CN114606227A

CN114606227A - 高精度腺嘌呤碱基编辑器及其应用

Info

Publication number: CN114606227A
Application number: CN202210163391.3A
Authority: CN
Inventors: 黄强; 朱海霞; 杜文豪; 姜欣怡; 秦琴
Original assignee: Fudan University
Current assignee: Fudan University
Priority date: 2022-02-22
Filing date: 2022-02-22
Publication date: 2022-06-10
Anticipated expiration: 2042-02-22
Also published as: CN114606227B

Abstract

本发明公开了一种高精度腺嘌呤碱基编辑器及其应用。本发明的腺嘌呤碱基编辑器是将野生型腺嘌呤碱基编辑器ABE8e的SpCas9(D10A)和脱氨酶TadA*之间的linker从头设计并替换后所得。这些高精度腺嘌呤碱基编辑器不仅大大缩短了linker的长度，而且在靶位点具有与野生型ABE8e相当甚至更高的基因编辑活性。并且这些腺嘌呤碱基编辑器大大缩小了野生型ABE8e的编辑窗口，有效提高了ABE8e的编辑精度，使之编辑更加准确。因而这六个高精度腺嘌呤碱基编辑器在生物医药的精准基因编辑方面具有更大的应用潜力。

Description

高精度腺嘌呤碱基编辑器及其应用

技术领域

本发明属于蛋白质工程技术领域，具体涉及腺嘌呤碱基编辑器及其应用。

背景技术

如何精确有效地进行基因编辑一直是生命科学领域的重要目标。近年来，人们发现CRISPR核酸酶可通过诱导靶双链DNA断裂(double-stranded DNA breaks,DSBs)刺激细胞修复来实现基因组修饰的目的(1)。但DSB引发的非同源末端连接(non-homologous endjoining，NHEJ)修复通常无法人为控制，可通过删除、复制或其他DNA重排导致基因插入缺失(Indels)(2)。加入供体DNA模板虽可以刺激同源定向修复(homology-directed repair，HDR)精确替换DNA等位基因，但这一过程主要限于分裂细胞(3)。因此，急需开发一种在不诱导DSB的情况下精确修饰DNA的编辑技术(4)。

为解决这一需求，David R.Liu等开发了碱基编辑，这是一种全新的基因组编辑技术，可直接转换目标碱基对，无需DSB、HDR或供体DNA模板，有效减少了非必要的Indels产物(5)。一般来说，碱基编辑器由保留单链切割活性的Cas蛋白(nickaseCas,nCas)和脱氨酶组成。在与DNA的目标位点结合后，引导RNA(single guide RNA，sgRNA)和靶DNA链(targetDNA,TS)之间的碱基配对导致“R环”中的一小段非靶DNA链(non-target DNA,NTS)被置换,接着该ssDNA内的碱基被脱氨酶修饰产生突变(6)。为了提高在真核细胞中的编辑效率，单链切口酶还在未编辑的DNA链中产生切口，诱导细胞使用编辑的链作为模板来修复未编辑的链(7)。

目前，已有两种类型的DNA碱基编辑器被表征：胞嘧啶碱基编辑器(CBE)，将C·G碱基对转换为T·A碱基对，以及腺嘌呤碱基编辑器(ABE)，将A·T碱基对转换为G·C碱基对(8,9)。然而，当前的CBE编辑效率较弱并具有高度位点依赖性，因此它们的应用受到了严重限制(7)。相比之下，ABE显得更为重要，因为它可以纠正ClinVar数据库中47％的疾病相关点突变(4)。为获得ABE系统,David R.Liu等通过定向进化将大肠杆菌tRNA腺苷脱氨酶(wtTadA)与Cas9的融合体进化为可催化腺苷脱氨的ABE8e，该系统编码TadA*-linker-SpCas9(D10A)复合物，其中TadA*为进化所得的wtTadA变体，负责对底物DNA编辑；linker为(SGGS)₂-XTEN-(SGGS)₂的接头序列，负责连接SpCas9(D10A)和TadA*两个结构域；SpCas9(D10A)保留了单链切口酶的活性，负责在sgRNA的引导下靶向目标序列(10)。作为一种高兼容性的碱基编辑工具，ABE8e具有广泛的应用前景。

虽然ABE8e是一个强大的基因编辑工具，但仍存在着许多有待解决的问题，其中限制其进一步应用的最大问题是ABE8e可以编辑底物DNA中四到五个核苷酸窗口中的任何A，即PAM远端的第四位到第八位核苷酸，然而编辑靶位点以外的A会引入非必要产物，从而导致有害突变(9)。为解决以上问题，我们对SpCas9(D10A)和TadA*之间的linker进行了从头设计，获得了编辑窗口明显缩小的六个ABE8e高精度突变体，即分别是4L-ABE8e、4P-ABE8e、6L-ABE8e、7L-ABE8e、7P-ABE8e和12EK-ABE8e，从而为基因编辑库提供了多个高精度基因编辑工具。

发明内容

本发明的目的是提供多个高精度、且具有高编辑活性的腺嘌呤碱基编辑器及其应用。

本发明提供的来源于高精度腺嘌呤碱基编辑器，其编辑窗口比野生型腺嘌呤碱基编辑器ABE8e更狭窄即编辑准确度更高，具体是将野生型腺嘌呤碱基编辑器ABE8e的32个氨基酸的linker即177位至208位氨基酸分别替换成ETRS、NKTYPP、TTEPPDN、PAPA、PAPAPAP、A(EAAAK)₂A后重组所得，分别记为4L-ABE8e、4P-ABE8e、6L-ABE8e、7L-ABE8e、7P-ABE8e和12EK-ABE8e，属于碱基编辑系统。

在同等反应条件下，所述4L-ABE8e、4P-ABE8e、6L-ABE8e、7L-ABE8e、7P-ABE8e和12EK-ABE8e与野生型ABE8e具有同等或更高的剪切活性。

在同等反应条件下，所述4L-ABE8e、4P-ABE8e、6L-ABE8e、7L-ABE8e、7P-ABE8e和12EK-ABE8e比野生型ABE8e的编辑窗口更小，编辑精度更高。

所述野生型ABE8e的核苷酸序列和氨基酸序列分别为SEQIDNO.1和SEQ ID NO.2所示；

所述高精度腺嘌呤碱基编辑器即4L-ABE8e、4P-ABE8e、6L-ABE8e、7L-ABE8e、7P-ABE8e和12EK-ABE8e的核苷酸序列分别为SEQ ID NO.3、SEQ ID NO.5、SEQ ID NO.7、SEQ IDNO.9、SEQ ID NO.11、SEQ ID NO.13所示，氨基酸序列分别为SEQ ID NO.4、SEQ ID NO.6、SEQ ID NO.8、SEQ ID NO.10、SEQ ID NO.12、SEQ ID NO.14所示，与野生型ABE8e的相似度达90％以上。

本发明还提供一种表达载体，其含有上述多核苷酸序列。

本发明还提供一种宿主细胞，可以用于转化上述表达载体。

本发明还提供所述高精度腺嘌呤碱基编辑器的制备方法，具体步骤包括：首先，构建所述高精度腺嘌呤碱基编辑器的多核苷酸序列表达载体；然后，将所述表达载体转化至宿主细胞，筛选并挑出单克隆；最后，将所述单克隆诱导表达，并通过亲和层析、分子筛方法从表达产物中分离出所述的高精度腺嘌呤碱基编辑器。

本发明提供的上述高精度腺嘌呤碱基编辑器、多核苷酸序列以及表达载体均可作为编辑基因组DNA的编辑工具，用于基因组DNA片段的相关编辑。

本发明中，所述的基因编辑是编辑位点大于两个的多位点编辑；所述编辑的手段是腺嘌呤(A)到鸟嘌呤(G)的碱基转换。

本发明中，所述高精度腺嘌呤碱基编辑器作为编辑工具包括与靶标DNA片段匹配的引导sgRNA。

所述的高精度腺嘌呤碱基编辑器能够与介导它的sgRNA组合，从而对目的基因进行编辑。

本发明中，所述含有编码高精度腺嘌呤碱基编辑器多核苷酸序列的载体和与之匹配的引导sgRNA一同转入宿主细胞，对基因进行编辑。

本发明中，所述多位点的腺嘌呤(A)到鸟嘌呤(G)的碱基转换，是利用腺嘌呤碱基编辑器对腺嘌呤(A)的脱氨作用，并通过宿主细胞的修复系统对双链之间的错配进行修复，从而将碱基从腺嘌呤(A)转换成鸟嘌呤(G)。

与现有技术相比，本发明的高精度腺嘌呤碱基编辑器，属于碱基编辑系统，具有高活性、高精度等特点，可实现对体内基因组特定位置的精准编辑，具有潜在的生物医学应用价值。

附图说明

图1为野生型ABE8e及六个高精度腺嘌呤碱基编辑器的结构域组成。

图2为单克隆筛选与细菌培养。

图3为镍柱亲和层析纯化蛋白原理

图4为目标蛋白纯化过程。

图5为目的蛋白的SDS-PAGE电泳结果。

图6为目的蛋白的体外编辑窗口和编辑活性检测结果。

具体实施方式

下面通过具体实施例进一步描述本发明。

下述实施例中所用的实验方法，如无特定说明，均为常规方法。

下述实施例中所用的材料、试剂等，如无特定说明，均为从商业途径获得。

一、高精度腺嘌呤碱基编辑器

本发明的六个高精度腺嘌呤碱基编辑器是将野生型腺嘌呤碱基编辑器ABE8e的32个氨基酸的linker即177位至208位氨基酸分别替换成ETRS、NKTYPP、TTEPPDN、PAPA、PAPAPAP、A(EAAAK)₂A后重组所得，分别记为4L-ABE8e、4P-ABE8e、6L-ABE8e、7L-ABE8e、7P-ABE8e和12EK-ABE8e，如图1所示。这六个高精度腺嘌呤碱基编辑器不仅具有与野生型ABE8e相当的基因编辑活性，而且具有更狭窄的编辑窗口、更高的编辑精度，因此能实现更精准的体内靶向编辑。

二、编码高精度腺嘌呤碱基编辑器的多核苷酸序列

转录和翻译所述的高精度腺嘌呤碱基编辑器的多核苷酸序列，包括DNA或RNA。DNA还可以细分为质粒DNA、基因组DNA或人工合成的DNA。

编码所述高精度腺嘌呤碱基编辑器的多核苷酸序列，可以利用该领域科研或技术人员所熟悉的相关分子生物学技术来制备，其不局限于重组DNA技术和化学合成方法。

三、表达载体

所述表达载体含有编码所述高精度腺嘌呤碱基编辑器的多核苷酸序列。该表达载体可以通过科研或技术人员所熟悉的分子生物学方法来构建，包括DNA重组技术和DNA合成技术等，主要将高精度腺嘌呤碱基编辑器的DNA有效连接到载体上的克隆位点中，然后通过转录翻译等过程表达目的蛋白4L-ABE8e、4P-ABE8e、6L-ABE8e、7L-ABE8e、7P-ABE8e和12EK-ABE8e。

四、宿主细胞

所述宿主细胞是用来转化表达高精度腺嘌呤碱基编辑器的重组质粒。主要包括原核细胞(如细菌)，低等真核细胞(如酵母)，高等真核细胞(如哺乳动物细胞)等。常用的宿主细胞如大肠杆菌DH5α、Rosetta(DE3)、HEK293、CHO、Hela细胞等。

五、高精度腺嘌呤碱基编辑器及其编码该酶的核苷酸序列和所述表达载体的用途

本发明的高精度腺嘌呤碱基编辑器及其编码该酶的多核苷酸序列和所述的表达载体能够用于基因组DNA片段的编辑或用于制备基因编辑工具。高精度腺嘌呤碱基编辑器编辑为多位点编辑，其编辑手段为腺嘌呤(A)到鸟嘌呤(G)的碱基转换。

六、基因编辑工具及其方法

本发明的基因编辑工具属于碱基编辑系统，高精度腺嘌呤碱基编辑器在特定的sgRNA的引导下可以在编辑窗口内将腺苷(A)脱氨成肌苷(I)，而DNA复制酶将肌苷(I)识别成鸟苷(G)，因此后续DNA复制则以鸟苷(G)为模板进行，从而完成A到G的碱基转换。该编辑过程可以在体外或体内进行。

如本发明的一些实施方式中所列举的，高精度腺嘌呤碱基编辑器在sgRNA的引导下，可以在体外对底物DNA(920bp)进行剪切。

在本发明中，高精度腺嘌呤碱基编辑器可作为4L-ABE8e、4P-ABE8e、6L-ABE8e、7L-ABE8e、7P-ABE8e和12EK-ABE8e的简称使用，其含义与4L-ABE8e、4P-ABE8e、6L-ABE8e、7L-ABE8e、7P-ABE8e和12EK-ABE8e相同。本发明的六个高精度腺嘌呤碱基编辑器是将野生型ABE8e的32个氨基酸的linker即177位至208位氨基酸分别替换成ETRS、NKTYPP、TTEPPDN、PAPA、PAPAPAP、A(EAAAK)₂A后重组所得。

在进一步描述本发明具体实施方式之前，应理解，本发明的保护范围并不局限为下述特定的具体实施方案，还应理解为，本发明实施例中的术语是为了描述特定的具体实施方案，而不是为了限制本发明的保护范围。下例实施例中未注明具体条件的试验方法，通常按照常规条件操作，或者按照各生产厂商所建议的条件操作。

在实施例给出的数值范围中，应理解，除非本发明另有说明，每个数值范围的两个端点以及两个端点之间任何一个数值均可选用。除非另外定义，本发明中使用的所有技术和科学术语与本技术领域技术人员通常理解的意义相同。除实施例中使用的具体方法、设备、材料外，根据本技术领域的技术人员对现有技术的掌握及本发明的记载，还可以使用与本发明实施例中所述的方法、设备、材料相似或等同的现有技术的任何方法、设备和材料来实现本发明。

除非另外说明，本发明中所公开的实验方法、检测方法、制备方法均采用本技术领域常规的分子生物学、生物化学、重组DNA技术及相关领域的常规技术。

实施例1，构建高精度腺嘌呤碱基编辑器的质粒

1.突变体的构建

以野生型ABE8e即碱基序列SEQ ID NO.1和氨基酸序列SEQ ID NO.2为模板，将氨基酸序列的177位至208位氨基酸分别替换成ETRS、NKTYPP、TTEPPDN、PAPA、PAPAPAP、A(EAAAK)₂A，从而分别生成4L-ABE8e、4P-ABE8e、6L-ABE8e、7L-ABE8e、7P-ABE8e和12EK-ABE8e，如图1所示。其详细步骤简述如下：

主要利用引物进行常规PCR扩增，然后将模板消化后获取。4L-ABE8e、4P-ABE8e、6L-ABE8e、7L-ABE8e、7P-ABE8e和12EK-ABE8e的引物分别为SEQ ID NO.15-16、SEQ IDNO.17-18、SEQ ID NO.19-20、SEQ ID NO.21-24、SEQ ID NO.25-28、SEQ ID NO.29-32，所有引物均从上海生工生物工程有限公司订购。

(1)使用

GXL DNA Polymerase进行PCR扩增，其反应体系和反应参数如下所示：

酶切反应体系:

反应条件：

PCR结束后需要加入试剂盒提供的消化酶来消化质粒模板，其反应体系如下：

充分混匀后，37℃下反应1h。最后将PCR产物转化至大肠杆菌DH5α，过夜培养后筛选单克隆。

(2)样品测序

在上海杰李生物技术有限公司通过一代测序对单克隆进行测序。测序成功后，将目的质粒转化至Rosetta(DE3)细胞，进行菌液的扩大培养和目的蛋白的表达纯化。

实施例2，制备高精度腺嘌呤碱基编辑器

2.蛋白表达与纯化

2.1蛋白表达

(1)打开超净台，用含75％酒精的棉球擦拭桌面以及各种器具耗材，开紫外灯照射20min，启动风机备用。

(2)移液枪吸取10μl菌液转至6ml含有双抗(Amp与Cm)的LB液体培养基中，37℃，200r/min振荡培养过夜。

(3)将过夜培养的菌液按照体积比为1:100转至800ml含双抗的LB(购于生工)液体培养基中，37℃，200r/min振荡培养。在培养过程中，随时检测菌液的OD值。

(4)当菌液的OD值接近0.8～1时，加入蛋白诱导剂IPTG，使其终浓度为0.1mM，然后20℃，200r/min振荡培养12h。

(5)收集菌液，5000r/min离心5min使菌体沉淀，弃上清，并称重菌体。

2.2蛋白纯化

所述蛋白纯化主要通过镍柱亲和层析技术，如图3所示；其纯化过程包括菌体破碎、蛋白样品离心收集、蛋白样品与镍柱介质共孵育以及目的蛋白的洗脱等，如图4所示。其详细步骤如下：

(1)向3g菌体中加入预先冰浴且PMSF终浓度为0.1mM的平衡液(20mM HEPES,500mMKCl,pH7.5)15ml，涡旋仪重悬使菌块分散混匀，细胞超声破碎仪破碎细胞，超声3sec停5sec，一次10min，超声两次，超声过程均在冰浴中进行。

(2)将破碎的菌液在4℃10000rpm条件下离心45～60min，收集上清。

(3)将上清与预先用平衡液处理的Qiagen Ni-NTA介质振荡孵育90min,然后静置30min，待Qiagen Ni-NTA沉降。

(4)将Qiagen Ni-NTA装载到重力柱中，在BioLogic LP系统的监测下，分别以流速为2ml/min的平衡液和洗脱液(20mM HEPES,500mM KCl,500mM咪唑,pH7.5)梯度(20、30、40、50、100、250、500mM咪唑)冲洗Qiagen Ni-NTA，并收集蛋白。

(5)将不同咪唑浓度洗脱下的蛋白溶液进行SDS-PAGE鉴定，观察目的蛋白的表达和挂柱效果。

所述蛋白野生型ABE8e、4L-ABE8e、4P-ABE8e、6L-ABE8e、7L-ABE8e、7P-ABE8e和12EK-ABE8e的纯化结果如图5所示，其显示了目的蛋白的纯化情况。如该图所示，杂蛋白的条带较少，表明目的蛋白较为纯净。

实施例3，检测高精度腺嘌呤碱基编辑器的编辑窗口和编辑活性

3.活性检测

为检测高精度腺嘌呤碱基编辑器的编辑窗口，我们设计了分别靶向PAM远端第四位到第八位腺嘌呤(A)的DNA(SEQ ID NO.33-SEQ ID NO.37)和sgRNA(SEQ ID NO.38-SEQID NO.42)

反应体系按ABE8e相关蛋白:sgRNA:DNA＝200nM:200nM:40nM比例混合，具体如下:

将反应体系置37℃孵育两个小时，然后使用超薄DNA产物纯化试剂盒(购自天根生化科技有限公司)对反应后的DNA进行纯化，接着按以下反应体系加入EndoV酶37℃孵育一个小时：

EndoV酶可特异性识别肌苷(I)并在其下游进行切割，当发生脱氨反应时，腺苷(A)脱氨成肌苷(I)，因此，加入EndoV酶后DNA便发生双链断裂。最后通过琼脂糖凝胶电泳检测高精度腺嘌呤碱基编辑器的体外编辑窗口和编辑活性。其结果如图6所示，与野生型ABE8e相比，4L-ABE8e在保留5A编辑活性的同时，降低对其他A位点的活性，因此大大缩小了ABE8e的编辑窗口，提高了编辑精度；6L-ABE8e、4P-ABE8e、7P-ABE8e和12EK-ABE8e在降低6A-8A位点编辑活性的同时，还提高了对4A-5A位点的编辑活性，因此不仅提高了编辑精度，还提高了编辑活性。综上所述，高精度腺嘌呤碱基编辑器在保留与野生型ABE8e相当的剪切活性的同时，编辑窗口更小，编辑精度更高，因此有望成为生物医学精准医疗领域中具有潜在应用价值的基因编辑工具。

参考文献

1.M.Jinek et al.,A programmable dual-RNA-guided DNA endonuclease inadaptive bacterial immunity.Science337,816-821(2012).

