile Leu His
20
<210> 523
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 523
Tyr Lys Cys Gly Gln Cys Gly Lys Phe Tyr Ser Gln Val Ser His Leu
1 5 10 15
Thr Arg His Gln Lys Ile His
20
<210> 524
<211> 24
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 524
Tyr Arg Cys Lys Tyr Cys Asp Arg Ser Phe Ser Ile Ser Ser Asn Leu
1 5 10 15
Gln Arg His Val Arg Asn Ile His
20
<210> 525
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 525
Phe Gln Cys Lys Thr Cys Gln Arg Lys Phe Ser Arg Ser Asp His Leu
1 5 10 15
Lys Thr His Thr Arg Thr His
20
<210> 526
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 526
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Glu Asp Asn Leu
1 5 10 15
His Thr His Gln Arg Thr His
20
<210> 527
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 527
Phe His Cys Gly Tyr Cys Glu Lys Ser Phe Ser Val Lys Asp Tyr Leu
1 5 10 15
Thr Lys His Ile Arg Thr His
20
<210> 528
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 528
Tyr Glu Cys Val Gln Cys Gly Lys Gly Phe Thr Gln Ser Ser Asn Leu
1 5 10 15
Ile Thr His Gln Arg Val His
20
<210> 529
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 529
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Thr His Leu Asp Leu
1 5 10 15
Ile Arg His Gln Arg Thr His
20
<210> 530
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 530
Tyr Glu Cys Asp His Cys Gly Lys Ser Phe Ser Gln Ser Ser His Leu
1 5 10 15
Asn Val His Lys Arg Thr His
20
<210> 531
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 531
Tyr Glu Cys Asn Tyr Cys Gly Lys Thr Phe Ser Val Ser Ser Thr Leu
1 5 10 15
Ile Arg His Gln Arg Ile His
20
<210> 532
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 532
Tyr Glu Cys His Asp Cys Gly Lys Ser Phe Arg Gln Ser Thr His Leu
1 5 10 15
Thr Gln His Arg Arg Ile His
20
<210> 533
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 533
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Glu Asp Asn Leu
1 5 10 15
His Thr His Gln Arg Thr His
20
<210> 534
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 534
Phe His Cys Gly Tyr Cys Glu Lys Ser Phe Ser Val Lys Asp Tyr Leu
1 5 10 15
Thr Lys His Ile Arg Thr His
20
<210> 535
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 535
Tyr Lys Cys Asp Glu Cys Gly Lys Asn Phe Thr Gln Ser Ser Asn Leu
1 5 10 15
