CN112010953B - 小麦白粉病抗性相关蛋白Pm24及其编码基因和应用 - Google Patents
小麦白粉病抗性相关蛋白Pm24及其编码基因和应用 Download PDFInfo
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- CN112010953B CN112010953B CN201910467997.4A CN201910467997A CN112010953B CN 112010953 B CN112010953 B CN 112010953B CN 201910467997 A CN201910467997 A CN 201910467997A CN 112010953 B CN112010953 B CN 112010953B
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Abstract
本发明公开了一种小麦白粉病抗性相关蛋白Pm24及其编码基因和应用。本发明提供的蛋白质，获自小麦，命名为Pm24蛋白，为序列表中序列3所示的蛋白质。编码Pm24蛋白的核酸分子也属于本发明的保护范围。本发明还保护Pm24蛋白的应用，为提高植物的白粉病抗病性。本发明还保护一种制备转基因植物的方法，包括如下步骤：在受体植物中导入编码Pm24蛋白的核酸分子，得到对白粉病抗性增强的转基因植物。本发明对于植物白粉病育种具有重大的应用推广价值。
Description
技术领域
本发明属于生物技术领域，具体涉及小麦白粉病抗性相关蛋白Pm24及其编码基因和应用。
背景技术
小麦是中国重要粮食作物之一，其生产与保障国家口粮绝对安全直接相关。小麦白粉病是由高度专化性寄生菌布氏禾白粉菌(Blumeriagraminisf.sp.tritici)引起的真菌性病害，是危害小麦生产的重要病害之一。近年来生产方式的转变、水肥条件的改善和主要推广品种缺乏有效的抗白粉病新基因，白粉病危害日益严重，成为影响和限制小麦稳产、高产的障碍。发掘和利用多样化抗白粉病基因培育抗病新品种是防治白粉病最经济、安全、有效的途径。
近年来，小麦白粉病在中国小麦主产区发病范围不断扩大，危害程度日益严重，已经成为影响中国小麦生产的主要病害。中国拥有丰富的小麦地方品种资源，其中蕴含丰富的抗白粉病基因，历经几千年的自然选择和人工选择，具有广谱和持久的抗性，是小麦抗病育种的重要资源。
发明内容
本发明的目的是提供一种植物白粉病抗性相关蛋白Pm24及其编码基因和应用。
本发明提供的蛋白质，获自小麦(Triticum aestivum L.)，命名为Pm24蛋白，为如下(A1)-(A4)任一所示：
(A1)由序列表中序列3所示的氨基酸序列组成的蛋白质；
(A2)在(A1)所述蛋白质的N端或/和C端连接标签得到的融合蛋白；
(A3)将序列表中序列3所示的氨基酸序列经过一个或几个氨基酸残基的取代和/或缺失和/或添加且功能相同的蛋白质；
(A4)与(A1)-(A3)中任一所限定的氨基酸序列具有99％以上、95％以上、90％以上、85％以上或者80％以上同源性且具有相同功能的蛋白质。
标签具体如表1所示。
表1标签的序列
| 标签 | 残基 | 序列 |
| Poly-Arg | 5-6(通常为5个) | RRRRR |
| PoN-His | 2-10(通常为6个) | HHHHHH |
| FLAG | 8 | DYKDDDDK |
| Strep-tag II | 8 | WSHPQFEK |
| c-myc | 10 | EQKLISEEDL |
| HA | 9 | YPYDVPDYA |
蛋白质可人工合成，也可先合成其编码基因，再进行生物表达得到。
本发明Pm24蛋白获自中国小麦地方品种葫芦头、齿牙糙和/或白葫芦。
编码Pm24蛋白的核酸分子也属于本发明的保护范围。
所述核酸分子如下(B1)-(B4)中任一所示：
(B1)序列表中序列1所示的DNA分子；
(B2)序列表中序列2所示的DNA分子；
(B3)在严格条件下与(B1)或(B2)限定的DNA分子杂交且编码权利要求1所述蛋白质的DNA分子；
(B4)与(B1)或(B2)限定的DNA序列至少具有70％、至少具有75％、至少具有80％、至少具有85％、至少具有90％、至少具有95％、至少具有96％、至少具有97％、至少具有98％或至少具有99％同源性且编码具有相同功能蛋白质的DNA分子。
所述严格条件是在2×SSC，0.1％SDS的溶液中，在68℃下杂交并洗膜2次，每次5min，又于0.5×SSC，0.1％SDS的溶液中，在68℃下杂交并洗膜2次，每次15min。
含有所述核酸分子的表达盒、重组载体或重组微生物均属于本发明的保护范围。
可用现有的表达载体构建含有所述核酸分子的重组表达载体。使用所述核酸分子构建重组表达载体时，可在其转录起始核苷酸前加上任何一种增强型、组成型、组织特异型或诱导型启动子，它们可单独使用或与其它的植物启动子结合使用；此外，使用所述核酸分子构建重组表达载体时，还可使用增强子，包括翻译增强子或转录增强子，这些增强子区域可以是ATG起始密码子或邻接区域起始密码子等，但必需与编码序列的阅读框相同，以保证整个序列的正确翻译。所述翻译控制信号和起始密码子的来源是广泛的，可以是天然的，也可以是合成的。翻译起始区域可以来自转录起始区域或结构基因。为了便于对转基因植物或转基因微生物进行鉴定及筛选，可对所用表达载体进行加工，如加入在植物或微生物中表达可产生颜色变化的酶或发光化合物的基因、具有抗性的抗生素标记物或是抗化学试剂标记基因等。从转基因安全性考虑，可不加任何选择性标记基因，直接以表型筛选转化植物或微生物。
所述重组载体具体可为重组表达载体。所述重组表达载体具体可为在现有植物表达载体中***所述核酸分子得到的重组质粒。现有植物表达载体具体可为pCAMBIA1300载体或pTCK303载体。
本发明还保护Pm24蛋白的应用，如下(C1)或(C2)：
(C1)调控植物对白粉病的抗性；
(C2)增强或降低植物对白粉病的抗性。
本发明还保护所述编码Pm24蛋白的核酸分子的应用，如下(D1)或(D2)：
(D1)培育对白粉病的抗性改变的转基因植物；
(D2)培育对白粉病的抗性增强或降低的转基因植物。
本发明还保护一种制备转基因植物的方法，包括如下步骤：在出发植物中导入编码Pm24蛋白的核酸分子，得到对白粉病的抗性增强的转基因植物。所述核酸分子具体可通过以上任一所述重组表达载体导入所述出发植物。携带有所述核酸分子的重组表达载体可通过Ti质粒、Ri质粒、植物病毒载体、直接DNA转化、显微注射、电导、农杆菌介导等常规生物学方法转化到出发植物中。
本发明还保护一种植物育种方法，包括如下步骤：增加目的植物中Pm24蛋白的活性和/或含量，使得目的植物对白粉病的抗性增强。
本发明还保护一种制备转基因植物的方法，包括如下步骤：抑制出发植物中编码Pm24蛋白的核酸分子的表达，得到对白粉病的抗性降低的转基因植物。
本发明还保护一种植物育种方法，包括如下步骤：降低目的植物中Pm24蛋白的含量和/或活性，从而降低目的植物对白粉病的抗性。
以上任一所述植物为双子叶植物或单子叶植物。所述单子叶植物可为禾本科植物。所述禾本科植物可为小麦属植物。所述小麦具体可为小麦Fielder。
以上任一所述白粉病可为白粉菌致病菌引起的白粉病。以上所述白粉病致病菌可为布氏白粉菌小麦专化型。所述白粉病致病菌具体为中国北方的小麦白粉菌菌株E09。
本发明获得的中国小麦地方品种中的抗白粉病基因Pm24，有助于解析抗病基因的结构、功能和抗性机理，也可以为小麦分子育种工作提供更加丰富高效的白粉病抗源，将Pm24蛋白的编码基因导入小麦中，在保证小麦产量和品质的前提下，提高小麦对白粉菌的抗性。本发明对于植物白粉病育种具有重大的应用推广价值。
附图说明
图1为葫芦头和石4185对白粉菌E09抗性鉴定结果；其中，a为白粉菌E09接种7天；b为白粉菌E09接种24h，DAB染色；c为白粉菌E09接种48h，台盼蓝染色。
图2为中国小麦地方品种葫芦头中的小麦白粉病抗性基因Pm24图位克隆；其中，a为小麦1D染色体；b为Pm24遗传图谱；c为粗山羊草AL8/78的Scaffold；d为粗山羊草AL8/78基因组序列上Pm24基因的候选区间基因注释；e为中国春基因组序列上Pm24基因的候选区间基因注释；f为候选基因WTK的结构及抗、感亲本的序列差异；g为候选基因CNL的结构及抗、感亲本的序列差异。
图3为互补载体pCPL-WTK、互补载体pCPL-CNL、过表达载体pOE-WTK结构图及通过转基因获得的转基因植株对白粉菌E09的抗性鉴定图；其中，a为互补载体pCPL-WTK；b为过表达载体pOE-WTK；c为互补载体pCPL-CNL；d为部分WTK的互补T0代转基因株系(WTK-CLP1、WTK-CLP2、WTK-CLP4)与小麦Fielder(Fielder)的抗性差异；e为部分WTK过表达T0代转基因株系(WTK-OE1、WTK-OE2、WTK-OE3)与小麦Fielder(Fielder)的抗性差异；f为部分CNL的互补T0代转基因株系(CNL-CLP1、CNL-CLP6、CNL-CLP15)与小麦Fielder(Fielder)的抗性差异。
图4为葫芦头EMS突变体及其对白粉菌E09的抗性鉴定图；其中，a为葫芦头EMS突变体在WTK基因上的点突变；b为葫芦头EMS突变体(Mut129、Mut410、Mut477、Mut614、Mut988、Mut1091、Mut1106、Mut1174、Mut1269、Mut1322、Mut1365)与对照(HLT)的抗性差异。
具体实施方式
以下的实施例便于更好地理解本发明，但并不限定本发明。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。
文中所述抗病或是感病即指白粉病。所述白粉病是指由小麦白粉菌即布氏白粉菌小麦专化型所引起的小麦白粉病，下文中如无特殊标识小麦白粉菌具体指广泛流行于中国北方的小麦白粉菌菌株E09(BgtE09)。
根据表型，植株的白粉病抗病性采用0-4级的分级标准。0为免疫(植株无白粉病相关症状)。0；为近免疫(出现过敏性坏死反应，失绿反应)。1为高度抗病(菌丝层较稀薄且病斑少，可透绿)。2为中度抗病(菌丝层较厚，不透绿，能产生一定量的孢子)。3为中度感病(菌丝层比较厚且病斑多，产孢量大，但菌丝不会连成片)。4为高度感病(菌丝层厚、菌丝致密且病斑多，产孢量大，菌丝连成片)。按大分类来说，0-2为抗病，3-4为感病。
pCAMBIA1300、pTPCK303、农杆菌菌株EHA105购自派瑞金(北京)生物科技有限公司。
齿牙糙记载在如下文献中：Huang，X.Q.，Hsam，S.L.K.&Zeller，F.J.Chromosomallocation of genes for resistance to powdery mildew in common wheat(Triticumaestivum L.em.Thell.)4.Gene Pm24 in Chinese landraceChiyacao.Theor.Appl.Genet.95，950-953(1997)；
白葫芦记载在如下文献中：Xue，F.et al.Molecular mapping of a powderymildew resistance gene in common wheat landrace Baihulu and its allelism withPm24.Theor.Appl.Genet.125，1425-1432(2012)；
葫芦头记载在如下文献中：Wang，Z.Z.et al.Genetic and physical mapping ofpowdery mildew resistance gene MlHLT in Chinese wheat landraceHulutou.Theor.Appl.Genet.128，365-373(2015)；
石4185记载在如下文献中：Wang，Z.Z.et al.Genetic and physical mapping ofpowdery mildew resistance gene MlHLT in Chinese wheat landraceHulutou.Theor.Appl.Genet.128，365-373(2015)；
Fielder记载在如下文献中：He，H.G.et al.Pm21，encoding a typical CC-NBS-LRR protein，confers broad-spectrum resistance to wheat powdery mildewdisease.Mol.Plant11，879-882(2018)。
实施例1葫芦头、齿牙糙和白葫芦的等位性检测和多小种鉴定