2.J.R.Chapman,M.R.Taylor,S.J.Boulton,Playing the end game:DNA double-strand break repair pathway choice.Mol.Cell47,497-510(2012).

3.D.Paquet et al.,Efficient introduction of specific homozygous andheterozygous mutations using CRISPR/Cas9.Nature533,125-129(2016).

4.M.J.Landrum et al.,ClinVar:public archive of relationships amongsequence variation and human phenotype.Nucleic Acids Res.42,D980-985(2014).

5.H.A.Rees,D.R.Liu,Base editing:precision chemistry on the genome andtranscriptome of living cells.Nat.Rev.Genet.19,770-788(2018).

6.T.P.Huang,G.A.Newby,D.R.Liu,Precision genome editing using cytosineand adenine base editors in mammalian cells.Nat.Protoc.16,1089-1128(2021).

7.K.Nishida et al.,Targeted nucleotide editing using hybridprokaryotic and vertebrate adaptive immune systems.Science353,6305(2016).

8.A.C.Komor,Y.B.Kim,M.S.Packer,J.A.Zuris,D.R.Liu,Programmable editingof a target base in genomic DNA without double-stranded DNAcleavage.Nature533,420-424(2016).

9.N.M.Gaudelli et al.,Programmable base editing of A·T to G·C ingenomic DNA without DNA cleavage.Nature551,464-471(2017).

10.M.F.Richter et al.,Phage-assisted evolution of an adenine baseeditor with improved Cas domain compatibility and activity.Nat.Biotechnol.38,883-891(2020)。

序列表

<110> 复旦大学

<120> 高精度腺嘌呤碱基编辑器及其应用

<160> 42

<170> SIPOSequenceListing 1.0

<210> 1

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atgggcagca gccatcatca tcatcatcac tcggaggtcg agttttcaca tgagtattgg 60

atgcgtcatg ccttgacgtt ggcgaaacgg gcgcgcgatg agcgtgaggt gcccgtggga 120

gcggtgttgg tactgaataa ccgggttata ggggaaggat ggaaccgggc tattgggtta 180

cacgacccaa cggcgcacgc cgagataatg gcactgcgcc aagggggctt agttatgcag 240

aattatcgcc ttatcgatgc tacactgtat gtaacctttg aaccctgcgt aatgtgtgcg 300

ggggctatga tccactcgag aatagggcgc gtggtattcg gcgtacgcaa cagcaaacgt 360

ggggctgcgg gctcgttgat gaacgttctg aactaccccg gaatgaatca cagagtagag 420

atcacggagg gaattttggc cgacgaatgt gcagctttac tgtgcgattt ttatcggatg 480

ccgcggcaag tcttcaacgc acagaagaag gctcaatctt ccattaactc aggtggctcg 540

agtggtggga gtagcggatc tgagacgcca ggcacatcag agagtgcaac ccccgagtca 600

tcgggtggga gttccggcgg atctgataag aaatactcaa taggcttagc tatcggcaca 660

aacagcgtcg gctgggccgt cattacggac gagtacaagg tgccgagcaa aaaattcaaa 720

gttctgggca ataccgatcg ccacagcata aagaagaacc tcattggcgc cctcctgttc 780

gactccgggg agacggccga agccacgcgg ctcaaaagaa cagcacggcg cagatatacc 840

cgcagaaaga atcggatctg ctacctgcag gagatcttta gtaatgagat ggctaaggtg 900

gatgactctt tcttccatag gctggaggag tcctttttgg tggaggagga taaaaagcac 960

gagcgccacc caatctttgg caatatcgtg gacgaggtgg cgtaccatga aaagtaccca 1020

accatatatc atctgaggaa gaagcttgta gacagtactg ataaggctga cttgcggttg 1080

atctatctcg cgctggcgca tatgatcaaa tttcggggac acttcctcat cgagggggac 1140

ctgaacccag acaacagcga tgtcgacaaa ctctttatcc aactggttca gacttacaat 1200

cagcttttcg aagagaaccc gatcaacgca tccggagttg acgccaaagc aatcctgagc 1260

gctaggctgt ccaaatcccg gcggctcgaa aacctcatcg cacagctccc tggggagaag 1320

aagaacggcc tgtttggtaa tcttatcgcc ctgtcactcg ggctgacccc caactttaaa 1380

tctaacttcg acctggccga agatgccaag cttcaactga gcaaagacac ctacgatgat 1440

gatctcgaca atctgctggc ccagatcggc gaccagtacg cagacctttt tttggcggca 1500

aagaacctgt cagacgccat tctgctgagt gatattctgc gagtgaacac ggagatcacc 1560

aaagctccgc tgagcgctag tatgatcaag cgctatgatg agcaccacca agacttgact 1620

ttgctgaagg cccttgtcag acagcaactg cctgagaagt acaaggaaat tttcttcgat 1680

cagtctaaaa atggctacgc cggatacatt gacggcggag caagccagga ggaattttac 1740

aaatttatta agcccatctt ggaaaaaatg gacggcaccg aggagctgct ggtaaagctt 1800

aacagagaag atctgttgcg caaacagcgc actttcgaca atggaagcat cccccaccag 1860

attcacctgg gcgaactgca cgctatcctc aggcggcaag aggatttcta cccctttttg 1920

aaagataaca gggaaaagat tgagaaaatc ctcacatttc ggatacccta ctatgtaggc 1980

cccctcgccc ggggaaattc cagattcgcg tggatgactc gcaaatcaga agagaccatc 2040

actccctgga acttcgagga agtcgtggat aagggggcct ctgcccagtc cttcatcgaa 2100

aggatgacta actttgataa aaatctgcct aacgaaaagg tgcttcctaa acactctctg 2160

ctgtacgagt acttcacagt ttataacgag ctcaccaagg tcaaatacgt cacagaaggg 2220

atgagaaagc cagcattcct gtctggagag cagaagaaag ctatcgtgga cctcctcttc 2280

aagacgaacc ggaaagttac cgtgaaacag ctcaaagaag actatttcaa aaagattgaa 2340

tgtttcgact ctgttgaaat cagcggagtg gaggatcgct tcaacgcatc cctgggaacg 2400

tatcacgatc tcctgaaaat cattaaagac aaggacttcc tggacaatga ggagaacgag 2460

gacattcttg aggacattgt cctcaccctt acgttgtttg aagataggga gatgattgaa 2520

gaacgcttga aaacttacgc tcatctcttc gacgacaaag tcatgaaaca gctcaagagg 2580

cgccgatata caggatgggg gcggctgtca agaaaactga tcaatgggat ccgagacaag 2640

cagagtggaa agacaatcct ggattttctt aagtccgatg gatttgccaa ccggaacttc 2700

atgcagttga tccatgatga ctctctcacc tttaaggagg acatccagaa agcacaagtt 2760

tctggccagg gggacagtct tcacgagcac atcgctaatc ttgcaggtag cccagctatc 2820

aaaaagggaa tactgcagac cgttaaggtc gtggatgaac tcgtcaaagt aatgggaagg 2880

cataagcccg agaatatcgt tatcgagatg gcccgagaga accaaactac ccagaaggga 2940

cagaagaaca gtagggaaag gatgaagagg attgaagagg gtataaaaga actggggtcc 3000

caaatcctta aggaacaccc agttgaaaac acccagcttc agaatgagaa gctctacctg 3060

tactacctgc agaacggcag ggacatgtac gtggatcagg aactggacat caatcggctc 3120

tccgactacg acgtggatca tatcgtgccc cagtcttttc tcaaagatga ttctattgat 3180

aataaagtgt tgacaagatc cgataaaaat agagggaaga gtgataacgt cccctcagaa 3240

gaagttgtca agaaaatgaa aaattattgg cggcagctgc tgaacgccaa actgatcaca 3300

caacggaagt tcgataatct gactaaggct gaacgaggtg gcctgtctga gttggataaa 3360

gcaggcttca tcaaaaggca gcttgttgag acacgccaga tcaccaagca cgtggcccaa 3420

attctcgatt cacgcatgaa caccaagtac gatgaaaatg acaaactgat tcgagaggtg 3480

aaagttatta ctctgaagtc taagctggtc tcagatttca gaaaggactt tcagttttat 3540

aaggtgagag agatcaacaa ttaccaccat gcgcatgatg cctacctgaa tgcagtggta 3600

ggcactgcac ttatcaaaaa atatcccaag cttgaatctg aatttgttta cggagactat 3660

aaagtgtacg atgttaggaa aatgatcgca aagtctgagc aggaaatagg caaggccacc 3720

gctaagtact tcttttacag caatattatg aattttttca agaccgagat tacactggcc 3780

aatggagaga ttcggaagcg accacttatc gaaacaaacg gagaaacagg agaaatcgtg 3840

tgggacaagg gtagggattt cgcgacagtc cggaaggtcc tgtccatgcc gcaggtgaac 3900

atcgttaaaa agaccgaagt acagaccgga ggcttctcca aggaaagtat cctcccgaaa 3960

aggaacagcg acaagctgat cgcacgcaaa aaagattggg accccaagaa atacggcgga 4020

ttcgattctc ctacagtcgc ttacagtgta ctggttgtgg ccaaagtgga gaaagggaag 4080

tctaaaaaac tcaaaagcgt caaggaactg ctgggcatca caatcatgga gcgatcaagc 4140

ttcgaaaaaa accccatcga ctttctcgag gcgaaaggat ataaagaggt caaaaaagac 4200

ctcatcatta agcttcccaa gtactctctc tttgagcttg aaaacggccg gaaacgaatg 4260

ctcgctagtg cgggcgagct gcagaaaggt aacgagctgg cactgccctc taaatacgtt 4320

aatttcttgt atctggccag ccactatgaa aagctcaaag ggtctcccga agataatgag 4380

cagaagcagc tgttcgtgga acaacacaaa cactaccttg atgagatcat cgagcaaata 4440

agcgaattct ccaaaagagt gatcctcgcc gacgctaacc tcgataaggt gctttctgct 4500

tacaataagc acagggataa gcccatcagg gagcaggcag aaaacattat ccacttgttt 4560

actctgacca acttgggcgc gcctgcagcc ttcaagtact tcgacaccac catagacaga 4620

aagcggtaca cctctacaaa ggaggtcctg gacgccacac tgattcatca gtcaattacg 4680

gggctctatg aaacaagaat cgacctctct cagctcggtg gagac 4725

<210> 2

<211> 1575

<212> PRT

<213> b

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Met Gly Ser Ser His His His His His His Ser Glu Val Glu Phe Ser