Ile Val His Lys Arg Ile His
20
<210> 536
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 536
Tyr Glu Cys Asp His Cys Gly Lys Ser Phe Ser Gln Ser Ser His Leu
1 5 10 15
Asn Val His Lys Arg Thr His
20
<210> 537
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 537
Tyr Glu Cys Asn Tyr Cys Gly Lys Thr Phe Ser Val Ser Ser Thr Leu
1 5 10 15
Ile Arg His Gln Arg Ile His
20
<210> 538
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 538
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Glu Asp Asn Leu
1 5 10 15
His Thr His Gln Arg Thr His
20
<210> 539
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 539
Tyr Lys Cys Pro Asp Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu
1 5 10 15
Ile Arg His Gln Arg Thr His
20
<210> 540
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 540
Tyr Glu Cys His Asp Cys Gly Lys Ser Phe Arg Gln Ser Thr His Leu
1 5 10 15
Thr Gln His Arg Arg Ile His
20
<210> 541
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 541
Tyr Lys Cys Pro Asp Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu
1 5 10 15
Ile Arg His Gln Arg Thr His
20
<210> 542
<211> 24
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 542
Tyr Arg Cys Lys Tyr Cys Asp Arg Ser Phe Ser Ile Ser Ser Asn Leu
1 5 10 15
Gln Arg His Val Arg Asn Ile His
20
<210> 543
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 543
Tyr Lys Cys Gly Gln Cys Gly Lys Phe Tyr Ser Gln Val Ser His Leu
1 5 10 15
Thr Arg His Gln Lys Ile His
20
<210> 544
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 544
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Glu Asp Asn Leu
1 5 10 15
His Thr His Gln Arg Thr His
20
<210> 545
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 545
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Ser Pro Ala Asp Leu
1 5 10 15
Thr Arg His Gln Arg Thr His
20
<210> 546
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 546
Tyr Thr Cys Ser Tyr Cys Gly Lys Ser Phe Thr Gln Ser Asn Thr Leu
1 5 10 15
Lys Gln His Thr Arg Ile His
20
<210> 547
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 547
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Ser Lys Lys Ala Leu
1 5 10 15
Thr Glu His Gln Arg Thr His
20
<210> 548
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 548
Phe Gln Cys Arg Ile Cys Met Arg Asn Phe Ser Asp Ser Gly Asn Leu