鉴于小麦1DS染色体上已经定位了来源于中国小麦地方品种的抗白粉病基因Pm24(齿牙糙)和Pm24b(白葫芦)，为明确来源于地方品种葫芦头的抗白粉病基因(命名为MlHLT)是否与Pm24和Pm24b位于相同的基因位点，分别配制了这3个地方品种间的杂交组合，对葫芦头×齿牙糙(4,517 F2单株)，葫芦头×白葫芦(4,318 F2单株)，齿牙糙×葫芦头(4,382F2单株)，齿牙糙×白葫芦(4,590 F2单株)和白葫芦×葫芦头(4,692 F2单株)的F2代单株进行苗期白粉病菌系E09人工接种鉴定，结果发现所有杂交组合的F2代均无感病单株分离出来，说明来源于地方品种葫芦头的MlHLT与Pm24和Pm24b是位于同一位点的抗白粉等位基因。
对这3个地方品种用36个白粉病菌系进行鉴定，结果表明葫芦头、白葫芦、齿牙糙对36个白粉菌生理小种均表现为抗病，证明MlHLT或者其等位基因是一个白粉病广谱抗性基因。
实施例2白粉病抗性基因的精细定位
对来源于中国陕西的小麦地方品种葫芦头和来源于中国河北的小麦品种石4185进行白粉病抗性鉴定，分别对接种24小时的和48小时的小麦叶片进行DAB染色和台盼蓝染色，以及接种7天后的小麦叶片进行观察，结果如图1所示，发现葫芦头对北京地区流行的BgtE09具有免疫级别的抗性，石4185对BgtE09表现为高感。构建葫芦头和石4185的杂交二代群体，对其310个F2∶3家系进行抗病性鉴定，71个F2∶3家系表现纯合抗病，150个F2∶3家系表现抗、感分离，而F2代89个感病单株的F2∶3家系均表现感病，根据对其F2∶3家系接种白粉菌后的抗感分离比(纯合抗病∶抗、感分离∶纯合感病＝1∶2∶1)，可判断葫芦头中存在一个显性的白粉病抗性位点。
根据实施例1可知葫芦头、齿牙糙和白葫芦的等位性检测证明MlHLT与Pm24和Pm24b是位于Pm24位点的抗白粉病基因。利用上述310个单株的F2群体，通过比较基因组学的方法将MlHLT(即Pm24)定位在1DS染色体上3.6cM范围内(图2中a所示)。为了精细定位抗白粉病基因Pm24，利用普通小麦D基因组遗传图谱上的分子标记以及根据D基因组供体粗山羊草AL8/78已经公布的scaffolds序列设计的标记，将此抗性位点定位在分子标记WGGB240和WGGB245之间，其中分子标记WGGB242和WGGB243与基因共分离(定位群体共3720个单株)，WGGB240和WGGB245之间的遗传距离为0.06cM(图2中b所示)，在粗山羊草AL8/78中对应的物理区间为380kb(包含Scaffold3178.1部分序列和Scaffold3268.1部分序列)(图2中c所示)。
实施例3白粉病抗性基因Pm24候选基因的确认
粗山羊草AL8/78的基因组序列和中国春的基因组序列在实施例2所述的物理区间内存在6个候选基因，包括2个编码受体激酶的基因(RLK1、RLK2)，1个编码串连激酶的基因(WTK)，1个编码CC-NBS-LRR结构域蛋白的基因(CNL)，1个编码假定蛋白的基因(HP)和1个编码50S核糖体蛋白的基因(RP)(图2中d和e所示)。对抗病小麦品种(葫芦头)和感病小麦品种(石4185)中6个候选基因(RLK1，WTK，RLK2，CNL，HP和RP)的基因组序列进行分析，结果显示只有WTK和CNL两个基因的基因组序列在两个小麦品种之间存在差异(图2中f和g所示)，将WTK和CNL初步确认为候选基因。
实施例4白粉病候选基因的转基因功能验证
为了确定中国小麦地方品种葫芦头对白粉病的抗性是由候选基因WTK决定的，还是由候选基因CNL决定的，进行候选基因的转基因功能验证。
一、互补载体pCPL-WTK、互补载体pCPL-CNL、过表达载体pOE-WTK的构建
一)、将如序列表中序列1所示的WTK基因包括起始密码子上游3378bp，WTK基因区域和终止密码子下游1811bp的基因组序列利用引物(WTK-CPLTransF1/R1、WTK-CPLTransF2/R2、WTK-CPLTransF3/R3和WTK-CPLTransF4/R4进行扩增)，然后按照北京全式金生物技术有限公司的
Seamless Cloning and Assembly Kit试剂盒的方法将候选基因重组到线性化的pCAMBIA1300上，成功构建WTK的互补载体pCPL-WTK，如图3中a所示。
其中，引物序列如下：
WTK-CPLTransF1：AATTCGAGCTCGGTACCCGGGTTGTTGGCCAATCGTTATCA(序列5)
WTK-CPLTransR1：AGACCCCTAGTTTCCTGATGATA(序列6)
WTK-CPLTransF2：TATCATCAGGAAACTAGGGGTCT(序列7)
WTK-CPLTransR2：GCATATATTCATAGCAGAGCAGTTG(序列8)
WTK-CPLTransF3：CAACTGCTCTGCTATGAATATATGC(序列9)
WTK-CPLTransR3：CTGTGATGCGTGGTATGTAGTTG(序列10)
WTK-CPLTransF4：CAACTACATACCACGCATCACAG(序列11)
WTK-CPLTransR4：TGCAGGTCGACTCTAGAGGATCGTCCTTCATTGATGTGTTCC(序列12)
二)、将如序列表中序列4所示的CNL基因包括起始密码子上游2903bp，CNL基因区域和终止密码子下游2519bp的基因组序列利用引物(CNL-CPLTransF/R)进行扩增，然后按照北京全式金生物技术有限公司的
Seamless Cloning and Assembly Kit试剂盒的方法将候选基因重组到线性化的pCAMBIA1300上，成功构建CNL的互补载体pCPL-CNL，如图3中c所示。
其中，引物序列如下：
CNL-CPLTransF：CGAATTCGAGCTCGGTACCCGGGGCAATTGTTGCTGTAATGTGGTA(序列13)
CNL-CPLTransR：AAACGACGGCCAGTGCCAAGCTCTTCCACTCCAAGGGTACTGACCT(序列14)
三)、以含有WTK基因全长cDNA的质粒为模板，引物WTK-OETransF/R扩增获得包含如序列表中序列2所示的2,682bp的WTK基因的ORF，然后将其与线性化的pTPCK303质粒进行同源重组，构建WTK的过表达载体pOE-WTK，其中以玉米的Ubiquitin作为WTK基因的启动子，如图3中b所示。
其中，引物序列如下：
WTK-OETransF：TCTAGAGGATCCCCGGGTACCATGGGCGGATACGAGTTCCA(序列15)
WTK-OETransR：TTCGAGCTCTCTAGAACTAGTTTACTTCTTCCATCCGAAGTGT(序列16)
二、T0代转基因植株的获得
利用质粒提取试剂盒(购自天根生化科技(北京)有限公司)提取并纯化互补载体pCPL-WTK、互补载体pCPL-CNL、过表达载体pOE-WTK的质粒，分别转入农杆菌菌株EHA105，通过农杆菌侵染的方法转入普通小麦Fielder，获得了稳定的7株WTK的互补T0代转基因株系、15株CNL的互补T0代转基因株系和12株WTK的过表达T0代转基因株系。
三、转基因植株的鉴定
经过分子鉴定和白粉病抗性鉴定发现：
1、如图3中d所示，7株WTK的互补T0代转基因株系(7株检测到WTK的序列)，全部都抗病，表型与Fielder存在显著的差异；收获T0代转基因株系的种子，培育获得T1代家系，对7个T1代家系进行抗病性鉴定，7个T1代家系中均存在抗病单株和感病单株，抗病单株中均检测到WTK的序列，而感病单株中均未检测到WTK的序列。
2、如图3中f所示，15株CNL的互补T0代转基因株系(14株检测到CNL的序列)，无论是否检测到CNL，全部都依然感病，表型无异于Fielder；收获T0代转基因株系的种子，培育获得T1代家系，对14个T1代家系进行鉴定，14个T1代家系中所有单株均表现为感病，表型与Fielder一致。
3、如图3中e所示，12株WTK的过表达T0代转基因植株(12株检测到WTK的序列)，全部都抗病，表型与Fielder存在显著的差异；收获T0代转基因株系的种子，培育获得T1代家系，对12个T1代家系进行抗病性鉴定，12个T1代家系中均存在抗病单株和感病单株，抗病单株中均检测到WTK的序列，而感病单株中均未检测到WTK的序列。
上述结果证明候选基因WTK就是中国小麦地方品种葫芦头中的白粉病抗性基因MlHLT。从齿牙糙和白葫芦中分别获得了WTK基因上游3378bp，WTK基因区域和终止密码子下游1811bp的基因组序列，序列比较分析表明葫芦头、齿牙糙和白葫芦中WTK基因序列完全一样，进一步表明MlHLT、Pm24和Pm24b均为抗白粉病基因Pm24。
实施例5白粉病基因的EMS感病突变体分析
利用化学诱变剂甲基磺酸乙酯(ethyl methane sulfonate，EMS)诱变处理抗病材料葫芦头种子15000粒，共获得1366份M2代材料；将M2代家系用白粉病菌系E09进行苗期温室人工接种抗病性鉴定，结果如图4所示，筛选得到的11个感白粉病的EMS突变体(命名为Mut129、Mut410、Mut477、Mut614、Mut988、Mut1091、Mut1106、Mut1174、Mut1269、Mut1322、Mut1365)均在WTK基因上发生了点突变，其中，2个EMS突变体(Mut410、Mut1091)的点突变导致了其编码的蛋白提前终止，1个EMS突变体(Mut1106)的点突变改变了WTK的剪切方式，其余8个EMS突变体(Mut129、Mut477、Mut614、Mut988、Mut1174、Mut1269、Mut1322、Mut1365)的点突变均导致了非同异氨基酸的变异，进一步证明了候选基因WTK就是中国小麦地方品种葫芦头中的白粉病抗性基因MlHLT，即Pm24，该基因的核苷酸序列如序列表中序列1所示，开放阅读框(ORF)如序列表中序列2所示；该基因编码的蛋白命名为Pm24蛋白，Pm24蛋白的氨基酸序列如序列表中序列3所示。
序列表
<110> 中国科学院遗传与发育生物学研究所
<120> 小麦白粉病抗性相关蛋白Pm24及其编码基因和应用
<130> CNYFY191056
<160> 16
<170> SIPOSequenceListing 1.0
<210> 1
<211> 15599
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 1
ttgttggcca atcgttatca ccggtactac caggggcgga gacagggagg ggcgagcagg 60
ggtctgaccc ctgccaatcg ctcgcccccc tcaacgattt gtagcaggta gtgcgtaaca 120
gccgcagcta atcacataat taacccattt ctttttgagc gtaacataag cccatataga 180
aacaattaac tggaccagca ccaaagcaga gaagcccacc caacaatcca tgtacgacgg 240
ttgtgcctag ctatttttta gctcggccca gtgcctaatc cgtcagtacg tacatgcgtg 300
ttctgcactt ctcctagggc gttcttcgcc gccgccgctt gcttgtactg ctagggtcta 360
gggtgtttgc cattgtctct ggccgctact cgacacgaca acaacaccgt ccaccactga 420
tttgcagtac gcagttgacg cacgggcatg ggcggacggc ttgacgctgg cggatctaga 480
tctcgggtga gcaggataga tgcaggtata ttacattgat agtagttatt actctctatg 540
ttctaaaata attgtagctc tacgtttgtc ctaagtaaaa ctaatttaag tttaatgatg 600
tttatagaaa aatatggtaa gatctacaaa attaaatata catagtgtgc atatttttta 660
aagattctaa taaagctatg gtgtttcaaa atattgatat atttttctac aaactggtaa 720
aacttgaaga agtttaactt agaataaatt tagagcttct attattttaa aacagaggga 780
gtgagacttt tctttggata aaaagacctt actatgctta attaatgttt catataaaac 840
agtaaattag tctagtaatt atgtgtctat ttgtactagt caaagatgaa gcggaagaat 900
caacaacaaa tggggtaaat tctttttcaa tccaattggt tcaatttcaa gctccaacat 960
tattactact aatagtatat tttttatagt tccaaacaat aattttatgt attatcatcg 1020
atgctatcgt gagacttaat atattgtgat attctgtcat ttgcgtcaac aatggtgtct 1080