1 5 10 15

His Glu Tyr Trp Met Arg His Ala Leu Thr Leu Ala Lys Arg Ala Arg

20 25 30

Asp Glu Arg Glu Val Pro Val Gly Ala Val Leu Val Leu Asn Asn Arg

35 40 45

Val Ile Gly Glu Gly Trp Asn Arg Ala Ile Gly Leu His Asp Pro Thr

50 55 60

Ala His Ala Glu Ile Met Ala Leu Arg Gln Gly Gly Leu Val Met Gln

65 70 75 80

Asn Tyr Arg Leu Ile Asp Ala Thr Leu Tyr Val Thr Phe Glu Pro Cys

85 90 95

Val Met Cys Ala Gly Ala Met Ile His Ser Arg Ile Gly Arg Val Val

100 105 110

Phe Gly Val Arg Asn Ser Lys Arg Gly Ala Ala Gly Ser Leu Met Asn

115 120 125

Val Leu Asn Tyr Pro Gly Met Asn His Arg Val Glu Ile Thr Glu Gly

130 135 140

Ile Leu Ala Asp Glu Cys Ala Ala Leu Leu Cys Asp Phe Tyr Arg Met

145 150 155 160

Pro Arg Gln Val Phe Asn Ala Gln Lys Lys Ala Gln Ser Ser Ile Asn

165 170 175

Ser Gly Gly Ser Ser Gly Gly Ser Ser Gly Ser Glu Thr Pro Gly Thr

180 185 190

Ser Glu Ser Ala Thr Pro Glu Ser Ser Gly Gly Ser Ser Gly Gly Ser

195 200 205

Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser Val Gly

210 215 220

Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe Lys

225 230 235 240

Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile Gly

245 250 255

Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu Lys

260 265 270

Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys Tyr

275 280 285

Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser Phe

290 295 300

Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys His

305 310 315 320

Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr His

325 330 335

Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp Ser

340 345 350

Thr Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His Met

355 360 365

Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro Asp

370 375 380

Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr Asn

385 390 395 400

Gln Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala Lys

405 410 415

Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn Leu

420 425 430

Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn Leu

435 440 445

Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe Asp

450 455 460

Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp Asp

465 470 475 480

Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp Leu

485 490 495

Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp Ile

500 505 510

Leu Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser Met

515 520 525

Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr Leu Leu Lys Ala

530 535 540

Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe Asp

545 550 555 560

Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser Gln

565 570 575

Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly

580 585 590

Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys

595 600 605

Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly

610 615 620

Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe Leu

625 630 635 640

Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro

645 650 655

Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met

660 665 670

Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu Val

675 680 685

Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn

690 695 700

Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser Leu

705 710 715 720

Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr

725 730 735

Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln Lys

740 745 750

Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val

755 760 765

Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser

770 775 780

Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr

785 790 795 800

Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn

805 810 815

Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu

820 825 830

Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His

835 840 845

Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr Thr

850 855 860

Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp Lys

865 870 875 880

Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ala

885 890 895

Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys

900 905 910

Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His

915 920 925

Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile

930 935 940

Leu Gln Thr Val Lys Val Val Asp Glu Leu Val Lys Val Met Gly Arg

945 950 955 960

His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln Thr

965 970 975

Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile Glu

980 985 990

Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro Val

995 1000 1005

Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu Gln

1010 1015 1020

Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg Leu

1025 1030 1035 1040

Ser Asp Tyr Asp Val Asp His Ile Val Pro Gln Ser Phe Leu Lys Asp

1045 1050 1055

Asp Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg Gly

1060 1065 1070

Lys Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys Lys Met Lys Asn

1075 1080 1085

Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys Phe

1090 1095 1100

Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp Lys

1105 1110 1115 1120

Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr Lys

1125 1130 1135

His Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp Glu

1140 1145 1150

Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser Lys

1155 1160 1165

Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg Glu

1170 1175 1180

Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu Asn Ala Val Val

1185 1190 1195 1200

Gly Thr Ala Leu Ile Lys Lys Tyr Pro Lys Leu Glu Ser Glu Phe Val

1205 1210 1215

Tyr Gly Asp Tyr Lys Val Tyr Asp Val Arg Lys Met Ile Ala Lys Ser

1220 1225 1230

Glu Gln Glu Ile Gly Lys Ala Thr Ala Lys Tyr Phe Phe Tyr Ser Asn

1235 1240 1245

Ile Met Asn Phe Phe Lys Thr Glu Ile Thr Leu Ala Asn Gly Glu Ile

1250 1255 1260

Arg Lys Arg Pro Leu Ile Glu Thr Asn Gly Glu Thr Gly Glu Ile Val

1265 1270 1275 1280

Trp Asp Lys Gly Arg Asp Phe Ala Thr Val Arg Lys Val Leu Ser Met

1285 1290 1295

Pro Gln Val Asn Ile Val Lys Lys Thr Glu Val Gln Thr Gly Gly Phe

1300 1305 1310

Ser Lys Glu Ser Ile Leu Pro Lys Arg Asn Ser Asp Lys Leu Ile Ala

1315 1320 1325

Arg Lys Lys Asp Trp Asp Pro Lys Lys Tyr Gly Gly Phe Asp Ser Pro

1330 1335 1340

Thr Val Ala Tyr Ser Val Leu Val Val Ala Lys Val Glu Lys Gly Lys

1345 1350 1355 1360

Ser Lys Lys Leu Lys Ser Val Lys Glu Leu Leu Gly Ile Thr Ile Met

1365 1370 1375

Glu Arg Ser Ser Phe Glu Lys Asn Pro Ile Asp Phe Leu Glu Ala Lys

1380 1385 1390

Gly Tyr Lys Glu Val Lys Lys Asp Leu Ile Ile Lys Leu Pro Lys Tyr

1395 1400 1405

Ser Leu Phe Glu Leu Glu Asn Gly Arg Lys Arg Met Leu Ala Ser Ala

1410 1415 1420

Gly Glu Leu Gln Lys Gly Asn Glu Leu Ala Leu Pro Ser Lys Tyr Val

1425 1430 1435 1440

Asn Phe Leu Tyr Leu Ala Ser His Tyr Glu Lys Leu Lys Gly Ser Pro

1445 1450 1455

Glu Asp Asn Glu Gln Lys Gln Leu Phe Val Glu Gln His Lys His Tyr

1460 1465 1470

Leu Asp Glu Ile Ile Glu Gln Ile Ser Glu Phe Ser Lys Arg Val Ile

1475 1480 1485

Leu Ala Asp Ala Asn Leu Asp Lys Val Leu Ser Ala Tyr Asn Lys His

1490 1495 1500

Arg Asp Lys Pro Ile Arg Glu Gln Ala Glu Asn Ile Ile His Leu Phe

1505 1510 1515 1520

Thr Leu Thr Asn Leu Gly Ala Pro Ala Ala Phe Lys Tyr Phe Asp Thr

1525 1530 1535

Thr Ile Asp Arg Lys Arg Tyr Thr Ser Thr Lys Glu Val Leu Asp Ala

1540 1545 1550

Thr Leu Ile His Gln Ser Ile Thr Gly Leu Tyr Glu Thr Arg Ile Asp

1555 1560 1565

Leu Ser Gln Leu Gly Gly Asp

1570 1575

<210> 3

<211> 4641

<212> DNA

<213> c

<400> 3

atgggcagca gccatcatca tcatcatcac tcggaggtcg agttttcaca tgagtattgg 60

atgcgtcatg ccttgacgtt ggcgaaacgg gcgcgcgatg agcgtgaggt gcccgtggga 120

gcggtgttgg tactgaataa ccgggttata ggggaaggat ggaaccgggc tattgggtta 180

cacgacccaa cggcgcacgc cgagataatg gcactgcgcc aagggggctt agttatgcag 240

aattatcgcc ttatcgatgc tacactgtat gtaacctttg aaccctgcgt aatgtgtgcg 300

ggggctatga tccactcgag aatagggcgc gtggtattcg gcgtacgcaa cagcaaacgt 360

ggggctgcgg gctcgttgat gaacgttctg aactaccccg gaatgaatca cagagtagag 420

atcacggagg gaattttggc cgacgaatgt gcagctttac tgtgcgattt ttatcggatg 480

ccgcggcaag tcttcaacgc acagaagaag gctcaatctt ccattaacga aacccgcagc 540

gataagaaat actcaatagg cttagctatc ggcacaaaca gcgtcggctg ggccgtcatt 600

acggacgagt acaaggtgcc gagcaaaaaa ttcaaagttc tgggcaatac cgatcgccac 660

agcataaaga agaacctcat tggcgccctc ctgttcgact ccggggagac ggccgaagcc 720

acgcggctca aaagaacagc acggcgcaga tatacccgca gaaagaatcg gatctgctac 780

ctgcaggaga tctttagtaa tgagatggct aaggtggatg actctttctt ccataggctg 840

gaggagtcct ttttggtgga ggaggataaa aagcacgagc gccacccaat ctttggcaat 900

atcgtggacg aggtggcgta ccatgaaaag tacccaacca tatatcatct gaggaagaag 960

cttgtagaca gtactgataa ggctgacttg cggttgatct atctcgcgct ggcgcatatg 1020

atcaaatttc ggggacactt cctcatcgag ggggacctga acccagacaa cagcgatgtc 1080

gacaaactct ttatccaact ggttcagact tacaatcagc ttttcgaaga gaacccgatc 1140

aacgcatccg gagttgacgc caaagcaatc ctgagcgcta ggctgtccaa atcccggcgg 1200

ctcgaaaacc tcatcgcaca gctccctggg gagaagaaga acggcctgtt tggtaatctt 1260

atcgccctgt cactcgggct gacccccaac tttaaatcta acttcgacct ggccgaagat 1320

gccaagcttc aactgagcaa agacacctac gatgatgatc tcgacaatct gctggcccag 1380

atcggcgacc agtacgcaga cctttttttg gcggcaaaga acctgtcaga cgccattctg 1440

ctgagtgata ttctgcgagt gaacacggag atcaccaaag ctccgctgag cgctagtatg 1500

atcaagcgct atgatgagca ccaccaagac ttgactttgc tgaaggccct tgtcagacag 1560

caactgcctg agaagtacaa ggaaattttc ttcgatcagt ctaaaaatgg ctacgccgga 1620

tacattgacg gcggagcaag ccaggaggaa ttttacaaat ttattaagcc catcttggaa 1680

aaaatggacg gcaccgagga gctgctggta aagcttaaca gagaagatct gttgcgcaaa 1740

cagcgcactt tcgacaatgg aagcatcccc caccagattc acctgggcga actgcacgct 1800

atcctcaggc ggcaagagga tttctacccc tttttgaaag ataacaggga aaagattgag 1860

aaaatcctca catttcggat accctactat gtaggccccc tcgcccgggg aaattccaga 1920

ttcgcgtgga tgactcgcaa atcagaagag accatcactc cctggaactt cgaggaagtc 1980

gtggataagg gggcctctgc ccagtccttc atcgaaagga tgactaactt tgataaaaat 2040

ctgcctaacg aaaaggtgct tcctaaacac tctctgctgt acgagtactt cacagtttat 2100

aacgagctca ccaaggtcaa atacgtcaca gaagggatga gaaagccagc attcctgtct 2160

ggagagcaga agaaagctat cgtggacctc ctcttcaaga cgaaccggaa agttaccgtg 2220

aaacagctca aagaagacta tttcaaaaag attgaatgtt tcgactctgt tgaaatcagc 2280

ggagtggagg atcgcttcaa cgcatccctg ggaacgtatc acgatctcct gaaaatcatt 2340

aaagacaagg acttcctgga caatgaggag aacgaggaca ttcttgagga cattgtcctc 2400

acccttacgt tgtttgaaga tagggagatg attgaagaac gcttgaaaac ttacgctcat 2460

ctcttcgacg acaaagtcat gaaacagctc aagaggcgcc gatatacagg atgggggcgg 2520

ctgtcaagaa aactgatcaa tgggatccga gacaagcaga gtggaaagac aatcctggat 2580

tttcttaagt ccgatggatt tgccaaccgg aacttcatgc agttgatcca tgatgactct 2640

ctcaccttta aggaggacat ccagaaagca caagtttctg gccaggggga cagtcttcac 2700

gagcacatcg ctaatcttgc aggtagccca gctatcaaaa agggaatact gcagaccgtt 2760

aaggtcgtgg atgaactcgt caaagtaatg ggaaggcata agcccgagaa tatcgttatc 2820

gagatggccc gagagaacca aactacccag aagggacaga agaacagtag ggaaaggatg 2880

aagaggattg aagagggtat aaaagaactg gggtcccaaa tccttaagga acacccagtt 2940

gaaaacaccc agcttcagaa tgagaagctc tacctgtact acctgcagaa cggcagggac 3000

atgtacgtgg atcaggaact ggacatcaat cggctctccg actacgacgt ggatcatatc 3060

gtgccccagt cttttctcaa agatgattct attgataata aagtgttgac aagatccgat 3120

aaaaatagag ggaagagtga taacgtcccc tcagaagaag ttgtcaagaa aatgaaaaat 3180

tattggcggc agctgctgaa cgccaaactg atcacacaac ggaagttcga taatctgact 3240

aaggctgaac gaggtggcct gtctgagttg gataaagcag gcttcatcaa aaggcagctt 3300

gttgagacac gccagatcac caagcacgtg gcccaaattc tcgattcacg catgaacacc 3360

aagtacgatg aaaatgacaa actgattcga gaggtgaaag ttattactct gaagtctaag 3420

ctggtctcag atttcagaaa ggactttcag ttttataagg tgagagagat caacaattac 3480

caccatgcgc atgatgccta cctgaatgca gtggtaggca ctgcacttat caaaaaatat 3540

cccaagcttg aatctgaatt tgtttacgga gactataaag tgtacgatgt taggaaaatg 3600

atcgcaaagt ctgagcagga aataggcaag gccaccgcta agtacttctt ttacagcaat 3660

attatgaatt ttttcaagac cgagattaca ctggccaatg gagagattcg gaagcgacca 3720

cttatcgaaa caaacggaga aacaggagaa atcgtgtggg acaagggtag ggatttcgcg 3780

acagtccgga aggtcctgtc catgccgcag gtgaacatcg ttaaaaagac cgaagtacag 3840

accggaggct tctccaagga aagtatcctc ccgaaaagga acagcgacaa gctgatcgca 3900

cgcaaaaaag attgggaccc caagaaatac ggcggattcg attctcctac agtcgcttac 3960

agtgtactgg ttgtggccaa agtggagaaa gggaagtcta aaaaactcaa aagcgtcaag 4020

gaactgctgg gcatcacaat catggagcga tcaagcttcg aaaaaaaccc catcgacttt 4080

ctcgaggcga aaggatataa agaggtcaaa aaagacctca tcattaagct tcccaagtac 4140

tctctctttg agcttgaaaa cggccggaaa cgaatgctcg ctagtgcggg cgagctgcag 4200

aaaggtaacg agctggcact gccctctaaa tacgttaatt tcttgtatct ggccagccac 4260

tatgaaaagc tcaaagggtc tcccgaagat aatgagcaga agcagctgtt cgtggaacaa 4320

cacaaacact accttgatga gatcatcgag caaataagcg aattctccaa aagagtgatc 4380

ctcgccgacg ctaacctcga taaggtgctt tctgcttaca ataagcacag ggataagccc 4440

atcagggagc aggcagaaaa cattatccac ttgtttactc tgaccaactt gggcgcgcct 4500

gcagccttca agtacttcga caccaccata gacagaaagc ggtacacctc tacaaaggag 4560

gtcctggacg ccacactgat tcatcagtca attacggggc tctatgaaac aagaatcgac 4620

ctctctcagc tcggtggaga c 4641

<210> 4

<211> 1547

<212> PRT

<213> d

<400> 4

Met Gly Ser Ser His His His His His His Ser Glu Val Glu Phe Ser

1 5 10 15

His Glu Tyr Trp Met Arg His Ala Leu Thr Leu Ala Lys Arg Ala Arg

20 25 30

Asp Glu Arg Glu Val Pro Val Gly Ala Val Leu Val Leu Asn Asn Arg

35 40 45

Val Ile Gly Glu Gly Trp Asn Arg Ala Ile Gly Leu His Asp Pro Thr

50 55 60

Ala His Ala Glu Ile Met Ala Leu Arg Gln Gly Gly Leu Val Met Gln

65 70 75 80

Asn Tyr Arg Leu Ile Asp Ala Thr Leu Tyr Val Thr Phe Glu Pro Cys

85 90 95

Val Met Cys Ala Gly Ala Met Ile His Ser Arg Ile Gly Arg Val Val

100 105 110

Phe Gly Val Arg Asn Ser Lys Arg Gly Ala Ala Gly Ser Leu Met Asn

115 120 125

Val Leu Asn Tyr Pro Gly Met Asn His Arg Val Glu Ile Thr Glu Gly

130 135 140

Ile Leu Ala Asp Glu Cys Ala Ala Leu Leu Cys Asp Phe Tyr Arg Met

145 150 155 160

Pro Arg Gln Val Phe Asn Ala Gln Lys Lys Ala Gln Ser Ser Ile Asn

165 170 175

Glu Thr Arg Ser Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr

180 185 190

Asn Ser Val Gly Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser

195 200 205

Lys Lys Phe Lys Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys

210 215 220

Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala

225 230 235 240

Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn

245 250 255

Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val

260 265 270

Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu

275 280 285

Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu

290 295 300

Val Ala Tyr His Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys

305 310 315 320

Leu Val Asp Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala

325 330 335

Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp

340 345 350

Leu Asn Pro Asp Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val

355 360 365

Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly

370 375 380

Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg

385 390 395 400

Leu Glu Asn Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu

405 410 415

Phe Gly Asn Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys

420 425 430

Ser Asn Phe Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp

435 440 445

Thr Tyr Asp Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln

450 455 460

Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu

465 470 475 480

Leu Ser Asp Ile Leu Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu

485 490 495

Ser Ala Ser Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr

500 505 510

Leu Leu Lys Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu

515 520 525

Ile Phe Phe Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly

530 535 540

Gly Ala Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu

545 550 555 560

Lys Met Asp Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp

565 570 575

Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln

580 585 590

Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe

595 600 605

Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr

610 615 620

Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg

625 630 635 640

Phe Ala Trp Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn

645 650 655

Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu

660 665 670

Arg Met Thr Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro

675 680 685

Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr

690 695 700

Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser

705 710 715 720

Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg

725 730 735

Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu

740 745 750

Cys Phe Asp Ser Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala

755 760 765

Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp

770 775 780

Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu

785 790 795 800

Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys

805 810 815

Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg

820 825 830

Arg Arg Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly

835 840 845

Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser

850 855 860

Asp Gly Phe Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser

865 870 875 880

Leu Thr Phe Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly

885 890 895

Asp Ser Leu His Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile

900 905 910

Lys Lys Gly Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu Val Lys

915 920 925

Val Met Gly Arg His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg

930 935 940

Glu Asn Gln Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met

945 950 955 960

Lys Arg Ile Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys

965 970 975

Glu His Pro Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu

980 985 990

Tyr Tyr Leu Gln Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp

995 1000 1005

Ile Asn Arg Leu Ser Asp Tyr Asp Val Asp His Ile Val Pro Gln Ser

1010 1015 1020

Phe Leu Lys Asp Asp Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp

1025 1030 1035 1040

Lys Asn Arg Gly Lys Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys

1045 1050 1055

Lys Met Lys Asn Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr

1060 1065 1070

Gln Arg Lys Phe Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser

1075 1080 1085

Glu Leu Asp Lys Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg

1090 1095 1100

Gln Ile Thr Lys His Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr

1105 1110 1115 1120

Lys Tyr Asp Glu Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr

1125 1130 1135

Leu Lys Ser Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr

1140 1145 1150

Lys Val Arg Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu

1155 1160 1165

Asn Ala Val Val Gly Thr Ala Leu Ile Lys Lys Tyr Pro Lys Leu Glu

1170 1175 1180

Ser Glu Phe Val Tyr Gly Asp Tyr Lys Val Tyr Asp Val Arg Lys Met

1185 1190 1195 1200

Ile Ala Lys Ser Glu Gln Glu Ile Gly Lys Ala Thr Ala Lys Tyr Phe

1205 1210 1215

Phe Tyr Ser Asn Ile Met Asn Phe Phe Lys Thr Glu Ile Thr Leu Ala

1220 1225 1230

Asn Gly Glu Ile Arg Lys Arg Pro Leu Ile Glu Thr Asn Gly Glu Thr

1235 1240 1245

Gly Glu Ile Val Trp Asp Lys Gly Arg Asp Phe Ala Thr Val Arg Lys

1250 1255 1260

Val Leu Ser Met Pro Gln Val Asn Ile Val Lys Lys Thr Glu Val Gln

1265 1270 1275 1280

Thr Gly Gly Phe Ser Lys Glu Ser Ile Leu Pro Lys Arg Asn Ser Asp

