CN113135985A - 一种生产l-谷氨酸的方法 - Google Patents

一种生产l-谷氨酸的方法 Download PDF

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CN113135985A
CN113135985A CN202010062047.6A CN202010062047A CN113135985A CN 113135985 A CN113135985 A CN 113135985A CN 202010062047 A CN202010062047 A CN 202010062047A CN 113135985 A CN113135985 A CN 113135985A
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朱蕾蕾
聂志华
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Abstract

本发明公开了一种生产L‑谷氨酸的方法。本发明通过定向进化的方法获得了谷氨酸外运能力提高的外运变体，并提供了一种生产L‑谷氨酸的方法，包括如下步骤：在受体菌中表达所述谷氨酸外运蛋白变体，得到重组菌；对所述重组菌进行发酵培养，从发酵液中获得谷氨酸。本发明为丰富谷氨酸外运的理解及提高生产中谷氨酸的产量奠定了基础。

Description

一种生产L-谷氨酸的方法
技术领域
本发明涉及基因工程技术领域，具体涉及一种生产L-谷氨酸的方法。
背景技术
谷氨酸棒杆菌的历史始于20世纪50年代，当时日本科学家Kinoshita和Udaka分离出这种细菌并发现了它们分泌大量谷氨酸的能力。后来，用细菌发酵来生产谷氨酸，从而完全用生物过程取代化学合成。随后，该细菌不断被科学家研究，并利用其独特的生理学特性来改善谷氨酸的生产。此外，谷氨酸棒杆菌现已成为生产各种氨基酸的工业主力菌。在氨基酸市场中，广泛用作增味剂的L-谷氨酸市场规模巨大，每年生产总量约250万吨，占氨基酸生产总量的60％-70％左右。因此，研究如何提高谷氨酸的产量是十分必要的。
但近年来随着分子生物学的快速发展，分子生物学技术在谷氨酸研究领域的应用越来越广泛，大量谷氨酸发酵的机理被不断揭示。发现负责谷氨酸外运是由外运蛋白负责的，如：mscCG基因编码的外运蛋白。目前对于这类外运蛋白的研究较少，未发现通过突变提高其转运能力的文献。所以，通过对外运蛋白进行基因改造，筛选出外运谷氨酸能力较强的L-谷氨酸棒杆菌菌株，从而提高谷氨酸产量是非常有必要的。
发明内容
为了克服现有技术的不足，本发明的目的是提供一种外运能力提高的谷氨酸外运蛋白变体及其编码基因和相关应用。
第一方面，本发明要求保护谷氨酸外运蛋白变体。
本发明所要求保护的谷氨酸外运蛋白变体，为将MscCG谷氨酸外运蛋白进行点突变后所得，突变位点含有(或为)如下中的部分或全部：自N端起第166位、第86位、第344位、第385位、第165位、第103位、第393位、第399位、第173位、第343位、第310位、第168位、第504位、第186位、第141位、第200位、第205位、第14位、第334位和第362位。
优选的，所述谷氨酸外运蛋白变体的氨基酸序列与仅含有上述各突变位点的序列相比具有95％以上的同一性。
进一步地，所述谷氨酸外运蛋白变体为将所述MscCG谷氨酸外运蛋白的至少如下位点的氨基酸残基进行点突变后得到的蛋白：自N端起第166位。
更进一步，所述谷氨酸外运蛋白变体为将所述MscCG谷氨酸外运蛋白的至少如下位点的氨基酸残基进行点突变后得到的蛋白：自N端起第166位、第86位、第344位、第385位。
进一步地，所述谷氨酸外运蛋白变体可为如下任一：
(A1)所述谷氨酸外运蛋白变体为将所述MscCG谷氨酸外运蛋白的至少如下位点(或如下位点)的氨基酸残基进行点突变后得到的蛋白：自N端起第86位、第166位、第344位、第385位和第141位(对应突变体M7)；
(A2)所述谷氨酸外运蛋白变体为将所述MscCG谷氨酸外运蛋白的至少如下位点(或如下位点)的氨基酸残基进行点突变后得到的蛋白：自N端起第86位、第166位、第344位、第385位、第200位和第205位(对应突变体M8)；
(A3)所述谷氨酸外运蛋白变体为将所述MscCG谷氨酸外运蛋白的至少如下位点(或如下位点)的氨基酸残基进行点突变后得到的蛋白：自N端起第86位、第166位、第344位、第385位、第168位和第504位(对应突变体M5)；
(A4)所述谷氨酸外运蛋白变体为将所述MscCG谷氨酸外运蛋白的至少如下位点(或如下位点)的氨基酸残基进行点突变后得到的蛋白：自N端起第86位、第166位、第344位、第385位和第186位(对应突变体M6)；
(A5)所述谷氨酸外运蛋白变体为将所述MscCG谷氨酸外运蛋白的至少如下位点(或如下位点)的氨基酸残基进行点突变后得到的蛋白：自N端起第86位、第166位、第344位、第385位和第14位(对应突变体M9)；
(A6)所述谷氨酸外运蛋白变体为将所述MscCG谷氨酸外运蛋白的至少如下位点(或如下位点)的氨基酸残基进行点突变后得到的蛋白：自N端起第166位和第310位(对应突变体M4)；
(A7)所述谷氨酸外运蛋白变体为将所述MscCG谷氨酸外运蛋白的至少如下位点(或如下位点)的氨基酸残基进行点突变后得到的蛋白：自N端起第334位和第362位(对应突变体M10)；
(A8)所述谷氨酸外运蛋白变体为将所述MscCG谷氨酸外运蛋白的至少如下位点(或如下位点)的氨基酸残基进行点突变后得到的蛋白：自N端起第165位(对应突变体M1)；
(A9)所述谷氨酸外运蛋白变体为将所述MscCG谷氨酸外运蛋白的至少如下位点(或如下位点)的氨基酸残基进行点突变后得到的蛋白：自N端起第173位和第343位(对应突变体M3)；
(A10)所述谷氨酸外运蛋白变体为将所述MscCG谷氨酸外运蛋白的至少如下位点(或如下位点)的氨基酸残基进行点突变后得到的蛋白：自N端起第103位、第393位和第399位(对应突变体M2)。
其中，所述MscCG谷氨酸外运蛋白可为来自于谷氨酸棒杆菌ATCC13032的MscCG谷氨酸外运蛋白。
所述来自于谷氨酸棒杆菌ATCC13032的MscCG谷氨酸外运蛋白的氨基酸序列具体如SEQ ID No.1所示。
在所述谷氨酸外运蛋白变体中，自N端起第166位的点突变具体为T166A、第86位的点突变具体为M86V、第344位的点突变具体为I344V、第385位的点突变具体为Y385C、第165位的点突变具体为E165G、第103位的点突变具体为A103T、第393位的点突变具体为I393V、第399位的点突变具体为R399C、第173位的点突变具体为T173A、第343位的点突变具体为V343E、第310位的点突变具体为S310P、第168位的点突变具体为I168T、第504位的点突变具体为Q504L、第186位的点突变具体为R186C、第141位的点突变具体为N141D、第200位的点突变具体为N200D、第205位的点突变具体为I205V、第14位的点突变具体为L14S、第334位的点突变具体为D334G、第362位的点突变具体为L362P。
对于氨基酸取代，使用下述命名法：原始氨基酸(野生型)，位置(即在SEQ ID No.1中的位置)，取代氨基酸。相应地，在SEQ ID No.1的第166位用丙氨酸取代原有的苏氨酸氨酸命名为“T166A”。
在本发明的具体实施方式中，所述谷氨酸外运蛋白变体具体为如下任一：
(a1)所述谷氨酸外运蛋白变体为将SEQ ID No.1所示的MscCG谷氨酸外运蛋白的如下位点的氨基酸残基进行点突变后得到的蛋白：M86V、T166A、I344V、Y385C、N141D(对应突变体M7，氨基酸序列如SEQ ID No.8所示)；
(a2)所述谷氨酸外运蛋白变体为将SEQ ID No.1所示的MscCG谷氨酸外运蛋白的如下位点的氨基酸残基进行点突变后得到的蛋白：M86V、T166A、I344V、Y385C、N200D、I205V(对应突变体M8，氨基酸序列如SEQ ID No.9所示)；
(a3)所述谷氨酸外运蛋白变体为将SEQ ID No.1所示的MscCG谷氨酸外运蛋白的如下位点的氨基酸残基进行点突变后得到的蛋白：M86V、T166A、I344V、Y385C、I168T、Q504L(对应突变体M5，氨基酸序列如SEQ ID No.6所示)；
(a4)所述谷氨酸外运蛋白变体为将SEQ ID No.1所示的MscCG谷氨酸外运蛋白的如下位点的氨基酸残基进行点突变后得到的蛋白：M86V、T166A、I344V、Y385C、R186C(对应突变体M6，氨基酸序列如SEQ ID No.7所示)；
(a5)所述谷氨酸外运蛋白变体为将SEQ ID No.1所示的MscCG谷氨酸外运蛋白的如下位点的氨基酸残基进行点突变后得到的蛋白：M86V、T166A、I344V、Y385C、L14S(对应突变体M9，氨基酸序列如SEQ ID No.10所示)；
(a6)所述谷氨酸外运蛋白变体为将SEQ ID No.1所示的MscCG谷氨酸外运蛋白的如下位点的氨基酸残基进行点突变后得到的蛋白：T166A、S310P(对应突变体M4，氨基酸序列如SEQ ID No.5所示)；
(a7)所述谷氨酸外运蛋白变体为将SEQ ID No.1所示的MscCG谷氨酸外运蛋白的如下位点的氨基酸残基进行点突变后得到的蛋白：D334G、L362P(对应突变体M10，氨基酸序列如SEQ ID No.11所示)；
(a8)所述谷氨酸外运蛋白变体为将SEQ ID No.1所示的MscCG谷氨酸外运蛋白的如下位点的氨基酸残基进行点突变后得到的蛋白：E165G(对应突变体M1，氨基酸序列如SEQID No.2所示)；
(a9)所述谷氨酸外运蛋白变体为将SEQ ID No.1所示的MscCG谷氨酸外运蛋白的如下位点的氨基酸残基进行点突变后得到的蛋白：T173A、V343E(对应突变体M3，氨基酸序列如SEQ ID No.4所示)；
(a10)所述谷氨酸外运蛋白变体为将SEQ ID No.1所示的MscCG谷氨酸外运蛋白的如下位点的氨基酸残基进行点突变后得到的蛋白：A103T、I393V、R399C(对应突变体M2，氨基酸序列如SEQ ID No.3所示)。
第二方面，本发明要求保护谷氨酸外运蛋白变体相关的生物材料。
本发明所要求保护的谷氨酸外运蛋白变体相关的生物材料，可为如下任一：
(I)编码所述谷氨酸外运蛋白变体的核酸分子；
(II)含有所述核酸分子的表达盒、重组载体、重组菌或转基因细胞系。
其中，编码所述来自于谷氨酸棒杆菌的MscCG谷氨酸外运蛋白的核酸分子为SEQID No.12所示DNA分子。
进一步地，编码所述谷氨酸外运蛋白变体的核酸分子具体为如下任一：
(B1)SEQ ID No.19所示DNA分子(对应突变体M7)；
(B2)SEQ ID No.20所示DNA分子(对应突变体M8)；
(B3)SEQ ID No.17所示DNA分子(对应突变体M5)；
(B4)SEQ ID No.18所示DNA分子(对应突变体M6)；
(B5)SEQ ID No.21所示DNA分子(对应突变体M9)；
(B6)SEQ ID No.16所示DNA分子(对应突变体M4)；
(B7)SEQ ID No.22所示DNA分子(对应突变体M10)；
(B8)SEQ ID No.13所示DNA分子(对应突变体M1)；
(B9)SEQ ID No.15所示DNA分子(对应突变体M3)；
(B10)SEQ ID No.14所示DNA分子(对应突变体M2)。
在本发明的具体实施方式中，所述重组载体为将所述“编码所述谷氨酸外运蛋白变体的核酸分子”克隆到pTRCmob载体的多克隆位点SacI和BamHI之间得到的重组质粒。
所述重组菌可为含有所述核酸分子的谷氨酸棒杆菌。
第三方面，本发明要求保护前文所述谷氨酸外运蛋白变体或生物材料在如下任一中的应用：
(C1)生产谷氨酸；
(C2)提高谷氨酸产量；
(C3)提高谷氨酸外运能力；
(C4)生产味精、香料或食品添加剂；
(C5)制备代盐剂、营养增补剂或生化试剂。
第四方面，本发明要求保护一种生产谷氨酸和/或提高谷氨酸产量和/或提高谷氨酸外运能力的方法。
本发明所要求保护的生产谷氨酸和/或提高谷氨酸产量的方法，可包括如下步骤：在受体菌中表达前文所述的谷氨酸外运蛋白变体，得到重组菌；对所述重组菌进行发酵培养，从发酵液中获得谷氨酸。
进一步地，在所述受体菌中表达所述谷氨酸外运蛋白变体，可通过像所述受体菌中导入前文所述的“编码所述谷氨酸外运蛋白变体的核酸分子”来实现。
更进一步地，所述“编码所述谷氨酸外运蛋白变体的核酸分子”可通过重组载体的形式导入所述受体菌中。
在本发明的具体实施方式中，所述重组载体具体为将所述“编码所述谷氨酸外运蛋白变体的核酸分子”克隆到pTRCmob载体的多克隆位点SacI和BamHI之间得到的重组质粒。
进一步地，所述受体菌为谷氨酸棒杆菌。
在所述方法中，所述发酵培养的培养基可为CGXII培养基，培养条件可为30℃、220r/min培养38h。
在本发明的具体实施方式中，是将所述重组菌接种于CGXII种子培养基中，以30℃、220r/min培养12h，然后以起始OD600为0.15的接种量接种于CGXII发酵培养基中，以30℃、220r/min培养38h。
在本发明中，所述谷氨酸具体为L-谷氨酸。
本发明通过定向进化的方法获得了谷氨酸外运能力提高的外运变体。相比于野生型，外运变体的外运能力提高。本发明所得谷氨酸外运蛋白变体更有利于提高谷氨酸的产量，为丰富谷氨酸外运的理解及提高生产中谷氨酸的产量奠定了基础。
附图说明
图1为在25mlCGXII发酵培养基中培养的外运变体与野生型谷氨酸产量对比图。
图2为在50mlCGXII发酵培养基中培养的外运变体与野生型谷氨酸产量对比图。
图3为在50mlCGXII发酵培养基中培养的外运变体与野生型谷氨酸产量对比图。
图1-3中标注MscCG的为野生型对照，纵坐标处“谷氨酸g/L”指的是每L发酵液中产L-谷氨酸的量。
具体实施方式
下述实施例中所使用的实验方法如无特殊说明，均为常规方法。
下述实施例中所用的材料、试剂等，如无特殊说明，均可从商业途径得到。
实施例1、谷氨酸外运蛋白的基因克隆
从谷氨酸棒杆菌的基因组中克隆出MscCG谷氨酸外运蛋白目的基因，基因序列如SEQ ID NO.12所示，然后连接到pTRCmob载体中，测序确定载体构建成功得到重组质粒，命名为pTRCmob-MscCG。
pTRCmob-MscCG的结构描述为：将SEQ ID NO.12所示DNA片段克隆到pTRCmob载体的酶切位点SacI和BamHI之间后得到的重组质粒。SEQ ID NO.12为野生型MscCG谷氨酸外运蛋白目的基因，编码SEQ ID NO.1所示的野生型MscCG谷氨酸外运蛋白。
实施例2、外运能力提高的谷氨酸外运蛋白突变体筛选
为了提高上述谷氨酸外运蛋白外运能力，以实施例1中构建得到的重组质粒pTRCmob-MscCG为模板，设计引物(上游引物：5′-ATGATTTTAGGCGTACCCAT-3′；下游引物：5′-CTAAGGGGTGGACGTCGGCG-3′)，加入Mn2+用PCR的方法对目的基因进行随机突变。将突变产物电转到谷氨酸棒杆菌感受态细胞中，30℃过夜培养得到重组菌之后，将长出的单克隆分别用无菌牙签接种到96孔板中(含有150μl LBG培养基，25mg/ml Kana)，30℃、800rpm摇床培养45h后检测谷氨酸的产量。对检测结果进行分析，最终筛选出10株谷氨酸外运量提高的突变菌株，分别命名为M1、M2、M3、M4、M5、M6、M7、M8、M9、M10。
对上述各突变菌株进行基因测序，其具体的突变氨基酸位点，以及突变后的具体氨基酸序列和基因序列详见表1。
表1突变氨基酸位点
Figure BDA0002374795210000061
Figure BDA0002374795210000071
注：表中的氨基酸取代，使用下述命名法：原始氨基酸(野生型)，位置(即在SEQ IDNo.1中的位置)，取代氨基酸。相应地，在SEQ ID No.1的第166位用丙氨酸取代原有的苏氨酸氨酸命名为“T166A”。
实施例3、谷氨酸外运蛋白及其外运变体在谷氨酸棒杆菌中的表达
接种环刮取野生型和实施例2获得的表1中所示10个突变体的阳性菌接种于10mLCGXII种子培养基中，以30℃、220rpm在恒温培养摇床(上海知楚仪器有限公司，上海，中国)培养12h，然后以起始OD600为0.15的接种量分别接种于25ml CGXII发酵培养基和50mlCGXII发酵培养基中(接种到500ml的锥形瓶中)，以30℃、220rpm在恒温培养摇床(上海知楚仪器有限公司，上海，中国)培养。培养38h后离心收集发酵液，用SBA-40D生物传感分析仪(山东省科学院生物研究所，山东，中国)测发酵液中L-谷氨酸的产量。
其中，CGXII种子培养基的溶剂为水，溶质及浓度如下：5g/L葡萄糖，20g/L硫酸铵，5g/L尿素，1g/L磷酸二氢钾，1.3g/L磷酸氢二钾，80g/L MOPS，0.01g/L氯化钙，0.25g/L硫酸镁，0.01g/L硫酸亚铁，0.01g/L硫酸锰，0.001g/L硫酸锌，0.2mg/L硫酸铜，0.02mg/L氯化镍，0.03g/L二羟基苯甲酸，2.5μg/L生物素，0.1mg/L Thiamine HCL VB1。
CGXII发酵培养基的溶剂为水，溶质及浓度如下：80g/L葡萄糖，20g/L硫酸铵，5g/L尿素，1g/L磷酸二氢钾，1.3g/L磷酸氢二钾，80g/L MOPS，0.01g/L氯化钙，0.25g/L硫酸镁，0.01g/L硫酸亚铁，0.01g/L硫酸锰，0.001g/L硫酸锌，0.2mg/L硫酸铜，0.02mg/L氯化镍，0.03g/L二羟基苯甲酸，1μg/L生物素，0.1mg/L Thiamine HCL VB1。