cgccctcctc atgctgaaat cctggctccc cccctgggta ctactacaat gaaatcatga 1140
cgcatgcaga acccgacgga caactgcttc ccttaatcga gagtgggaca cttgcaaact 1200
cgcaacttta cccctagacc caccaaatta cgacgcttac aaacacacgg gattcagaaa 1260
cttttactga agcaaaatgc tctgattttg catattttga agccgatgca catttaccgt 1320
cctagcaatc ccatgctcag attgcgtata ctgaagcaaa caagctgtcg ggggaggagg 1380
cagcgagaag ctccataaaa cacaaaaata gaatcgccca aacgagcaga acacgctgct 1440
ggcaacagtt tggttggata atggtgaggt gatcggcagc aggaacttgc agtggccatg 1500
gaaaaaacta agtaagagag aaaaagaaga agaaaagtca gttagttcat acggtgttac 1560
agcctggtaa ggataaggtc agcaggatga taaggcacat gtggtggtgg agaagtgggc 1620
acttgttgct tggctacgta cacacctaga cgtacatgag aaagaacatc tcaaaaataa 1680
tcaaacaaac atgtgatgta ttaaatcctg tttcctcaag tgggacagag gatttgacca 1740
cattgtttgg atttcttgtg ataattcaag tcttttttct attagccatg ttcaagaatt 1800
cgcacgataa ttaaccagtt aacttgcgat taatccttat ccgtagggtc accaagtagt 1860
caataaactg ataaatcatt taattaatta attaattaaa tggccattaa cttactgatt 1920
agcctattaa ttccctactc accagccaac cgagcagcca ctggcgtacc aacgggtgtg 1980
ccaggtgttg aaaaatattt ttgcaccatg taaaaattag ctgattttta aggcccatgc 2040
aaggtaattg aaatagactt catttctttt aaatgtgcac accgttcatt ttatttctag 2100
attcgccatt gcgagcagct accagttaac gatttcctta acaatgatga ttagtatatg 2160
tgttactaaa gagccacaaa gttgagatga ctgtgctgtg tgtgaaaaaa aaatatgaat 2220
cgggtaaatt cgtggcatga aaaagccgca atattaaaat gcaagattga tcctaaaacc 2280
aattaagtga atggttgtct caatacaacc ttaaatatat taaaacaagt ttcattatca 2340
aaatatatat taaaaagttc cctttcttcc tcaaatttga gcaggcacct gtgcttctcc 2400
aaagattatt tcgcctcttc tcgcccatgg cgtctcctct tcccacagac gttgtcgcag 2460
ccccattcac cccctcagtc atgcacatgc gccccgttga tgtctggccg ccgagtacat 2520
aaattcataa ataaattcaa cgagacggat ccgatgatct cctcaagagc gtgcttttaa 2580
aatgcaggct gccgcgaacc aatctgtgat tggatagtta gaaggtctgc ggtaacccca 2640
tccaaccaga ttcatccagt cccaaactta atattgatgc tcacattttt ctagatttat 2700
ttcaggcttt cgtcaatctc aacatgctat gtcggctcag actctcgaag gtatttatag 2760
ggtagggtat acgtgtatgt gttcacagag gtgtgagtgc tcgtttatgt gagcgacttc 2820
gactgtgtcg tgttaaaaaa aatgcaagct gcctctgtcc ccgccgtttt tttaattatt 2880
aatctggcat gtgtatggta attcaacaag ttgatcgggt cttaggaaat gcaaggtccc 2940
tcggtcacca ggccgaccgg tctttttttt ttcagacaaa gggccgaccg atcatttatt 3000
agtactctgc ttatcgggcc attggcacag ctctagagct ccggatctgg atcgacttgt 3060
tatgctcccc ccggtcaacg cctttgcctg ctgagcctgg gaatgattga tccgctcatg 3120
atctacaaga cgagaagcaa gcccagtttt gctcctagct agctgctttc tcctcctcct 3180
ccaatctcca ggtgagtaat taagatgcaa ctaggtagct gctttctggt aagactttag 3240
ctacctactt atcttctgct tgatttgctt tgtgcagcaa cgaatgaagg agaagctggc 3300
aaccatattc atatccataa tccatatcta tatgcgagct attgcttgct agaaagcttc 3360
ttgctcgcat actcggaaat gggcggatac gagttccaga gggcggagct agatgcactg 3420
gaaggcgtcg tacgcgatcc aactgcggag ccaatgagtc tgacgttgcc gcttctcagg 3480
cacataacaa atgatttctc ccctgaattt gaaattagta aagatgattc tgcagtggtt 3540
tacctggtac gactaaactc atatctcgtt tattattaat ataagcaaat cctatctcgt 3600
taattagtac tctgcttatg tggtcgaata tatatgtagg gggtgcttcc aagtgggttc 3660
cgtgttgctg tcaagaagtc tcactttcgt ttttgcttgg atgatgaaga tgcattcaca 3720
aatgaagttt ctattgcaat gaaggctgct cataagaaca cagtgcgagt cataggctac 3780
tgtcatcaca cgcatgagca aattgccgaa tacgaaggaa aacaagtttt cgcagaggtc 3840
agagaaaggt tgatctgtac cgagtatgtg cctaacggac cccttagtgg acatatcgaa 3900
ggtaagatat gtgcgcaaat ggatggatac gagttccaga gggcagaact agatgcacta 3960
gaacgcgtcg tacgcgatac aagtgcggag ccaatgagtc tgacgttgcc gcttctcagg 4020
cacataacaa atgatttctc cgatgaatct cgaattggcc gaggtggatt cgcagtggtt 4080
tacctggtat gattaaactc atacagtatc tccttagata ttaagcaatt cctatcttgt 4140
caattactaa cttcttattg tggtcaaata tacagggggt gcttccaagt gggttacgta 4200
ttgctgttaa gaggcttagc aatattgctt atatgaacga aagtgcattt caaaatgaag 4260
tgtttatcac aatgaaggcc actcacaaga acacagtgcg attcatgggc tactgtagtc 4320
aaatacaagg taaactcatc gaacacgacg ggcaacatgt tttcgcacag ctcgaggaaa 4380
ggttgatctg tgtggaatat gcgcctaaag gaacccttga tgcacatatc ggtaagatcg 4440
ataacacggc atgcccgccc tagcaataat tccctctctt tattttggtg tttctgtttg 4500
tgtggctgtt ttttactttg tgctcacaga acaaagaatg aataccagtg gaacaaaaaa 4560
gtagactata attttagcgc aactgaaacc aacatgacca gtcgtagaac acaaatttag 4620
acaaagtgga ccaaatcata agaaatatta aaacattaat tttttagaac ataaggcaca 4680
aaattatagt gctcgtgaat caaaacttgg ctacaagtgg aacacctatt gtaataaatg 4740
caagaaaact attagaggct tgtgcaataa aatttcggtg cccatggaac aaatcaaaat 4800
tctagtggaa caaaagtttt aactaagtga atgagcaaag tatatcactt gcaggaacaa 4860
attgaaacta ctactgctgc cacttcacca cttcttgtgc tgactttgcc atcctcacgt 4920
ggtaccaatt tccttcagtt ttcgtgtgtg ccaatgaatt cagatttcca aacaagtgga 4980
ggttttttct tccttcccca atatccctcg ttctacccaa ctatataatg gggcagagaa 5040
ggataagcaa agtaggcggt agaacaaaaa attaatgtaa ttgggacaaa atttaatcag 5100
taaatgctca ctagtctgag gataaacgaa agcatgaaac aaggaaaaga tgaggatgag 5160
caactattgg aataaaagca aaattaaact agcagagcaa agtaaagttg caaccaaatc 5220
atcagcgagc accacatcag cgcgctagtc tcccctcccg gtggtaaccg ctattggtgg 5280
ctcaccggcg tctaaggccc ctaggcggat gaagactaga tggctttcct tgccgatcta 5340
catgacatct ggcccttggc ttattggtgg cgatttcaac atgatcgtct ctgaggcgga 5400
caagaataac ccgcacgtga accggcggat catgcatagc tttcgatgct tccttgtcga 5460
cgaggaactt catgacattt gtatgcatgg taggtgctac acatggtcca gcgagcgcaa 5520
gattcctact cttgtcaaga acaaccgcat cgtctcctcc tcctcgaagg atgcgcatcc 5580
ccactgcctg ctgtagtgcc tctattcagc tgcttcagag cactggccgc tccttcttga 5640
ttgtgtcccg tgtacccatg ggatcaagcg gttccacttc gagtgatttt ggccgaagct 5700
tgatgatttc caactagtcg cctctgaggc ttggaactcc attgagcatg cccctgatgc 5760
atttcgccat atcgtcgtgc gccttaagtc taccacacgt cgtctccaga gctggagtgc 5820