1285 1290 1295

Lys Leu Ile Ala Arg Lys Lys Asp Trp Asp Pro Lys Lys Tyr Gly Gly

1300 1305 1310

Phe Asp Ser Pro Thr Val Ala Tyr Ser Val Leu Val Val Ala Lys Val

1315 1320 1325

Glu Lys Gly Lys Ser Lys Lys Leu Lys Ser Val Lys Glu Leu Leu Gly

1330 1335 1340

Ile Thr Ile Met Glu Arg Ser Ser Phe Glu Lys Asn Pro Ile Asp Phe

1345 1350 1355 1360

Leu Glu Ala Lys Gly Tyr Lys Glu Val Lys Lys Asp Leu Ile Ile Lys

1365 1370 1375

Leu Pro Lys Tyr Ser Leu Phe Glu Leu Glu Asn Gly Arg Lys Arg Met

1380 1385 1390

Leu Ala Ser Ala Gly Glu Leu Gln Lys Gly Asn Glu Leu Ala Leu Pro

1395 1400 1405

Ser Lys Tyr Val Asn Phe Leu Tyr Leu Ala Ser His Tyr Glu Lys Leu

1410 1415 1420

Lys Gly Ser Pro Glu Asp Asn Glu Gln Lys Gln Leu Phe Val Glu Gln

1425 1430 1435 1440

His Lys His Tyr Leu Asp Glu Ile Ile Glu Gln Ile Ser Glu Phe Ser

1445 1450 1455

Lys Arg Val Ile Leu Ala Asp Ala Asn Leu Asp Lys Val Leu Ser Ala

1460 1465 1470

Tyr Asn Lys His Arg Asp Lys Pro Ile Arg Glu Gln Ala Glu Asn Ile

1475 1480 1485

Ile His Leu Phe Thr Leu Thr Asn Leu Gly Ala Pro Ala Ala Phe Lys

1490 1495 1500

Tyr Phe Asp Thr Thr Ile Asp Arg Lys Arg Tyr Thr Ser Thr Lys Glu

1505 1510 1515 1520

Val Leu Asp Ala Thr Leu Ile His Gln Ser Ile Thr Gly Leu Tyr Glu

1525 1530 1535

Thr Arg Ile Asp Leu Ser Gln Leu Gly Gly Asp

1540 1545

<210> 5

<211> 4641

<212> DNA

<213> e

<400> 5

atgggcagca gccatcatca tcatcatcac tcggaggtcg agttttcaca tgagtattgg 60

atgcgtcatg ccttgacgtt ggcgaaacgg gcgcgcgatg agcgtgaggt gcccgtggga 120

gcggtgttgg tactgaataa ccgggttata ggggaaggat ggaaccgggc tattgggtta 180

cacgacccaa cggcgcacgc cgagataatg gcactgcgcc aagggggctt agttatgcag 240

aattatcgcc ttatcgatgc tacactgtat gtaacctttg aaccctgcgt aatgtgtgcg 300

ggggctatga tccactcgag aatagggcgc gtggtattcg gcgtacgcaa cagcaaacgt 360

ggggctgcgg gctcgttgat gaacgttctg aactaccccg gaatgaatca cagagtagag 420

atcacggagg gaattttggc cgacgaatgt gcagctttac tgtgcgattt ttatcggatg 480

ccgcggcaag tcttcaacgc acagaagaag gctcaatctt ccattaaccc ggcgccggcg 540

gataagaaat actcaatagg cttagctatc ggcacaaaca gcgtcggctg ggccgtcatt 600

acggacgagt acaaggtgcc gagcaaaaaa ttcaaagttc tgggcaatac cgatcgccac 660

agcataaaga agaacctcat tggcgccctc ctgttcgact ccggggagac ggccgaagcc 720

acgcggctca aaagaacagc acggcgcaga tatacccgca gaaagaatcg gatctgctac 780

ctgcaggaga tctttagtaa tgagatggct aaggtggatg actctttctt ccataggctg 840

gaggagtcct ttttggtgga ggaggataaa aagcacgagc gccacccaat ctttggcaat 900

atcgtggacg aggtggcgta ccatgaaaag tacccaacca tatatcatct gaggaagaag 960

cttgtagaca gtactgataa ggctgacttg cggttgatct atctcgcgct ggcgcatatg 1020

atcaaatttc ggggacactt cctcatcgag ggggacctga acccagacaa cagcgatgtc 1080

gacaaactct ttatccaact ggttcagact tacaatcagc ttttcgaaga gaacccgatc 1140

aacgcatccg gagttgacgc caaagcaatc ctgagcgcta ggctgtccaa atcccggcgg 1200

ctcgaaaacc tcatcgcaca gctccctggg gagaagaaga acggcctgtt tggtaatctt 1260

atcgccctgt cactcgggct gacccccaac tttaaatcta acttcgacct ggccgaagat 1320

gccaagcttc aactgagcaa agacacctac gatgatgatc tcgacaatct gctggcccag 1380

atcggcgacc agtacgcaga cctttttttg gcggcaaaga acctgtcaga cgccattctg 1440

ctgagtgata ttctgcgagt gaacacggag atcaccaaag ctccgctgag cgctagtatg 1500

atcaagcgct atgatgagca ccaccaagac ttgactttgc tgaaggccct tgtcagacag 1560

caactgcctg agaagtacaa ggaaattttc ttcgatcagt ctaaaaatgg ctacgccgga 1620

tacattgacg gcggagcaag ccaggaggaa ttttacaaat ttattaagcc catcttggaa 1680

aaaatggacg gcaccgagga gctgctggta aagcttaaca gagaagatct gttgcgcaaa 1740

cagcgcactt tcgacaatgg aagcatcccc caccagattc acctgggcga actgcacgct 1800

atcctcaggc ggcaagagga tttctacccc tttttgaaag ataacaggga aaagattgag 1860

aaaatcctca catttcggat accctactat gtaggccccc tcgcccgggg aaattccaga 1920

ttcgcgtgga tgactcgcaa atcagaagag accatcactc cctggaactt cgaggaagtc 1980

gtggataagg gggcctctgc ccagtccttc atcgaaagga tgactaactt tgataaaaat 2040

ctgcctaacg aaaaggtgct tcctaaacac tctctgctgt acgagtactt cacagtttat 2100

aacgagctca ccaaggtcaa atacgtcaca gaagggatga gaaagccagc attcctgtct 2160

ggagagcaga agaaagctat cgtggacctc ctcttcaaga cgaaccggaa agttaccgtg 2220

aaacagctca aagaagacta tttcaaaaag attgaatgtt tcgactctgt tgaaatcagc 2280

ggagtggagg atcgcttcaa cgcatccctg ggaacgtatc acgatctcct gaaaatcatt 2340

aaagacaagg acttcctgga caatgaggag aacgaggaca ttcttgagga cattgtcctc 2400

acccttacgt tgtttgaaga tagggagatg attgaagaac gcttgaaaac ttacgctcat 2460

ctcttcgacg acaaagtcat gaaacagctc aagaggcgcc gatatacagg atgggggcgg 2520

ctgtcaagaa aactgatcaa tgggatccga gacaagcaga gtggaaagac aatcctggat 2580

tttcttaagt ccgatggatt tgccaaccgg aacttcatgc agttgatcca tgatgactct 2640

ctcaccttta aggaggacat ccagaaagca caagtttctg gccaggggga cagtcttcac 2700

gagcacatcg ctaatcttgc aggtagccca gctatcaaaa agggaatact gcagaccgtt 2760

aaggtcgtgg atgaactcgt caaagtaatg ggaaggcata agcccgagaa tatcgttatc 2820

gagatggccc gagagaacca aactacccag aagggacaga agaacagtag ggaaaggatg 2880

aagaggattg aagagggtat aaaagaactg gggtcccaaa tccttaagga acacccagtt 2940

gaaaacaccc agcttcagaa tgagaagctc tacctgtact acctgcagaa cggcagggac 3000

atgtacgtgg atcaggaact ggacatcaat cggctctccg actacgacgt ggatcatatc 3060

gtgccccagt cttttctcaa agatgattct attgataata aagtgttgac aagatccgat 3120

aaaaatagag ggaagagtga taacgtcccc tcagaagaag ttgtcaagaa aatgaaaaat 3180

tattggcggc agctgctgaa cgccaaactg atcacacaac ggaagttcga taatctgact 3240

aaggctgaac gaggtggcct gtctgagttg gataaagcag gcttcatcaa aaggcagctt 3300

gttgagacac gccagatcac caagcacgtg gcccaaattc tcgattcacg catgaacacc 3360

aagtacgatg aaaatgacaa actgattcga gaggtgaaag ttattactct gaagtctaag 3420

ctggtctcag atttcagaaa ggactttcag ttttataagg tgagagagat caacaattac 3480

caccatgcgc atgatgccta cctgaatgca gtggtaggca ctgcacttat caaaaaatat 3540

cccaagcttg aatctgaatt tgtttacgga gactataaag tgtacgatgt taggaaaatg 3600

atcgcaaagt ctgagcagga aataggcaag gccaccgcta agtacttctt ttacagcaat 3660

attatgaatt ttttcaagac cgagattaca ctggccaatg gagagattcg gaagcgacca 3720

cttatcgaaa caaacggaga aacaggagaa atcgtgtggg acaagggtag ggatttcgcg 3780

acagtccgga aggtcctgtc catgccgcag gtgaacatcg ttaaaaagac cgaagtacag 3840

accggaggct tctccaagga aagtatcctc ccgaaaagga acagcgacaa gctgatcgca 3900

cgcaaaaaag attgggaccc caagaaatac ggcggattcg attctcctac agtcgcttac 3960

agtgtactgg ttgtggccaa agtggagaaa gggaagtcta aaaaactcaa aagcgtcaag 4020

gaactgctgg gcatcacaat catggagcga tcaagcttcg aaaaaaaccc catcgacttt 4080

ctcgaggcga aaggatataa agaggtcaaa aaagacctca tcattaagct tcccaagtac 4140

tctctctttg agcttgaaaa cggccggaaa cgaatgctcg ctagtgcggg cgagctgcag 4200

aaaggtaacg agctggcact gccctctaaa tacgttaatt tcttgtatct ggccagccac 4260

tatgaaaagc tcaaagggtc tcccgaagat aatgagcaga agcagctgtt cgtggaacaa 4320

cacaaacact accttgatga gatcatcgag caaataagcg aattctccaa aagagtgatc 4380

ctcgccgacg ctaacctcga taaggtgctt tctgcttaca ataagcacag ggataagccc 4440

atcagggagc aggcagaaaa cattatccac ttgtttactc tgaccaactt gggcgcgcct 4500

gcagccttca agtacttcga caccaccata gacagaaagc ggtacacctc tacaaaggag 4560

gtcctggacg ccacactgat tcatcagtca attacggggc tctatgaaac aagaatcgac 4620

ctctctcagc tcggtggaga c 4641

<210> 6

<211> 1547

<212> PRT

<213> f

<400> 6

Met Gly Ser Ser His His His His His His Ser Glu Val Glu Phe Ser

1 5 10 15

His Glu Tyr Trp Met Arg His Ala Leu Thr Leu Ala Lys Arg Ala Arg

20 25 30

Asp Glu Arg Glu Val Pro Val Gly Ala Val Leu Val Leu Asn Asn Arg

35 40 45

Val Ile Gly Glu Gly Trp Asn Arg Ala Ile Gly Leu His Asp Pro Thr

50 55 60

Ala His Ala Glu Ile Met Ala Leu Arg Gln Gly Gly Leu Val Met Gln

65 70 75 80

Asn Tyr Arg Leu Ile Asp Ala Thr Leu Tyr Val Thr Phe Glu Pro Cys

85 90 95

Val Met Cys Ala Gly Ala Met Ile His Ser Arg Ile Gly Arg Val Val

100 105 110

Phe Gly Val Arg Asn Ser Lys Arg Gly Ala Ala Gly Ser Leu Met Asn

115 120 125

Val Leu Asn Tyr Pro Gly Met Asn His Arg Val Glu Ile Thr Glu Gly

130 135 140

Ile Leu Ala Asp Glu Cys Ala Ala Leu Leu Cys Asp Phe Tyr Arg Met

145 150 155 160

Pro Arg Gln Val Phe Asn Ala Gln Lys Lys Ala Gln Ser Ser Ile Asn

165 170 175

Pro Ala Pro Ala Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile Gly Thr

180 185 190

Asn Ser Val Gly Trp Ala Val Ile Thr Asp Glu Tyr Lys Val Pro Ser

195 200 205

Lys Lys Phe Lys Val Leu Gly Asn Thr Asp Arg His Ser Ile Lys Lys

210 215 220

Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala Glu Ala

225 230 235 240

Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg Lys Asn

245 250 255

Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met Ala Lys Val

260 265 270

Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val Glu Glu

275 280 285

Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Ile Val Asp Glu

290 295 300

Val Ala Tyr His Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg Lys Lys

305 310 315 320

Leu Val Asp Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile Tyr Leu Ala

325 330 335

Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu Gly Asp

340 345 350

Leu Asn Pro Asp Asn Ser Asp Val Asp Lys Leu Phe Ile Gln Leu Val

355 360 365

Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Ala Ser Gly

370 375 380

Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser Arg Arg

385 390 395 400

Leu Glu Asn Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn Gly Leu

405 410 415

Phe Gly Asn Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn Phe Lys

420 425 430

Ser Asn Phe Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser Lys Asp

435 440 445

Thr Tyr Asp Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly Asp Gln

450 455 460

Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala Ile Leu

465 470 475 480

Leu Ser Asp Ile Leu Arg Val Asn Thr Glu Ile Thr Lys Ala Pro Leu

485 490 495

Ser Ala Ser Met Ile Lys Arg Tyr Asp Glu His His Gln Asp Leu Thr

500 505 510

Leu Leu Lys Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr Lys Glu

515 520 525

Ile Phe Phe Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile Asp Gly

530 535 540

Gly Ala Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile Leu Glu

545 550 555 560

Lys Met Asp Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg Glu Asp

565 570 575

Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro His Gln

580 585 590

Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu Asp Phe

595 600 605

Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile Leu Thr

610 615 620

Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn Ser Arg

625 630 635 640

Phe Ala Trp Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro Trp Asn

645 650 655

Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe Ile Glu

660 665 670

Arg Met Thr Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys Val Leu Pro

675 680 685

Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu Leu Thr

690 695 700

Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro Ala Phe Leu Ser

705 710 715 720

Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr Asn Arg

725 730 735

Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys Ile Glu

740 745 750

Cys Phe Asp Ser Val Glu Ile Ser Gly Val Glu Asp Arg Phe Asn Ala

755 760 765

Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp Lys Asp

770 775 780

Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile Val Leu

785 790 795 800

Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg Leu Lys

805 810 815

Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys Gln Leu Lys Arg

820 825 830

Arg Arg Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile Asn Gly

835 840 845

Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu Lys Ser

850 855 860

Asp Gly Phe Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp Asp Ser

865 870 875 880

Leu Thr Phe Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly Gln Gly

885 890 895

Asp Ser Leu His Glu His Ile Ala Asn Leu Ala Gly Ser Pro Ala Ile

900 905 910

Lys Lys Gly Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu Val Lys

915 920 925

Val Met Gly Arg His Lys Pro Glu Asn Ile Val Ile Glu Met Ala Arg

930 935 940

Glu Asn Gln Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu Arg Met

945 950 955 960

Lys Arg Ile Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile Leu Lys

965 970 975

Glu His Pro Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu Tyr Leu

980 985 990

Tyr Tyr Leu Gln Asn Gly Arg Asp Met Tyr Val Asp Gln Glu Leu Asp

995 1000 1005

Ile Asn Arg Leu Ser Asp Tyr Asp Val Asp His Ile Val Pro Gln Ser

1010 1015 1020

Phe Leu Lys Asp Asp Ser Ile Asp Asn Lys Val Leu Thr Arg Ser Asp

1025 1030 1035 1040

Lys Asn Arg Gly Lys Ser Asp Asn Val Pro Ser Glu Glu Val Val Lys

1045 1050 1055

Lys Met Lys Asn Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu Ile Thr

1060 1065 1070

Gln Arg Lys Phe Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly Leu Ser

1075 1080 1085

Glu Leu Asp Lys Ala Gly Phe Ile Lys Arg Gln Leu Val Glu Thr Arg

1090 1095 1100

Gln Ile Thr Lys His Val Ala Gln Ile Leu Asp Ser Arg Met Asn Thr

1105 1110 1115 1120

Lys Tyr Asp Glu Asn Asp Lys Leu Ile Arg Glu Val Lys Val Ile Thr

1125 1130 1135

Leu Lys Ser Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln Phe Tyr

1140 1145 1150

Lys Val Arg Glu Ile Asn Asn Tyr His His Ala His Asp Ala Tyr Leu

1155 1160 1165

Asn Ala Val Val Gly Thr Ala Leu Ile Lys Lys Tyr Pro Lys Leu Glu

1170 1175 1180

Ser Glu Phe Val Tyr Gly Asp Tyr Lys Val Tyr Asp Val Arg Lys Met

1185 1190 1195 1200

Ile Ala Lys Ser Glu Gln Glu Ile Gly Lys Ala Thr Ala Lys Tyr Phe

1205 1210 1215

Phe Tyr Ser Asn Ile Met Asn Phe Phe Lys Thr Glu Ile Thr Leu Ala

1220 1225 1230

Asn Gly Glu Ile Arg Lys Arg Pro Leu Ile Glu Thr Asn Gly Glu Thr

1235 1240 1245

Gly Glu Ile Val Trp Asp Lys Gly Arg Asp Phe Ala Thr Val Arg Lys

1250 1255 1260

Val Leu Ser Met Pro Gln Val Asn Ile Val Lys Lys Thr Glu Val Gln

1265 1270 1275 1280

Thr Gly Gly Phe Ser Lys Glu Ser Ile Leu Pro Lys Arg Asn Ser Asp

1285 1290 1295

Lys Leu Ile Ala Arg Lys Lys Asp Trp Asp Pro Lys Lys Tyr Gly Gly

1300 1305 1310

Phe Asp Ser Pro Thr Val Ala Tyr Ser Val Leu Val Val Ala Lys Val

1315 1320 1325

Glu Lys Gly Lys Ser Lys Lys Leu Lys Ser Val Lys Glu Leu Leu Gly

1330 1335 1340

Ile Thr Ile Met Glu Arg Ser Ser Phe Glu Lys Asn Pro Ile Asp Phe

1345 1350 1355 1360

Leu Glu Ala Lys Gly Tyr Lys Glu Val Lys Lys Asp Leu Ile Ile Lys

1365 1370 1375

Leu Pro Lys Tyr Ser Leu Phe Glu Leu Glu Asn Gly Arg Lys Arg Met

1380 1385 1390

Leu Ala Ser Ala Gly Glu Leu Gln Lys Gly Asn Glu Leu Ala Leu Pro

1395 1400 1405

Ser Lys Tyr Val Asn Phe Leu Tyr Leu Ala Ser His Tyr Glu Lys Leu

1410 1415 1420

Lys Gly Ser Pro Glu Asp Asn Glu Gln Lys Gln Leu Phe Val Glu Gln

1425 1430 1435 1440

His Lys His Tyr Leu Asp Glu Ile Ile Glu Gln Ile Ser Glu Phe Ser

1445 1450 1455

Lys Arg Val Ile Leu Ala Asp Ala Asn Leu Asp Lys Val Leu Ser Ala

1460 1465 1470

Tyr Asn Lys His Arg Asp Lys Pro Ile Arg Glu Gln Ala Glu Asn Ile

1475 1480 1485

Ile His Leu Phe Thr Leu Thr Asn Leu Gly Ala Pro Ala Ala Phe Lys

1490 1495 1500

Tyr Phe Asp Thr Thr Ile Asp Arg Lys Arg Tyr Thr Ser Thr Lys Glu

1505 1510 1515 1520

Val Leu Asp Ala Thr Leu Ile His Gln Ser Ile Thr Gly Leu Tyr Glu

1525 1530 1535

Thr Arg Ile Asp Leu Ser Gln Leu Gly Gly Asp

1540 1545

<210> 7

<211> 4647

<212> DNA

<213> g

<400> 7

atgggcagca gccatcatca tcatcatcac tcggaggtcg agttttcaca tgagtattgg 60

atgcgtcatg ccttgacgtt ggcgaaacgg gcgcgcgatg agcgtgaggt gcccgtggga 120

gcggtgttgg tactgaataa ccgggttata ggggaaggat ggaaccgggc tattgggtta 180

cacgacccaa cggcgcacgc cgagataatg gcactgcgcc aagggggctt agttatgcag 240

aattatcgcc ttatcgatgc tacactgtat gtaacctttg aaccctgcgt aatgtgtgcg 300

ggggctatga tccactcgag aatagggcgc gtggtattcg gcgtacgcaa cagcaaacgt 360

ggggctgcgg gctcgttgat gaacgttctg aactaccccg gaatgaatca cagagtagag 420

atcacggagg gaattttggc cgacgaatgt gcagctttac tgtgcgattt ttatcggatg 480

ccgcggcaag tcttcaacgc acagaagaag gctcaatctt ccattaacaa caaaacctat 540

ccgccggata agaaatactc aataggctta gctatcggca caaacagcgt cggctgggcc 600

gtcattacgg acgagtacaa ggtgccgagc aaaaaattca aagttctggg caataccgat 660

cgccacagca taaagaagaa cctcattggc gccctcctgt tcgactccgg ggagacggcc 720

gaagccacgc ggctcaaaag aacagcacgg cgcagatata cccgcagaaa gaatcggatc 780

tgctacctgc aggagatctt tagtaatgag atggctaagg tggatgactc tttcttccat 840

aggctggagg agtccttttt ggtggaggag gataaaaagc acgagcgcca cccaatcttt 900

ggcaatatcg tggacgaggt ggcgtaccat gaaaagtacc caaccatata tcatctgagg 960

aagaagcttg tagacagtac tgataaggct gacttgcggt tgatctatct cgcgctggcg 1020

catatgatca aatttcgggg acacttcctc atcgaggggg acctgaaccc agacaacagc 1080

gatgtcgaca aactctttat ccaactggtt cagacttaca atcagctttt cgaagagaac 1140

ccgatcaacg catccggagt tgacgccaaa gcaatcctga gcgctaggct gtccaaatcc 1200

cggcggctcg aaaacctcat cgcacagctc cctggggaga agaagaacgg cctgtttggt 1260

aatcttatcg ccctgtcact cgggctgacc cccaacttta aatctaactt cgacctggcc 1320

gaagatgcca agcttcaact gagcaaagac acctacgatg atgatctcga caatctgctg 1380

gcccagatcg gcgaccagta cgcagacctt tttttggcgg caaagaacct gtcagacgcc 1440

attctgctga gtgatattct gcgagtgaac acggagatca ccaaagctcc gctgagcgct 1500

agtatgatca agcgctatga tgagcaccac caagacttga ctttgctgaa ggcccttgtc 1560

agacagcaac tgcctgagaa gtacaaggaa attttcttcg atcagtctaa aaatggctac 1620

gccggataca ttgacggcgg agcaagccag gaggaatttt acaaatttat taagcccatc 1680

ttggaaaaaa tggacggcac cgaggagctg ctggtaaagc ttaacagaga agatctgttg 1740

cgcaaacagc gcactttcga caatggaagc atcccccacc agattcacct gggcgaactg 1800

cacgctatcc tcaggcggca agaggatttc tacccctttt tgaaagataa cagggaaaag 1860

attgagaaaa tcctcacatt tcggataccc tactatgtag gccccctcgc ccggggaaat 1920

tccagattcg cgtggatgac tcgcaaatca gaagagacca tcactccctg gaacttcgag 1980

gaagtcgtgg ataagggggc ctctgcccag tccttcatcg aaaggatgac taactttgat 2040

aaaaatctgc ctaacgaaaa ggtgcttcct aaacactctc tgctgtacga gtacttcaca 2100

gtttataacg agctcaccaa ggtcaaatac gtcacagaag ggatgagaaa gccagcattc 2160

ctgtctggag agcagaagaa agctatcgtg gacctcctct tcaagacgaa ccggaaagtt 2220

accgtgaaac agctcaaaga agactatttc aaaaagattg aatgtttcga ctctgttgaa 2280

atcagcggag tggaggatcg cttcaacgca tccctgggaa cgtatcacga tctcctgaaa 2340

atcattaaag acaaggactt cctggacaat gaggagaacg aggacattct tgaggacatt 2400

gtcctcaccc ttacgttgtt tgaagatagg gagatgattg aagaacgctt gaaaacttac 2460

gctcatctct tcgacgacaa agtcatgaaa cagctcaaga ggcgccgata tacaggatgg 2520

gggcggctgt caagaaaact gatcaatggg atccgagaca agcagagtgg aaagacaatc 2580

ctggattttc ttaagtccga tggatttgcc aaccggaact tcatgcagtt gatccatgat 2640

gactctctca cctttaagga ggacatccag aaagcacaag tttctggcca gggggacagt 2700

cttcacgagc acatcgctaa tcttgcaggt agcccagcta tcaaaaaggg aatactgcag 2760

accgttaagg tcgtggatga actcgtcaaa gtaatgggaa ggcataagcc cgagaatatc 2820

gttatcgaga tggcccgaga gaaccaaact acccagaagg gacagaagaa cagtagggaa 2880

aggatgaaga ggattgaaga gggtataaaa gaactggggt cccaaatcct taaggaacac 2940

ccagttgaaa acacccagct tcagaatgag aagctctacc tgtactacct gcagaacggc 3000

agggacatgt acgtggatca ggaactggac atcaatcggc tctccgacta cgacgtggat 3060

catatcgtgc cccagtcttt tctcaaagat gattctattg ataataaagt gttgacaaga 3120

tccgataaaa atagagggaa gagtgataac gtcccctcag aagaagttgt caagaaaatg 3180

aaaaattatt ggcggcagct gctgaacgcc aaactgatca cacaacggaa gttcgataat 3240

ctgactaagg ctgaacgagg tggcctgtct gagttggata aagcaggctt catcaaaagg 3300

cagcttgttg agacacgcca gatcaccaag cacgtggccc aaattctcga ttcacgcatg 3360

aacaccaagt acgatgaaaa tgacaaactg attcgagagg tgaaagttat tactctgaag 3420

tctaagctgg tctcagattt cagaaaggac tttcagtttt ataaggtgag agagatcaac 3480

aattaccacc atgcgcatga tgcctacctg aatgcagtgg taggcactgc acttatcaaa 3540

aaatatccca agcttgaatc tgaatttgtt tacggagact ataaagtgta cgatgttagg 3600

aaaatgatcg caaagtctga gcaggaaata ggcaaggcca ccgctaagta cttcttttac 3660

agcaatatta tgaatttttt caagaccgag attacactgg ccaatggaga gattcggaag 3720

cgaccactta tcgaaacaaa cggagaaaca ggagaaatcg tgtgggacaa gggtagggat 3780

ttcgcgacag tccggaaggt cctgtccatg ccgcaggtga acatcgttaa aaagaccgaa 3840