检测结果如图1、图2、图3所示。图1是在25ml CGXII发酵培养基中培养的，结果表明外运变体M2、M6、M7、M10谷氨酸的产量分别比野生型谷氨酸的产量高33.3％、37.5％、33.3％、25％。图2是在50ml CGXII发酵培养基中培养的，结果表明外运变体M5、M8、M9谷氨酸的产量分别是野生型谷氨酸产量的4.4倍、6.6倍、5.2倍。图3是在50ml CGXII发酵培养基中培养的，结果表明外运变体M1、M3、M4、谷氨酸的产量分别比野生型谷氨酸的产量高50％、50％、25％。
虽然，上文中已经用一般性说明及具体实施方案对本发明作了详尽的描述，但在本发明基础上，可以对之作一些修改或改进，这对本领域技术人员而言是显而易见的。因此，在不偏离本发明精神的基础上所做的修改或改进，均属于本发明要求保护的范围。
<110> 中国科学院天津工业生物技术研究所
<120> 一种生产L-谷氨酸的方法
<130> GNCLN200292
<160> 22
<170> PatentIn version 3.5
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Met Ile Leu Gly Val Pro Ile Gln Tyr Leu Leu Tyr Ser Leu Trp Asn
1 5 10 15
Trp Ile Val Asp Thr Gly Phe Asp Val Ala Ile Ile Leu Val Leu Ala
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Phe Leu Ile Pro Arg Ile Gly Arg Leu Ala Met Arg Ile Ile Lys Arg
35 40 45
Arg Val Glu Ser Ala Ala Asp Ala Asp Thr Thr Lys Asn Gln Leu Ala
50 55 60
Phe Ala Gly Val Gly Val Tyr Ile Ala Gln Ile Val Ala Phe Phe Met
65 70 75 80
Leu Ala Val Ser Ala Met Gln Ala Phe Gly Phe Ser Leu Ala Gly Ala
85 90 95
Ala Ile Pro Ala Thr Ile Ala Ser Ala Ala Ile Gly Leu Gly Ala Gln
100 105 110
Ser Ile Val Ala Asp Phe Leu Ala Gly Phe Phe Ile Leu Thr Glu Lys
115 120 125
Gln Phe Gly Val Gly Asp Trp Val Arg Phe Glu Gly Asn Gly Ile Val
130 135 140
Val Glu Gly Thr Val Ile Glu Ile Thr Met Arg Ala Thr Lys Ile Arg
145 150 155 160
Thr Ile Ala Gln Glu Thr Val Ile Ile Pro Asn Ser Thr Ala Lys Val
165 170 175
Cys Ile Asn Asn Ser Asn Asn Trp Ser Arg Ala Val Val Val Ile Pro
180 185 190
Ile Pro Met Leu Gly Ser Glu Asn Ile Thr Asp Val Ile Ala Arg Ser
195 200 205
Glu Ala Ala Thr Arg Arg Ala Leu Gly Gln Glu Lys Ile Ala Pro Glu
210 215 220
Ile Leu Gly Glu Leu Asp Val His Pro Ala Thr Glu Val Thr Pro Pro
225 230 235 240
Thr Val Val Gly Met Pro Trp Met Val Thr Met Arg Phe Leu Val Gln
245 250 255
Val Thr Ala Gly Asn Gln Trp Leu Val Glu Arg Ala Ile Arg Thr Glu
260 265 270
Ile Ile Ser Glu Phe Trp Glu Glu Tyr Gly Ser Ala Thr Thr Thr Ser
275 280 285
Gly Thr Leu Ile Asp Ser Leu His Val Glu His Glu Glu Pro Lys Thr
290 295 300
Ser Leu Ile Asp Ala Ser Pro Gln Ala Leu Lys Glu Pro Lys Pro Glu
305 310 315 320
Ala Ala Ala Thr Val Ala Ser Leu Ala Ala Ser Ser Asn Asp Asp Ala
325 330 335
Asp Asn Ala Asp Ala Ser Val Ile Asn Ala Gly Asn Pro Glu Lys Glu
340 345 350
Leu Asp Ser Asp Val Leu Glu Gln Glu Leu Ser Ser Glu Glu Pro Glu
355 360 365
Glu Thr Ala Lys Pro Asp His Ser Leu Arg Gly Phe Phe Arg Thr Asp
370 375 380
Tyr Tyr Pro Asn Arg Trp Gln Lys Ile Leu Ser Phe Gly Gly Arg Val
385 390 395 400
Arg Met Ser Thr Ser Leu Leu Leu Gly Ala Leu Leu Leu Leu Ser Leu
405 410 415
Phe Lys Val Met Thr Val Glu Pro Ser Glu Asn Trp Gln Asn Ser Ser
420 425 430
Gly Trp Leu Ser Pro Ser Thr Ala Thr Ser Thr Ala Val Thr Thr Ser
435 440 445
Glu Thr Ser Ala Pro Val Ser Thr Pro Ser Met Thr Val Pro Thr Thr
450 455 460
Val Glu Glu Thr Pro Thr Met Glu Ser Asn Val Glu Thr Gln Gln Glu
465 470 475 480
Thr Ser Thr Pro Ala Thr Ala Thr Pro Gln Arg Ala Asp Thr Ile Glu
485 490 495
Pro Thr Glu Glu Ala Thr Ser Gln Glu Glu Thr Thr Ala Ser Gln Thr
500 505 510
Gln Ser Pro Ala Val Glu Ala Pro Thr Ala Val Gln Glu Thr Val Ala
515 520 525
Pro Thr Ser Thr Pro
530
<210> 2
<211> 533
<212> PRT
<213> Artificial sequence
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Met Ile Leu Gly Val Pro Ile Gln Tyr Leu Leu Tyr Ser Leu Trp Asn
1 5 10 15
Trp Ile Val Asp Thr Gly Phe Asp Val Ala Ile Ile Leu Val Leu Ala
20 25 30
Phe Leu Ile Pro Arg Ile Gly Arg Leu Ala Met Arg Ile Ile Lys Arg
35 40 45
Arg Val Glu Ser Ala Ala Asp Ala Asp Thr Thr Lys Asn Gln Leu Ala
50 55 60
Phe Ala Gly Val Gly Val Tyr Ile Ala Gln Ile Val Ala Phe Phe Met
65 70 75 80
Leu Ala Val Ser Ala Met Gln Ala Phe Gly Phe Ser Leu Ala Gly Ala
85 90 95
Ala Ile Pro Ala Thr Ile Ala Ser Ala Ala Ile Gly Leu Gly Ala Gln
100 105 110
Ser Ile Val Ala Asp Phe Leu Ala Gly Phe Phe Ile Leu Thr Glu Lys
115 120 125
Gln Phe Gly Val Gly Asp Trp Val Arg Phe Glu Gly Asn Gly Ile Val
130 135 140
Val Glu Gly Thr Val Ile Glu Ile Thr Met Arg Ala Thr Lys Ile Arg
145 150 155 160
Thr Ile Ala Gln Gly Thr Val Ile Ile Pro Asn Ser Thr Ala Lys Val
165 170 175
Cys Ile Asn Asn Ser Asn Asn Trp Ser Arg Ala Val Val Val Ile Pro
180 185 190
Ile Pro Met Leu Gly Ser Glu Asn Ile Thr Asp Val Ile Ala Arg Ser
195 200 205
Glu Ala Ala Thr Arg Arg Ala Leu Gly Gln Glu Lys Ile Ala Pro Glu
210 215 220
Ile Leu Gly Glu Leu Asp Val His Pro Ala Thr Glu Val Thr Pro Pro
225 230 235 240
Thr Val Val Gly Met Pro Trp Met Val Thr Met Arg Phe Leu Val Gln
245 250 255
Val Thr Ala Gly Asn Gln Trp Leu Val Glu Arg Ala Ile Arg Thr Glu
260 265 270
Ile Ile Ser Glu Phe Trp Glu Glu Tyr Gly Ser Ala Thr Thr Thr Ser
275 280 285
Gly Thr Leu Ile Asp Ser Leu His Val Glu His Glu Glu Pro Lys Thr
290 295 300
Ser Leu Ile Asp Ala Ser Pro Gln Ala Leu Lys Glu Pro Lys Pro Glu
305 310 315 320
Ala Ala Ala Thr Val Ala Ser Leu Ala Ala Ser Ser Asn Asp Asp Ala
325 330 335
Asp Asn Ala Asp Ala Ser Val Ile Asn Ala Gly Asn Pro Glu Lys Glu
340 345 350
Leu Asp Ser Asp Val Leu Glu Gln Glu Leu Ser Ser Glu Glu Pro Glu
355 360 365
Glu Thr Ala Lys Pro Asp His Ser Leu Arg Gly Phe Phe Arg Thr Asp
370 375 380
Tyr Tyr Pro Asn Arg Trp Gln Lys Ile Leu Ser Phe Gly Gly Arg Val
385 390 395 400
Arg Met Ser Thr Ser Leu Leu Leu Gly Ala Leu Leu Leu Leu Ser Leu
405 410 415
Phe Lys Val Met Thr Val Glu Pro Ser Glu Asn Trp Gln Asn Ser Ser
420 425 430
Gly Trp Leu Ser Pro Ser Thr Ala Thr Ser Thr Ala Val Thr Thr Ser
435 440 445
Glu Thr Ser Ala Pro Val Ser Thr Pro Ser Met Thr Val Pro Thr Thr
450 455 460
Val Glu Glu Thr Pro Thr Met Glu Ser Asn Val Glu Thr Gln Gln Glu
465 470 475 480
Thr Ser Thr Pro Ala Thr Ala Thr Pro Gln Arg Ala Asp Thr Ile Glu
485 490 495
Pro Thr Glu Glu Ala Thr Ser Gln Glu Glu Thr Thr Ala Ser Gln Thr
500 505 510
Gln Ser Pro Ala Val Glu Ala Pro Thr Ala Val Gln Glu Thr Val Ala
515 520 525
Pro Thr Ser Thr Pro
530
<210> 3
<211> 533
<212> PRT
<213> Artificial sequence
<400> 3
Met Ile Leu Gly Val Pro Ile Gln Tyr Leu Leu Tyr Ser Leu Trp Asn
1 5 10 15
Trp Ile Val Asp Thr Gly Phe Asp Val Ala Ile Ile Leu Val Leu Ala
20 25 30
Phe Leu Ile Pro Arg Ile Gly Arg Leu Ala Met Arg Ile Ile Lys Arg
35 40 45
Arg Val Glu Ser Ala Ala Asp Ala Asp Thr Thr Lys Asn Gln Leu Ala
50 55 60
Phe Ala Gly Val Gly Val Tyr Ile Ala Gln Ile Val Ala Phe Phe Met
65 70 75 80
Leu Ala Val Ser Ala Met Gln Ala Phe Gly Phe Ser Leu Ala Gly Ala
85 90 95
Ala Ile Pro Ala Thr Ile Thr Ser Ala Ala Ile Gly Leu Gly Ala Gln
100 105 110
Ser Ile Val Ala Asp Phe Leu Ala Gly Phe Phe Ile Leu Thr Glu Lys
115 120 125
Gln Phe Gly Val Gly Asp Trp Val Arg Phe Glu Gly Asn Gly Ile Val
130 135 140
Val Glu Gly Thr Val Ile Glu Ile Thr Met Arg Ala Thr Lys Ile Arg
145 150 155 160
Thr Ile Ala Gln Glu Thr Val Ile Ile Pro Asn Ser Thr Ala Lys Val
165 170 175
Cys Ile Asn Asn Ser Asn Asn Trp Ser Arg Ala Val Val Val Ile Pro
180 185 190
Ile Pro Met Leu Gly Ser Glu Asn Ile Thr Asp Val Ile Ala Arg Ser
195 200 205
Glu Ala Ala Thr Arg Arg Ala Leu Gly Gln Glu Lys Ile Ala Pro Glu
210 215 220
Ile Leu Gly Glu Leu Asp Val His Pro Ala Thr Glu Val Thr Pro Pro
225 230 235 240
Thr Val Val Gly Met Pro Trp Met Val Thr Met Arg Phe Leu Val Gln
245 250 255
Val Thr Ala Gly Asn Gln Trp Leu Val Glu Arg Ala Ile Arg Thr Glu
260 265 270
Ile Ile Ser Glu Phe Trp Glu Glu Tyr Gly Ser Ala Thr Thr Thr Ser
275 280 285
Gly Thr Leu Ile Asp Ser Leu His Val Glu His Glu Glu Pro Lys Thr
290 295 300
Ser Leu Ile Asp Ala Ser Pro Gln Ala Leu Lys Glu Pro Lys Pro Glu
305 310 315 320
Ala Ala Ala Thr Val Ala Ser Leu Ala Ala Ser Ser Asn Asp Asp Ala
325 330 335
Asp Asn Ala Asp Ala Ser Val Ile Asn Ala Gly Asn Pro Glu Lys Glu
340 345 350
Leu Asp Ser Asp Val Leu Glu Gln Glu Leu Ser Ser Glu Glu Pro Glu
355 360 365