caagtctatt ggccaaatct cggtgcagct tcagatcgcg tgtgagctca ttgctcattt 5880
ggacgccgct cagcactttc ggccgctctc gacctcggag gcctggttgc gacataggct 5940
gaaggccgcc taccttggcc ttgcctcgct cgagtgttcc atcattcggc aacgcacgag 6000
gtttgcctac gaggtttgcc tgtctcaggg aggctgatgc tggccctacc ttcttcaaca 6060
tgcatgcatg cctcgtacca taagcagaaa aaccgcatct tcgagctcaa ggtgggtgat 6120
agagttgtct ccacgccgga ggcgtttgcc actgcgggat tttgatcact tcaccggtgc 6180
ccttgagacc tccgaggacc gcgacttctc catctccctt cgggatctgc atgctggtgc 6240
tttcgatatc aaggcccttg atgagccatt tttttaggcc gaaatttggc gcgttgtcaa 6300
ggctctaccc tccagtaaga cgccggggcc atccctcaat gcgtgaggct ttcaaaaact 6360
caacgaggca tacattaccc tgctccccaa acggcatgcc gacgcaacca cactcttcta 6420
cttttggtcc attagcctca tccatctgat ggcgaagttg ttcgccaagg cgctctctct 6480
ctctctctct ctctctcgct ctctctctct ctctgcaaac ggcgtgtgtt ctgcacaatc 6540
tcaaggttcc tcgggtgatg ctgaagcttg acatcgttag ggccttcgac tcggtgtctt 6600
agccctttct tctggcccga atgtccctct cgtccgcaga aagggcgtgt gattcttggg 6660
caatgtcaag gcgaaggata acgagaaggg ctgcatgaag atgcaacttt gccttggaga 6720
agaggccctt actccatctc gccaagtgcc accccgtttt cctgagctta tggaaaagcc 6780
gctggcaagg ctccgtgtgc tcgcatggct catcccaagc ctttgacacg atgtcaaaga 6840
agccggggag acaggacaga aaattctcaa atttgaaaca tcttggtcgc ttgggcccct 6900
tgtcatcagc gaggaggaga gggcagtgat ccgaaagtga ataggagatg gagtggagca 6960
catgggagtt aaaatgcatg tcccaatcgg catttcggaa catcaagtca agcttgcaaa 7020
gagttgggtt ttgacgctcg ttgctccaag tgaatcgtat gttttgcatt accatgcctt 7080
ctatttttat ctcaagcacg atagatttgg ttgaagtcgc ccatgacaat ccatttagtg 7140
ccattgttgg gtttttgtga gattagctcg ttaaagaagt caccctttct gtcggaggcc 7200
gtgggaacat agacactagt gatcatgaag taggtatcgg tgtgaatcac attgaccatt 7260
gccgagaggc agaactcagt aaacttgacg tctgaaatac ggacgacact atcgtcccaa 7320
agcatgagta ggccccctct ggttccaata gccggacgct gacaaaactc gagagcctat 7380
tgccgccgag gtaggaggcg atgaatcggt ccactgattc catcttggac tcctgaaagc 7440
aaacaatgtg gcaagaggag gaggcaatcg tctcatgaac cgtcgcccgt ctacctgggt 7500
tgccaaattt tgataattgg tttgatgcca aaaattggca tgccaaattt tggcaactgc 7560
caaatattgg ctagagattg attgcttgcc aattttcatt gccaatctca gaaaaagttg 7620
tcaaatgtga agttgccaac atttgacaat aaaaattggc aagcaaccaa tctctagtta 7680
atatttggca cttgccaaaa tttggcatgc caacttttag catcaaacca attatgctct 7740
taacccttgc agatttcaac tcaaaaaaaa ttatgttttg ttctatcatc aggaaactag 7800
gggtctcgtg gcactcacaa ttagagcccg tcggggttgc atgaagtgtg cttgctgaac 7860
cagctctttg cacaacaaag agagaaataa tctggaagct gcgtggagtt tgaggtcagg 7920
gtggctgctg tctagagtag aggttgcaac ttgcaagcga tctttgcaca accatgatct 7980
tcattaagca ctggcgacga tgaacaccca agttacaacc aacaggaacg ttacagcaac 8040
tagttcctct tggttaaggg ataacagaga gctttgctaa catcagggca actagtgctt 8100
tggctctgtg aagtggggag aaacaatgga ggcacttgcc caccaattga ggggggattt 8160
gaggccgcct gtgaggagct ggaggggggg tttggcaggc caggggaggg gaggggaggt 8220
tgtcgctgcg tccagccagg ctccaggggg catcctgtac ttcctctgca aggcgatgcg 8280
cgggggcggc gtttgcagcg gaagacccgg ggcggggttg ccgcccgggc attgatggcg 8340
gcctcctctt gaatgcgttg cagagacgcg tcgggggcga cgatgatgga agcgacgcgc 8400
acgcctcccc ctcgaaggac aggcggcgtc tccagaacct cgtcagcaag ctcagctgcc 8460
tggatccgga gacgtgcagc ggccgatcca gaccgggcac cagtggcacg acagtggaca 8520
cagggataac aaccggggaa aaaaaattct ccggcaagcg gggcggcggc ggtggcagct 8580
catgttgtcc ggcgagcccc gaatggcacg cgtcgggagg ggctggtggg tgggtcgccc 8640
actcatgttg tccggtgatt tttccagctg tgccagctaa gtttttcaag tgtttgcgat 8700
ttaggaattt tgagaggata catgcttcgc tgtttgatct atttctatct atctcatgta 8760
agcctttcga gatgacaagc agatcaactt catgacatta tcaatacttc tgcaggtgac 8820
tatggtgaac ttgactggaa ccagcgttat caaattctaa aaggaatttg tcaaggtttg 8880
catcatctcc atgacgaaat gcacgttttt catggagata tcaaaccagc caatatatta 8940
ataggggata accttgtgcc taaaatctat gacttcggtc tctcccagat gtttgaagaa 9000
gaagaaacgg aacgtattgt tgaaaatatc gccggaacat tgtaagctaa cccgttcctc 9060
tttgttattt tgttttgttt ctgctgacac aggtacttcg ataaccaaaa tgcattagag 9120
ttagtcggcc gattgcctca catctaacat atgtgcagcg gatatatggc accggagttt 9180
tgtactaata atatggtgtc atttaaggct gagatataca gtttgggcgt tgtgatcggg 9240
gagttattga tcgggaagtg gtttgatgag gatgtgagaa aactatttgt acagcaactt 9300
aagggtttga gaaaaacatt ggtaaaagaa ggagcgtttt catcatggga aaacaaatac 9360
caccaagtta gaacatgtat ggagattggg caggactgca tagaccccaa cccacataaa 9420
aggcccactt tgttggagat tatccagcgg cttaatgaag cggaagatat gaactattct 9480
gcagcatcac tttggcaggt atggaaaatc gaaagcataa ggaaacaact gttctttatc 9540
taccatttca ctttagctaa cgtactgttt gagaagaatc agtcttagcc aagcttacat 9600
cctcacagac acagtacaga agtacatttc ttttctttag ccattacgat taacataaag 9660
agttcacaaa gccatctcca acgaaagata tgtgctaatt aaactaaatt ggaagagaaa 9720
atacataaca aagcgaagac accatgtaaa ttagagaaga aacaaactct tggtactcct 9780
tcagcgaggg aagacacctt gtcaacttag cagaaacaaa ctcttggtac tccttcagcg 9840
agggaagaca ccttgtcaac ttagcagaaa ccttacgact ctagtcctac atgacatgtc 9900
taagagaata ttgatgtcat gcattgcata atgcttacat ggtaagttat atataagtgc 9960
agtcaggaga cgaggaatcc gatttatcgg atacagaagc tttggagaca gagacaacat 10020
ccgagtttct tccaagtgac gaagaacccg cctctgtggg caagaccgga gaaacaagca 10080
cacaggagcc tgataaaccg gacctaataa gtaagttgcc agcatcggtg gacctgtctg 10140
acctaaaagt cctggagaaa atcacagatg atttttcaca cgaaagaata gttgggaagg 10200
acggtacatt caaaggttgt cataaggcat ttgtttataa ggtacacata gttgtaagtt 10260
actatgcagt gctttgcttt gctgatgtcc tttgagttca acatattctt ccatcttaat 10320
tatactccta acacacacac gcatactaaa tgtacattta tttgaacagg gtgacattcc 10380
acttagagaa atgatagccg tgaagaggtt aattggagtg gagattccat ttgaaaagtt 10440
taagagggaa gcagaacagt tcattagtct cgatcataag aatatagtaa aggttgccag 10500
ctactgccac gaccagtcta gaggacatag actggtacag ttcaaaggaa aaccgctacc 10560