gtacagaccg gaggcttctc caaggaaagt atcctcccga aaaggaacag cgacaagctg 3900

atcgcacgca aaaaagattg ggaccccaag aaatacggcg gattcgattc tcctacagtc 3960

gcttacagtg tactggttgt ggccaaagtg gagaaaggga agtctaaaaa actcaaaagc 4020

gtcaaggaac tgctgggcat cacaatcatg gagcgatcaa gcttcgaaaa aaaccccatc 4080

gactttctcg aggcgaaagg atataaagag gtcaaaaaag acctcatcat taagcttccc 4140

aagtactctc tctttgagct tgaaaacggc cggaaacgaa tgctcgctag tgcgggcgag 4200

ctgcagaaag gtaacgagct ggcactgccc tctaaatacg ttaatttctt gtatctggcc 4260

agccactatg aaaagctcaa agggtctccc gaagataatg agcagaagca gctgttcgtg 4320

gaacaacaca aacactacct tgatgagatc atcgagcaaa taagcgaatt ctccaaaaga 4380

gtgatcctcg ccgacgctaa cctcgataag gtgctttctg cttacaataa gcacagggat 4440

aagcccatca gggagcaggc agaaaacatt atccacttgt ttactctgac caacttgggc 4500

gcgcctgcag ccttcaagta cttcgacacc accatagaca gaaagcggta cacctctaca 4560

aaggaggtcc tggacgccac actgattcat cagtcaatta cggggctcta tgaaacaaga 4620

atcgacctct ctcagctcgg tggagac 4647

<210> 8

<211> 1549

<212> PRT

<213> h

<400> 8

Met Gly Ser Ser His His His His His His Ser Glu Val Glu Phe Ser

1 5 10 15

His Glu Tyr Trp Met Arg His Ala Leu Thr Leu Ala Lys Arg Ala Arg

20 25 30

Asp Glu Arg Glu Val Pro Val Gly Ala Val Leu Val Leu Asn Asn Arg

35 40 45

Val Ile Gly Glu Gly Trp Asn Arg Ala Ile Gly Leu His Asp Pro Thr

50 55 60

Ala His Ala Glu Ile Met Ala Leu Arg Gln Gly Gly Leu Val Met Gln

65 70 75 80

Asn Tyr Arg Leu Ile Asp Ala Thr Leu Tyr Val Thr Phe Glu Pro Cys

85 90 95

Val Met Cys Ala Gly Ala Met Ile His Ser Arg Ile Gly Arg Val Val

100 105 110

Phe Gly Val Arg Asn Ser Lys Arg Gly Ala Ala Gly Ser Leu Met Asn

115 120 125

Val Leu Asn Tyr Pro Gly Met Asn His Arg Val Glu Ile Thr Glu Gly

130 135 140

Ile Leu Ala Asp Glu Cys Ala Ala Leu Leu Cys Asp Phe Tyr Arg Met

145 150 155 160

Pro Arg Gln Val Phe Asn Ala Gln Lys Lys Ala Gln Ser Ser Ile Asn

165 170 175

Asn Lys Thr Tyr Pro Pro Asp Lys Lys Tyr Ser Ile Gly Leu Ala Ile

180 185 190

Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Glu Tyr Lys Val

195 200 205

Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asp Arg His Ser Ile

210 215 220

Lys Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr Ala

225 230 235 240

Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg Arg

245 250 255

Lys Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met Ala

260 265 270

Lys Val Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu Val

275 280 285

Glu Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Ile Val

290 295 300

Asp Glu Val Ala Tyr His Glu Lys Tyr Pro Thr Ile Tyr His Leu Arg

305 310 315 320

Lys Lys Leu Val Asp Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile Tyr

325 330 335

Leu Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile Glu

340 345 350

Gly Asp Leu Asn Pro Asp Asn Ser Asp Val Asp Lys Leu Phe Ile Gln

355 360 365

Leu Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn Ala

370 375 380

Ser Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys Ser

385 390 395 400

Arg Arg Leu Glu Asn Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys Asn

405 410 415

Gly Leu Phe Gly Asn Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro Asn

420 425 430

Phe Lys Ser Asn Phe Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu Ser

435 440 445

Lys Asp Thr Tyr Asp Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile Gly

450 455 460

Asp Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp Ala

465 470 475 480

Ile Leu Leu Ser Asp Ile Leu Arg Val Asn Thr Glu Ile Thr Lys Ala

485 490 495

Pro Leu Ser Ala Ser Met Ile Lys Arg Tyr Asp Glu His His Gln Asp

500 505 510

Leu Thr Leu Leu Lys Ala Leu Val Arg Gln Gln Leu Pro Glu Lys Tyr

515 520 525

Lys Glu Ile Phe Phe Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr Ile

530 535 540

Asp Gly Gly Ala Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro Ile

545 550 555 560

Leu Glu Lys Met Asp Gly Thr Glu Glu Leu Leu Val Lys Leu Asn Arg

565 570 575

Glu Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile Pro

580 585 590

His Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg Gln Glu

595 600 605

Asp Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys Ile

610 615 620

Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly Asn

625 630 635 640

Ser Arg Phe Ala Trp Met Thr Arg Lys Ser Glu Glu Thr Ile Thr Pro

645 650 655

Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln Ser Phe

660 665 670

Ile Glu Arg Met Thr Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys Val

675 680 685

Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn Glu

690 695 700

Leu Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro Ala Phe

705 710 715 720

Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe Lys Thr

725 730 735

Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys Lys

740 745 750

Ile Glu Cys Phe Asp Ser Val Glu Ile Ser Gly Val Glu Asp Arg Phe

755 760 765

Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile Lys Asp

770 775 780

Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp Ile

785 790 795 800

Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu Glu Arg

805 810 815

Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys Gln Leu

820 825 830

Lys Arg Arg Arg Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu Ile

835 840 845

Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe Leu

850 855 860

Lys Ser Asp Gly Phe Ala Asn Arg Asn Phe Met Gln Leu Ile His Asp

865 870 875 880

Asp Ser Leu Thr Phe Lys Glu Asp Ile Gln Lys Ala Gln Val Ser Gly

885 890 895

Gln Gly Asp Ser Leu His Glu His Ile Ala Asn Leu Ala Gly Ser Pro

900 905 910

Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Val Val Asp Glu Leu

915 920 925

Val Lys Val Met Gly Arg His Lys Pro Glu Asn Ile Val Ile Glu Met

930 935 940

Ala Arg Glu Asn Gln Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg Glu

945 950 955 960

Arg Met Lys Arg Ile Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln Ile

965 970 975

Leu Lys Glu His Pro Val Glu Asn Thr Gln Leu Gln Asn Glu Lys Leu

980 985 990

Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg Asp Met Tyr Val Asp Gln Glu

995 1000 1005

Leu Asp Ile Asn Arg Leu Ser Asp Tyr Asp Val Asp His Ile Val Pro

1010 1015 1020

Gln Ser Phe Leu Lys Asp Asp Ser Ile Asp Asn Lys Val Leu Thr Arg

1025 1030 1035 1040

Ser Asp Lys Asn Arg Gly Lys Ser Asp Asn Val Pro Ser Glu Glu Val

1045 1050 1055

Val Lys Lys Met Lys Asn Tyr Trp Arg Gln Leu Leu Asn Ala Lys Leu

1060 1065 1070

Ile Thr Gln Arg Lys Phe Asp Asn Leu Thr Lys Ala Glu Arg Gly Gly

1075 1080 1085

Leu Ser Glu Leu Asp Lys Ala Gly Phe Ile Lys Arg Gln Leu Val Glu

1090 1095 1100

Thr Arg Gln Ile Thr Lys His Val Ala Gln Ile Leu Asp Ser Arg Met

1105 1110 1115 1120

Asn Thr Lys Tyr Asp Glu Asn Asp Lys Leu Ile Arg Glu Val Lys Val

1125 1130 1135

Ile Thr Leu Lys Ser Lys Leu Val Ser Asp Phe Arg Lys Asp Phe Gln

1140 1145 1150

Phe Tyr Lys Val Arg Glu Ile Asn Asn Tyr His His Ala His Asp Ala

1155 1160 1165

Tyr Leu Asn Ala Val Val Gly Thr Ala Leu Ile Lys Lys Tyr Pro Lys

1170 1175 1180

Leu Glu Ser Glu Phe Val Tyr Gly Asp Tyr Lys Val Tyr Asp Val Arg

1185 1190 1195 1200

Lys Met Ile Ala Lys Ser Glu Gln Glu Ile Gly Lys Ala Thr Ala Lys

1205 1210 1215

Tyr Phe Phe Tyr Ser Asn Ile Met Asn Phe Phe Lys Thr Glu Ile Thr

1220 1225 1230

Leu Ala Asn Gly Glu Ile Arg Lys Arg Pro Leu Ile Glu Thr Asn Gly

1235 1240 1245

Glu Thr Gly Glu Ile Val Trp Asp Lys Gly Arg Asp Phe Ala Thr Val

1250 1255 1260

Arg Lys Val Leu Ser Met Pro Gln Val Asn Ile Val Lys Lys Thr Glu

1265 1270 1275 1280

Val Gln Thr Gly Gly Phe Ser Lys Glu Ser Ile Leu Pro Lys Arg Asn

1285 1290 1295

Ser Asp Lys Leu Ile Ala Arg Lys Lys Asp Trp Asp Pro Lys Lys Tyr

1300 1305 1310

Gly Gly Phe Asp Ser Pro Thr Val Ala Tyr Ser Val Leu Val Val Ala

1315 1320 1325

Lys Val Glu Lys Gly Lys Ser Lys Lys Leu Lys Ser Val Lys Glu Leu

1330 1335 1340

Leu Gly Ile Thr Ile Met Glu Arg Ser Ser Phe Glu Lys Asn Pro Ile

1345 1350 1355 1360

Asp Phe Leu Glu Ala Lys Gly Tyr Lys Glu Val Lys Lys Asp Leu Ile

1365 1370 1375

Ile Lys Leu Pro Lys Tyr Ser Leu Phe Glu Leu Glu Asn Gly Arg Lys

1380 1385 1390

Arg Met Leu Ala Ser Ala Gly Glu Leu Gln Lys Gly Asn Glu Leu Ala

1395 1400 1405

Leu Pro Ser Lys Tyr Val Asn Phe Leu Tyr Leu Ala Ser His Tyr Glu

1410 1415 1420

Lys Leu Lys Gly Ser Pro Glu Asp Asn Glu Gln Lys Gln Leu Phe Val

1425 1430 1435 1440

Glu Gln His Lys His Tyr Leu Asp Glu Ile Ile Glu Gln Ile Ser Glu

1445 1450 1455

Phe Ser Lys Arg Val Ile Leu Ala Asp Ala Asn Leu Asp Lys Val Leu

1460 1465 1470

Ser Ala Tyr Asn Lys His Arg Asp Lys Pro Ile Arg Glu Gln Ala Glu

1475 1480 1485

Asn Ile Ile His Leu Phe Thr Leu Thr Asn Leu Gly Ala Pro Ala Ala

1490 1495 1500

Phe Lys Tyr Phe Asp Thr Thr Ile Asp Arg Lys Arg Tyr Thr Ser Thr

1505 1510 1515 1520

Lys Glu Val Leu Asp Ala Thr Leu Ile His Gln Ser Ile Thr Gly Leu

1525 1530 1535

Tyr Glu Thr Arg Ile Asp Leu Ser Gln Leu Gly Gly Asp

1540 1545

<210> 9

<211> 4650

<212> DNA

<213> i

<400> 9

atgggcagca gccatcatca tcatcatcac tcggaggtcg agttttcaca tgagtattgg 60

atgcgtcatg ccttgacgtt ggcgaaacgg gcgcgcgatg agcgtgaggt gcccgtggga 120

gcggtgttgg tactgaataa ccgggttata ggggaaggat ggaaccgggc tattgggtta 180

cacgacccaa cggcgcacgc cgagataatg gcactgcgcc aagggggctt agttatgcag 240

aattatcgcc ttatcgatgc tacactgtat gtaacctttg aaccctgcgt aatgtgtgcg 300

ggggctatga tccactcgag aatagggcgc gtggtattcg gcgtacgcaa cagcaaacgt 360

ggggctgcgg gctcgttgat gaacgttctg aactaccccg gaatgaatca cagagtagag 420

atcacggagg gaattttggc cgacgaatgt gcagctttac tgtgcgattt ttatcggatg 480

ccgcggcaag tcttcaacgc acagaagaag gctcaatctt ccattaacac caccgaaccg 540

ccggataacg ataagaaata ctcaataggc ttagctatcg gcacaaacag cgtcggctgg 600

gccgtcatta cggacgagta caaggtgccg agcaaaaaat tcaaagttct gggcaatacc 660

gatcgccaca gcataaagaa gaacctcatt ggcgccctcc tgttcgactc cggggagacg 720

gccgaagcca cgcggctcaa aagaacagca cggcgcagat atacccgcag aaagaatcgg 780

atctgctacc tgcaggagat ctttagtaat gagatggcta aggtggatga ctctttcttc 840

cataggctgg aggagtcctt tttggtggag gaggataaaa agcacgagcg ccacccaatc 900

tttggcaata tcgtggacga ggtggcgtac catgaaaagt acccaaccat atatcatctg 960

aggaagaagc ttgtagacag tactgataag gctgacttgc ggttgatcta tctcgcgctg 1020

gcgcatatga tcaaatttcg gggacacttc ctcatcgagg gggacctgaa cccagacaac 1080

agcgatgtcg acaaactctt tatccaactg gttcagactt acaatcagct tttcgaagag 1140

aacccgatca acgcatccgg agttgacgcc aaagcaatcc tgagcgctag gctgtccaaa 1200

tcccggcggc tcgaaaacct catcgcacag ctccctgggg agaagaagaa cggcctgttt 1260

ggtaatctta tcgccctgtc actcgggctg acccccaact ttaaatctaa cttcgacctg 1320

gccgaagatg ccaagcttca actgagcaaa gacacctacg atgatgatct cgacaatctg 1380

ctggcccaga tcggcgacca gtacgcagac ctttttttgg cggcaaagaa cctgtcagac 1440

gccattctgc tgagtgatat tctgcgagtg aacacggaga tcaccaaagc tccgctgagc 1500

gctagtatga tcaagcgcta tgatgagcac caccaagact tgactttgct gaaggccctt 1560

gtcagacagc aactgcctga gaagtacaag gaaattttct tcgatcagtc taaaaatggc 1620

tacgccggat acattgacgg cggagcaagc caggaggaat tttacaaatt tattaagccc 1680

atcttggaaa aaatggacgg caccgaggag ctgctggtaa agcttaacag agaagatctg 1740

ttgcgcaaac agcgcacttt cgacaatgga agcatccccc accagattca cctgggcgaa 1800

ctgcacgcta tcctcaggcg gcaagaggat ttctacccct ttttgaaaga taacagggaa 1860

aagattgaga aaatcctcac atttcggata ccctactatg taggccccct cgcccgggga 1920

aattccagat tcgcgtggat gactcgcaaa tcagaagaga ccatcactcc ctggaacttc 1980

gaggaagtcg tggataaggg ggcctctgcc cagtccttca tcgaaaggat gactaacttt 2040

gataaaaatc tgcctaacga aaaggtgctt cctaaacact ctctgctgta cgagtacttc 2100

acagtttata acgagctcac caaggtcaaa tacgtcacag aagggatgag aaagccagca 2160

ttcctgtctg gagagcagaa gaaagctatc gtggacctcc tcttcaagac gaaccggaaa 2220

gttaccgtga aacagctcaa agaagactat ttcaaaaaga ttgaatgttt cgactctgtt 2280

gaaatcagcg gagtggagga tcgcttcaac gcatccctgg gaacgtatca cgatctcctg 2340

aaaatcatta aagacaagga cttcctggac aatgaggaga acgaggacat tcttgaggac 2400

attgtcctca cccttacgtt gtttgaagat agggagatga ttgaagaacg cttgaaaact 2460

tacgctcatc tcttcgacga caaagtcatg aaacagctca agaggcgccg atatacagga 2520

tgggggcggc tgtcaagaaa actgatcaat gggatccgag acaagcagag tggaaagaca 2580

atcctggatt ttcttaagtc cgatggattt gccaaccgga acttcatgca gttgatccat 2640

gatgactctc tcacctttaa ggaggacatc cagaaagcac aagtttctgg ccagggggac 2700

agtcttcacg agcacatcgc taatcttgca ggtagcccag ctatcaaaaa gggaatactg 2760

cagaccgtta aggtcgtgga tgaactcgtc aaagtaatgg gaaggcataa gcccgagaat 2820

atcgttatcg agatggcccg agagaaccaa actacccaga agggacagaa gaacagtagg 2880

gaaaggatga agaggattga agagggtata aaagaactgg ggtcccaaat ccttaaggaa 2940

cacccagttg aaaacaccca gcttcagaat gagaagctct acctgtacta cctgcagaac 3000

ggcagggaca tgtacgtgga tcaggaactg gacatcaatc ggctctccga ctacgacgtg 3060

gatcatatcg tgccccagtc ttttctcaaa gatgattcta ttgataataa agtgttgaca 3120

agatccgata aaaatagagg gaagagtgat aacgtcccct cagaagaagt tgtcaagaaa 3180

atgaaaaatt attggcggca gctgctgaac gccaaactga tcacacaacg gaagttcgat 3240

aatctgacta aggctgaacg aggtggcctg tctgagttgg ataaagcagg cttcatcaaa 3300

aggcagcttg ttgagacacg ccagatcacc aagcacgtgg cccaaattct cgattcacgc 3360

atgaacacca agtacgatga aaatgacaaa ctgattcgag aggtgaaagt tattactctg 3420

aagtctaagc tggtctcaga tttcagaaag gactttcagt tttataaggt gagagagatc 3480

aacaattacc accatgcgca tgatgcctac ctgaatgcag tggtaggcac tgcacttatc 3540

aaaaaatatc ccaagcttga atctgaattt gtttacggag actataaagt gtacgatgtt 3600

aggaaaatga tcgcaaagtc tgagcaggaa ataggcaagg ccaccgctaa gtacttcttt 3660

tacagcaata ttatgaattt tttcaagacc gagattacac tggccaatgg agagattcgg 3720

aagcgaccac ttatcgaaac aaacggagaa acaggagaaa tcgtgtggga caagggtagg 3780

gatttcgcga cagtccggaa ggtcctgtcc atgccgcagg tgaacatcgt taaaaagacc 3840

gaagtacaga ccggaggctt ctccaaggaa agtatcctcc cgaaaaggaa cagcgacaag 3900

ctgatcgcac gcaaaaaaga ttgggacccc aagaaatacg gcggattcga ttctcctaca 3960

gtcgcttaca gtgtactggt tgtggccaaa gtggagaaag ggaagtctaa aaaactcaaa 4020

agcgtcaagg aactgctggg catcacaatc atggagcgat caagcttcga aaaaaacccc 4080

atcgactttc tcgaggcgaa aggatataaa gaggtcaaaa aagacctcat cattaagctt 4140

cccaagtact ctctctttga gcttgaaaac ggccggaaac gaatgctcgc tagtgcgggc 4200

gagctgcaga aaggtaacga gctggcactg ccctctaaat acgttaattt cttgtatctg 4260

gccagccact atgaaaagct caaagggtct cccgaagata atgagcagaa gcagctgttc 4320

gtggaacaac acaaacacta ccttgatgag atcatcgagc aaataagcga attctccaaa 4380

agagtgatcc tcgccgacgc taacctcgat aaggtgcttt ctgcttacaa taagcacagg 4440

gataagccca tcagggagca ggcagaaaac attatccact tgtttactct gaccaacttg 4500

ggcgcgcctg cagccttcaa gtacttcgac accaccatag acagaaagcg gtacacctct 4560

acaaaggagg tcctggacgc cacactgatt catcagtcaa ttacggggct ctatgaaaca 4620

agaatcgacc tctctcagct cggtggagac 4650

<210> 10

<211> 1550

<212> PRT

<213> j

<400> 10

Met Gly Ser Ser His His His His His His Ser Glu Val Glu Phe Ser

1 5 10 15

His Glu Tyr Trp Met Arg His Ala Leu Thr Leu Ala Lys Arg Ala Arg

20 25 30

Asp Glu Arg Glu Val Pro Val Gly Ala Val Leu Val Leu Asn Asn Arg

35 40 45

Val Ile Gly Glu Gly Trp Asn Arg Ala Ile Gly Leu His Asp Pro Thr

50 55 60

Ala His Ala Glu Ile Met Ala Leu Arg Gln Gly Gly Leu Val Met Gln

65 70 75 80

Asn Tyr Arg Leu Ile Asp Ala Thr Leu Tyr Val Thr Phe Glu Pro Cys

85 90 95

Val Met Cys Ala Gly Ala Met Ile His Ser Arg Ile Gly Arg Val Val

100 105 110

Phe Gly Val Arg Asn Ser Lys Arg Gly Ala Ala Gly Ser Leu Met Asn

115 120 125

Val Leu Asn Tyr Pro Gly Met Asn His Arg Val Glu Ile Thr Glu Gly

130 135 140

Ile Leu Ala Asp Glu Cys Ala Ala Leu Leu Cys Asp Phe Tyr Arg Met

145 150 155 160

Pro Arg Gln Val Phe Asn Ala Gln Lys Lys Ala Gln Ser Ser Ile Asn

165 170 175

Thr Thr Glu Pro Pro Asp Asn Asp Lys Lys Tyr Ser Ile Gly Leu Ala

180 185 190

Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Glu Tyr Lys

195 200 205

Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asp Arg His Ser

210 215 220

Ile Lys Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr

225 230 235 240

Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg

245 250 255

Arg Lys Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met

260 265 270

Ala Lys Val Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu

275 280 285

Val Glu Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Ile

290 295 300

Val Asp Glu Val Ala Tyr His Glu Lys Tyr Pro Thr Ile Tyr His Leu

305 310 315 320

Arg Lys Lys Leu Val Asp Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile

325 330 335

Tyr Leu Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile

340 345 350

Glu Gly Asp Leu Asn Pro Asp Asn Ser Asp Val Asp Lys Leu Phe Ile

355 360 365

Gln Leu Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn

370 375 380

Ala Ser Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys

385 390 395 400

Ser Arg Arg Leu Glu Asn Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys

405 410 415

Asn Gly Leu Phe Gly Asn Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro

420 425 430

Asn Phe Lys Ser Asn Phe Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu

435 440 445

Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile

450 455 460

Gly Asp Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp

465 470 475 480

Ala Ile Leu Leu Ser Asp Ile Leu Arg Val Asn Thr Glu Ile Thr Lys

485 490 495

Ala Pro Leu Ser Ala Ser Met Ile Lys Arg Tyr Asp Glu His His Gln

500 505 510

Asp Leu Thr Leu Leu Lys Ala Leu Val Arg Gln Gln Leu Pro Glu Lys

515 520 525

Tyr Lys Glu Ile Phe Phe Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr

530 535 540

Ile Asp Gly Gly Ala Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro

545 550 555 560

Ile Leu Glu Lys Met Asp Gly Thr Glu Glu Leu Leu Val Lys Leu Asn

565 570 575

Arg Glu Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile

580 585 590

Pro His Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg Gln

595 600 605

Glu Asp Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys

610 615 620

Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly

625 630 635 640

Asn Ser Arg Phe Ala Trp Met Thr Arg Lys Ser Glu Glu Thr Ile Thr

645 650 655

Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln Ser

660 665 670

Phe Ile Glu Arg Met Thr Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys

675 680 685

Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn

690 695 700

Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro Ala

705 710 715 720

Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe Lys

725 730 735

Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys

740 745 750

Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ser Gly Val Glu Asp Arg

755 760 765

Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile Lys

770 775 780

Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp

785 790 795 800

Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu Glu

805 810 815

Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys Gln

820 825 830

Leu Lys Arg Arg Arg Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu

835 840 845

Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe

850 855 860

Leu Lys Ser Asp Gly Phe Ala Asn Arg Asn Phe Met Gln Leu Ile His

865 870 875 880

Asp Asp Ser Leu Thr Phe Lys Glu Asp Ile Gln Lys Ala Gln Val Ser

885 890 895

Gly Gln Gly Asp Ser Leu His Glu His Ile Ala Asn Leu Ala Gly Ser

900 905 910

Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Val Val Asp Glu

915 920 925

Leu Val Lys Val Met Gly Arg His Lys Pro Glu Asn Ile Val Ile Glu

930 935 940

Met Ala Arg Glu Asn Gln Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg

945 950 955 960

Glu Arg Met Lys Arg Ile Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln

965 970 975

Ile Leu Lys Glu His Pro Val Glu Asn Thr Gln Leu Gln Asn Glu Lys

980 985 990

Leu Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg Asp Met Tyr Val Asp Gln