1 5 10 15
Arg Val His Ile Arg Thr His
20
<210> 549
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 549
Tyr Glu Cys Asp His Cys Gly Lys Ala Phe Ser Val Ser Ser Asn Leu
1 5 10 15
Asn Val His Arg Arg Ile His
20
<210> 550
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 550
Tyr Lys Cys Gly Gln Cys Gly Lys Phe Tyr Ser Gln Val Ser His Leu
1 5 10 15
Thr Arg His Gln Lys Ile His
20
<210> 551
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 551
Tyr Gly Cys His Leu Cys Gly Lys Ala Phe Ser Lys Ser Ser Asn Leu
1 5 10 15
Arg Arg His Glu Met Ile His
20
<210> 552
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 552
Tyr Thr Cys Ser Asp Cys Gly Lys Ala Phe Arg Asp Lys Ser Cys Leu
1 5 10 15
Asn Arg His Arg Arg Thr His
20
<210> 553
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 553
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Asp Pro Gly His Leu
1 5 10 15
Val Arg His Gln Arg Thr His
20
<210> 554
<211> 24
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 554
Tyr Arg Cys Lys Tyr Cys Asp Arg Ser Phe Ser Ile Ser Ser Asn Leu
1 5 10 15
Gln Arg His Val Arg Asn Ile His
20
<210> 555
<211> 24
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 555
Tyr Arg Cys Lys Tyr Cys Asp Arg Ser Phe Ser Ile Ser Ser Asn Leu
1 5 10 15
Gln Arg His Val Arg Asn Ile His
20
<210> 556
<211> 24
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 556
Tyr Arg Cys Lys Tyr Cys Asp Arg Ser Phe Ser Ile Ser Ser Asn Leu
1 5 10 15
Gln Arg His Val Arg Asn Ile His
20
<210> 557
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 557
Tyr Lys Cys Gly Gln Cys Gly Lys Phe Tyr Ser Gln Val Ser His Leu
1 5 10 15
Thr Arg His Gln Lys Ile His
20
<210> 558
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 558
Tyr Lys Cys Pro Asp Cys Gly Lys Ser Phe Ser Gln Ser Ser Ser Leu
1 5 10 15
Ile Arg His Gln Arg Thr His
20
<210> 559
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 559
Tyr Glu Cys Asp His Cys Gly Lys Ser Phe Ser Gln Ser Ser His Leu
1 5 10 15
Asn Val His Lys Arg Thr His
20
<210> 560
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 560
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Glu Asp Asn Leu
1 5 10 15
His Thr His Gln Arg Thr His
20
<210> 561
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 561