acaactcttt aacggtcccg aacaactgct ctgctatgaa tatatgcaca acggaagcct 10620
tcgcgactat cttatgggta atgataccga tcactaatca actactgttt tctttaacat 10680
ccgtaaacaa ttactgtttt ctttaacatc tgtaatcaac tagtgacaat tgctttcgat 10740
ggaagagagc tgcacatcca gtacaaatca tatatatgga tgcaaatatt taggaactgc 10800
ttgcaatgga agccaggtta gaagaaccct agcgcttgga aggcagcatc aaacctctcc 10860
cctcccctgg gtcgcccccg cgcggcgacc agggggaacc ctagcgccgc caagccctcg 10920
gctccctctc ccacctctcc tgcccgccgc cgccggggcg cgccgtcggg caaagcccgg 10980
ttggcgtcgg cggcggcggg atctctcctc cccgcagcgg ccggacctgg cgcgggtggc 11040
gttcggcggc ggttggcggc gcgttggtga cggggcgcgc cggcgagggc tcagggggtg 11100
acggcaggag ctgcttcccc cggtggtcgc gtggggggcc attcggggcg cgcctatggt 11160
ggccatggtc gatctgtttt agatctgacc gccggtgcct cgggcggcgc ggggatctca 11220
gggcggcgac ctcgatcgat ggcgcgccgt gcggcctgcc tggcgggcgg cacccgcggg 11280
tgcagatggg gggccttcca cggctgtgtg ggcggtgtgg aggtggcggc gctacgacgg 11340
ctcctcgacg gccgcccgga gtcccatggg aggtgtcgtc tcagactcga tctactccgg 11400
ttcgtgctgg ctacagtagc gactacggtc gacaccaata ctgcctggac ggatctggcg 11460
ggtgggcatg gatccggggg aaactccagg ccggcgtggc ggccgcacca aagtcaacgc 11520
ccttggcgcc gattaccttc ctggaggctt cggtgtggat catacgcccc ccacctctac 11580
catgagcgca agcgaaagcc tctgttcctc tgtttgggcg acgatgacac tttggtgtca 11640
tgttccttcc tgaaggcatc ggccgaggac cgctcagggt ggtggagttg ctggtagttc 11700
ggagtcgaca cgagatggca tgtaggtgga gcagtgtgcc atcaacagta tcatcgacgg 11760
tgggtctcgg cggcatggcg cagtggagtc tcggcgtccg atgcgcggag atggactcgc 11820
gcaggaggag gtagctgtct ggcgtcgtgg tggcgtcgac ggtagctaga ccgtgcaagg 11880
gagatgcaac agtacaactc tgaagatgga ttggtggcag gtggctgcgg cagcctcata 11940
cccggcaggc gtcctggttg aggagtgcgc tggactggta gcaggtgccc catacccggc 12000
aggcgtcctg gttgaggagt gcgctggact ggtagcaggt gccccatacc cggcaggcgt 12060
cctggttggg acctcaggtc ttagatgttt aggtttggct gcgatgtctg tttggtatta 12120
ggcccagact atcagtgccc cttcatcaat tggataggtg tagcgacaat tattgcttag 12180
acggtggctt tagtcttgct gttgtatgac tttgtaaggt cttatgagaa taattaataa 12240
agtggccgta tgcatcgccc agatacagag gccgggggtc atcctccttt tcttaaaaaa 12300
aggaactgct tgcaaaaata aaaataaaat tgaacaattt aaagttcaat aagctagctc 12360
cttgctcatt gatctctcca tgtctacttg ttgtgttact aggtcaagga tctcgtgtaa 12420
ttgattggca aatgcgctac aaattgatca aagggacttg cgcaggctta cattaccttc 12480
acaagggccg tgcaggttgt ccaattgttc atttgaattt aagcccgtca aatgtattgc 12540
tggaccacaa ctacatacca cgcatcacag ggttcgattt ttcgaagctc attggtgaaa 12600
agaacaccaa atcagtggta cttaagctga atggacccat gtatgtatgc tctcagtgct 12660
actggtttaa aattagctct tgaatttgct catgatacat gtctatgttt tggaaaaatg 12720
tcttatgttt tgtggcagat cggtttagct agtctttttt ctgtgagaaa cgtttagcaa 12780
gtctattgac cgtaatatgc tttaattaaa ctgatgtgtt catatatttt tgcagagcgt 12840
acctgccacc ggatttcttc cattcgaagg gtactgatct taaatatctt gctacggtag 12900
atatatacag cttgggtctt atgattttag aaatcgcaac acaacaagag atcaaaggca 12960
tccatggagt gcttattaag agtgtaagac aaatttatta accctctgat catgagatag 13020
cttttattgc ttgacataaa agaacaaaaa aattcaacct tttaaaactt tgatgatcaa 13080
atatactcta cagatagagg aaaactggag ggaggagtca caaataacac ggctgtatac 13140
ctcactaggg gccgacgagc tgcggcaagt aaaaatgtgc attgatattg gcctagactg 13200
tgtcaagtca aaccctgaaa agagacctac agctggggcc atcatgctct ggcttgacaa 13260
agagagcaaa ccggtcccag tttcaagggc aggtgcagga gtgctgccaa gacctccggt 13320
ccctactaat atcaaccatg caggtcgcat ccaaggtatt ccgattgaga agtctcaagt 13380
gttaattcct tcgtttgcat gcaatgaata ttcttcggtg aaaatacaaa atcttcatct 13440
acttctgttt atgtattact aagcagccgc aagctattcc ctacttctca attgttaatt 13500
ccttcattag catgcagtga atattctttg gtgaaaattg catctatgaa aggaaattaa 13560
aatacagtgt cttcatctat tactatttct attcatccaa gctgatcagt aaaagcatcc 13620
acatgttata agggcaatct atatgtggta tttaaaatgt tatttctgtc gcttttcaca 13680
tcgtatatgc aatctaagat aatgctctcc ctctcactgt aaataaatat gttaccagaa 13740
aaggagaagg cgggattcct gaaacgacac ttcggatgga agaagtaacc acctgcgctg 13800
gcttgtattc ctgtgaatat tatagatggt tcgacaaagc tcgtggttaa ccgaaggagg 13860
atatttgaga gcttttgtgt tagaacatgg ccatcttgtg cgtccgcagc agccggggtc 13920
atcaggaggg agtgctcact gcatggacca gatcacgctc tgtcagccat gggagtcgtg 13980
cgcacatagt gccacacatg taacgtttag ttgtgtgatc tgaactgtta tgtgtcgtgt 14040
aacctacctc atgtgtgtgt atgataaaga aagaaccaca tcagtggaag agttaaacca 14100
atcgtatggt atgacaggtg agacgtcaca tatgtggaag caaaaaagag ttggaggtaa 14160
ggtggttttc gtacatactg gttctgatcc ctctcatatt tttgtatcct tattatctac 14220
tccctctgtc ccataatgta agagtgtttt tgacattagt gtcaaaaact gtgtcaaaaa 14280
cgctcttaca ttactggacg gagggagcat gtaagtttcg tattaatcgt actatgcgcc 14340
atcttgctct atgaatccct atgcgagact tctttcttct cacaattgat ttattatttt 14400
atggtaatct tgtttcacat ttattgacgt gattttatca ccatatcttg gtgttctttc 14460
cttaaacacc caaacatatc cgaaaatggt ttcccacgaa aacaggcaaa agaaaggtgc 14520
acaagcacgg taaaatccgg gaaagcttac tacgtaggca caaaacaaag ggtaacaaca 14580
tcataaaatt tggactcatc gcaactcacg aggggctaag tgctagcatg cctcgtgcac 14640
aagatggggc tgcttctggt ccacctagct agctccaaca atagtagcct tggctcacat 14700
ctacactact attaaaaaaa caaacataaa ctccctacag ctacaccaac aactgtacac 14760
gcgattatca agaacgtcag accaatctaa ttaaatcaaa tccgagagtc tatattatat 14820
caatagtctg ccatacaagt taacggtcca gattaaatgt tctgccgggg cagaccacct 14880
cgcgcacctc tttgtccaca attaatgggg atatatggta accaccacgc gcacctcttg 14940
gtactcaatt agtgaggata ttaatcccgt aattttctct ctggcctttg ccttagttgt 15000
tgttaataag tgccttgcgg ttactcctct gatgccccaa ggaccgctgc cgcctacttc 15060
tcctcctctg ttgccgagtc aattgcaaga aaccaccaca tttgcggcta ggtttgcaga 15120
aaaccaccaa ctcgttaatc cgttgcagaa aacaccgaaa aatttgttaa gtcattgcaa 15180
aaagcactga tcggatgatt tggcccgttt aatcactttc ttacaagtgg ggccagattg 15240