995 1000 1005

Glu Leu Asp Ile Asn Arg Leu Ser Asp Tyr Asp Val Asp His Ile Val

1010 1015 1020

Pro Gln Ser Phe Leu Lys Asp Asp Ser Ile Asp Asn Lys Val Leu Thr

1025 1030 1035 1040

Arg Ser Asp Lys Asn Arg Gly Lys Ser Asp Asn Val Pro Ser Glu Glu

1045 1050 1055

Val Val Lys Lys Met Lys Asn Tyr Trp Arg Gln Leu Leu Asn Ala Lys

1060 1065 1070

Leu Ile Thr Gln Arg Lys Phe Asp Asn Leu Thr Lys Ala Glu Arg Gly

1075 1080 1085

Gly Leu Ser Glu Leu Asp Lys Ala Gly Phe Ile Lys Arg Gln Leu Val

1090 1095 1100

Glu Thr Arg Gln Ile Thr Lys His Val Ala Gln Ile Leu Asp Ser Arg

1105 1110 1115 1120

Met Asn Thr Lys Tyr Asp Glu Asn Asp Lys Leu Ile Arg Glu Val Lys

1125 1130 1135

Val Ile Thr Leu Lys Ser Lys Leu Val Ser Asp Phe Arg Lys Asp Phe

1140 1145 1150

Gln Phe Tyr Lys Val Arg Glu Ile Asn Asn Tyr His His Ala His Asp

1155 1160 1165

Ala Tyr Leu Asn Ala Val Val Gly Thr Ala Leu Ile Lys Lys Tyr Pro

1170 1175 1180

Lys Leu Glu Ser Glu Phe Val Tyr Gly Asp Tyr Lys Val Tyr Asp Val

1185 1190 1195 1200

Arg Lys Met Ile Ala Lys Ser Glu Gln Glu Ile Gly Lys Ala Thr Ala

1205 1210 1215

Lys Tyr Phe Phe Tyr Ser Asn Ile Met Asn Phe Phe Lys Thr Glu Ile

1220 1225 1230

Thr Leu Ala Asn Gly Glu Ile Arg Lys Arg Pro Leu Ile Glu Thr Asn

1235 1240 1245

Gly Glu Thr Gly Glu Ile Val Trp Asp Lys Gly Arg Asp Phe Ala Thr

1250 1255 1260

Val Arg Lys Val Leu Ser Met Pro Gln Val Asn Ile Val Lys Lys Thr

1265 1270 1275 1280

Glu Val Gln Thr Gly Gly Phe Ser Lys Glu Ser Ile Leu Pro Lys Arg

1285 1290 1295

Asn Ser Asp Lys Leu Ile Ala Arg Lys Lys Asp Trp Asp Pro Lys Lys

1300 1305 1310

Tyr Gly Gly Phe Asp Ser Pro Thr Val Ala Tyr Ser Val Leu Val Val

1315 1320 1325

Ala Lys Val Glu Lys Gly Lys Ser Lys Lys Leu Lys Ser Val Lys Glu

1330 1335 1340

Leu Leu Gly Ile Thr Ile Met Glu Arg Ser Ser Phe Glu Lys Asn Pro

1345 1350 1355 1360

Ile Asp Phe Leu Glu Ala Lys Gly Tyr Lys Glu Val Lys Lys Asp Leu

1365 1370 1375

Ile Ile Lys Leu Pro Lys Tyr Ser Leu Phe Glu Leu Glu Asn Gly Arg

1380 1385 1390

Lys Arg Met Leu Ala Ser Ala Gly Glu Leu Gln Lys Gly Asn Glu Leu

1395 1400 1405

Ala Leu Pro Ser Lys Tyr Val Asn Phe Leu Tyr Leu Ala Ser His Tyr

1410 1415 1420

Glu Lys Leu Lys Gly Ser Pro Glu Asp Asn Glu Gln Lys Gln Leu Phe

1425 1430 1435 1440

Val Glu Gln His Lys His Tyr Leu Asp Glu Ile Ile Glu Gln Ile Ser

1445 1450 1455

Glu Phe Ser Lys Arg Val Ile Leu Ala Asp Ala Asn Leu Asp Lys Val

1460 1465 1470

Leu Ser Ala Tyr Asn Lys His Arg Asp Lys Pro Ile Arg Glu Gln Ala

1475 1480 1485

Glu Asn Ile Ile His Leu Phe Thr Leu Thr Asn Leu Gly Ala Pro Ala

1490 1495 1500

Ala Phe Lys Tyr Phe Asp Thr Thr Ile Asp Arg Lys Arg Tyr Thr Ser

1505 1510 1515 1520

Thr Lys Glu Val Leu Asp Ala Thr Leu Ile His Gln Ser Ile Thr Gly

1525 1530 1535

Leu Tyr Glu Thr Arg Ile Asp Leu Ser Gln Leu Gly Gly Asp

1540 1545 1550

<210> 11

<211> 4650

<212> DNA

<213> k

<400> 11

atgggcagca gccatcatca tcatcatcac tcggaggtcg agttttcaca tgagtattgg 60

atgcgtcatg ccttgacgtt ggcgaaacgg gcgcgcgatg agcgtgaggt gcccgtggga 120

gcggtgttgg tactgaataa ccgggttata ggggaaggat ggaaccgggc tattgggtta 180

cacgacccaa cggcgcacgc cgagataatg gcactgcgcc aagggggctt agttatgcag 240

aattatcgcc ttatcgatgc tacactgtat gtaacctttg aaccctgcgt aatgtgtgcg 300

ggggctatga tccactcgag aatagggcgc gtggtattcg gcgtacgcaa cagcaaacgt 360

ggggctgcgg gctcgttgat gaacgttctg aactaccccg gaatgaatca cagagtagag 420

atcacggagg gaattttggc cgacgaatgt gcagctttac tgtgcgattt ttatcggatg 480

ccgcggcaag tcttcaacgc acagaagaag gctcaatctt ccattaaccc ggcgccggcg 540

ccggcgccgg ataagaaata ctcaataggc ttagctatcg gcacaaacag cgtcggctgg 600

gccgtcatta cggacgagta caaggtgccg agcaaaaaat tcaaagttct gggcaatacc 660

gatcgccaca gcataaagaa gaacctcatt ggcgccctcc tgttcgactc cggggagacg 720

gccgaagcca cgcggctcaa aagaacagca cggcgcagat atacccgcag aaagaatcgg 780

atctgctacc tgcaggagat ctttagtaat gagatggcta aggtggatga ctctttcttc 840

cataggctgg aggagtcctt tttggtggag gaggataaaa agcacgagcg ccacccaatc 900

tttggcaata tcgtggacga ggtggcgtac catgaaaagt acccaaccat atatcatctg 960

aggaagaagc ttgtagacag tactgataag gctgacttgc ggttgatcta tctcgcgctg 1020

gcgcatatga tcaaatttcg gggacacttc ctcatcgagg gggacctgaa cccagacaac 1080

agcgatgtcg acaaactctt tatccaactg gttcagactt acaatcagct tttcgaagag 1140

aacccgatca acgcatccgg agttgacgcc aaagcaatcc tgagcgctag gctgtccaaa 1200

tcccggcggc tcgaaaacct catcgcacag ctccctgggg agaagaagaa cggcctgttt 1260

ggtaatctta tcgccctgtc actcgggctg acccccaact ttaaatctaa cttcgacctg 1320

gccgaagatg ccaagcttca actgagcaaa gacacctacg atgatgatct cgacaatctg 1380

ctggcccaga tcggcgacca gtacgcagac ctttttttgg cggcaaagaa cctgtcagac 1440

gccattctgc tgagtgatat tctgcgagtg aacacggaga tcaccaaagc tccgctgagc 1500

gctagtatga tcaagcgcta tgatgagcac caccaagact tgactttgct gaaggccctt 1560

gtcagacagc aactgcctga gaagtacaag gaaattttct tcgatcagtc taaaaatggc 1620

tacgccggat acattgacgg cggagcaagc caggaggaat tttacaaatt tattaagccc 1680

atcttggaaa aaatggacgg caccgaggag ctgctggtaa agcttaacag agaagatctg 1740

ttgcgcaaac agcgcacttt cgacaatgga agcatccccc accagattca cctgggcgaa 1800

ctgcacgcta tcctcaggcg gcaagaggat ttctacccct ttttgaaaga taacagggaa 1860

aagattgaga aaatcctcac atttcggata ccctactatg taggccccct cgcccgggga 1920

aattccagat tcgcgtggat gactcgcaaa tcagaagaga ccatcactcc ctggaacttc 1980

gaggaagtcg tggataaggg ggcctctgcc cagtccttca tcgaaaggat gactaacttt 2040

gataaaaatc tgcctaacga aaaggtgctt cctaaacact ctctgctgta cgagtacttc 2100

acagtttata acgagctcac caaggtcaaa tacgtcacag aagggatgag aaagccagca 2160

ttcctgtctg gagagcagaa gaaagctatc gtggacctcc tcttcaagac gaaccggaaa 2220

gttaccgtga aacagctcaa agaagactat ttcaaaaaga ttgaatgttt cgactctgtt 2280

gaaatcagcg gagtggagga tcgcttcaac gcatccctgg gaacgtatca cgatctcctg 2340

aaaatcatta aagacaagga cttcctggac aatgaggaga acgaggacat tcttgaggac 2400

attgtcctca cccttacgtt gtttgaagat agggagatga ttgaagaacg cttgaaaact 2460

tacgctcatc tcttcgacga caaagtcatg aaacagctca agaggcgccg atatacagga 2520

tgggggcggc tgtcaagaaa actgatcaat gggatccgag acaagcagag tggaaagaca 2580

atcctggatt ttcttaagtc cgatggattt gccaaccgga acttcatgca gttgatccat 2640

gatgactctc tcacctttaa ggaggacatc cagaaagcac aagtttctgg ccagggggac 2700

agtcttcacg agcacatcgc taatcttgca ggtagcccag ctatcaaaaa gggaatactg 2760

cagaccgtta aggtcgtgga tgaactcgtc aaagtaatgg gaaggcataa gcccgagaat 2820

atcgttatcg agatggcccg agagaaccaa actacccaga agggacagaa gaacagtagg 2880

gaaaggatga agaggattga agagggtata aaagaactgg ggtcccaaat ccttaaggaa 2940

cacccagttg aaaacaccca gcttcagaat gagaagctct acctgtacta cctgcagaac 3000

ggcagggaca tgtacgtgga tcaggaactg gacatcaatc ggctctccga ctacgacgtg 3060

gatcatatcg tgccccagtc ttttctcaaa gatgattcta ttgataataa agtgttgaca 3120

agatccgata aaaatagagg gaagagtgat aacgtcccct cagaagaagt tgtcaagaaa 3180

atgaaaaatt attggcggca gctgctgaac gccaaactga tcacacaacg gaagttcgat 3240

aatctgacta aggctgaacg aggtggcctg tctgagttgg ataaagcagg cttcatcaaa 3300

aggcagcttg ttgagacacg ccagatcacc aagcacgtgg cccaaattct cgattcacgc 3360

atgaacacca agtacgatga aaatgacaaa ctgattcgag aggtgaaagt tattactctg 3420

aagtctaagc tggtctcaga tttcagaaag gactttcagt tttataaggt gagagagatc 3480

aacaattacc accatgcgca tgatgcctac ctgaatgcag tggtaggcac tgcacttatc 3540

aaaaaatatc ccaagcttga atctgaattt gtttacggag actataaagt gtacgatgtt 3600

aggaaaatga tcgcaaagtc tgagcaggaa ataggcaagg ccaccgctaa gtacttcttt 3660

tacagcaata ttatgaattt tttcaagacc gagattacac tggccaatgg agagattcgg 3720

aagcgaccac ttatcgaaac aaacggagaa acaggagaaa tcgtgtggga caagggtagg 3780

gatttcgcga cagtccggaa ggtcctgtcc atgccgcagg tgaacatcgt taaaaagacc 3840

gaagtacaga ccggaggctt ctccaaggaa agtatcctcc cgaaaaggaa cagcgacaag 3900

ctgatcgcac gcaaaaaaga ttgggacccc aagaaatacg gcggattcga ttctcctaca 3960

gtcgcttaca gtgtactggt tgtggccaaa gtggagaaag ggaagtctaa aaaactcaaa 4020

agcgtcaagg aactgctggg catcacaatc atggagcgat caagcttcga aaaaaacccc 4080

atcgactttc tcgaggcgaa aggatataaa gaggtcaaaa aagacctcat cattaagctt 4140

cccaagtact ctctctttga gcttgaaaac ggccggaaac gaatgctcgc tagtgcgggc 4200

gagctgcaga aaggtaacga gctggcactg ccctctaaat acgttaattt cttgtatctg 4260

gccagccact atgaaaagct caaagggtct cccgaagata atgagcagaa gcagctgttc 4320

gtggaacaac acaaacacta ccttgatgag atcatcgagc aaataagcga attctccaaa 4380

agagtgatcc tcgccgacgc taacctcgat aaggtgcttt ctgcttacaa taagcacagg 4440

gataagccca tcagggagca ggcagaaaac attatccact tgtttactct gaccaacttg 4500

ggcgcgcctg cagccttcaa gtacttcgac accaccatag acagaaagcg gtacacctct 4560

acaaaggagg tcctggacgc cacactgatt catcagtcaa ttacggggct ctatgaaaca 4620

agaatcgacc tctctcagct cggtggagac 4650

<210> 12

<211> 1550

<212> PRT

<213> l

<400> 12

Met Gly Ser Ser His His His His His His Ser Glu Val Glu Phe Ser

1 5 10 15

His Glu Tyr Trp Met Arg His Ala Leu Thr Leu Ala Lys Arg Ala Arg

20 25 30

Asp Glu Arg Glu Val Pro Val Gly Ala Val Leu Val Leu Asn Asn Arg

35 40 45

Val Ile Gly Glu Gly Trp Asn Arg Ala Ile Gly Leu His Asp Pro Thr

50 55 60

Ala His Ala Glu Ile Met Ala Leu Arg Gln Gly Gly Leu Val Met Gln

65 70 75 80

Asn Tyr Arg Leu Ile Asp Ala Thr Leu Tyr Val Thr Phe Glu Pro Cys

85 90 95

Val Met Cys Ala Gly Ala Met Ile His Ser Arg Ile Gly Arg Val Val

100 105 110

Phe Gly Val Arg Asn Ser Lys Arg Gly Ala Ala Gly Ser Leu Met Asn

115 120 125

Val Leu Asn Tyr Pro Gly Met Asn His Arg Val Glu Ile Thr Glu Gly

130 135 140

Ile Leu Ala Asp Glu Cys Ala Ala Leu Leu Cys Asp Phe Tyr Arg Met

145 150 155 160

Pro Arg Gln Val Phe Asn Ala Gln Lys Lys Ala Gln Ser Ser Ile Asn

165 170 175

Pro Ala Pro Ala Pro Ala Pro Asp Lys Lys Tyr Ser Ile Gly Leu Ala

180 185 190

Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile Thr Asp Glu Tyr Lys

195 200 205

Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn Thr Asp Arg His Ser

210 215 220

Ile Lys Lys Asn Leu Ile Gly Ala Leu Leu Phe Asp Ser Gly Glu Thr

225 230 235 240

Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg Arg Arg Tyr Thr Arg

245 250 255

Arg Lys Asn Arg Ile Cys Tyr Leu Gln Glu Ile Phe Ser Asn Glu Met

260 265 270

Ala Lys Val Asp Asp Ser Phe Phe His Arg Leu Glu Glu Ser Phe Leu

275 280 285

Val Glu Glu Asp Lys Lys His Glu Arg His Pro Ile Phe Gly Asn Ile

290 295 300

Val Asp Glu Val Ala Tyr His Glu Lys Tyr Pro Thr Ile Tyr His Leu

305 310 315 320

Arg Lys Lys Leu Val Asp Ser Thr Asp Lys Ala Asp Leu Arg Leu Ile

325 330 335

Tyr Leu Ala Leu Ala His Met Ile Lys Phe Arg Gly His Phe Leu Ile

340 345 350

Glu Gly Asp Leu Asn Pro Asp Asn Ser Asp Val Asp Lys Leu Phe Ile

355 360 365

Gln Leu Val Gln Thr Tyr Asn Gln Leu Phe Glu Glu Asn Pro Ile Asn

370 375 380

Ala Ser Gly Val Asp Ala Lys Ala Ile Leu Ser Ala Arg Leu Ser Lys

385 390 395 400

Ser Arg Arg Leu Glu Asn Leu Ile Ala Gln Leu Pro Gly Glu Lys Lys

405 410 415

Asn Gly Leu Phe Gly Asn Leu Ile Ala Leu Ser Leu Gly Leu Thr Pro

420 425 430

Asn Phe Lys Ser Asn Phe Asp Leu Ala Glu Asp Ala Lys Leu Gln Leu

435 440 445

Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Asn Leu Leu Ala Gln Ile

450 455 460

Gly Asp Gln Tyr Ala Asp Leu Phe Leu Ala Ala Lys Asn Leu Ser Asp

465 470 475 480

Ala Ile Leu Leu Ser Asp Ile Leu Arg Val Asn Thr Glu Ile Thr Lys

485 490 495

Ala Pro Leu Ser Ala Ser Met Ile Lys Arg Tyr Asp Glu His His Gln

500 505 510

Asp Leu Thr Leu Leu Lys Ala Leu Val Arg Gln Gln Leu Pro Glu Lys

515 520 525

Tyr Lys Glu Ile Phe Phe Asp Gln Ser Lys Asn Gly Tyr Ala Gly Tyr

530 535 540

Ile Asp Gly Gly Ala Ser Gln Glu Glu Phe Tyr Lys Phe Ile Lys Pro

545 550 555 560

Ile Leu Glu Lys Met Asp Gly Thr Glu Glu Leu Leu Val Lys Leu Asn

565 570 575

Arg Glu Asp Leu Leu Arg Lys Gln Arg Thr Phe Asp Asn Gly Ser Ile

580 585 590

Pro His Gln Ile His Leu Gly Glu Leu His Ala Ile Leu Arg Arg Gln

595 600 605

Glu Asp Phe Tyr Pro Phe Leu Lys Asp Asn Arg Glu Lys Ile Glu Lys

610 615 620

Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly Pro Leu Ala Arg Gly

625 630 635 640

Asn Ser Arg Phe Ala Trp Met Thr Arg Lys Ser Glu Glu Thr Ile Thr

645 650 655

Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly Ala Ser Ala Gln Ser

660 665 670

Phe Ile Glu Arg Met Thr Asn Phe Asp Lys Asn Leu Pro Asn Glu Lys

675 680 685

Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr Phe Thr Val Tyr Asn

690 695 700

Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Gly Met Arg Lys Pro Ala

705 710 715 720

Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val Asp Leu Leu Phe Lys

725 730 735

Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys Glu Asp Tyr Phe Lys

740 745 750

Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ser Gly Val Glu Asp Arg

755 760 765

Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu Leu Lys Ile Ile Lys

770 775 780

Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu Asp Ile Leu Glu Asp

785 790 795 800

Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg Glu Met Ile Glu Glu

805 810 815

Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp Lys Val Met Lys Gln

820 825 830

Leu Lys Arg Arg Arg Tyr Thr Gly Trp Gly Arg Leu Ser Arg Lys Leu

835 840 845

Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys Thr Ile Leu Asp Phe

850 855 860

Leu Lys Ser Asp Gly Phe Ala Asn Arg Asn Phe Met Gln Leu Ile His

865 870 875 880

Asp Asp Ser Leu Thr Phe Lys Glu Asp Ile Gln Lys Ala Gln Val Ser

885 890 895

Gly Gln Gly Asp Ser Leu His Glu His Ile Ala Asn Leu Ala Gly Ser

900 905 910

Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr Val Lys Val Val Asp Glu

915 920 925

Leu Val Lys Val Met Gly Arg His Lys Pro Glu Asn Ile Val Ile Glu

930 935 940

Met Ala Arg Glu Asn Gln Thr Thr Gln Lys Gly Gln Lys Asn Ser Arg

945 950 955 960

Glu Arg Met Lys Arg Ile Glu Glu Gly Ile Lys Glu Leu Gly Ser Gln

965 970 975

Ile Leu Lys Glu His Pro Val Glu Asn Thr Gln Leu Gln Asn Glu Lys

980 985 990

Leu Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg Asp Met Tyr Val Asp Gln