Glu Thr Ala Lys Pro Asp His Ser Leu Arg Gly Phe Phe Arg Thr Asp
370 375 380
Tyr Tyr Pro Asn Arg Trp Gln Lys Val Leu Ser Phe Gly Gly Cys Val
385 390 395 400
Arg Met Ser Thr Ser Leu Leu Leu Gly Ala Leu Leu Leu Leu Ser Leu
405 410 415
Phe Lys Val Met Thr Val Glu Pro Ser Glu Asn Trp Gln Asn Ser Ser
420 425 430
Gly Trp Leu Ser Pro Ser Thr Ala Thr Ser Thr Ala Val Thr Thr Ser
435 440 445
Glu Thr Ser Ala Pro Val Ser Thr Pro Ser Met Thr Val Pro Thr Thr
450 455 460
Val Glu Glu Thr Pro Thr Met Glu Ser Asn Val Glu Thr Gln Gln Glu
465 470 475 480
Thr Ser Thr Pro Ala Thr Ala Thr Pro Gln Arg Ala Asp Thr Ile Glu
485 490 495
Pro Thr Glu Glu Ala Thr Ser Gln Glu Glu Thr Thr Ala Ser Gln Thr
500 505 510
Gln Ser Pro Ala Val Glu Ala Pro Thr Ala Val Gln Glu Thr Val Ala
515 520 525
Pro Thr Ser Thr Pro
530
<210> 4
<211> 533
<212> PRT
<213> Artificial sequence
<400> 4
Met Ile Leu Gly Val Pro Ile Gln Tyr Leu Leu Tyr Ser Leu Trp Asn
1 5 10 15
Trp Ile Val Asp Thr Gly Phe Asp Val Ala Ile Ile Leu Val Leu Ala
20 25 30
Phe Leu Ile Pro Arg Ile Gly Arg Leu Ala Met Arg Ile Ile Lys Arg
35 40 45
Arg Val Glu Ser Ala Ala Asp Ala Asp Thr Thr Lys Asn Gln Leu Ala
50 55 60
Phe Ala Gly Val Gly Val Tyr Ile Ala Gln Ile Val Ala Phe Phe Met
65 70 75 80
Leu Ala Val Ser Ala Met Gln Ala Phe Gly Phe Ser Leu Ala Gly Ala
85 90 95
Ala Ile Pro Ala Thr Ile Ala Ser Ala Ala Ile Gly Leu Gly Ala Gln
100 105 110
Ser Ile Val Ala Asp Phe Leu Ala Gly Phe Phe Ile Leu Thr Glu Lys
115 120 125
Gln Phe Gly Val Gly Asp Trp Val Arg Phe Glu Gly Asn Gly Ile Val
130 135 140
Val Glu Gly Thr Val Ile Glu Ile Thr Met Arg Ala Thr Lys Ile Arg
145 150 155 160
Thr Ile Ala Gln Glu Thr Val Ile Ile Pro Asn Ser Ala Ala Lys Val
165 170 175
Cys Ile Asn Asn Ser Asn Asn Trp Ser Arg Ala Val Val Val Ile Pro
180 185 190
Ile Pro Met Leu Gly Ser Glu Asn Ile Thr Asp Val Ile Ala Arg Ser
195 200 205
Glu Ala Ala Thr Arg Arg Ala Leu Gly Gln Glu Lys Ile Ala Pro Glu
210 215 220
Ile Leu Gly Glu Leu Asp Val His Pro Ala Thr Glu Val Thr Pro Pro
225 230 235 240
Thr Val Val Gly Met Pro Trp Met Val Thr Met Arg Phe Leu Val Gln
245 250 255
Val Thr Ala Gly Asn Gln Trp Leu Val Glu Arg Ala Ile Arg Thr Glu
260 265 270
Ile Ile Ser Glu Phe Trp Glu Glu Tyr Gly Ser Ala Thr Thr Thr Ser
275 280 285
Gly Thr Leu Ile Asp Ser Leu His Val Glu His Glu Glu Pro Lys Thr
290 295 300
Ser Leu Ile Asp Ala Ser Pro Gln Ala Leu Lys Glu Pro Lys Pro Glu
305 310 315 320
Ala Ala Ala Thr Val Ala Ser Leu Ala Ala Ser Ser Asn Asp Asp Ala
325 330 335
Asp Asn Ala Asp Ala Ser Glu Ile Asn Ala Gly Asn Pro Glu Lys Glu
340 345 350
Leu Asp Ser Asp Val Leu Glu Gln Glu Leu Ser Ser Glu Glu Pro Glu
355 360 365
Glu Thr Ala Lys Pro Asp His Ser Leu Arg Gly Phe Phe Arg Thr Asp
370 375 380
Tyr Tyr Pro Asn Arg Trp Gln Lys Ile Leu Ser Phe Gly Gly Arg Val
385 390 395 400
Arg Met Ser Thr Ser Leu Leu Leu Gly Ala Leu Leu Leu Leu Ser Leu
405 410 415
Phe Lys Val Met Thr Val Glu Pro Ser Glu Asn Trp Gln Asn Ser Ser
420 425 430
Gly Trp Leu Ser Pro Ser Thr Ala Thr Ser Thr Ala Val Thr Thr Ser
435 440 445
Glu Thr Ser Ala Pro Val Ser Thr Pro Ser Met Thr Val Pro Thr Thr
450 455 460
Val Glu Glu Thr Pro Thr Met Glu Ser Asn Val Glu Thr Gln Gln Glu
465 470 475 480
Thr Ser Thr Pro Ala Thr Ala Thr Pro Gln Arg Ala Asp Thr Ile Glu
485 490 495
Pro Thr Glu Glu Ala Thr Ser Gln Glu Glu Thr Thr Ala Ser Gln Thr
500 505 510
Gln Ser Pro Ala Val Glu Ala Pro Thr Ala Val Gln Glu Thr Val Ala
515 520 525
Pro Thr Ser Thr Pro
530
<210> 5
<211> 533
<212> PRT
<213> Artificial sequence
<400> 5
Met Ile Leu Gly Val Pro Ile Gln Tyr Leu Leu Tyr Ser Leu Trp Asn
1 5 10 15
Trp Ile Val Asp Thr Gly Phe Asp Val Ala Ile Ile Leu Val Leu Ala
20 25 30
Phe Leu Ile Pro Arg Ile Gly Arg Leu Ala Met Arg Ile Ile Lys Arg
35 40 45
Arg Val Glu Ser Ala Ala Asp Ala Asp Thr Thr Lys Asn Gln Leu Ala
50 55 60
Phe Ala Gly Val Gly Val Tyr Ile Ala Gln Ile Val Ala Phe Phe Met
65 70 75 80
Leu Ala Val Ser Ala Met Gln Ala Phe Gly Phe Ser Leu Ala Gly Ala
85 90 95
Ala Ile Pro Ala Thr Ile Ala Ser Ala Ala Ile Gly Leu Gly Ala Gln
100 105 110
Ser Ile Val Ala Asp Phe Leu Ala Gly Phe Phe Ile Leu Thr Glu Lys
115 120 125
Gln Phe Gly Val Gly Asp Trp Val Arg Phe Glu Gly Asn Gly Ile Val
130 135 140
Val Glu Gly Thr Val Ile Glu Ile Thr Met Arg Ala Thr Lys Ile Arg
145 150 155 160
Thr Ile Ala Gln Glu Ala Val Ile Ile Pro Asn Ser Thr Ala Lys Val
165 170 175
Cys Ile Asn Asn Ser Asn Asn Trp Ser Arg Ala Val Val Val Ile Pro
180 185 190
Ile Pro Met Leu Gly Ser Glu Asn Ile Thr Asp Val Ile Ala Arg Ser
195 200 205
Glu Ala Ala Thr Arg Arg Ala Leu Gly Gln Glu Lys Ile Ala Pro Glu
210 215 220
Ile Leu Gly Glu Leu Asp Val His Pro Ala Thr Glu Val Thr Pro Pro
225 230 235 240
Thr Val Val Gly Met Pro Trp Met Val Thr Met Arg Phe Leu Val Gln
245 250 255
Val Thr Ala Gly Asn Gln Trp Leu Val Glu Arg Ala Ile Arg Thr Glu
260 265 270
Ile Ile Ser Glu Phe Trp Glu Glu Tyr Gly Ser Ala Thr Thr Thr Ser
275 280 285
Gly Thr Leu Ile Asp Ser Leu His Val Glu His Glu Glu Pro Lys Thr
290 295 300
Ser Leu Ile Asp Ala Pro Pro Gln Ala Leu Lys Glu Pro Lys Pro Glu
305 310 315 320
Ala Ala Ala Thr Val Ala Ser Leu Ala Ala Ser Ser Asn Asp Asp Ala
325 330 335
Asp Asn Ala Asp Ala Ser Val Ile Asn Ala Gly Asn Pro Glu Lys Glu
340 345 350
Leu Asp Ser Asp Val Leu Glu Gln Glu Leu Ser Ser Glu Glu Pro Glu
355 360 365
Glu Thr Ala Lys Pro Asp His Ser Leu Arg Gly Phe Phe Arg Thr Asp
370 375 380
Tyr Tyr Pro Asn Arg Trp Gln Lys Ile Leu Ser Phe Gly Gly Arg Val
385 390 395 400
Arg Met Ser Thr Ser Leu Leu Leu Gly Ala Leu Leu Leu Leu Ser Leu
405 410 415
Phe Lys Val Met Thr Val Glu Pro Ser Glu Asn Trp Gln Asn Ser Ser
420 425 430
Gly Trp Leu Ser Pro Ser Thr Ala Thr Ser Thr Ala Val Thr Thr Ser
435 440 445
Glu Thr Ser Ala Pro Val Ser Thr Pro Ser Met Thr Val Pro Thr Thr
450 455 460
Val Glu Glu Thr Pro Thr Met Glu Ser Asn Val Glu Thr Gln Gln Glu
465 470 475 480
Thr Ser Thr Pro Ala Thr Ala Thr Pro Gln Arg Ala Asp Thr Ile Glu
485 490 495
Pro Thr Glu Glu Ala Thr Ser Gln Glu Glu Thr Thr Ala Ser Gln Thr
500 505 510
Gln Ser Pro Ala Val Glu Ala Pro Thr Ala Val Gln Glu Thr Val Ala
515 520 525
Pro Thr Ser Thr Pro
530
<210> 6
<211> 533
<212> PRT
<213> Artificial sequence
<400> 6
Met Ile Leu Gly Val Pro Ile Gln Tyr Leu Leu Tyr Ser Leu Trp Asn
1 5 10 15
Trp Ile Val Asp Thr Gly Phe Asp Val Ala Ile Ile Leu Val Leu Ala
20 25 30
Phe Leu Ile Pro Arg Ile Gly Arg Leu Ala Met Arg Ile Ile Lys Arg
35 40 45
Arg Val Glu Ser Ala Ala Asp Ala Asp Thr Thr Lys Asn Gln Leu Ala
50 55 60
Phe Ala Gly Val Gly Val Tyr Ile Ala Gln Ile Val Ala Phe Phe Met
65 70 75 80
Leu Ala Val Ser Ala Val Gln Ala Phe Gly Phe Ser Leu Ala Gly Ala
85 90 95
Ala Ile Pro Ala Thr Ile Ala Ser Ala Ala Ile Gly Leu Gly Ala Gln
100 105 110
Ser Ile Val Ala Asp Phe Leu Ala Gly Phe Phe Ile Leu Thr Glu Lys
115 120 125
Gln Phe Gly Val Gly Asp Trp Val Arg Phe Glu Gly Asn Gly Ile Val
130 135 140
Val Glu Gly Thr Val Ile Glu Ile Thr Met Arg Ala Thr Lys Ile Arg
145 150 155 160
Thr Ile Ala Gln Glu Ala Val Thr Ile Pro Asn Ser Thr Ala Lys Val
165 170 175
Cys Ile Asn Asn Ser Asn Asn Trp Ser Arg Ala Val Val Val Ile Pro
180 185 190
Ile Pro Met Leu Gly Ser Glu Asn Ile Thr Asp Val Ile Ala Arg Ser
195 200 205
Glu Ala Ala Thr Arg Arg Ala Leu Gly Gln Glu Lys Ile Ala Pro Glu
210 215 220
Ile Leu Gly Glu Leu Asp Val His Pro Ala Thr Glu Val Thr Pro Pro
225 230 235 240
Thr Val Val Gly Met Pro Trp Met Val Thr Met Arg Phe Leu Val Gln
245 250 255
Val Thr Ala Gly Asn Gln Trp Leu Val Glu Arg Ala Ile Arg Thr Glu
260 265 270
Ile Ile Ser Glu Phe Trp Glu Glu Tyr Gly Ser Ala Thr Thr Thr Ser
275 280 285
Gly Thr Leu Ile Asp Ser Leu His Val Glu His Glu Glu Pro Lys Thr
290 295 300
Ser Leu Ile Asp Ala Ser Pro Gln Ala Leu Lys Glu Pro Lys Pro Glu
305 310 315 320
Ala Ala Ala Thr Val Ala Ser Leu Ala Ala Ser Ser Asn Asp Asp Ala