taaggaattg acttagcaaa aaaataacac atacacccct agatctaaaa aacaaaagca 15300
atcagaccct cccccgtgga acactgacgc cttgtacact acgcgtcgcc gcggcgctgc 15360
tgctatccgc cgccgctatg ctccgggccg ggaggatgag ggaggaggag cccagctggt 15420
tcgggcgctt cgacgaggac ctgccgtcgc cggacgagct cattccgctc tcacactggc 15480
tcatcacccg cgatctcccc gccgccttca acatccccac gcacggggcc ggccgctcag 15540
cgatcttggc aaaggaaaga gggagcgagg cgtcgccggg gaacacatca atgaaggac 15599
<210> 2
<211> 2682
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 2
atgggcggat acgagttcca gagggcggag ctagatgcac tggaaggcgt cgtacgcgat 60
ccaactgcgg agccaatgag tctgacgttg ccgcttctca ggcacataac aaatgatttc 120
tcccctgaat ttgaaattag taaagatgat tctgcagtgg tttacctggg ggtgcttcca 180
agtgggttcc gtgttgctgt caagaagtct cactttcgtt tttgcttgga tgatgaagat 240
gcattcacaa atgaagtttc tattgcaatg aaggctgctc ataagaacac agtgcgagtc 300
ataggctact gtcatcacac gcatgagcaa attgccgaat acgaaggaaa acaagttttc 360
gcagaggtca gagaaaggtt gatctgtacc gagtatgtgc ctaacggacc ccttagtgga 420
catatcgaag gtaagatatg tgcgcaaatg gatggatacg agttccagag ggcagaacta 480
gatgcactag aacgcgtcgt acgcgataca agtgcggagc caatgagtct gacgttgccg 540
cttctcaggc acataacaaa tgatttctcc gatgaatctc gaattggccg aggtggattc 600
gcagtggttt acctgggggt gcttccaagt gggttacgta ttgctgttaa gaggcttagc 660
aatattgctt atatgaacga aagtgcattt caaaatgaag tgtttatcac aatgaaggcc 720
actcacaaga acacagtgcg attcatgggc tactgtagtc aaatacaagg taaactcatc 780
gaacacgacg ggcaacatgt tttcgcacag ctcgaggaaa ggttgatctg tgtggaatat 840
gcgcctaaag gaacccttga tgcacatatc ggtgactatg gtgaacttga ctggaaccag 900
cgttatcaaa ttctaaaagg aatttgtcaa ggtttgcatc atctccatga cgaaatgcac 960
gtttttcatg gagatatcaa accagccaat atattaatag gggataacct tgtgcctaaa 1020
atctatgact tcggtctctc ccagatgttt gaagaagaag aaacggaacg tattgttgaa 1080
aatatcgccg gaacattcgg atatatggca ccggagtttt gtactaataa tatggtgtca 1140
tttaaggctg agatatacag tttgggcgtt gtgatcgggg agttattgat cgggaagtgg 1200
tttgatgagg atgtgagaaa actatttgta cagcaactta agggtttgag aaaaacattg 1260
gtaaaagaag gagcgttttc atcatgggaa aacaaatacc accaagttag aacatgtatg 1320
gagattgggc aggactgcat agaccccaac ccacataaaa ggcccacttt gttggagatt 1380
atccagcggc ttaatgaagc ggaagatatg aactattctg cagcatcact ttggcagtca 1440
ggagacgagg aatccgattt atcggataca gaagctttgg agacagagac aacatccgag 1500
tttcttccaa gtgacgaaga acccgcctct gtgggcaaga ccggagaaac aagcacacag 1560
gagcctgata aaccggacct aataagtaag ttgccagcat cggtggacct gtctgaccta 1620
aaagtcctgg agaaaatcac agatgatttt tcacacgaaa gaatagttgg gaaggacggt 1680
acattcaaag gttgtcataa ggcatttgtt tataagggtg acattccact tagagaaatg 1740
atagccgtga agaggttaat tggagtggag attccatttg aaaagtttaa gagggaagca 1800
gaacagttca ttagtctcga tcataagaat atagtaaagg ttgccagcta ctgccacgac 1860
cagtctagag gacatagact ggtacagttc aaaggaaaac cgctaccaca actctttaac 1920
ggtcccgaac aactgctctg ctatgaatat atgcacaacg gaagccttcg cgactatctt 1980
atgggtcaag gatctcgtgt aattgattgg caaatgcgct acaaattgat caaagggact 2040
tgcgcaggct tacattacct tcacaagggc cgtgcaggtt gtccaattgt tcatttgaat 2100
ttaagcccgt caaatgtatt gctggaccac aactacatac cacgcatcac agggttcgat 2160
ttttcgaagc tcattggtga aaagaacacc aaatcagtgg tacttaagct gaatggaccc 2220
atagcgtacc tgccaccgga tttcttccat tcgaagggta ctgatcttaa atatcttgct 2280
acggtagata tatacagctt gggtcttatg attttagaaa tcgcaacaca acaagagatc 2340
aaaggcatcc atggagtgct tattaagagt atagaggaaa actggaggga ggagtcacaa 2400
ataacacggc tgtatacctc actaggggcc gacgagctgc ggcaagtaaa aatgtgcatt 2460
gatattggcc tagactgtgt caagtcaaac cctgaaaaga gacctacagc tggggccatc 2520
atgctctggc ttgacaaaga gagcaaaccg gtcccagttt caagggcagg tgcaggagtg 2580
ctgccaagac ctccggtccc tactaatatc aaccatgcag gtcgcatcca agaaaaggag 2640
aaggcgggat tcctgaaacg acacttcgga tggaagaagt aa 2682
<210> 3
<211> 893
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 3
Met Gly Gly Tyr Glu Phe Gln Arg Ala Glu Leu Asp Ala Leu Glu Gly
1 5 10 15
Val Val Arg Asp Pro Thr Ala Glu Pro Met Ser Leu Thr Leu Pro Leu
20 25 30
Leu Arg His Ile Thr Asn Asp Phe Ser Pro Glu Phe Glu Ile Ser Lys
35 40 45
Asp Asp Ser Ala Val Val Tyr Leu Gly Val Leu Pro Ser Gly Phe Arg
50 55 60
Val Ala Val Lys Lys Ser His Phe Arg Phe Cys Leu Asp Asp Glu Asp
65 70 75 80
Ala Phe Thr Asn Glu Val Ser Ile Ala Met Lys Ala Ala His Lys Asn
85 90 95
Thr Val Arg Val Ile Gly Tyr Cys His His Thr His Glu Gln Ile Ala
100 105 110
Glu Tyr Glu Gly Lys Gln Val Phe Ala Glu Val Arg Glu Arg Leu Ile
115 120 125
Cys Thr Glu Tyr Val Pro Asn Gly Pro Leu Ser Gly His Ile Glu Gly
130 135 140
Lys Ile Cys Ala Gln Met Asp Gly Tyr Glu Phe Gln Arg Ala Glu Leu
145 150 155 160
Asp Ala Leu Glu Arg Val Val Arg Asp Thr Ser Ala Glu Pro Met Ser
165 170 175
Leu Thr Leu Pro Leu Leu Arg His Ile Thr Asn Asp Phe Ser Asp Glu
180 185 190
Ser Arg Ile Gly Arg Gly Gly Phe Ala Val Val Tyr Leu Gly Val Leu
195 200 205
Pro Ser Gly Leu Arg Ile Ala Val Lys Arg Leu Ser Asn Ile Ala Tyr
210 215 220
Met Asn Glu Ser Ala Phe Gln Asn Glu Val Phe Ile Thr Met Lys Ala
225 230 235 240
Thr His Lys Asn Thr Val Arg Phe Met Gly Tyr Cys Ser Gln Ile Gln
245 250 255
Gly Lys Leu Ile Glu His Asp Gly Gln His Val Phe Ala Gln Leu Glu
260 265 270
Glu Arg Leu Ile Cys Val Glu Tyr Ala Pro Lys Gly Thr Leu Asp Ala
275 280 285
His Ile Gly Asp Tyr Gly Glu Leu Asp Trp Asn Gln Arg Tyr Gln Ile
290 295 300
Leu Lys Gly Ile Cys Gln Gly Leu His His Leu His Asp Glu Met His
305 310 315 320
Val Phe His Gly Asp Ile Lys Pro Ala Asn Ile Leu Ile Gly Asp Asn
325 330 335
Leu Val Pro Lys Ile Tyr Asp Phe Gly Leu Ser Gln Met Phe Glu Glu
340 345 350
Glu Glu Thr Glu Arg Ile Val Glu Asn Ile Ala Gly Thr Phe Gly Tyr
355 360 365
Met Ala Pro Glu Phe Cys Thr Asn Asn Met Val Ser Phe Lys Ala Glu
370 375 380
Ile Tyr Ser Leu Gly Val Val Ile Gly Glu Leu Leu Ile Gly Lys Trp
385 390 395 400