995 1000 1005

Glu Leu Asp Ile Asn Arg Leu Ser Asp Tyr Asp Val Asp His Ile Val

1010 1015 1020

Pro Gln Ser Phe Leu Lys Asp Asp Ser Ile Asp Asn Lys Val Leu Thr

1025 1030 1035 1040

Arg Ser Asp Lys Asn Arg Gly Lys Ser Asp Asn Val Pro Ser Glu Glu

1045 1050 1055

Val Val Lys Lys Met Lys Asn Tyr Trp Arg Gln Leu Leu Asn Ala Lys

1060 1065 1070

Leu Ile Thr Gln Arg Lys Phe Asp Asn Leu Thr Lys Ala Glu Arg Gly

1075 1080 1085

Gly Leu Ser Glu Leu Asp Lys Ala Gly Phe Ile Lys Arg Gln Leu Val

1090 1095 1100

Glu Thr Arg Gln Ile Thr Lys His Val Ala Gln Ile Leu Asp Ser Arg

1105 1110 1115 1120

Met Asn Thr Lys Tyr Asp Glu Asn Asp Lys Leu Ile Arg Glu Val Lys

1125 1130 1135

Val Ile Thr Leu Lys Ser Lys Leu Val Ser Asp Phe Arg Lys Asp Phe

1140 1145 1150

Gln Phe Tyr Lys Val Arg Glu Ile Asn Asn Tyr His His Ala His Asp

1155 1160 1165

Ala Tyr Leu Asn Ala Val Val Gly Thr Ala Leu Ile Lys Lys Tyr Pro

1170 1175 1180

Lys Leu Glu Ser Glu Phe Val Tyr Gly Asp Tyr Lys Val Tyr Asp Val

1185 1190 1195 1200

Arg Lys Met Ile Ala Lys Ser Glu Gln Glu Ile Gly Lys Ala Thr Ala

1205 1210 1215

Lys Tyr Phe Phe Tyr Ser Asn Ile Met Asn Phe Phe Lys Thr Glu Ile

1220 1225 1230

Thr Leu Ala Asn Gly Glu Ile Arg Lys Arg Pro Leu Ile Glu Thr Asn

1235 1240 1245

Gly Glu Thr Gly Glu Ile Val Trp Asp Lys Gly Arg Asp Phe Ala Thr

1250 1255 1260

Val Arg Lys Val Leu Ser Met Pro Gln Val Asn Ile Val Lys Lys Thr

1265 1270 1275 1280

Glu Val Gln Thr Gly Gly Phe Ser Lys Glu Ser Ile Leu Pro Lys Arg

1285 1290 1295

Asn Ser Asp Lys Leu Ile Ala Arg Lys Lys Asp Trp Asp Pro Lys Lys

1300 1305 1310

Tyr Gly Gly Phe Asp Ser Pro Thr Val Ala Tyr Ser Val Leu Val Val

1315 1320 1325

Ala Lys Val Glu Lys Gly Lys Ser Lys Lys Leu Lys Ser Val Lys Glu

1330 1335 1340

Leu Leu Gly Ile Thr Ile Met Glu Arg Ser Ser Phe Glu Lys Asn Pro

1345 1350 1355 1360

Ile Asp Phe Leu Glu Ala Lys Gly Tyr Lys Glu Val Lys Lys Asp Leu

1365 1370 1375

Ile Ile Lys Leu Pro Lys Tyr Ser Leu Phe Glu Leu Glu Asn Gly Arg

1380 1385 1390

Lys Arg Met Leu Ala Ser Ala Gly Glu Leu Gln Lys Gly Asn Glu Leu

1395 1400 1405

Ala Leu Pro Ser Lys Tyr Val Asn Phe Leu Tyr Leu Ala Ser His Tyr

1410 1415 1420

Glu Lys Leu Lys Gly Ser Pro Glu Asp Asn Glu Gln Lys Gln Leu Phe

1425 1430 1435 1440

Val Glu Gln His Lys His Tyr Leu Asp Glu Ile Ile Glu Gln Ile Ser

1445 1450 1455

Glu Phe Ser Lys Arg Val Ile Leu Ala Asp Ala Asn Leu Asp Lys Val

1460 1465 1470

Leu Ser Ala Tyr Asn Lys His Arg Asp Lys Pro Ile Arg Glu Gln Ala

1475 1480 1485

Glu Asn Ile Ile His Leu Phe Thr Leu Thr Asn Leu Gly Ala Pro Ala

1490 1495 1500

Ala Phe Lys Tyr Phe Asp Thr Thr Ile Asp Arg Lys Arg Tyr Thr Ser

1505 1510 1515 1520

Thr Lys Glu Val Leu Asp Ala Thr Leu Ile His Gln Ser Ile Thr Gly

1525 1530 1535

Leu Tyr Glu Thr Arg Ile Asp Leu Ser Gln Leu Gly Gly Asp

1540 1545 1550

<210> 13

<211> 4665

<212> DNA

<213> m

<400> 13

atgggcagca gccatcatca tcatcatcac tcggaggtcg agttttcaca tgagtattgg 60

atgcgtcatg ccttgacgtt ggcgaaacgg gcgcgcgatg agcgtgaggt gcccgtggga 120

gcggtgttgg tactgaataa ccgggttata ggggaaggat ggaaccgggc tattgggtta 180

cacgacccaa cggcgcacgc cgagataatg gcactgcgcc aagggggctt agttatgcag 240

aattatcgcc ttatcgatgc tacactgtat gtaacctttg aaccctgcgt aatgtgtgcg 300

ggggctatga tccactcgag aatagggcgc gtggtattcg gcgtacgcaa cagcaaacgt 360

ggggctgcgg gctcgttgat gaacgttctg aactaccccg gaatgaatca cagagtagag 420

atcacggagg gaattttggc cgacgaatgt gcagctttac tgtgcgattt ttatcggatg 480

ccgcggcaag tcttcaacgc acagaagaag gctcaatctt ccattaacgc ggaagcggcg 540

gcgaaagaag cggcggcgaa agcggataag aaatactcaa taggcttagc tatcggcaca 600

aacagcgtcg gctgggccgt cattacggac gagtacaagg tgccgagcaa aaaattcaaa 660

gttctgggca ataccgatcg ccacagcata aagaagaacc tcattggcgc cctcctgttc 720

gactccgggg agacggccga agccacgcgg ctcaaaagaa cagcacggcg cagatatacc 780

cgcagaaaga atcggatctg ctacctgcag gagatcttta gtaatgagat ggctaaggtg 840

gatgactctt tcttccatag gctggaggag tcctttttgg tggaggagga taaaaagcac 900

gagcgccacc caatctttgg caatatcgtg gacgaggtgg cgtaccatga aaagtaccca 960

accatatatc atctgaggaa gaagcttgta gacagtactg ataaggctga cttgcggttg 1020

atctatctcg cgctggcgca tatgatcaaa tttcggggac acttcctcat cgagggggac 1080

ctgaacccag acaacagcga tgtcgacaaa ctctttatcc aactggttca gacttacaat 1140

cagcttttcg aagagaaccc gatcaacgca tccggagttg acgccaaagc aatcctgagc 1200

gctaggctgt ccaaatcccg gcggctcgaa aacctcatcg cacagctccc tggggagaag 1260

aagaacggcc tgtttggtaa tcttatcgcc ctgtcactcg ggctgacccc caactttaaa 1320

tctaacttcg acctggccga agatgccaag cttcaactga gcaaagacac ctacgatgat 1380

gatctcgaca atctgctggc ccagatcggc gaccagtacg cagacctttt tttggcggca 1440

aagaacctgt cagacgccat tctgctgagt gatattctgc gagtgaacac ggagatcacc 1500

aaagctccgc tgagcgctag tatgatcaag cgctatgatg agcaccacca agacttgact 1560

ttgctgaagg cccttgtcag acagcaactg cctgagaagt acaaggaaat tttcttcgat 1620

cagtctaaaa atggctacgc cggatacatt gacggcggag caagccagga ggaattttac 1680

aaatttatta agcccatctt ggaaaaaatg gacggcaccg aggagctgct ggtaaagctt 1740

aacagagaag atctgttgcg caaacagcgc actttcgaca atggaagcat cccccaccag 1800

attcacctgg gcgaactgca cgctatcctc aggcggcaag aggatttcta cccctttttg 1860

aaagataaca gggaaaagat tgagaaaatc ctcacatttc ggatacccta ctatgtaggc 1920

cccctcgccc ggggaaattc cagattcgcg tggatgactc gcaaatcaga agagaccatc 1980

actccctgga acttcgagga agtcgtggat aagggggcct ctgcccagtc cttcatcgaa 2040

aggatgacta actttgataa aaatctgcct aacgaaaagg tgcttcctaa acactctctg 2100

ctgtacgagt acttcacagt ttataacgag ctcaccaagg tcaaatacgt cacagaaggg 2160

atgagaaagc cagcattcct gtctggagag cagaagaaag ctatcgtgga cctcctcttc 2220

aagacgaacc ggaaagttac cgtgaaacag ctcaaagaag actatttcaa aaagattgaa 2280

tgtttcgact ctgttgaaat cagcggagtg gaggatcgct tcaacgcatc cctgggaacg 2340

tatcacgatc tcctgaaaat cattaaagac aaggacttcc tggacaatga ggagaacgag 2400

gacattcttg aggacattgt cctcaccctt acgttgtttg aagataggga gatgattgaa 2460

gaacgcttga aaacttacgc tcatctcttc gacgacaaag tcatgaaaca gctcaagagg 2520

cgccgatata caggatgggg gcggctgtca agaaaactga tcaatgggat ccgagacaag 2580

cagagtggaa agacaatcct ggattttctt aagtccgatg gatttgccaa ccggaacttc 2640

atgcagttga tccatgatga ctctctcacc tttaaggagg acatccagaa agcacaagtt 2700

tctggccagg gggacagtct tcacgagcac atcgctaatc ttgcaggtag cccagctatc 2760

aaaaagggaa tactgcagac cgttaaggtc gtggatgaac tcgtcaaagt aatgggaagg 2820

cataagcccg agaatatcgt tatcgagatg gcccgagaga accaaactac ccagaaggga 2880

cagaagaaca gtagggaaag gatgaagagg attgaagagg gtataaaaga actggggtcc 2940

caaatcctta aggaacaccc agttgaaaac acccagcttc agaatgagaa gctctacctg 3000

tactacctgc agaacggcag ggacatgtac gtggatcagg aactggacat caatcggctc 3060

tccgactacg acgtggatca tatcgtgccc cagtcttttc tcaaagatga ttctattgat 3120

aataaagtgt tgacaagatc cgataaaaat agagggaaga gtgataacgt cccctcagaa 3180

gaagttgtca agaaaatgaa aaattattgg cggcagctgc tgaacgccaa actgatcaca 3240

caacggaagt tcgataatct gactaaggct gaacgaggtg gcctgtctga gttggataaa 3300

gcaggcttca tcaaaaggca gcttgttgag acacgccaga tcaccaagca cgtggcccaa 3360

attctcgatt cacgcatgaa caccaagtac gatgaaaatg acaaactgat tcgagaggtg 3420

aaagttatta ctctgaagtc taagctggtc tcagatttca gaaaggactt tcagttttat 3480

aaggtgagag agatcaacaa ttaccaccat gcgcatgatg cctacctgaa tgcagtggta 3540

ggcactgcac ttatcaaaaa atatcccaag cttgaatctg aatttgttta cggagactat 3600

aaagtgtacg atgttaggaa aatgatcgca aagtctgagc aggaaatagg caaggccacc 3660

gctaagtact tcttttacag caatattatg aattttttca agaccgagat tacactggcc 3720

aatggagaga ttcggaagcg accacttatc gaaacaaacg gagaaacagg agaaatcgtg 3780

tgggacaagg gtagggattt cgcgacagtc cggaaggtcc tgtccatgcc gcaggtgaac 3840

atcgttaaaa agaccgaagt acagaccgga ggcttctcca aggaaagtat cctcccgaaa 3900

aggaacagcg acaagctgat cgcacgcaaa aaagattggg accccaagaa atacggcgga 3960

ttcgattctc ctacagtcgc ttacagtgta ctggttgtgg ccaaagtgga gaaagggaag 4020

tctaaaaaac tcaaaagcgt caaggaactg ctgggcatca caatcatgga gcgatcaagc 4080

ttcgaaaaaa accccatcga ctttctcgag gcgaaaggat ataaagaggt caaaaaagac 4140

ctcatcatta agcttcccaa gtactctctc tttgagcttg aaaacggccg gaaacgaatg 4200

ctcgctagtg cgggcgagct gcagaaaggt aacgagctgg cactgccctc taaatacgtt 4260

aatttcttgt atctggccag ccactatgaa aagctcaaag ggtctcccga agataatgag 4320

cagaagcagc tgttcgtgga acaacacaaa cactaccttg atgagatcat cgagcaaata 4380

agcgaattct ccaaaagagt gatcctcgcc gacgctaacc tcgataaggt gctttctgct 4440

tacaataagc acagggataa gcccatcagg gagcaggcag aaaacattat ccacttgttt 4500

actctgacca acttgggcgc gcctgcagcc ttcaagtact tcgacaccac catagacaga 4560

aagcggtaca cctctacaaa ggaggtcctg gacgccacac tgattcatca gtcaattacg 4620

gggctctatg aaacaagaat cgacctctct cagctcggtg gagac 4665

<210> 14

<211> 1555

<212> PRT

<213> n

<400> 14

Met Gly Ser Ser His His His His His His Ser Glu Val Glu Phe Ser

1 5 10 15

His Glu Tyr Trp Met Arg His Ala Leu Thr Leu Ala Lys Arg Ala Arg

20 25 30

Asp Glu Arg Glu Val Pro Val Gly Ala Val Leu Val Leu Asn Asn Arg

35 40 45

Val Ile Gly Glu Gly Trp Asn Arg Ala Ile Gly Leu His Asp Pro Thr

50 55 60

Ala His Ala Glu Ile Met Ala Leu Arg Gln Gly Gly Leu Val Met Gln

65 70 75 80

Asn Tyr Arg Leu Ile Asp Ala Thr Leu Tyr Val Thr Phe Glu Pro Cys

85 90 95

Val Met Cys Ala Gly Ala Met Ile His Ser Arg Ile Gly Arg Val Val

100 105 110

Phe Gly Val Arg Asn Ser Lys Arg Gly Ala Ala Gly Ser Leu Met Asn

115 120 125

Val Leu Asn Tyr Pro Gly Met Asn His Arg Val Glu Ile Thr Glu Gly

130 135 140

Ile Leu Ala Asp Glu Cys Ala Ala Leu Leu Cys Asp Phe Tyr Arg Met

145 150 155 160

Pro Arg Gln Val Phe Asn Ala Gln Lys Lys Ala Gln Ser Ser Ile Asn

165 170 175

Ala Glu Ala Ala Ala Lys Glu Ala Ala Ala Lys Ala Asp Lys Lys Tyr

180 185 190

Ser Ile Gly Leu Ala Ile Gly Thr Asn Ser Val Gly Trp Ala Val Ile

195 200 205

Thr Asp Glu Tyr Lys Val Pro Ser Lys Lys Phe Lys Val Leu Gly Asn

210 215 220

Thr Asp Arg His Ser Ile Lys Lys Asn Leu Ile Gly Ala Leu Leu Phe

225 230 235 240

Asp Ser Gly Glu Thr Ala Glu Ala Thr Arg Leu Lys Arg Thr Ala Arg

245 250 255

Arg Arg Tyr Thr Arg Arg Lys Asn Arg Ile Cys Tyr Leu Gln Glu Ile

260 265 270

Phe Ser Asn Glu Met Ala Lys Val Asp Asp Ser Phe Phe His Arg Leu

275 280 285

Glu Glu Ser Phe Leu Val Glu Glu Asp Lys Lys His Glu Arg His Pro

290 295 300

Ile Phe Gly Asn Ile Val Asp Glu Val Ala Tyr His Glu Lys Tyr Pro

305 310 315 320

Thr Ile Tyr His Leu Arg Lys Lys Leu Val Asp Ser Thr Asp Lys Ala

325 330 335

Asp Leu Arg Leu Ile Tyr Leu Ala Leu Ala His Met Ile Lys Phe Arg

340 345 350

Gly His Phe Leu Ile Glu Gly Asp Leu Asn Pro Asp Asn Ser Asp Val

355 360 365

Asp Lys Leu Phe Ile Gln Leu Val Gln Thr Tyr Asn Gln Leu Phe Glu

370 375 380

Glu Asn Pro Ile Asn Ala Ser Gly Val Asp Ala Lys Ala Ile Leu Ser

385 390 395 400

Ala Arg Leu Ser Lys Ser Arg Arg Leu Glu Asn Leu Ile Ala Gln Leu

405 410 415

Pro Gly Glu Lys Lys Asn Gly Leu Phe Gly Asn Leu Ile Ala Leu Ser

420 425 430

Leu Gly Leu Thr Pro Asn Phe Lys Ser Asn Phe Asp Leu Ala Glu Asp

435 440 445

Ala Lys Leu Gln Leu Ser Lys Asp Thr Tyr Asp Asp Asp Leu Asp Asn

450 455 460

Leu Leu Ala Gln Ile Gly Asp Gln Tyr Ala Asp Leu Phe Leu Ala Ala

465 470 475 480

Lys Asn Leu Ser Asp Ala Ile Leu Leu Ser Asp Ile Leu Arg Val Asn

485 490 495

Thr Glu Ile Thr Lys Ala Pro Leu Ser Ala Ser Met Ile Lys Arg Tyr

500 505 510

Asp Glu His His Gln Asp Leu Thr Leu Leu Lys Ala Leu Val Arg Gln

515 520 525

Gln Leu Pro Glu Lys Tyr Lys Glu Ile Phe Phe Asp Gln Ser Lys Asn

530 535 540

Gly Tyr Ala Gly Tyr Ile Asp Gly Gly Ala Ser Gln Glu Glu Phe Tyr

545 550 555 560

Lys Phe Ile Lys Pro Ile Leu Glu Lys Met Asp Gly Thr Glu Glu Leu

565 570 575

Leu Val Lys Leu Asn Arg Glu Asp Leu Leu Arg Lys Gln Arg Thr Phe

580 585 590

Asp Asn Gly Ser Ile Pro His Gln Ile His Leu Gly Glu Leu His Ala

595 600 605

Ile Leu Arg Arg Gln Glu Asp Phe Tyr Pro Phe Leu Lys Asp Asn Arg

610 615 620

Glu Lys Ile Glu Lys Ile Leu Thr Phe Arg Ile Pro Tyr Tyr Val Gly

625 630 635 640

Pro Leu Ala Arg Gly Asn Ser Arg Phe Ala Trp Met Thr Arg Lys Ser

645 650 655

Glu Glu Thr Ile Thr Pro Trp Asn Phe Glu Glu Val Val Asp Lys Gly

660 665 670

Ala Ser Ala Gln Ser Phe Ile Glu Arg Met Thr Asn Phe Asp Lys Asn

675 680 685

Leu Pro Asn Glu Lys Val Leu Pro Lys His Ser Leu Leu Tyr Glu Tyr

690 695 700

Phe Thr Val Tyr Asn Glu Leu Thr Lys Val Lys Tyr Val Thr Glu Gly

705 710 715 720

Met Arg Lys Pro Ala Phe Leu Ser Gly Glu Gln Lys Lys Ala Ile Val

725 730 735

Asp Leu Leu Phe Lys Thr Asn Arg Lys Val Thr Val Lys Gln Leu Lys

740 745 750

Glu Asp Tyr Phe Lys Lys Ile Glu Cys Phe Asp Ser Val Glu Ile Ser

755 760 765

Gly Val Glu Asp Arg Phe Asn Ala Ser Leu Gly Thr Tyr His Asp Leu

770 775 780

Leu Lys Ile Ile Lys Asp Lys Asp Phe Leu Asp Asn Glu Glu Asn Glu

785 790 795 800

Asp Ile Leu Glu Asp Ile Val Leu Thr Leu Thr Leu Phe Glu Asp Arg

805 810 815

Glu Met Ile Glu Glu Arg Leu Lys Thr Tyr Ala His Leu Phe Asp Asp

820 825 830

Lys Val Met Lys Gln Leu Lys Arg Arg Arg Tyr Thr Gly Trp Gly Arg