325 330 335
Asp Asn Ala Asp Ala Ser Val Val Asn Ala Gly Asn Pro Glu Lys Glu
340 345 350
Leu Asp Ser Asp Val Leu Glu Gln Glu Leu Ser Ser Glu Glu Pro Glu
355 360 365
Glu Thr Ala Lys Pro Asp His Ser Leu Arg Gly Phe Phe Arg Thr Asp
370 375 380
Cys Tyr Pro Asn Arg Trp Gln Lys Ile Leu Ser Phe Gly Gly Arg Val
385 390 395 400
Arg Met Ser Thr Ser Leu Leu Leu Gly Ala Leu Leu Leu Leu Ser Leu
405 410 415
Phe Lys Val Met Thr Val Glu Pro Ser Glu Asn Trp Gln Asn Ser Ser
420 425 430
Gly Trp Leu Ser Pro Ser Thr Ala Thr Ser Thr Ala Val Thr Thr Ser
435 440 445
Glu Thr Ser Ala Pro Val Ser Thr Pro Ser Met Thr Val Pro Thr Thr
450 455 460
Val Glu Glu Thr Pro Thr Met Glu Ser Asn Val Glu Thr Gln Gln Glu
465 470 475 480
Thr Ser Thr Pro Ala Thr Ala Thr Pro Gln Arg Ala Asp Thr Ile Glu
485 490 495
Pro Thr Glu Glu Ala Thr Ser Leu Glu Glu Thr Thr Ala Ser Gln Thr
500 505 510
Gln Ser Pro Ala Val Glu Ala Pro Thr Ala Val Gln Glu Thr Val Ala
515 520 525
Pro Thr Ser Thr Pro
530
<210> 7
<211> 533
<212> PRT
<213> Artificial sequence
<400> 7
Met Ile Leu Gly Val Pro Ile Gln Tyr Leu Leu Tyr Ser Leu Trp Asn
1 5 10 15
Trp Ile Val Asp Thr Gly Phe Asp Val Ala Ile Ile Leu Val Leu Ala
20 25 30
Phe Leu Ile Pro Arg Ile Gly Arg Leu Ala Met Arg Ile Ile Lys Arg
35 40 45
Arg Val Glu Ser Ala Ala Asp Ala Asp Thr Thr Lys Asn Gln Leu Ala
50 55 60
Phe Ala Gly Val Gly Val Tyr Ile Ala Gln Ile Val Ala Phe Phe Met
65 70 75 80
Leu Ala Val Ser Ala Val Gln Ala Phe Gly Phe Ser Leu Ala Gly Ala
85 90 95
Ala Ile Pro Ala Thr Ile Ala Ser Ala Ala Ile Gly Leu Gly Ala Gln
100 105 110
Ser Ile Val Ala Asp Phe Leu Ala Gly Phe Phe Ile Leu Thr Glu Lys
115 120 125
Gln Phe Gly Val Gly Asp Trp Val Arg Phe Glu Gly Asn Gly Ile Val
130 135 140
Val Glu Gly Thr Val Ile Glu Ile Thr Met Arg Ala Thr Lys Ile Arg
145 150 155 160
Thr Ile Ala Gln Glu Ala Val Ile Ile Pro Asn Ser Thr Ala Lys Val
165 170 175
Cys Ile Asn Asn Ser Asn Asn Trp Ser Cys Ala Val Val Val Ile Pro
180 185 190
Ile Pro Met Leu Gly Ser Glu Asn Ile Thr Asp Val Ile Ala Arg Ser
195 200 205
Glu Ala Ala Thr Arg Arg Ala Leu Gly Gln Glu Lys Ile Ala Pro Glu
210 215 220
Ile Leu Gly Glu Leu Asp Val His Pro Ala Thr Glu Val Thr Pro Pro
225 230 235 240
Thr Val Val Gly Met Pro Trp Met Val Thr Met Arg Phe Leu Val Gln
245 250 255
Val Thr Ala Gly Asn Gln Trp Leu Val Glu Arg Ala Ile Arg Thr Glu
260 265 270
Ile Ile Ser Glu Phe Trp Glu Glu Tyr Gly Ser Ala Thr Thr Thr Ser
275 280 285
Gly Thr Leu Ile Asp Ser Leu His Val Glu His Glu Glu Pro Lys Thr
290 295 300
Ser Leu Ile Asp Ala Ser Pro Gln Ala Leu Lys Glu Pro Lys Pro Glu
305 310 315 320
Ala Ala Ala Thr Val Ala Ser Leu Ala Ala Ser Ser Asn Asp Asp Ala
325 330 335
Asp Asn Ala Asp Ala Ser Val Val Asn Ala Gly Asn Pro Glu Lys Glu
340 345 350
Leu Asp Ser Asp Val Leu Glu Gln Glu Leu Ser Ser Glu Glu Pro Glu
355 360 365
Glu Thr Ala Lys Pro Asp His Ser Leu Arg Gly Phe Phe Arg Thr Asp
370 375 380
Cys Tyr Pro Asn Arg Trp Gln Lys Ile Leu Ser Phe Gly Gly Arg Val
385 390 395 400
Arg Met Ser Thr Ser Leu Leu Leu Gly Ala Leu Leu Leu Leu Ser Leu
405 410 415
Phe Lys Val Met Thr Val Glu Pro Ser Glu Asn Trp Gln Asn Ser Ser
420 425 430
Gly Trp Leu Ser Pro Ser Thr Ala Thr Ser Thr Ala Val Thr Thr Ser
435 440 445
Glu Thr Ser Ala Pro Val Ser Thr Pro Ser Met Thr Val Pro Thr Thr
450 455 460
Val Glu Glu Thr Pro Thr Met Glu Ser Asn Val Glu Thr Gln Gln Glu
465 470 475 480
Thr Ser Thr Pro Ala Thr Ala Thr Pro Gln Arg Ala Asp Thr Ile Glu
485 490 495
Pro Thr Glu Glu Ala Thr Ser Gln Glu Glu Thr Thr Ala Ser Gln Thr
500 505 510
Gln Ser Pro Ala Val Glu Ala Pro Thr Ala Val Gln Glu Thr Val Ala
515 520 525
Pro Thr Ser Thr Pro
530
<210> 8
<211> 533
<212> PRT
<213> Artificial sequence
<400> 8
Met Ile Leu Gly Val Pro Ile Gln Tyr Leu Leu Tyr Ser Leu Trp Asn
1 5 10 15
Trp Ile Val Asp Thr Gly Phe Asp Val Ala Ile Ile Leu Val Leu Ala
20 25 30
Phe Leu Ile Pro Arg Ile Gly Arg Leu Ala Met Arg Ile Ile Lys Arg
35 40 45
Arg Val Glu Ser Ala Ala Asp Ala Asp Thr Thr Lys Asn Gln Leu Ala
50 55 60
Phe Ala Gly Val Gly Val Tyr Ile Ala Gln Ile Val Ala Phe Phe Met
65 70 75 80
Leu Ala Val Ser Ala Val Gln Ala Phe Gly Phe Ser Leu Ala Gly Ala
85 90 95
Ala Ile Pro Ala Thr Ile Ala Ser Ala Ala Ile Gly Leu Gly Ala Gln
100 105 110
Ser Ile Val Ala Asp Phe Leu Ala Gly Phe Phe Ile Leu Thr Glu Lys
115 120 125
Gln Phe Gly Val Gly Asp Trp Val Arg Phe Glu Gly Asp Gly Ile Val
130 135 140
Val Glu Gly Thr Val Ile Glu Ile Thr Met Arg Ala Thr Lys Ile Arg
145 150 155 160
Thr Ile Ala Gln Glu Ala Val Ile Ile Pro Asn Ser Thr Ala Lys Val
165 170 175
Cys Ile Asn Asn Ser Asn Asn Trp Ser Arg Ala Val Val Val Ile Pro
180 185 190
Ile Pro Met Leu Gly Ser Glu Asn Ile Thr Asp Val Ile Ala Arg Ser
195 200 205
Glu Ala Ala Thr Arg Arg Ala Leu Gly Gln Glu Lys Ile Ala Pro Glu
210 215 220
Ile Leu Gly Glu Leu Asp Val His Pro Ala Thr Glu Val Thr Pro Pro
225 230 235 240
Thr Val Val Gly Met Pro Trp Met Val Thr Met Arg Phe Leu Val Gln
245 250 255
Val Thr Ala Gly Asn Gln Trp Leu Val Glu Arg Ala Ile Arg Thr Glu
260 265 270
Ile Ile Ser Glu Phe Trp Glu Glu Tyr Gly Ser Ala Thr Thr Thr Ser
275 280 285
Gly Thr Leu Ile Asp Ser Leu His Val Glu His Glu Glu Pro Lys Thr
290 295 300
Ser Leu Ile Asp Ala Ser Pro Gln Ala Leu Lys Glu Pro Lys Pro Glu
305 310 315 320
Ala Ala Ala Thr Val Ala Ser Leu Ala Ala Ser Ser Asn Asp Asp Ala
325 330 335
Asp Asn Ala Asp Ala Ser Val Val Asn Ala Gly Asn Pro Glu Lys Glu
340 345 350
Leu Asp Ser Asp Val Leu Glu Gln Glu Leu Ser Ser Glu Glu Pro Glu
355 360 365
Glu Thr Ala Lys Pro Asp His Ser Leu Arg Gly Phe Phe Arg Thr Asp
370 375 380
Cys Tyr Pro Asn Arg Trp Gln Lys Ile Leu Ser Phe Gly Gly Arg Val
385 390 395 400
Arg Met Ser Thr Ser Leu Leu Leu Gly Ala Leu Leu Leu Leu Ser Leu
405 410 415
Phe Lys Val Met Thr Val Glu Pro Ser Glu Asn Trp Gln Asn Ser Ser
420 425 430
Gly Trp Leu Ser Pro Ser Thr Ala Thr Ser Thr Ala Val Thr Thr Ser
435 440 445
Glu Thr Ser Ala Pro Val Ser Thr Pro Ser Met Thr Val Pro Thr Thr
450 455 460
Val Glu Glu Thr Pro Thr Met Glu Ser Asn Val Glu Thr Gln Gln Glu
465 470 475 480
Thr Ser Thr Pro Ala Thr Ala Thr Pro Gln Arg Ala Asp Thr Ile Glu
485 490 495
Pro Thr Glu Glu Ala Thr Ser Gln Glu Glu Thr Thr Ala Ser Gln Thr
500 505 510
Gln Ser Pro Ala Val Glu Ala Pro Thr Ala Val Gln Glu Thr Val Ala
515 520 525
Pro Thr Ser Thr Pro
530
<210> 9
<211> 533
<212> PRT
<213> Artificial sequence
<400> 9
Met Ile Leu Gly Val Pro Ile Gln Tyr Leu Leu Tyr Ser Leu Trp Asn
1 5 10 15
Trp Ile Val Asp Thr Gly Phe Asp Val Ala Ile Ile Leu Val Leu Ala
20 25 30
Phe Leu Ile Pro Arg Ile Gly Arg Leu Ala Met Arg Ile Ile Lys Arg
35 40 45
Arg Val Glu Ser Ala Ala Asp Ala Asp Thr Thr Lys Asn Gln Leu Ala
50 55 60
Phe Ala Gly Val Gly Val Tyr Ile Ala Gln Ile Val Ala Phe Phe Met
65 70 75 80
Leu Ala Val Ser Ala Val Gln Ala Phe Gly Phe Ser Leu Ala Gly Ala
85 90 95
Ala Ile Pro Ala Thr Ile Ala Ser Ala Ala Ile Gly Leu Gly Ala Gln
100 105 110
Ser Ile Val Ala Asp Phe Leu Ala Gly Phe Phe Ile Leu Thr Glu Lys
115 120 125
Gln Phe Gly Val Gly Asp Trp Val Arg Phe Glu Gly Asn Gly Ile Val
130 135 140
Val Glu Gly Thr Val Ile Glu Ile Thr Met Arg Ala Thr Lys Ile Arg
145 150 155 160
Thr Ile Ala Gln Glu Ala Val Ile Ile Pro Asn Ser Thr Ala Lys Val
165 170 175
Cys Ile Asn Asn Ser Asn Asn Trp Ser Arg Ala Val Val Val Ile Pro
180 185 190
Ile Pro Met Leu Gly Ser Glu Asp Ile Thr Asp Val Val Ala Arg Ser
195 200 205
Glu Ala Ala Thr Arg Arg Ala Leu Gly Gln Glu Lys Ile Ala Pro Glu
210 215 220
Ile Leu Gly Glu Leu Asp Val His Pro Ala Thr Glu Val Thr Pro Pro
225 230 235 240
Thr Val Val Gly Met Pro Trp Met Val Thr Met Arg Phe Leu Val Gln
245 250 255
Val Thr Ala Gly Asn Gln Trp Leu Val Glu Arg Ala Ile Arg Thr Glu
260 265 270
Ile Ile Ser Glu Phe Trp Glu Glu Tyr Gly Ser Ala Thr Thr Thr Ser
275 280 285