Phe Asp Glu Asp Val Arg Lys Leu Phe Val Gln Gln Leu Lys Gly Leu
405 410 415
Arg Lys Thr Leu Val Lys Glu Gly Ala Phe Ser Ser Trp Glu Asn Lys
420 425 430
Tyr His Gln Val Arg Thr Cys Met Glu Ile Gly Gln Asp Cys Ile Asp
435 440 445
Pro Asn Pro His Lys Arg Pro Thr Leu Leu Glu Ile Ile Gln Arg Leu
450 455 460
Asn Glu Ala Glu Asp Met Asn Tyr Ser Ala Ala Ser Leu Trp Gln Ser
465 470 475 480
Gly Asp Glu Glu Ser Asp Leu Ser Asp Thr Glu Ala Leu Glu Thr Glu
485 490 495
Thr Thr Ser Glu Phe Leu Pro Ser Asp Glu Glu Pro Ala Ser Val Gly
500 505 510
Lys Thr Gly Glu Thr Ser Thr Gln Glu Pro Asp Lys Pro Asp Leu Ile
515 520 525
Ser Lys Leu Pro Ala Ser Val Asp Leu Ser Asp Leu Lys Val Leu Glu
530 535 540
Lys Ile Thr Asp Asp Phe Ser His Glu Arg Ile Val Gly Lys Asp Gly
545 550 555 560
Thr Phe Lys Gly Cys His Lys Ala Phe Val Tyr Lys Gly Asp Ile Pro
565 570 575
Leu Arg Glu Met Ile Ala Val Lys Arg Leu Ile Gly Val Glu Ile Pro
580 585 590
Phe Glu Lys Phe Lys Arg Glu Ala Glu Gln Phe Ile Ser Leu Asp His
595 600 605
Lys Asn Ile Val Lys Val Ala Ser Tyr Cys His Asp Gln Ser Arg Gly
610 615 620
His Arg Leu Val Gln Phe Lys Gly Lys Pro Leu Pro Gln Leu Phe Asn
625 630 635 640
Gly Pro Glu Gln Leu Leu Cys Tyr Glu Tyr Met His Asn Gly Ser Leu
645 650 655
Arg Asp Tyr Leu Met Gly Gln Gly Ser Arg Val Ile Asp Trp Gln Met
660 665 670
Arg Tyr Lys Leu Ile Lys Gly Thr Cys Ala Gly Leu His Tyr Leu His
675 680 685
Lys Gly Arg Ala Gly Cys Pro Ile Val His Leu Asn Leu Ser Pro Ser
690 695 700
Asn Val Leu Leu Asp His Asn Tyr Ile Pro Arg Ile Thr Gly Phe Asp
705 710 715 720
Phe Ser Lys Leu Ile Gly Glu Lys Asn Thr Lys Ser Val Val Leu Lys
725 730 735
Leu Asn Gly Pro Ile Ala Tyr Leu Pro Pro Asp Phe Phe His Ser Lys
740 745 750
Gly Thr Asp Leu Lys Tyr Leu Ala Thr Val Asp Ile Tyr Ser Leu Gly
755 760 765
Leu Met Ile Leu Glu Ile Ala Thr Gln Gln Glu Ile Lys Gly Ile His
770 775 780
Gly Val Leu Ile Lys Ser Ile Glu Glu Asn Trp Arg Glu Glu Ser Gln
785 790 795 800
Ile Thr Arg Leu Tyr Thr Ser Leu Gly Ala Asp Glu Leu Arg Gln Val
805 810 815
Lys Met Cys Ile Asp Ile Gly Leu Asp Cys Val Lys Ser Asn Pro Glu
820 825 830
Lys Arg Pro Thr Ala Gly Ala Ile Met Leu Trp Leu Asp Lys Glu Ser
835 840 845
Lys Pro Val Pro Val Ser Arg Ala Gly Ala Gly Val Leu Pro Arg Pro
850 855 860
Pro Val Pro Thr Asn Ile Asn His Ala Gly Arg Ile Gln Glu Lys Glu
865 870 875 880
Lys Ala Gly Phe Leu Lys Arg His Phe Gly Trp Lys Lys
885 890
<210> 4
<211> 9704
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 4
gcaattgttg ctgtaatgtg gtactatgtt tccatagagt gccacttatg atgtggtctg 60
tagctacaac ttttcacttg gcatgcaaca gttctgtagg ccatgctact gctgctttgg 120
catgcatttc acttgttgca atgctgctgc tgctgctgtg cttgccatgc tgctgttgtt 180
gtgcactact ctagatagca gcagtagtag atattcaatt ggcagttaac atttaaattc 240
ataaaacctc agtgcactga atttttgcta aatagataac tgaatttttg ttaacattta 300
aattcagtta actgaagatt cagttaactg aatttgtagt taactgaaga tatctcattt 360
aaattcaatt aactaaagat tcagataact aaatttgtag ttaactgaag ataactcatt 420
taaattcagt taactgaaga ttcagataac tcagtttgta gttaactgaa gatttctcat 480
ttaaatttag ttaactgaat cttcagataa ctgaacttca gttaactaca aactaagtta 540
attgacactg taaaaattgc acttatctgg agacattcag ataactagta acactagaag 600
cagtcataca gatttagtta actaaaagca gtcatacaaa ttcagtgcat tacataatac 660
tggcattaca ccaacaaatt cagtacaact aattaataga tctccttcaa tgaattactc 720
aaattcatct aagatctcct tcaccctcct tatcttgctc ttggtctctt ccttgtgcct 780
gaacaaatca ccaatcatgt actctagttt cttcttctcc ttcttcagct catccctgtg 840
tgacaccact ttgtccatct cttccttcat cttgtccatc tgttgcttca tctcatccct 900
ctctttctct gccttagccc tcacaacctg atgaatgcta gtgttagatt tatcagacaa 960
cttcttcttc tcaagctctt tctttaggtc agaaagagca agtcttgcat tgcccaactc 1020
agtaattgcc tgctccttgt cagccaccat cttagcaaaa tctagcttaa aaaacctcag 1080
atctttttcc atcttttcct tctctctcac aaccctaaca ttttcttccg caagaaccaa 1140
attctccctg atccttagct tgttctcatc atcatacatg ccccaaaccc tagctaaact 1200
catcttcaga gtggcaggcc attcagggtc aatccactcc acaaagttgc atttaggtat 1260
ttcctaccaa ataagaaaga aaacaacttt agataaatga aaaagcttgg agcagtattg 1320
attcagttaa attcatttaa attcagttaa gttcacttaa ctgaatttaa atctagttaa 1380
gttcacttaa ctgaatttaa atccagttaa gttcagttat ttcatttcaa ttcagttaag 1440
ttcagttatt tcatttaaat tcagttaagt tcagttagtt catttaaatt cagttatgtt 1500
cacaaacaag atgctgttaa ctcaaaaatt cagttaagtt cacaaactag atgctcttaa 1560
ctgaaaatgc actcctccac agttacatgg cacactctta tacagatcat agtaagatga 1620
cacattgcaa gtttggattt aactgtagat ctactaacct tttgtgcaca agcaagatac 1680
ctccttccac tatcaactga ttcaaaggac acaaacttct cacacacaca atggtgttca 1740
catctagctg caagatctgg agcaaggcct ctccactcag agcagaaaat ggtggcagga 1800
gcaggagcct acaacattgt gtacaacata ttctagtcag caaagaggca ggcatcgcat 1860
acctaactac tgcatcatct atgctgaaat tttgctactc aagaacatac ccccttcccc 1920
atttttgctc aatccctaac tgccagaggc atagtcaaga actaacctca cttgtcacag 1980
agtagccgtc ggacgacgag ctggatcctt cactccagga taccatgggg gcgagctgga 2040
gggcggcgtt gagctggagg cggcggcgag ctggacggcg gcggcgagct tgctggcggc 2100
gtggagaagc tgctgctgga gctcaggggg gcagggggta gatggggaac gagtggagct 2160
ggagctggag ctagaccgga cgccaactca attccgacag atttaattct ggtgttgtgt 2220
actgcgggtt gattagagga aacagcaggg gggtttctgc aaaagtgtgt gcgacgaccg 2280
ggccgtccac ctggctgcca gttgtccagt tgccattatc ctgggactaa aatgtcacat 2340
tgactgcaag ttagggtatg gtggggttga gttttgtaag tttgggatta aattatcaca 2400
ttctgggcaa gttagggtac ctggagtgct tttacctcat aataaattgc ttctttgcta 2460
ctccctccgt tccaaaatag atggctcgac tttgtactaa ctttagtaca aagttagtat 2520
aaagttgggt catctatttc atcaattttt tttaaagctg gagctagctg aaaacatttg 2580
aataaaatga atttgatggc atcgagctga ttttagagca atcctaacca atcccttagg 2640
gcatctatgt tagtcttgtt gataaaatat tcatatcatc aaccaacaag ctattataca 2700