835 840 845

Leu Ser Arg Lys Leu Ile Asn Gly Ile Arg Asp Lys Gln Ser Gly Lys

850 855 860

Thr Ile Leu Asp Phe Leu Lys Ser Asp Gly Phe Ala Asn Arg Asn Phe

865 870 875 880

Met Gln Leu Ile His Asp Asp Ser Leu Thr Phe Lys Glu Asp Ile Gln

885 890 895

Lys Ala Gln Val Ser Gly Gln Gly Asp Ser Leu His Glu His Ile Ala

900 905 910

Asn Leu Ala Gly Ser Pro Ala Ile Lys Lys Gly Ile Leu Gln Thr Val

915 920 925

Lys Val Val Asp Glu Leu Val Lys Val Met Gly Arg His Lys Pro Glu

930 935 940

Asn Ile Val Ile Glu Met Ala Arg Glu Asn Gln Thr Thr Gln Lys Gly

945 950 955 960

Gln Lys Asn Ser Arg Glu Arg Met Lys Arg Ile Glu Glu Gly Ile Lys

965 970 975

Glu Leu Gly Ser Gln Ile Leu Lys Glu His Pro Val Glu Asn Thr Gln

980 985 990

Leu Gln Asn Glu Lys Leu Tyr Leu Tyr Tyr Leu Gln Asn Gly Arg Asp

995 1000 1005

Met Tyr Val Asp Gln Glu Leu Asp Ile Asn Arg Leu Ser Asp Tyr Asp

1010 1015 1020

Val Asp His Ile Val Pro Gln Ser Phe Leu Lys Asp Asp Ser Ile Asp

1025 1030 1035 1040

Asn Lys Val Leu Thr Arg Ser Asp Lys Asn Arg Gly Lys Ser Asp Asn

1045 1050 1055

Val Pro Ser Glu Glu Val Val Lys Lys Met Lys Asn Tyr Trp Arg Gln

1060 1065 1070

Leu Leu Asn Ala Lys Leu Ile Thr Gln Arg Lys Phe Asp Asn Leu Thr

1075 1080 1085

Lys Ala Glu Arg Gly Gly Leu Ser Glu Leu Asp Lys Ala Gly Phe Ile

1090 1095 1100

Lys Arg Gln Leu Val Glu Thr Arg Gln Ile Thr Lys His Val Ala Gln

1105 1110 1115 1120

Ile Leu Asp Ser Arg Met Asn Thr Lys Tyr Asp Glu Asn Asp Lys Leu

1125 1130 1135

Ile Arg Glu Val Lys Val Ile Thr Leu Lys Ser Lys Leu Val Ser Asp

1140 1145 1150

Phe Arg Lys Asp Phe Gln Phe Tyr Lys Val Arg Glu Ile Asn Asn Tyr

1155 1160 1165

His His Ala His Asp Ala Tyr Leu Asn Ala Val Val Gly Thr Ala Leu

1170 1175 1180

Ile Lys Lys Tyr Pro Lys Leu Glu Ser Glu Phe Val Tyr Gly Asp Tyr

1185 1190 1195 1200

Lys Val Tyr Asp Val Arg Lys Met Ile Ala Lys Ser Glu Gln Glu Ile

1205 1210 1215

Gly Lys Ala Thr Ala Lys Tyr Phe Phe Tyr Ser Asn Ile Met Asn Phe

1220 1225 1230

Phe Lys Thr Glu Ile Thr Leu Ala Asn Gly Glu Ile Arg Lys Arg Pro

1235 1240 1245

Leu Ile Glu Thr Asn Gly Glu Thr Gly Glu Ile Val Trp Asp Lys Gly

1250 1255 1260

Arg Asp Phe Ala Thr Val Arg Lys Val Leu Ser Met Pro Gln Val Asn

1265 1270 1275 1280

Ile Val Lys Lys Thr Glu Val Gln Thr Gly Gly Phe Ser Lys Glu Ser

1285 1290 1295

Ile Leu Pro Lys Arg Asn Ser Asp Lys Leu Ile Ala Arg Lys Lys Asp

1300 1305 1310

Trp Asp Pro Lys Lys Tyr Gly Gly Phe Asp Ser Pro Thr Val Ala Tyr

1315 1320 1325

Ser Val Leu Val Val Ala Lys Val Glu Lys Gly Lys Ser Lys Lys Leu

1330 1335 1340

Lys Ser Val Lys Glu Leu Leu Gly Ile Thr Ile Met Glu Arg Ser Ser

1345 1350 1355 1360

Phe Glu Lys Asn Pro Ile Asp Phe Leu Glu Ala Lys Gly Tyr Lys Glu

1365 1370 1375

Val Lys Lys Asp Leu Ile Ile Lys Leu Pro Lys Tyr Ser Leu Phe Glu

1380 1385 1390

Leu Glu Asn Gly Arg Lys Arg Met Leu Ala Ser Ala Gly Glu Leu Gln

1395 1400 1405

Lys Gly Asn Glu Leu Ala Leu Pro Ser Lys Tyr Val Asn Phe Leu Tyr

1410 1415 1420

Leu Ala Ser His Tyr Glu Lys Leu Lys Gly Ser Pro Glu Asp Asn Glu

1425 1430 1435 1440

Gln Lys Gln Leu Phe Val Glu Gln His Lys His Tyr Leu Asp Glu Ile

1445 1450 1455

Ile Glu Gln Ile Ser Glu Phe Ser Lys Arg Val Ile Leu Ala Asp Ala

1460 1465 1470

Asn Leu Asp Lys Val Leu Ser Ala Tyr Asn Lys His Arg Asp Lys Pro

1475 1480 1485

Ile Arg Glu Gln Ala Glu Asn Ile Ile His Leu Phe Thr Leu Thr Asn

1490 1495 1500

Leu Gly Ala Pro Ala Ala Phe Lys Tyr Phe Asp Thr Thr Ile Asp Arg

1505 1510 1515 1520

Lys Arg Tyr Thr Ser Thr Lys Glu Val Leu Asp Ala Thr Leu Ile His

1525 1530 1535

Gln Ser Ile Thr Gly Leu Tyr Glu Thr Arg Ile Asp Leu Ser Gln Leu

1540 1545 1550

Gly Gly Asp

1555

<210> 15

<211> 37

<212> DNA

<213> of

<400> 15

gaaacccgca gcgataagaa atactcaata ggcttag 37

<210> 16

<211> 37

<212> DNA

<213> or

<400> 16

gctgcgggtt tcgttaatgg aagattgagc cttcttc 37

<210> 17

<211> 37

<212> DNA

<213> pf

<400> 17

ccggcgccgg cggataagaa atactcaata ggcttag 37

<210> 18

<211> 37

<212> DNA

<213> or

<400> 18

cgccggcgcc gggttaatgg aagattgagc cttcttc 37

<210> 19

<211> 43

<212> DNA

<213> pf

<400> 19

aacaaaacct atccgccgga taagaaatac tcaataggct tag 43

<210> 20

<211> 43

<212> DNA

<213> pr

<400> 20

cggcggatag gttttgttgt taatggaaga ttgagccttc ttc 43

<210> 21

<211> 36

<212> DNA

<213> qf

<400> 21

accaccgaac cgataagaaa tactcaatag gcttag 36

<210> 22

<211> 36

<212> DNA

<213> qr

<400> 22

ggttcggtgg tgttaatgga agattgagcc ttcttc 36

<210> 23

<211> 36

<212> DNA

<213> qff

<400> 23

gccggataac gataagaaat actcaatagg cttagc 36

<210> 24

<211> 36

<212> DNA

<213> qrr

<400> 24

gttatccggc ggttcggtgg tgttaatgga agattg 36

<210> 25

<211> 36

<212> DNA

<213> rf

<400> 25

ccggcgccgg cgataagaaa tactcaatag gcttag 36

<210> 26

<211> 36

<212> DNA

<213> rr

<400> 26

gccggcgccg ggttaatgga agattgagcc ttcttc 36

<210> 27

<211> 35

<212> DNA

<213> rff

<400> 27

gccggcgccg gataagaaat actcaatagg cttag 35

<210> 28

<211> 35

<212> DNA

<213> rrr

<400> 28

cggcgccggc gccggcgccg ggttaatgga agatt 35

<210> 29

<211> 43

<212> DNA

<213> sf

<400> 29

gaagcggcgg cgaaagcgga taagaaatac tcaataggct tag 43

<210> 30

<211> 43

<212> DNA

<213> sr

<400> 30

cgctttcgcc gccgcttcgt taatggaaga ttgagccttc ttc 43

<210> 31

<211> 42

<212> DNA

<213> sff

<400> 31

gcggaagcgg cggcgaaaga agcggcggcg aaagcggata ag 42

<210> 32

<211> 43

<212> DNA

<213> srr

<400> 32

tttcgccgcc gcttccgcgt taatggaaga ttgagccttc ttc 43

<210> 33

<211> 920

<212> DNA

<213> t

<400> 33

tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg 60

ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg 120

ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat 180

taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc 240

agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc 300

gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa 360

cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc 420

ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga 480

ccatgattac gccaagctcg aaattaaccc tcactaaagg gaacaaaagc tggagctcca 540

ccgcggtggc ggccgctcta gaactagtgg atcccccggg ctgcaggaat tcgatatcaa 600

gcttatcgat gttacccttt ctttgtctgt aggttagagc tagaaatagc aagttaaaat 660

aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctctc gagggggggc 720

ccggtaccca attcgcccta tagtgagtcg tattacaatt cactggccgt cgttttacaa 780

cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct 840

ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc 900

agcctgaatg gcgaatggaa 920

<210> 34

<211> 920

<212> DNA

<213> u

<400> 34

tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg 60

ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg 120

ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat 180

taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc 240

agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc 300

gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa 360

cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc 420

ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga 480

ccatgattac gccaagctcg aaattaaccc tcactaaagg gaacaaaagc tggagctcca 540

ccgcggtggc ggccgctcta gaactagtgg atcccccggg ctgcaggaat tcgatatcaa 600

gcttatcgat gttcaccttt ctttgtctgt aggttagagc tagaaatagc aagttaaaat 660

aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctctc gagggggggc 720

ccggtaccca attcgcccta tagtgagtcg tattacaatt cactggccgt cgttttacaa 780

cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct 840

ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc 900

agcctgaatg gcgaatggaa 920

<210> 35

<211> 920

<212> DNA

<213> v

<400> 35

tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg 60

ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg 120

ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat 180

taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc 240

agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc 300

gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa 360

cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc 420

ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga 480

ccatgattac gccaagctcg aaattaaccc tcactaaagg gaacaaaagc tggagctcca 540

ccgcggtggc ggccgctcta gaactagtgg atcccccggg ctgcaggaat tcgatatcaa 600

gcttatcgat gttccacttt ctttgtctgt aggttagagc tagaaatagc aagttaaaat 660

aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctctc gagggggggc 720

ccggtaccca attcgcccta tagtgagtcg tattacaatt cactggccgt cgttttacaa 780

cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct 840

ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc 900

agcctgaatg gcgaatggaa 920

<210> 36

<211> 920

<212> DNA

<213> w

<400> 36

tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg 60

ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg 120

ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat 180

taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc 240

agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc 300

gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa 360

cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc 420

ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga 480

ccatgattac gccaagctcg aaattaaccc tcactaaagg gaacaaaagc tggagctcca 540

ccgcggtggc ggccgctcta gaactagtgg atcccccggg ctgcaggaat tcgatatcaa 600

gcttatcgat gttcccattt ctttgtctgt aggttagagc tagaaatagc aagttaaaat 660

aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctctc gagggggggc 720

ccggtaccca attcgcccta tagtgagtcg tattacaatt cactggccgt cgttttacaa 780

cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct 840

ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc 900

agcctgaatg gcgaatggaa 920

<210> 37

<211> 920

<212> DNA

<213> x

<400> 37

tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg 60

ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg 120

ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat 180

taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc 240

agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc 300

gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa 360

cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc 420

ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga 480

ccatgattac gccaagctcg aaattaaccc tcactaaagg gaacaaaagc tggagctcca 540

ccgcggtggc ggccgctcta gaactagtgg atcccccggg ctgcaggaat tcgatatcaa 600

gcttatcgat gttctccatt ctttgtctgt aggttagagc tagaaatagc aagttaaaat 660

aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctctc gagggggggc 720

ccggtaccca attcgcccta tagtgagtcg tattacaatt cactggccgt cgttttacaa 780

cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct 840

ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc 900

agcctgaatg gcgaatggaa 920

<210> 38

<211> 115

<212> DNA

<213> y

<400> 38

uaauacgacu cacuauaggg uuacccuuuc uuugucugug uuuuagagcu agaaauagca 60

aguuaaaaua aggcuagucc guuaucaacu ugaaaaagug gcaccgaguc ggugc 115

<210> 39

<211> 115

<212> DNA

<213> yy

<400> 39

uaauacgacu cacuauaggg uucaccuuuc uuugucugug uuuuagagcu agaaauagca 60

aguuaaaaua aggcuagucc guuaucaacu ugaaaaagug gcaccgaguc ggugc 115

<210> 40

<211> 115

<212> DNA

<213> yyy

<400> 40

uaauacgacu cacuauaggg uuccacuuuc uuugucugug uuuuagagcu agaaauagca 60

aguuaaaaua aggcuagucc guuaucaacu ugaaaaagug gcaccgaguc ggugc 115

<210> 41

<211> 115

<212> DNA

<213> yyyy

<400> 41

uaauacgacu cacuauaggg uucccauuuc uuugucugug uuuuagagcu agaaauagca 60

aguuaaaaua aggcuagucc guuaucaacu ugaaaaagug gcaccgaguc ggugc 115

<210> 42

<211> 115

<212> DNA

<213> yyyyy

<400> 42

uaauacgacu cacuauaggg uucuccauuc uuugucugug uuuuagagcu agaaauagca 60

aguuaaaaua aggcuagucc guuaucaacu ugaaaaagug gcaccgaguc ggugc 115

Claims

1.一种高精度腺嘌呤碱基编辑器，其特征在于，是将野生型ABE8e的32个氨基酸的linker即177位至208位氨基酸分别替换成ETRS、NKTYPP、TTEPPDN、PAPA、PAPAPAP、A(EAAAK)₂A后重组所得，依次记为4L-ABE8e、4P-ABE8e、6L-ABE8e、7L-ABE8e、7P-ABE8e和12EK-ABE8e；其核苷酸序列分别为SEQ ID NO.3、SEQ ID NO.5、SEQ ID NO.7、SEQ ID NO.9、SEQ ID NO.11、SEQ ID NO.13所示，氨基酸序列分别为SEQ ID NO.4、SEQ ID NO.6、SEQ IDNO.8、SEQ ID NO.10、SEQ ID NO.12、SEQ ID NO.14所示。

2.一种表达载体，其含有如权利要求1所述的多核苷酸序列。

3.一种宿主细胞，用于转化如权利要求2所述的表达载体。

4.如权利要求1所述的六个高精度腺嘌呤碱基编辑器的制备方法，其特征在于，具体步骤包括：首先，构建如权利要求2所述的多核苷酸序列表达载体；然后，将所述表达载体转化至如权利要求3所述的宿主细胞，筛选并挑出单克隆；最后，将所述单克隆诱导表达，并通过亲和层析、分子筛方法从表达产物中分离出所述的六个高精度腺嘌呤碱基编辑器。

5.一种如权利要求1所述的高精度腺嘌呤碱基编辑器或者如权利要求2所述的表达载体作为编辑基因组DNA的编辑工具在基因组DNA片段的相关编辑中的应用。

6.根据权利要求5所述的应用，其特征在于，所述基因编辑是编辑位点大于两个的多位点编辑；所述编辑的手段是腺嘌呤(A)到鸟嘌呤(G)的碱基转换。

7.根据权利要求6所述的应用，其特性在于，所述的六个高精度腺嘌呤碱基编辑器即4L-ABE8e、4P-ABE8e、6L-ABE8e、7L-ABE8e、7P-ABE8e和12EK-ABE8e作为编辑工具，包括与靶标DNA片段匹配的引导sgRNA；所述的六个高精度腺嘌呤碱基编辑器能够与介导它的sgRNA组合，对目的基因进行编辑。

8.根据权利要求7所述的应用，其特性在于，所述的表达载体和与之匹配的引导sgRNA一同转入宿主细胞，对基因进行编辑。

9.根据权利要求7所述的应用，其特性在于，所述多位点的腺嘌呤(A)到鸟嘌呤(G)的碱基转换，是利用腺嘌呤碱基编辑器对腺嘌呤(A)的脱氨作用，并通过宿主细胞的修复系统对双链之间的错配进行修复，从而将碱基从腺嘌呤(A)转换成鸟嘌呤(G)。