Gly Thr Leu Ile Asp Ser Leu His Val Glu His Glu Glu Pro Lys Thr
290 295 300
Ser Leu Ile Asp Ala Ser Pro Gln Ala Leu Lys Glu Pro Lys Pro Glu
305 310 315 320
Ala Ala Ala Thr Val Ala Ser Leu Ala Ala Ser Ser Asn Asp Asp Ala
325 330 335
Asp Asn Ala Asp Ala Ser Val Val Asn Ala Gly Asn Pro Glu Lys Glu
340 345 350
Leu Asp Ser Asp Val Leu Glu Gln Glu Leu Ser Ser Glu Glu Pro Glu
355 360 365
Glu Thr Ala Lys Pro Asp His Ser Leu Arg Gly Phe Phe Arg Thr Asp
370 375 380
Cys Tyr Pro Asn Arg Trp Gln Lys Ile Leu Ser Phe Gly Gly Arg Val
385 390 395 400
Arg Met Ser Thr Ser Leu Leu Leu Gly Ala Leu Leu Leu Leu Ser Leu
405 410 415
Phe Lys Val Met Thr Val Glu Pro Ser Glu Asn Trp Gln Asn Ser Ser
420 425 430
Gly Trp Leu Ser Pro Ser Thr Ala Thr Ser Thr Ala Val Thr Thr Ser
435 440 445
Glu Thr Ser Ala Pro Val Ser Thr Pro Ser Met Thr Val Pro Thr Thr
450 455 460
Val Glu Glu Thr Pro Thr Met Glu Ser Asn Val Glu Thr Gln Gln Glu
465 470 475 480
Thr Ser Thr Pro Ala Thr Ala Thr Pro Gln Arg Ala Asp Thr Ile Glu
485 490 495
Pro Thr Glu Glu Ala Thr Ser Gln Glu Glu Thr Thr Ala Ser Gln Thr
500 505 510
Gln Ser Pro Ala Val Glu Ala Pro Thr Ala Val Gln Glu Thr Val Ala
515 520 525
Pro Thr Ser Thr Pro
530
<210> 10
<211> 533
<212> PRT
<213> Artificial sequence
<400> 10
Met Ile Leu Gly Val Pro Ile Gln Tyr Leu Leu Tyr Ser Ser Trp Asn
1 5 10 15
Trp Ile Val Asp Thr Gly Phe Asp Val Ala Ile Ile Leu Val Leu Ala
20 25 30
Phe Leu Ile Pro Arg Ile Gly Arg Leu Ala Met Arg Ile Ile Lys Arg
35 40 45
Arg Val Glu Ser Ala Ala Asp Ala Asp Thr Thr Lys Asn Gln Leu Ala
50 55 60
Phe Ala Gly Val Gly Val Tyr Ile Ala Gln Ile Val Ala Phe Phe Met
65 70 75 80
Leu Ala Val Ser Ala Val Gln Ala Phe Gly Phe Ser Leu Ala Gly Ala
85 90 95
Ala Ile Pro Ala Thr Ile Ala Ser Ala Ala Ile Gly Leu Gly Ala Gln
100 105 110
Ser Ile Val Ala Asp Phe Leu Ala Gly Phe Phe Ile Leu Thr Glu Lys
115 120 125
Gln Phe Gly Val Gly Asp Trp Val Arg Phe Glu Gly Asn Gly Ile Val
130 135 140
Val Glu Gly Thr Val Ile Glu Ile Thr Met Arg Ala Thr Lys Ile Arg
145 150 155 160
Thr Ile Ala Gln Glu Ala Val Ile Ile Pro Asn Ser Thr Ala Lys Val
165 170 175
Cys Ile Asn Asn Ser Asn Asn Trp Ser Arg Ala Val Val Val Ile Pro
180 185 190
Ile Pro Met Leu Gly Ser Glu Asn Ile Thr Asp Val Ile Ala Arg Ser
195 200 205
Glu Ala Ala Thr Arg Arg Ala Leu Gly Gln Glu Lys Ile Ala Pro Glu
210 215 220
Ile Leu Gly Glu Leu Asp Val His Pro Ala Thr Glu Val Thr Pro Pro
225 230 235 240
Thr Val Val Gly Met Pro Trp Met Val Thr Met Arg Phe Leu Val Gln
245 250 255
Val Thr Ala Gly Asn Gln Trp Leu Val Glu Arg Ala Ile Arg Thr Glu
260 265 270
Ile Ile Ser Glu Phe Trp Glu Glu Tyr Gly Ser Ala Thr Thr Thr Ser
275 280 285
Gly Thr Leu Ile Asp Ser Leu His Val Glu His Glu Glu Pro Lys Thr
290 295 300
Ser Leu Ile Asp Ala Ser Pro Gln Ala Leu Lys Glu Pro Lys Pro Glu
305 310 315 320
Ala Ala Ala Thr Val Ala Ser Leu Ala Ala Ser Ser Asn Asp Asp Ala
325 330 335
Asp Asn Ala Asp Ala Ser Val Val Asn Ala Gly Asn Pro Glu Lys Glu
340 345 350
Leu Asp Ser Asp Val Leu Glu Gln Glu Leu Ser Ser Glu Glu Pro Glu
355 360 365
Glu Thr Ala Lys Pro Asp His Ser Leu Arg Gly Phe Phe Arg Thr Asp
370 375 380
Cys Tyr Pro Asn Arg Trp Gln Lys Ile Leu Ser Phe Gly Gly Arg Val
385 390 395 400
Arg Met Ser Thr Ser Leu Leu Leu Gly Ala Leu Leu Leu Leu Ser Leu
405 410 415
Phe Lys Val Met Thr Val Glu Pro Ser Glu Asn Trp Gln Asn Ser Ser
420 425 430
Gly Trp Leu Ser Pro Ser Thr Ala Thr Ser Thr Ala Val Thr Thr Ser
435 440 445
Glu Thr Ser Ala Pro Val Ser Thr Pro Ser Met Thr Val Pro Thr Thr
450 455 460
Val Glu Glu Thr Pro Thr Met Glu Ser Asn Val Glu Thr Gln Gln Glu
465 470 475 480
Thr Ser Thr Pro Ala Thr Ala Thr Pro Gln Arg Ala Asp Thr Ile Glu
485 490 495
Pro Thr Glu Glu Ala Thr Ser Gln Glu Glu Thr Thr Ala Ser Gln Thr
500 505 510
Gln Ser Pro Ala Val Glu Ala Pro Thr Ala Val Gln Glu Thr Val Ala
515 520 525
Pro Thr Ser Thr Pro
530
<210> 11
<211> 533
<212> PRT
<213> Artificial sequence
<400> 11
Met Ile Leu Gly Val Pro Ile Gln Tyr Leu Leu Tyr Ser Leu Trp Asn
1 5 10 15
Trp Ile Val Asp Thr Gly Phe Asp Val Ala Ile Ile Leu Val Leu Ala
20 25 30
Phe Leu Ile Pro Arg Ile Gly Arg Leu Ala Met Arg Ile Ile Lys Arg
35 40 45
Arg Val Glu Ser Ala Ala Asp Ala Asp Thr Thr Lys Asn Gln Leu Ala
50 55 60
Phe Ala Gly Val Gly Val Tyr Ile Ala Gln Ile Val Ala Phe Phe Met
65 70 75 80
Leu Ala Val Ser Ala Met Gln Ala Phe Gly Phe Ser Leu Ala Gly Ala
85 90 95
Ala Ile Pro Ala Thr Ile Ala Ser Ala Ala Ile Gly Leu Gly Ala Gln
100 105 110
Ser Ile Val Ala Asp Phe Leu Ala Gly Phe Phe Ile Leu Thr Glu Lys
115 120 125
Gln Phe Gly Val Gly Asp Trp Val Arg Phe Glu Gly Asn Gly Ile Val
130 135 140
Val Glu Gly Thr Val Ile Glu Ile Thr Met Arg Ala Thr Lys Ile Arg
145 150 155 160
Thr Ile Ala Gln Glu Thr Val Ile Ile Pro Asn Ser Thr Ala Lys Val
165 170 175
Cys Ile Asn Asn Ser Asn Asn Trp Ser Arg Ala Val Val Val Ile Pro
180 185 190
Ile Pro Met Leu Gly Ser Glu Asn Ile Thr Asp Val Ile Ala Arg Ser
195 200 205
Glu Ala Ala Thr Arg Arg Ala Leu Gly Gln Glu Lys Ile Ala Pro Glu
210 215 220
Ile Leu Gly Glu Leu Asp Val His Pro Ala Thr Glu Val Thr Pro Pro
225 230 235 240
Thr Val Val Gly Met Pro Trp Met Val Thr Met Arg Phe Leu Val Gln
245 250 255
Val Thr Ala Gly Asn Gln Trp Leu Val Glu Arg Ala Ile Arg Thr Glu
260 265 270
Ile Ile Ser Glu Phe Trp Glu Glu Tyr Gly Ser Ala Thr Thr Thr Ser
275 280 285
Gly Thr Leu Ile Asp Ser Leu His Val Glu His Glu Glu Pro Lys Thr
290 295 300
Ser Leu Ile Asp Ala Ser Pro Gln Ala Leu Lys Glu Pro Lys Pro Glu
305 310 315 320
Ala Ala Ala Thr Val Ala Ser Leu Ala Ala Ser Ser Asn Gly Asp Ala
325 330 335
Asp Asn Ala Asp Ala Ser Val Ile Asn Ala Gly Asn Pro Glu Lys Glu
340 345 350
Leu Asp Ser Asp Val Leu Glu Gln Glu Pro Ser Ser Glu Glu Pro Glu
355 360 365
Glu Thr Ala Lys Pro Asp His Ser Leu Arg Gly Phe Phe Arg Thr Asp
370 375 380
Tyr Tyr Pro Asn Arg Trp Gln Lys Ile Leu Ser Phe Gly Gly Arg Val
385 390 395 400
Arg Met Ser Thr Ser Leu Leu Leu Gly Ala Leu Leu Leu Leu Ser Leu
405 410 415
Phe Lys Val Met Thr Val Glu Pro Ser Glu Asn Trp Gln Asn Ser Ser
420 425 430
Gly Trp Leu Ser Pro Ser Thr Ala Thr Ser Thr Ala Val Thr Thr Ser
435 440 445
Glu Thr Ser Ala Pro Val Ser Thr Pro Ser Met Thr Val Pro Thr Thr
450 455 460
Val Glu Glu Thr Pro Thr Met Glu Ser Asn Val Glu Thr Gln Gln Glu
465 470 475 480
Thr Ser Thr Pro Ala Thr Ala Thr Pro Gln Arg Ala Asp Thr Ile Glu
485 490 495
Pro Thr Glu Glu Ala Thr Ser Gln Glu Glu Thr Thr Ala Ser Gln Thr
500 505 510
Gln Ser Pro Ala Val Glu Ala Pro Thr Ala Val Gln Glu Thr Val Ala
515 520 525
Pro Thr Ser Thr Pro
530
<210> 12
<211> 1602
<212> DNA
<213> Artificial sequence
<400> 12
atgattttag gcgtacccat tcaatatttg ctctattcat tgtggaattg gattgtcgat 60
accggttttg atgtagcaat tatcctggtc ttggcgtttt tgattccacg tatcggccga 120
ctggccatgc gtattatcaa gcgccgagtg gagtctgcag ccgatgcgga caccactaag 180
aaccagctcg cgttcgccgg cgttggcgtt tatatcgcgc aaattgtggc gtttttcatg 240
cttgccgtct ccgcgatgca ggcttttggt ttctctctcg cgggcgctgc gattccggca 300
accattgcgt cagctgccat tggccttggt gcgcagtcga ttgttgcgga cttcttggcc 360
ggatttttca tcctgacgga aaagcaattc ggcgtgggtg actgggtgcg ttttgagggc 420
aacggcatcg ttgtcgaagg caccgtcatt gagatcacca tgcgcgcgac caaaattcgc 480
acgattgcac aagagaccgt gatcatcccc aactccacgg cgaaagtgtg catcaacaat 540
tctaataact ggtcgcgtgc ggttgtcgtt attccgatcc ccatgttggg ttctgaaaac 600
atcacagatg tcatcgcgcg ctctgaagct gcgactcgtc gcgcacttgg ccaggagaaa 660
atcgcaccgg aaatcctcgg tgaactcgat gtgcacccag ccacggaagt cacgccgcca 720
acggtggtcg gcatgccgtg gatggtcacc atgcgtttcc tcgtgcaagt caccgccggc 780
aatcaatggc tggtcgaacg cgccatccgc acagaaatca tcagcgaatt ctgggaagaa 840
tacggcagcg caaccactac atcgggaacc ctcattgatt ccttacacgt tgagcatgaa 900
gagccaaaga cctcgcttat cgacgcctcc ccccaggctc ttaaggaacc gaagccggag 960
gctgcggcga cggttgcatc gctagctgca tcctctaacg acgatgcaga caatgcagac 1020