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Glu Asp Asn Leu
1 5 10 15
His Thr His Gln Arg Thr His
20
<210> 562
<211> 24
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 562
Tyr Arg Cys Lys Tyr Cys Asp Arg Ser Phe Ser Asp Ser Ser Asn Leu
1 5 10 15
Gln Arg His Val Arg Asn Ile His
20
<210> 563
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 563
Phe Gln Cys Arg Ile Cys Met Arg Asn Phe Ser Asp Ser Gly Asn Leu
1 5 10 15
Arg Val His Ile Arg Thr His
20
<210> 564
<211> 24
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 564
Tyr Arg Cys Lys Tyr Cys Asp Arg Ser Phe Ser Ile Ser Ser Asn Leu
1 5 10 15
Gln Arg His Val Arg Asn Ile His
20
<210> 565
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 565
Phe Gln Cys Arg Ile Cys Met Arg Asn Phe Ser Asp Ser Gly Asn Leu
1 5 10 15
Arg Val His Ile Arg Thr His
20
<210> 566
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 566
Tyr Glu Cys Asp His Cys Gly Lys Ser Phe Ser Gln Ser Ser His Leu
1 5 10 15
Asn Val His Lys Arg Thr His
20
<210> 567
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 567
Tyr Glu Cys Asp His Cys Gly Lys Ala Phe Ser Val Ser Ser Asn Leu
1 5 10 15
Asn Val His Arg Arg Ile His
20
<210> 568
<211> 25
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 568
Tyr His Cys Asp Trp Asp Gly Cys Gly Trp Lys Phe Ala Arg Ser Asp
1 5 10 15
Glu Leu Thr Arg His Tyr Arg Lys His
20 25
<210> 569
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 569
Tyr Lys Cys Pro Glu Cys Gly Lys Ser Phe Ser Arg Glu Asp Asn Leu
1 5 10 15
His Thr His Gln Arg Thr His
20
<210> 570
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 570
Tyr Thr Cys Ser Tyr Cys Gly Lys Ser Phe Thr Gln Ser Asn Thr Leu
1 5 10 15
Lys Gln His Thr Arg Ile His
20
<210> 571
<211> 23
<212> PRT
<213> Artificial Sequence
<220>
<223> Synthetic Sequence
<400> 571
Tyr Glu Cys Asp His Cys Gly Lys Ser Phe Ser Gln Ser Ser His Leu
1 5 10 15
Asn Val His Lys Arg Thr His
20

Claims (54)

1.一种融合蛋白，所述融合蛋白包含：
(i)DNA结合蛋白；以及
(ii)衍生自胞嘧啶脱氨酶或其变体的第一分裂物和第二分裂物，
其中所述第一分裂物和所述第二分裂物中的每一个与所述DNA结合蛋白融合。
2.一种融合蛋白，所述融合蛋白包含：
(i)DNA结合蛋白；以及
(ii)衍生自胞嘧啶脱氨酶或其变体的无毒全长胞嘧啶脱氨酶。
3.根据权利要求1所述的融合蛋白，其中所述第一分裂物和所述第二分裂物中的每一个不具有胞嘧啶脱氨酶活性。
4.根据权利要求1所述的融合蛋白，其中所述第一分裂物包含从SEQ ID NO:1的N末端残基开始至选自G33、G44、A54、N68、G82、N98和G108的至少一个残基的氨基酸序列。
5.根据权利要求1或2所述的融合蛋白，其中所述胞嘧啶脱氨酶衍生自双链DNA脱氨酶(DddA)或其直系同源物。
6.根据权利要求1所述的融合蛋白，其中所述第二分裂物包含从选自SEQ ID NO:1的G34、P45、G55、N69、T83、A99和A109的至少一个残基开始至C末端残基的氨基酸序列。
7.根据权利要求1所述的融合蛋白，其中所述胞嘧啶脱氨酶的变体在包含从SEQ IDNO:1的N末端残基开始至G44的氨基酸序列的第一分裂物中的残基3、5、10、11、13、14、15、16、17、18、19、28、30和31处的至少一个氨基酸被不同的氨基酸取代。
8.根据权利要求1所述的融合蛋白，其中所述胞嘧啶脱氨酶的变体在包含从SEQ IDNO:1的P45开始至C末端残基的氨基酸序列的第二分裂物中的残基13、16、17、20、21、28、29、30、31、32、33、56、57、58和60处的至少一个氨基酸被不同的氨基酸取代。
9.根据权利要求1所述的融合蛋白，其中所述胞嘧啶脱氨酶的变体在包含从SEQ IDNO:1的N末端残基开始至G108的氨基酸序列的第一分裂物中的残基87、88、91、92、95、100、101、102和103处的至少一个氨基酸被不同的氨基酸取代。
10.根据权利要求1所述的融合蛋白，其中所述胞嘧啶脱氨酶的变体在包含从SEQ IDNO:1的A109开始至C末端残基的氨基酸序列的第二分裂物中的残基13、14、15和16处的至少一个氨基酸被不同的氨基酸取代。
11.根据权利要求2所述的融合蛋白，其中所述无毒全长胞嘧啶脱氨酶在SEQ ID NO:1的野生型胞嘧啶脱氨酶中的残基37、59、109和129处的至少一个氨基酸被不同的氨基酸取代。
12.根据权利要求7至11中任一项所述的融合蛋白，其中所述不同的氨基酸是丙氨酸。
13.根据权利要求2所述的融合蛋白，其中所述无毒全长胞嘧啶脱氨酶是选自SEQ IDNO:12至22的至少一种。
14.根据权利要求1或2所述的融合蛋白，其中所述DNA结合蛋白是锌指蛋白、TALE蛋白或CRISPR相关核酸酶。
15.根据权利要求1或2所述的融合蛋白，其中所述DNA结合蛋白通过包含2至40个氨基酸残基的肽接头与胞嘧啶脱氨酶或其变体融合。
16.根据权利要求15所述的融合蛋白，其中所述接头包括：
2a.a接头：GS；
5a.a接头：TGEKQ(SEQ ID NO:8)；
10a.a接头：SGAQGSTLDF(SEQ ID NO:9)；
16a.a接头：SGSETPGTSESATPES(SEQ ID NO:10)；
24a.a接头：SGTPHEVGVYTLSGTPHEVGVYTL(SEQ ID NO:115)；或
32a.a接头：GSGGSSGGSSGSETPGTSESATPESSGGSSGGS(SEQ ID NO:11)。
17.根据权利要求1所述的融合蛋白，其中所述第一分裂物和所述第二分裂物中的每一个与锌指蛋白的N末端或C末端融合。
18.根据权利要求1或2所述的融合蛋白，其中单个TALE阵列或第一TALE阵列和第二TALE阵列中的每一个与所述胞嘧啶脱氨酶融合。
19.根据权利要求1或2所述的融合蛋白，所述融合蛋白进一步包含(iii)腺嘌呤脱氨酶。
20.根据权利要求19所述的融合蛋白，其中所述腺嘌呤脱氨酶是作为大肠杆菌TadA的变体的脱氧腺嘌呤脱氨酶。
21.根据权利要求19所述的融合蛋白，其中所述腺嘌呤脱氨酶与所述DNA结合蛋白或所述胞嘧啶脱氨酶或其变体的N末端或C末端融合。
22.一种核酸，所述核酸编码根据权利要求1至21中任一项所述的融合蛋白。
23.根据权利要求22所述的核酸，所述核酸是核糖核酸或DNA。
24.一种用于碱基编辑的组合物，所述组合物包含根据权利要求1至21中任一项所述的融合蛋白或根据权利要求22所述的核酸。
25.根据权利要求24所述的组合物，所述组合物进一步包含尿嘧啶糖基化酶抑制剂(UGI)。
26.一种用于在真核细胞中进行碱基编辑的组合物，所述组合物包含根据权利要求1至21中任一项所述的融合蛋白或根据权利要求22所述的核酸。
27.一种用于在植物细胞中进行碱基编辑的组合物，所述组合物包含根据权利要求1至21中任一项所述的融合蛋白或根据权利要求22所述的核酸以及核定位信号(NLS)肽或编码其的核酸。
28.一种用于在植物细胞中进行碱基编辑的组合物，所述组合物包含根据权利要求1至21中任一项所述的融合蛋白或根据权利要求22所述的核酸以及叶绿体转运肽或编码其的核酸。
29.一种用于在植物细胞中进行碱基编辑的组合物，所述组合物包含根据权利要求1至21中任一项所述的融合蛋白或根据权利要求22所述的核酸以及线粒体靶向信号(MTS)或编码其的核酸。
30.根据权利要求29所述的组合物，所述组合物进一步包含核输出信号(NES)或编码其的核酸。
31.根据权利要求27至30中任一项所述的组合物，其中将所述融合蛋白通过使用基因枪的注射(轰击)、PEG介导的原生质体转染、通过电穿孔的原生质体转染或通过显微注射的原生质体注射递送到植物细胞。
32.根据权利要求31所述的组合物，其中将所述核酸通过使用土壤杆菌属(例如根癌土壤杆菌或发根土壤杆菌)的转化、病毒转染、使用基因枪的注射(轰击)、PEG介导的原生质体转染、通过电穿孔的原生质体转染或通过显微注射的原生质体注射递送到植物细胞。
33.根据权利要求27至30中任一项所述的组合物，所述组合物用于在植物的线粒体、叶绿体或质体(白色体、色质体)中进行碱基编辑。