gcctcggtga tcaatgcagg caatccagag aaggaacttg attccgatgt gctggaacaa 1080
gaactctcca gcgaagaacc ggaagaaaca gcaaaaccag atcactctct ccgaggcttc 1140
ttccgcactg attactaccc aaatcggtgg cagaagatcc tgtcgtttgg cggacgtgtc 1200
cgcatgagca cgtccctgtt gttgggtgcg ctgctcttgc tgtcactatt taaggtcatg 1260
actgtggaac caagtgagaa ttggcaaaac tccagtggat ggctgtcacc aagcactgcc 1320
acctcaactg cggtgaccac ctccgaaact tccgcgccag taagcacgcc ttcgatgaca 1380
gtgcccacta cggtggagga gaccccaacg atggaatcta acgtcgaaac gcagcaggaa 1440
acctcaaccc ctgcaaccgc aacgccccag cgagccgaca ccatcgaacc gaccgaggaa 1500
gccacgtcgc aggaggaaac gactgcgtcg cagacgcagt ctccagcagt ggaagcacca 1560
accgcggtcc aagagacagt tgcgccgacg tccacccctt ag 1602
<210> 13
<211> 1602
<212> DNA
<213> Artificial sequence
<400> 13
atgattttag gcgtacccat tcaatatttg ctctattcat tgtggaattg gattgtcgat 60
accggttttg atgtagcaat tatcctggtc ttggcgtttt tgattccacg tatcggccga 120
ctggccatgc gtattatcaa gcgccgagtg gagtctgcag ccgatgcgga caccactaag 180
aaccagctcg cgttcgccgg cgttggcgtt tatatcgcgc aaattgtggc gtttttcatg 240
cttgccgtct ccgcgatgca ggcttttggt ttctctctcg cgggcgctgc gattccggca 300
accattgcgt cagctgccat tggccttggt gcgcagtcga ttgttgcgga cttcttggcc 360
ggatttttca tcctgacgga aaagcaattc ggcgtgggtg actgggtgcg ttttgagggc 420
aacggcatcg ttgtcgaagg caccgtcatt gagatcacca tgcgcgcgac caaaattcgc 480
acgattgcac aagggaccgt gatcatcccc aactccacgg cgaaagtgtg catcaacaat 540
tctaataact ggtcgcgtgc ggttgtcgtt attccgatcc ccatgttggg ttctgaaaac 600
atcacagatg tcatcgcgcg ctctgaagct gcgactcgtc gcgcacttgg ccaggagaaa 660
atcgcaccgg aaatcctcgg tgaactcgat gtgcacccag ccacggaagt cacgccgcca 720
acggtggtcg gcatgccgtg gatggtcacc atgcgtttcc tcgtgcaagt caccgccggc 780
aatcaatggc tggtcgaacg cgccatccgc acagaaatca tcagcgaatt ctgggaagaa 840
tacggcagcg caaccactac atcgggaacc ctcattgatt ccttacacgt tgagcatgaa 900
gagccaaaga cctcgcttat cgacgcctcc ccccaggctc ttaaggaacc gaagccggag 960
gctgcggcga cggttgcatc gctagctgca tcctctaacg acgatgcaga caatgcagac 1020
gcctcggtga tcaatgcagg caatccagag aaggaacttg attccgatgt gctggaacaa 1080
gaactctcca gcgaagaacc ggaagaaaca gcaaaaccag atcactctct ccgaggcttc 1140
ttccgcactg attactaccc aaatcggtgg cagaagatcc tgtcgtttgg cggacgtgtc 1200
cgcatgagca cgtccctgtt gttgggtgcg ctgctcttgc tgtcactatt taaggtcatg 1260
actgtggaac caagtgagaa ttggcaaaac tccagtggat ggctgtcacc aagcactgcc 1320
acctcaactg cggtgaccac ctccgaaact tccgcgccag taagcacgcc ttcgatgaca 1380
gtgcccacta cggtggagga gaccccaacg atggaatcta acgtcgaaac gcagcaggaa 1440
acctcaaccc ctgcaaccgc aacgccccag cgagccgaca ccatcgaacc gaccgaggaa 1500
gccacgtcgc aggaggaaac gactgcgtcg cagacgcagt ctccagcagt ggaagcacca 1560
accgcggtcc aagagacagt tgcgccgacg tccacccctt ag 1602
<210> 14
<211> 1602
<212> DNA
<213> Artificial sequence
<400> 14
atgattttag gcgtacccat tcaatatttg ctctattcat tgtggaattg gattgtcgat 60
accggttttg atgtagcaat tatcctggtc ttggcgtttt tgattccacg tatcggccga 120
ctggccatgc gtattatcaa gcgccgagtg gagtctgcag ccgatgcgga caccactaag 180
aaccagctcg cgttcgccgg cgttggcgtt tatatcgcgc aaattgtggc gtttttcatg 240
cttgccgtct ccgcgatgca ggcttttggt ttctctctcg cgggcgctgc gattccggca 300
accattacgt cagctgccat tggccttggt gcgcagtcga ttgttgcgga cttcttggcc 360
ggatttttca tcctgacgga aaagcaattc ggcgtgggtg actgggtgcg ttttgagggc 420
aacggcatcg ttgtcgaagg caccgtcatt gagatcacca tgcgcgcgac caaaattcgc 480
acgattgcac aagagaccgt gatcatcccc aactccacgg cgaaagtgtg catcaacaat 540
tctaataact ggtcgcgtgc ggttgtcgtt attccgatcc ccatgttggg ttctgaaaac 600
atcacagatg tcatcgcgcg ctctgaagct gcgactcgtc gcgcacttgg ccaggagaaa 660
atcgcaccgg aaatcctcgg tgaactcgat gtgcacccag ccacggaagt cacgccgcca 720
acggtggtcg gcatgccgtg gatggtcacc atgcgtttcc tcgtgcaagt caccgccggc 780
aatcaatggc tggtcgaacg cgccatccgc acagaaatca tcagcgaatt ctgggaagaa 840
tacggcagcg caaccactac atcgggaacc ctcattgatt ccttacacgt tgagcatgaa 900
gagccaaaga cctcgcttat cgacgcctcc ccccaggctc ttaaggaacc gaagccggag 960
gctgcggcga cggttgcatc gctagctgca tcctctaacg acgatgcaga caatgcagac 1020
gcctcggtga tcaatgcagg caatccagag aaggaacttg attccgatgt gctggaacaa 1080
gaactctcca gcgaagaacc ggaagaaaca gcaaaaccag atcactctct ccgaggcttc 1140
ttccgcactg attactaccc aaatcggtgg cagaaggtcc tgtcgtttgg cggatgtgtc 1200
cgcatgagca cgtccctgtt gttgggtgcg ctgctcttgc tgtcactatt taaggtcatg 1260
actgtggaac caagtgagaa ttggcaaaac tccagtggat ggctgtcacc aagcactgcc 1320
acctcaactg cggtgaccac ctccgaaact tccgcgccag taagcacgcc ttcgatgaca 1380
gtgcccacta cggtggagga gaccccaacg atggaatcta acgtcgaaac gcagcaggaa 1440
acctcaaccc ctgcaaccgc aacgccccag cgagccgaca ccatcgaacc gaccgaggaa 1500
gccacgtcgc aggaggaaac gactgcgtcg cagacgcagt ctccagcagt ggaagcacca 1560
accgcggtcc aagagacagt tgcgccgacg tccacccctt ag 1602
<210> 15
<211> 1602
<212> DNA
<213> Artificial sequence
<400> 15
atgattttag gcgtacccat tcaatatttg ctctattcat tgtggaattg gattgtcgat 60
accggttttg atgtagcaat tatcctggtc ttggcgtttt tgattccacg tatcggccga 120
ctggccatgc gtattatcaa gcgccgagtg gagtctgcag ccgatgcgga caccactaag 180
aaccagctcg cgttcgccgg cgttggcgtt tatatcgcgc aaattgtggc gtttttcatg 240
cttgccgtct ccgcgatgca ggcttttggt ttctctctcg cgggcgctgc gattccggca 300
accattgcgt cagctgccat tggccttggt gcgcagtcga ttgttgcgga cttcttggcc 360
ggatttttca tcctgacgga aaagcaattc ggcgtgggtg actgggtgcg ttttgagggc 420
aacggcatcg ttgtcgaagg caccgtcatt gagatcacca tgcgcgcgac caaaattcgc 480
acgattgcac aagagaccgt gatcatcccc aactccgcgg cgaaagtgtg catcaacaat 540
tctaataact ggtcgcgtgc ggttgtcgtt attccgatcc ccatgttggg ttctgaaaac 600
atcacagatg tcatcgcgcg ctctgaagct gcgactcgtc gcgcacttgg ccaggagaaa 660
atcgcaccgg aaatcctcgg tgaactcgat gtgcacccag ccacggaagt cacgccgcca 720
acggtggtcg gcatgccgtg gatggtcacc atgcgtttcc tcgtgcaagt caccgccggc 780
aatcaatggc tggtcgaacg cgccatccgc acagaaatca tcagcgaatt ctgggaagaa 840
tacggcagcg caaccactac atcgggaacc ctcattgatt ccttacacgt tgagcatgaa 900
gagccaaaga cctcgcttat cgacgcctcc ccccaggctc ttaaggaacc gaagccggag 960
gctgcggcga cggttgcatc gctagctgca tcctctaacg acgatgcaga caatgcagac 1020
gcctcggaga tcaatgcagg caatccagag aaggaacttg attccgatgt gctggaacaa 1080
gaactctcca gcgaagaacc ggaagaaaca gcaaaaccag atcactctct ccgaggcttc 1140
ttccgcactg attactaccc aaatcggtgg cagaagatcc tgtcgtttgg cggacgtgtc 1200
cgcatgagca cgtccctgtt gttgggtgcg ctgctcttgc tgtcactatt taaggtcatg 1260
actgtggaac caagtgagaa ttggcaaaac tccagtggat ggctgtcacc aagcactgcc 1320
acctcaactg cggtgaccac ctccgaaact tccgcgccag taagcacgcc ttcgatgaca 1380
gtgcccacta cggtggagga gaccccaacg atggaatcta acgtcgaaac gcagcaggaa 1440
acctcaaccc ctgcaaccgc aacgccccag cgagccgaca ccatcgaacc gaccgaggaa 1500
gccacgtcgc aggaggaaac gactgcgtcg cagacgcagt ctccagcagt ggaagcacca 1560
accgcggtcc aagagacagt tgcgccgacg tccacccctt ag 1602
<210> 16
<211> 1602
<212> DNA
<213> Artificial sequence
<400> 16
atgattttag gcgtacccat tcaatatttg ctctattcat tgtggaattg gattgtcgat 60
accggttttg atgtagcaat tatcctggtc ttggcgtttt tgattccacg tatcggccga 120
ctggccatgc gtattatcaa gcgccgagtg gagtctgcag ccgatgcgga caccactaag 180
aaccagctcg cgttcgccgg cgttggcgtt tatatcgcgc aaattgtggc gtttttcatg 240
cttgccgtct ccgcgatgca ggcttttggt ttctctctcg cgggcgctgc gattccggca 300
accattgcgt cagctgccat tggccttggt gcgcagtcga ttgttgcgga cttcttggcc 360
ggatttttca tcctgacgga aaagcaattc ggcgtgggtg actgggtgcg ttttgagggc 420
aacggcatcg ttgtcgaagg caccgtcatt gagatcacca tgcgcgcgac caaaattcgc 480
acgattgcac aagaggccgt gatcatcccc aactccacgg cgaaagtgtg catcaacaat 540
tctaataact ggtcgcgtgc ggttgtcgtt attccgatcc ccatgttggg ttctgaaaac 600
atcacagatg tcatcgcgcg ctctgaagct gcgactcgtc gcgcacttgg ccaggagaaa 660
atcgcaccgg aaatcctcgg tgaactcgat gtgcacccag ccacggaagt cacgccgcca 720
acggtggtcg gcatgccgtg gatggtcacc atgcgtttcc tcgtgcaagt caccgccggc 780
aatcaatggc tggtcgaacg cgccatccgc acagaaatca tcagcgaatt ctgggaagaa 840