34.根据权利要求27至30中任一项所述的组合物，所述组合物进一步包含切割野生型DNA序列但不切割编辑的碱基序列的转录激活因子样效应物(TALE)-FokI核酸酶或锌指核酸酶(ZFN)或编码其的核酸。
35.一种用于在真核生物细胞核、线粒体或质体DNA中进行碱基编辑的方法，所述方法包括用根据权利要求27至30中任一项所述的组合物进行处理。
36.根据权利要求35所述的方法，其中碱基编辑效率通过进一步包含切割野生型DNA序列但不切割编辑的碱基序列的TALEN或ZFN或编码其的核酸来提高。
37.一种用于在植物细胞中进行碱基编辑的方法，所述方法包括用根据权利要求27至30中任一项所述的组合物处理植物细胞。
38.一种用于在植物细胞中进行碱基编辑的方法，所述方法包括用根据权利要求1至21中任一项所述的融合蛋白或根据权利要求22所述的核酸以及核定位信号(NLS)肽或编码其的核酸处理植物细胞。
39.一种用于在植物细胞中进行碱基编辑的方法，所述方法包括用根据权利要求1至21中任一项所述的融合蛋白或根据权利要求22所述的核酸以及叶绿体转运肽或编码其的核酸处理植物细胞。
40.一种用于在植物细胞中进行碱基编辑的方法，所述方法包括用根据权利要求1至21中任一项所述的融合蛋白或根据权利要求22所述的核酸以及线粒体靶向信号(MTS)或编码其的核酸处理植物细胞。
41.一种用于在动物细胞中进行碱基编辑的组合物，所述组合物包含根据权利要求1至21中任一项所述的融合蛋白或根据权利要求22所述的核酸以及核定位信号(NLS)肽或编码其的核酸。
42.一种用于在动物细胞中进行碱基编辑的组合物，所述组合物包含根据权利要求1至21中任一项所述的融合蛋白或根据权利要求22所述的核酸以及线粒体靶向信号(MTS)或编码其的核酸。
43.根据权利要求42所述的组合物，所述组合物进一步包含核输出信号或编码其的核酸。
44.根据权利要求42所述的组合物，所述组合物进一步包含切割野生型DNA序列但不切割编辑的碱基序列的转录激活因子样效应物(TALE)-FokI核酸酶或ZFN或编码其的核酸。
45.一种用于在动物细胞中进行碱基编辑的方法，所述方法包括用根据权利要求41或42所述的组合物处理动物细胞。
46.一种用于在动物细胞中进行碱基编辑的方法，所述方法包括用根据权利要求1至21中任一项所述的融合蛋白或根据权利要求22所述的核酸以及核定位信号(NLS)肽或编码其的核酸处理动物细胞。
47.一种用于在动物细胞中进行碱基编辑的方法，所述方法包括用根据权利要求1至21中任一项所述的融合蛋白或根据权利要求22所述的核酸以及线粒体靶向信号(MTS)或编码其的核酸处理动物细胞。
48.根据权利要求46或47所述的方法，其中碱基编辑效率通过进一步包含切割野生型DNA序列但不切割编辑的碱基序列的TALEN或ZFN或编码其的核酸来提高。
49.一种用于在原核细胞或真核细胞中进行A至G碱基编辑的组合物，所述组合物包含根据权利要求19所述的融合蛋白或编码其的核酸，其中DNA结合蛋白是锌指蛋白、TALE蛋白或CRISPR相关核酸酶，并且所述融合蛋白的胞嘧啶脱氨酶或其变体衍生自细菌并且对双链DNA具有特异性。
50.一种用于在原核细胞或真核细胞中进行A至G碱基编辑的组合物，所述组合物包含根据权利要求19所述的融合蛋白或编码其的核酸，其中DNA结合蛋白是锌指蛋白、TALE蛋白或CRISPR相关核酸酶，所述融合蛋白的胞嘧啶脱氨酶或其变体衍生自细菌并且对双链DNA具有特异性，所述DNA结合蛋白与所述胞嘧啶脱氨酶或其变体的N末端融合，并且所述DNA结合蛋白与所述融合蛋白的腺嘌呤脱氨酶的C末端融合。
51.一种用于在原核细胞或真核细胞中进行C至T碱基编辑的组合物，所述组合物包含根据权利要求19所述的融合蛋白或编码其的核酸和尿嘧啶糖基化酶抑制剂(UGI)，其中DNA结合蛋白是锌指蛋白、TALE蛋白或CRISPR相关核酸酶，并且所述融合蛋白的胞嘧啶脱氨酶或其变体衍生自细菌并且对双链DNA具有特异性。
52.一种用于在原核细胞或真核细胞中进行A至G碱基编辑的方法，所述方法包括用根据权利要求19所述的融合蛋白或编码其的核酸处理原核细胞或真核细胞，其中DNA结合蛋白是锌指蛋白、TALE蛋白或CRISPR相关核酸酶，并且所述融合蛋白的胞嘧啶脱氨酶或其变体衍生自细菌并且对双链DNA具有特异性。
53.一种用于在原核细胞或真核细胞中进行A至G碱基编辑的方法，所述方法包括用根据权利要求19所述的融合蛋白或编码其的核酸处理原核细胞或真核细胞，其中DNA结合蛋白是锌指蛋白、TALE蛋白或CRISPR相关核酸酶，所述融合蛋白的胞嘧啶脱氨酶或其变体衍生自细菌并且对双链DNA具有特异性，所述DNA结合蛋白与所述胞嘧啶脱氨酶或其变体的N末端融合，并且所述DNA结合蛋白与所述融合蛋白的腺嘌呤脱氨酶的C末端融合。
54.一种用于在原核细胞或真核细胞中进行C至T碱基编辑的方法，所述方法包括用根据权利要求19所述的融合蛋白或编码其的核酸和尿嘧啶糖基化酶抑制剂(UGI)处理原核细胞或真核细胞，其中DNA结合蛋白是锌指蛋白、TALE蛋白或CRISPR相关核酸酶，并且所述融合蛋白的胞嘧啶脱氨酶或其变体衍生自细菌并且对双链DNA具有特异性。
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