tacggcagcg caaccactac atcgggaacc ctcattgatt ccttacacgt tgagcatgaa 900
gagccaaaga cctcgcttat cgacgccccc ccccaggctc ttaaggaacc gaagccggag 960
gctgcggcga cggttgcatc gctagctgca tcctctaacg acgatgcaga caatgcagac 1020
gcctcggtga tcaatgcagg caatccagag aaggaacttg attccgatgt gctggaacaa 1080
gaactctcca gcgaagaacc ggaagaaaca gcaaaaccag atcactctct ccgaggcttc 1140
ttccgcactg attactaccc aaatcggtgg cagaagatcc tgtcgtttgg cggacgtgtc 1200
cgcatgagca cgtccctgtt gttgggtgcg ctgctcttgc tgtcactatt taaggtcatg 1260
actgtggaac caagtgagaa ttggcaaaac tccagtggat ggctgtcacc aagcactgcc 1320
acctcaactg cggtgaccac ctccgaaact tccgcgccag taagcacgcc ttcgatgaca 1380
gtgcccacta cggtggagga gaccccaacg atggaatcta acgtcgaaac gcagcaggaa 1440
acctcaaccc ctgcaaccgc aacgccccag cgagccgaca ccatcgaacc gaccgaggaa 1500
gccacgtcgc aggaggaaac gactgcgtcg cagacgcagt ctccagcagt ggaagcacca 1560
accgcggtcc aagagacagt tgcgccgacg tccacccctt ag 1602
<210> 17
<211> 1602
<212> DNA
<213> Artificial sequence
<400> 17
atgattttag gcgtacccat tcaatatttg ctctattcat tgtggaattg gattgtcgat 60
accggttttg atgtagcaat tatcctggtc ttggcgtttt tgattccacg tatcggccga 120
ctggccatgc gtattatcaa gcgccgagtg gagtctgcag ccgatgcgga caccactaag 180
aaccagctcg cgttcgccgg cgttggcgtt tatatcgcgc aaattgtggc gtttttcatg 240
cttgccgtct ccgcggtgca ggcttttggt ttctctctcg cgggcgctgc gattccggca 300
accattgcgt cagctgccat tggccttggt gcgcagtcga ttgttgcgga cttcttggcc 360
ggatttttca tcctgacgga aaagcaattc ggcgtgggtg actgggtgcg ttttgagggc 420
aacggcatcg ttgtcgaagg caccgtcatt gagatcacca tgcgcgcgac caaaattcgc 480
acgattgcac aagaggccgt gaccatcccc aactccacgg cgaaagtgtg catcaacaat 540
tctaataact ggtcgcgtgc ggttgtcgtt attccgatcc ccatgttggg ttctgaaaac 600
atcacagatg tcatcgcgcg ctctgaagct gcgactcgtc gcgcacttgg ccaggagaaa 660
atcgcaccgg aaatcctcgg tgaactcgat gtgcacccag ccacggaagt cacgccgcca 720
acggtggtcg gcatgccgtg gatggtcacc atgcgtttcc tcgtgcaagt caccgccggc 780
aatcaatggc tggtcgaacg cgccatccgc acagaaatca tcagcgaatt ctgggaagaa 840
tacggcagcg caaccactac atcgggaacc ctcattgatt ccttacacgt tgagcatgaa 900
gagccaaaga cctcgcttat cgacgcctcc ccccaggctc ttaaggaacc gaagccggag 960
gctgcggcga cggttgcatc gctagctgca tcctctaacg acgatgcaga caatgcagac 1020
gcctcggtgg tcaatgcagg caatccagag aaggaacttg attccgatgt gctggaacaa 1080
gaactctcca gcgaagaacc ggaagaaaca gcaaaaccag atcactctct ccgaggcttc 1140
ttccgcactg attgctaccc aaatcggtgg cagaagatcc tgtcgtttgg cggacgtgtc 1200
cgcatgagca cgtccctgtt gttgggtgcg ctgctcttgc tgtcactatt taaggtcatg 1260
actgtggaac caagtgagaa ttggcaaaac tccagtggat ggctgtcacc aagcactgcc 1320
acctcaactg cggtgaccac ctccgaaact tccgcgccag taagcacgcc ttcgatgaca 1380
gtgcccacta cggtggagga gaccccaacg atggaatcta acgtcgaaac gcagcaggaa 1440
acctcaaccc ctgcaaccgc aacgccccag cgagccgaca ccatcgaacc gaccgaggaa 1500
gccacgtcgc tggaggaaac gactgcgtcg cagacgcagt ctccagcagt ggaagcacca 1560
accgcggtcc aagagacagt tgcgccgacg tccacccctt ag 1602
<210> 18
<211> 1602
<212> DNA
<213> Artificial sequence
<400> 18
atgattttag gcgtacccat tcaatatttg ctctattcat tgtggaattg gattgtcgat 60
accggttttg atgtagcaat tatcctggtc ttggcgtttt tgattccacg tatcggccga 120
ctggccatgc gtattatcaa gcgccgagtg gagtctgcag ccgatgcgga caccactaag 180
aaccagctcg cgttcgccgg cgttggcgtt tatatcgcgc aaattgtggc gtttttcatg 240
cttgccgtct ccgcggtgca ggcttttggt ttctctctcg cgggcgctgc gattccggca 300
accattgcgt cagctgccat tggccttggt gcgcagtcga ttgttgcgga cttcttggcc 360
ggatttttca tcctgacgga aaagcaattc ggcgtgggtg actgggtgcg ttttgagggc 420
aacggcatcg ttgtcgaagg caccgtcatt gagatcacca tgcgcgcgac caaaattcgc 480
acgattgcac aagaggccgt gatcatcccc aactccacgg cgaaagtgtg catcaacaat 540
tctaataact ggtcgtgtgc ggttgtcgtt attccgatcc ccatgttggg ttctgaaaac 600
atcacagatg tcatcgcgcg ctctgaagct gcgactcgtc gcgcacttgg ccaggagaaa 660
atcgcaccgg aaatcctcgg tgaactcgat gtgcacccag ccacggaagt cacgccgcca 720
acggtggtcg gcatgccgtg gatggtcacc atgcgtttcc tcgtgcaagt caccgccggc 780
aatcaatggc tggtcgaacg cgccatccgc acagaaatca tcagcgaatt ctgggaagaa 840
tacggcagcg caaccactac atcgggaacc ctcattgatt ccttacacgt tgagcatgaa 900
gagccaaaga cctcgcttat cgacgcctcc ccccaggctc ttaaggaacc gaagccggag 960
gctgcggcga cggttgcatc gctagctgca tcctctaacg acgatgcaga caatgcagac 1020
gcctcggtgg tcaatgcagg caatccagag aaggaacttg attccgatgt gctggaacaa 1080
gaactctcca gcgaagaacc ggaagaaaca gcaaaaccag atcactctct ccgaggcttc 1140
ttccgcactg attgctaccc aaatcggtgg cagaagatcc tgtcgtttgg cggacgtgtc 1200
cgcatgagca cgtccctgtt gttgggtgcg ctgctcttgc tgtcactatt taaggtcatg 1260
actgtggaac caagtgagaa ttggcaaaac tccagtggat ggctgtcacc aagcactgcc 1320
acctcaactg cggtgaccac ctccgaaact tccgcgccag taagcacgcc ttcgatgaca 1380
gtgcccacta cggtggagga gaccccaacg atggaatcta acgtcgaaac gcagcaggaa 1440
acctcaaccc ctgcaaccgc aacgccccag cgagccgaca ccatcgaacc gaccgaggaa 1500
gccacgtcgc aggaggaaac gactgcgtcg cagacgcagt ctccagcagt ggaagcacca 1560
accgcggtcc aagagacagt tgcgccgacg tccacccctt ag 1602
<210> 19
<211> 1602
<212> DNA
<213> Artificial sequence
<400> 19
atgattttag gcgtacccat tcaatatttg ctctattcat tgtggaattg gattgtcgat 60
accggttttg atgtagcaat tatcctggtc ttggcgtttt tgattccacg tatcggccga 120
ctggccatgc gtattatcaa gcgccgagtg gagtctgcag ccgatgcgga caccactaag 180
aaccagctcg cgttcgccgg cgttggcgtt tatatcgcgc aaattgtggc gtttttcatg 240
cttgccgtct ccgcggtgca ggcttttggt ttctctctcg cgggcgctgc gattccggca 300
accattgcgt cagctgccat tggccttggt gcgcagtcga ttgttgcgga cttcttggcc 360
ggatttttca tcctgacgga aaagcaattc ggcgtgggtg actgggtgcg ttttgagggc 420
gacggcatcg ttgtcgaagg caccgtcatt gagatcacca tgcgcgcgac caaaattcgc 480
acgattgcac aagaggccgt gatcatcccc aactccacgg cgaaagtgtg catcaacaat 540
tctaataact ggtcgcgtgc ggttgtcgtt attccgatcc ccatgttggg ttctgaaaac 600
atcacagatg tcatcgcgcg ctctgaagct gcgactcgtc gcgcacttgg ccaggagaaa 660
atcgcaccgg aaatcctcgg tgaactcgat gtgcacccag ccacggaagt cacgccgcca 720
acggtggtcg gcatgccgtg gatggtcacc atgcgtttcc tcgtgcaagt caccgccggc 780
aatcaatggc tggtcgaacg cgccatccgc acagaaatca tcagcgaatt ctgggaagaa 840
tacggcagcg caaccactac atcgggaacc ctcattgatt ccttacacgt tgagcatgaa 900
gagccaaaga cctcgcttat cgacgcctcc ccccaggctc ttaaggaacc gaagccggag 960
gctgcggcga cggttgcatc gctagctgca tcctctaacg acgatgcaga caatgcagac 1020
gcctcggtgg tcaatgcagg caatccagag aaggaacttg attccgatgt gctggaacaa 1080
gaactctcca gcgaagaacc ggaagaaaca gcaaaaccag atcactctct ccgaggcttc 1140
ttccgcactg attgctaccc aaatcggtgg cagaagatcc tgtcgtttgg cggacgtgtc 1200
cgcatgagca cgtccctgtt gttgggtgcg ctgctcttgc tgtcactatt taaggtcatg 1260
actgtggaac caagtgagaa ttggcaaaac tccagtggat ggctgtcacc aagcactgcc 1320
acctcaactg cggtgaccac ctccgaaact tccgcgccag taagcacgcc ttcgatgaca 1380
gtgcccacta cggtggagga gaccccaacg atggaatcta acgtcgaaac gcagcaggaa 1440
acctcaaccc ctgcaaccgc aacgccccag cgagccgaca ccatcgaacc gaccgaggaa 1500
gccacgtcgc aggaggaaac gactgcgtcg cagacgcagt ctccagcagt ggaagcacca 1560
accgcggtcc aagagacagt tgcgccgacg tccacccctt ag 1602
<210> 20
<211> 1602
<212> DNA
<213> Artificial sequence
<400> 20
atgattttag gcgtacccat tcaatatttg ctctattcat tgtggaattg gattgtcgat 60
accggttttg atgtagcaat tatcctggtc ttggcgtttt tgattccacg tatcggccga 120
ctggccatgc gtattatcaa gcgccgagtg gagtctgcag ccgatgcgga caccactaag 180
aaccagctcg cgttcgccgg cgttggcgtt tatatcgcgc aaattgtggc gtttttcatg 240
cttgccgtct ccgcggtgca ggcttttggt ttctctctcg cgggcgctgc gattccggca 300
accattgcgt cagctgccat tggccttggt gcgcagtcga ttgttgcgga cttcttggcc 360
ggatttttca tcctgacgga aaagcaattc ggcgtgggtg actgggtgcg ttttgagggc 420
aacggcatcg ttgtcgaagg caccgtcatt gagatcacca tgcgcgcgac caaaattcgc 480
acgattgcac aagaggccgt gatcatcccc aactccacgg cgaaagtgtg catcaacaat 540
tctaataact ggtcgcgtgc ggttgtcgtt attccgatcc ccatgttggg ttctgaagac 600
atcacagatg tcgtcgcgcg ctctgaagct gcgactcgtc gcgcacttgg ccaggagaaa 660
atcgcaccgg aaatcctcgg tgaactcgat gtgcacccag ccacggaagt cacgccgcca 720
acggtggtcg gcatgccgtg gatggtcacc atgcgtttcc tcgtgcaagt caccgccggc 780
aatcaatggc tggtcgaacg cgccatccgc acagaaatca tcagcgaatt ctgggaagaa 840
tacggcagcg caaccactac atcgggaacc ctcattgatt ccttacacgt tgagcatgaa 900
gagccaaaga cctcgcttat cgacgcctcc ccccaggctc ttaaggaacc gaagccggag 960
gctgcggcga cggttgcatc gctagctgca tcctctaacg acgatgcaga caatgcagac 1020
gcctcggtgg tcaatgcagg caatccagag aaggaacttg attccgatgt gctggaacaa 1080
gaactctcca gcgaagaacc ggaagaaaca gcaaaaccag atcactctct ccgaggcttc 1140
ttccgcactg attgctaccc aaatcggtgg cagaagatcc tgtcgtttgg cggacgtgtc 1200
cgcatgagca cgtccctgtt gttgggtgcg ctgctcttgc tgtcactatt taaggtcatg 1260
actgtggaac caagtgagaa ttggcaaaac tccagtggat ggctgtcacc aagcactgcc 1320
acctcaactg cggtgaccac ctccgaaact tccgcgccag taagcacgcc ttcgatgaca 1380
gtgcccacta cggtggagga gaccccaacg atggaatcta acgtcgaaac gcagcaggaa 1440
acctcaaccc ctgcaaccgc aacgccccag cgagccgaca ccatcgaacc gaccgaggaa 1500
gccacgtcgc aggaggaaac gactgcgtcg cagacgcagt ctccagcagt ggaagcacca 1560
accgcggtcc aagagacagt tgcgccgacg tccacccctt ag 1602
<210> 21
<211> 1602
<212> DNA
<213> Artificial sequence
<400> 21
atgattttag gcgtacccat tcaatatttg ctctattcat cgtggaattg gattgtcgat 60
accggttttg atgtagcaat tatcctggtc ttggcgtttt tgattccacg tatcggccga 120
ctggccatgc gtattatcaa gcgccgagtg gagtctgcag ccgatgcgga caccactaag 180
aaccagctcg cgttcgccgg cgttggcgtt tatatcgcgc aaattgtggc gtttttcatg 240
cttgccgtct ccgcggtgca ggcttttggt ttctctctcg cgggcgctgc gattccggca 300
accattgcgt cagctgccat tggccttggt gcgcagtcga ttgttgcgga cttcttggcc 360
ggatttttca tcctgacgga aaagcaattc ggcgtgggtg actgggtgcg ttttgagggc 420
aacggcatcg ttgtcgaagg caccgtcatt gagatcacca tgcgcgcgac caaaattcgc 480
acgattgcac aagaggccgt gatcatcccc aactccacgg cgaaagtgtg catcaacaat 540
tctaataact ggtcgcgtgc ggttgtcgtt attccgatcc ccatgttggg ttctgaaaac 600
atcacagatg tcatcgcgcg ctctgaagct gcgactcgtc gcgcacttgg ccaggagaaa 660
atcgcaccgg aaatcctcgg tgaactcgat gtgcacccag ccacggaagt cacgccgcca 720
acggtggtcg gcatgccgtg gatggtcacc atgcgtttcc tcgtgcaagt caccgccggc 780
aatcaatggc tggtcgaacg cgccatccgc acagaaatca tcagcgaatt ctgggaagaa 840
tacggcagcg caaccactac atcgggaacc ctcattgatt ccttacacgt tgagcatgaa 900
gagccaaaga cctcgcttat cgacgcctcc ccccaggctc ttaaggaacc gaagccggag 960
gctgcggcga cggttgcatc gctagctgca tcctctaacg acgatgcaga caatgcagac 1020
gcctcggtgg tcaatgcagg caatccagag aaggaacttg attccgatgt gctggaacaa 1080
gaactctcca gcgaagaacc ggaagaaaca gcaaaaccag atcactctct ccgaggcttc 1140
ttccgcactg attgctaccc aaatcggtgg cagaagatcc tgtcgtttgg cggacgtgtc 1200
cgcatgagca cgtccctgtt gttgggtgcg ctgctcttgc tgtcactatt taaggtcatg 1260
actgtggaac caagtgagaa ttggcaaaac tccagtggat ggctgtcacc aagcactgcc 1320
acctcaactg cggtgaccac ctccgaaact tccgcgccag taagcacgcc ttcgatgaca 1380
gtgcccacta cggtggagga gaccccaacg atggaatcta acgtcgaaac gcagcaggaa 1440
acctcaaccc ctgcaaccgc aacgccccag cgagccgaca ccatcgaacc gaccgaggaa 1500
gccacgtcgc aggaggaaac gactgcgtcg cagacgcagt ctccagcagt ggaagcacca 1560
accgcggtcc aagagacagt tgcgccgacg tccacccctt ag 1602
<210> 22
<211> 1602
<212> DNA
<213> Artificial sequence
<400> 22
atgattttag gcgtacccat tcaatatttg ctctattcat tgtggaattg gattgtcgat 60
accggttttg atgtagcaat tatcctggtc ttggcgtttt tgattccacg tatcggccga 120
ctggccatgc gtattatcaa gcgccgagtg gagtctgcag ccgatgcgga caccactaag 180
aaccagctcg cgttcgccgg cgttggcgtt tatatcgcgc aaattgtggc gtttttcatg 240
cttgccgtct ccgcgatgca ggcttttggt ttctctctcg cgggcgctgc gattccggca 300
accattgcgt cagctgccat tggccttggt gcgcagtcga ttgttgcgga cttcttggcc 360
ggatttttca tcctgacgga aaagcaattc ggcgtgggtg actgggtgcg ttttgagggc 420
aacggcatcg ttgtcgaagg caccgtcatt gagatcacca tgcgcgcgac caaaattcgc 480
acgattgcac aagagaccgt gatcatcccc aactccacgg cgaaagtgtg catcaacaat 540
tctaataact ggtcgcgtgc ggttgtcgtt attccgatcc ccatgttggg ttctgaaaac 600
atcacagatg tcatcgcgcg ctctgaagct gcgactcgtc gcgcacttgg ccaggagaaa 660
atcgcaccgg aaatcctcgg tgaactcgat gtgcacccag ccacggaagt cacgccgcca 720
acggtggtcg gcatgccgtg gatggtcacc atgcgtttcc tcgtgcaagt caccgccggc 780
aatcaatggc tggtcgaacg cgccatccgc acagaaatca tcagcgaatt ctgggaagaa 840
tacggcagcg caaccactac atcgggaacc ctcattgatt ccttacacgt tgagcatgaa 900
gagccaaaga cctcgcttat cgacgcctcc ccccaggctc ttaaggaacc gaagccggag 960
gctgcggcga cggttgcatc gctagctgca tcctctaacg gcgatgcaga caatgcagac 1020
gcctcggtga tcaatgcagg caatccagag aaggaacttg attccgatgt gctggaacaa 1080
gaaccctcca gcgaagaacc ggaagaaaca gcaaaaccag atcactctct ccgaggcttc 1140
ttccgcactg attactaccc aaatcggtgg cagaagatcc tgtcgtttgg cggacgtgtc 1200
cgcatgagca cgtccctgtt gttgggtgcg ctgctcttgc tgtcactatt taaggtcatg 1260
actgtggaac caagtgagaa ttggcaaaac tccagtggat ggctgtcacc aagcactgcc 1320
acctcaactg cggtgaccac ctccgaaact tccgcgccag taagcacgcc ttcgatgaca 1380
gtgcccacta cggtggagga gaccccaacg atggaatcta acgtcgaaac gcagcaggaa 1440
acctcaaccc ctgcaaccgc aacgccccag cgagccgaca ccatcgaacc gaccgaggaa 1500
gccacgtcgc aggaggaaac gactgcgtcg cagacgcagt ctccagcagt ggaagcacca 1560
accgcggtcc aagagacagt tgcgccgacg tccacccctt ag 1602

Claims (10)

1.谷氨酸外运蛋白变体，为将MscCG谷氨酸外运蛋白进行点突变后所得，突变位点含有如下中的部分或全部：自N端起第166位、第86位、第344位、第385位、第165位、第103位、第393位、第399位、第173位、第343位、第310位、第168位、第504位、第186位、第141位、第200位、第205位、第14位、第334位和第362位。
2.根据权利要求1所述的谷氨酸外运蛋白变体，其特征在于：所述谷氨酸外运蛋白变体为将所述MscCG谷氨酸外运蛋白的至少如下位点的氨基酸残基进行点突变后得到的蛋白：自N端起第166位；
进一步地，所述谷氨酸外运蛋白变体为将所述MscCG谷氨酸外运蛋白的至少如下位点的氨基酸残基进行点突变后得到的蛋白：自N端起第166位、第86位、第344位、第385位。
3.根据权利要求1或2所述的谷氨酸外运蛋白变体，其特征在于：所述谷氨酸外运蛋白变体为如下任一：
(A1)所述谷氨酸外运蛋白变体为将所述MscCG谷氨酸外运蛋白的至少如下位点的氨基酸残基进行点突变后得到的蛋白：自N端起第86位、第166位、第344位、第385位和第141位；
(A2)所述谷氨酸外运蛋白变体为将所述MscCG谷氨酸外运蛋白的至少如下位点的氨基酸残基进行点突变后得到的蛋白：自N端起第86位、第166位、第344位、第385位、第200位和第205位；
(A3)所述谷氨酸外运蛋白变体为将所述MscCG谷氨酸外运蛋白的至少如下位点的氨基酸残基进行点突变后得到的蛋白：自N端起第86位、第166位、第344位、第385位、第168位和第504位；
(A4)所述谷氨酸外运蛋白变体为将所述MscCG谷氨酸外运蛋白的至少如下位点的氨基酸残基进行点突变后得到的蛋白：自N端起第86位、第166位、第344位、第385位和第186位；
(A5)所述谷氨酸外运蛋白变体为将所述MscCG谷氨酸外运蛋白的至少如下位点的氨基酸残基进行点突变后得到的蛋白：自N端起第86位、第166位、第344位、第385位和第14位；
(A6)所述谷氨酸外运蛋白变体为将所述MscCG谷氨酸外运蛋白的至少如下位点的氨基酸残基进行点突变后得到的蛋白：自N端起第166位和第310位；
(A7)所述谷氨酸外运蛋白变体为将所述MscCG谷氨酸外运蛋白的至少如下位点的氨基酸残基进行点突变后得到的蛋白：自N端起第334位和第362位；
(A8)所述谷氨酸外运蛋白变体为将所述MscCG谷氨酸外运蛋白的至少如下位点的氨基酸残基进行点突变后得到的蛋白：自N端起第165位；
(A9)所述谷氨酸外运蛋白变体为将所述MscCG谷氨酸外运蛋白的至少如下位点的氨基酸残基进行点突变后得到的蛋白：自N端起第173位和第343位；
(A10)所述谷氨酸外运蛋白变体为将所述MscCG谷氨酸外运蛋白的至少如下位点的氨基酸残基进行点突变后得到的蛋白：自N端起第103位、第393位和第399位。
4.根据权利要求1-3中任一所述的谷氨酸外运蛋白变体，其特征在于：所述MscCG谷氨酸外运蛋白为来自于谷氨酸棒杆菌ATCC13032的MscCG谷氨酸外运蛋白。
5.根据权利要求4所述的谷氨酸外运蛋白变体，其特征在于：所述来自于谷氨酸棒杆菌ATCC13032的MscCG谷氨酸外运蛋白的氨基酸序列如SEQ ID No.1所示。
6.根据权利要求1-5中任一所述的谷氨酸外运蛋白变体，其特征在于：在所述谷氨酸外运蛋白变体中，自N端起第166位的点突变为T166A、第86位的点突变为M86V、第344位的点突变为I344V、第385位的点突变为Y385C、第165位的点突变为E165G、第103位的点突变为A103T、第393位的点突变为I393V、第399位的点突变为R399C、第173位的点突变为T173A、第343位的点突变为V343E、第310位的点突变为S310P、第168位的点突变为I168T、第504位的点突变为Q504L、第186位的点突变为R186C、第141位的点突变为N141D、第200位的点突变为N200D、第205位的点突变为I205V、第14位的点突变为L14S、第334位的点突变为D334G、第362位的点突变为L362P。
7.谷氨酸外运蛋白变体相关的生物材料，为如下任一：
(I)编码权利要求1-6中任一所述的谷氨酸外运蛋白变体的核酸分子；
(II)含有所述核酸分子的表达盒、重组载体、重组菌或转基因细胞系。
8.根据权利要求7所述的生物材料，其特征在于：编码所述来自于谷氨酸棒杆菌ATCC13032的MscCG谷氨酸外运蛋白的核酸分子为SEQ ID No.12所示DNA分子；
进一步地，编码所述谷氨酸外运蛋白变体的核酸分子为如下任一：
(B1)SEQ ID No.19所示DNA分子；
(B2)SEQ ID No.20所示DNA分子；
(B3)SEQ ID No.17所示DNA分子；
(B4)SEQ ID No.18所示DNA分子；
(B5)SEQ ID No.21所示DNA分子；
(B6)SEQ ID No.16所示DNA分子；
(B7)SEQ ID No.22所示DNA分子；
(B8)SEQ ID No.13所示DNA分子；
(B9)SEQ ID No.15所示DNA分子；
(B10)SEQ ID No.14所示DNA分子；
和/或
所述重组菌为含有所述核酸分子的谷氨酸棒杆菌。
9.权利要求1-8中任一所述的谷氨酸外运蛋白变体或生物材在如下任一中的应用：
(C1)生产谷氨酸；
(C2)提高谷氨酸产量；
(C3)提高谷氨酸外运能力；
(C4)生产味精、香料或食品添加剂；
(C5)制备代盐剂、营养增补剂或生化试剂。
10.一种生产谷氨酸和/或提高谷氨酸产量和/或提高谷氨酸外运能力的方法，包括如下步骤：在受体菌中表达权利要求1-6中任一所述的谷氨酸外运蛋白变体，得到重组菌；对所述重组菌进行发酵培养，从发酵液中获得谷氨酸；
进一步地，在所述受体菌中表达所述谷氨酸外运蛋白变体，是通过像所述受体菌中导入权利要求7或8中所述的“编码所述谷氨酸外运蛋白变体的核酸分子”来实现的；和/或
进一步地，所述受体菌为谷氨酸棒杆菌。
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