accattccaa tagattgtat ctaagctatc caatagatga tgagaaaata aatgtgattg 2760
ctctttattt caccttggag cttgtgcaaa ggttgttggt tattttacat ataactttgc 2820
cctctcttca catttattac atgccacgtt atcactatgt cctagatggc caaatttaca 2880
gacacctatt ttacaactgt tggagatgcc cttaatccca tctcaacgga cgtcgcctat 2940
gtgagaggca ctaattagca gacctactct gctctctgag tcttgattga gccattgacc 3000
acgaacacac acacgggaca ggataaggca gagaagctgg tgagcaggcg cgagggcggc 3060
agcaatggag gagatcgtgc ttggactgtc caagacggtg gtggagggga cgctggtcaa 3120
ggtgaaggcc gcgattgacg aggaggcgaa gctgaagctt gccgtgcaga gcgacctggt 3180
gttcatcacg ggggaattcg agatgatgca gtcgttcctc aacgtcgccg acgcggagcg 3240
catcaagaac aacgccgtca agacgtgggt gcggcagctc cgcgacctgg cctacgacac 3300
cgaagactgc atcgagctgg tggtccacct ggatcccaag cccagatggt ggcgccgcct 3360
gctggtcctg ccttgcctgc cggcggtgtc gctgcccatg gacaacgcgg cggcggagat 3420
aaaggagctc aaggaccggg tggagttcgt gagccagagg aacatgcgct acaagctcat 3480
caccgacttc ggcggcgcca agtccgtcgc gcagcagcag cttgaccggg ccgccggagc 3540
tctcgacatc gccgtggagg aggcggcgaa gaaaggtcgt agcttggtgg atctgaccga 3600
gttgatcccc agaatggaag accgccctga gctcggcgtg atctcggtgt gggggaccgg 3660
cggcgacctt ggggtggcgt ccatcgtcag gaaggcctac gacgaccctg aaatttgcaa 3720
gaacttccat agtcgtggct gggccaagtt gacccaccct ttcgatccgc ggaagatcct 3780
ccggagcttg ttcattcagt tctgcactaa cactgctgct ccacagaagc aaggggaaac 3840
tctggatgta gattcccttc taaggatgga gaagacggtg gtggaagaag gggagctggt 3900
caagcagttt gtgagtcatg tgagcaccca caggttcctc gttgtcctcg aaggcgtgtc 3960
aaccatggcg gaatgggatg ccctccggat gtacctgcca gacatgggta acgggagcca 4020
gatcatcgta tccacacagc attttgacat cgcgagcttg tgcaccgggc agctgcacaa 4080
agtttcagag ttgagaaaat tcacacctga ccactctgtt tgtgtctttt tcaaggaagt 4140
atgtaattaa gcacaccgtg acactttatt cacttgtcat ttattcattg tctgaagaaa 4200
tgcaacgctg gaggtgtaac ttttgttgat ttgtcaggtt tccggaggtg atgatcctgg 4260
aaatgcaatg ttggagggag atgacgaaac tgaacttatt gggcgcgaca cagaagagga 4320
cgagcttatg gagcagattg atgaggtttc ccgtgtgatc tccgtatggg ggattgctgg 4380
tgtaggaaaa tcggcacttg tcagaagtgc ctactgcaag tatctgcagg accgccccaa 4440
cttcgccggg ttaaatctga cctacataca cgggcgcgtg aacgtgcccc atcccttcaa 4500
tctgagggag ttctgccgca gcttgctcct gaacttgtcc tctgaacctg ctaaaaccaa 4560
agaggacgcc gtcgctgaac tggcaaacat gaccaatcca attgatgtct gccgtaagct 4620
tcttagcgac gatcaacagc agaagcagtg ccttattgtt attgatgacc tgcagttcac 4680
tgaagaatgg gacagaataa aacaagcctt gtcatttggt aaagaatgtt gtatcatagt 4740
tgttacaaat gattcaagtg ttgccaaata ttgttcaggg caagatcaga aactcatgtt 4800
caacgtcaaa ggtcttggtg acgagcatgc ccttgacctc ttcaacacgg tactcgctgc 4860
tctaaaataa ccattcctgc aaattcaact ccttattaga gtcctatatt tactttttct 4920
gtataccaca gcacggatct tcagtcccag atgtagctat agagcatgtt tggttcatag 4980
tcatactcat accgtgtcta attttgccac tttcttgtat agcagcctgt atcctcagcc 5040
agtctcacac tttcgcaaaa ccgggggcaa agtattaact gccaacttct ttgtcggact 5100
gaataaaaat gtatcaaact taagtcatgc tttttggaac acggcaaaaa caacatggta 5160
tacatacaac cagtacagga acttatgtga tggttagcag gatagtatag gaataatgaa 5220
ggcatgaatt agccagaagc ggtgtttgaa tgcttcaggg gaaaaatagt tacatatttg 5280
taaagtacct tccaaaattt atagaaaata attcttctat gcagttgtac atttcaaaga 5340
ccgtgactaa ctatgtcaag tttggccact tttcaataaa attccggtag ctagggaacc 5400
ctctctatcc cccatgacca ttcggaaaga aaaacgaccc tagctatttg gaactctcct 5460
tatgactgaa ttctcaaatt ataaacaatg ctcattgctc gtaattaaca tatcaaggtc 5520
agatgccaag aaataagatt aggataccag cgttactctc tctgttctaa aatagattat 5580
tcattctaag atgactttat caacgattca atgttgcacg attttcacta cacaactttg 5640
attgttatca tttggcaagg ctatgtaaca gaaaaatagt tttcattaaa aaactaatga 5700
tatcattctg tgtgatgaga agtgtttttt tattaatcag tggtcataga tagagaagtt 5760
tagttggaca attcctaaaa ccacagttta ttttcaggta tatgacggcg tgggtaacca 5820
catcacacgg gatgaggaca tgatcaatca gtcgaagcta atgttaaaca aatgcggcgg 5880
actacccaag gtgatagttg caattggttg tttcttggcc aagagtctga attgggaggc 5940
catgaatact aactttattc accagttgga aaacaacccg gagctggcta gtgtacatag 6000
catgtttagt tggctagact cctacttcca caattgtcct gacgaactca agccatgcat 6060
cttctatttg tcaatcttcc ctcgacgcca tggcattcgt cgaaggcgtc tggtgaggcg 6120
gtggattgcc gagggctact caagagacac cgacgggaat ctcgcggaag tgaacgggga 6180
aaattacttc tccaggctcg tcaacctcag catgctcaca gatgctgagc gtgatgtcaa 6240
cgtggctggg acagcgactg gggctggtac tgggaggaga atggccatgt gcaacgtcaa 6300
cgatttcttc cgggagtaca ttgtgtcgcg gcgaatggag gagaaccatg tgtttgcact 6360
ggaggggagg tgcagccaga ccacgcggcg cactggacgg cacctggtca tcgacaaaag 6420
ctgggacggg gacgagagcg tgttcaacag gatcgagttc tcgcgactgc ggtcggtgac 6480
ggtgttcgga ccctggaagc cattcctggc ctccgacaag atgagggtgc tccgcgtgct 6540
tgatctggaa ggcactgagg gtctgacaga caacgacatc aagaacatcg tcaaacggct 6600
gcctcgacta aagttcctct ccctgcgagg atgcaagcac atctttcggc tgccaaaatc 6660
cctgggtcgc ctgaggcagc tccagaccct ggatgtgagg cacactgccg tagccgggct 6720
gccagcgacc gtcgtgaagc tacagaagct gcagtatgtt cgtgccggca acacaccaga 6780
ggagtatgcc gtcagcgaca gcacggaatc tagcacctca gtacccacct gtttctcatg 6840
tggccctgtt ggcaatcaca ttgtcggcgt tgaggtgcct cgcggggtcg agaatctgac 6900
caacctgcac acactgggcg tcatcaaagc cactgtggca ggactcaagg agctcaagaa 6960
gcttacccag ctgcgcaagc tcggcgtgtc tggcataaac cggaggaacc acaaagagct 7020
ctgcgccgtc atctcgggcc acggccatct ggaatctctg tcgatatggt ttgacaagga 7080
caccgagagg attgctgacc cttgttcgcc gagcaatgat gacttcaagc ctccagagaa 7140
gctacggagg ctcaaactcc atgggcacac tgacaacctg ccagcatggg ttagcactct 7200
caaaatattt gtcactcaag atcaatgacc tagctcttct gcgtcgacga agaagtctca 7260
ctatgctgca ttaggtatta agttctcttt tgttggcttt ctagcgactg cctatatgtg 7320
ttcctaaact tgaatgtatt tggtgttagt aggtttttgc cctgtagagt gcttcgacat 7380
caattgcact cacagcggtt tcctgccagt gcctcaactc agagctctct tttcgatctt 7440
cttttgctgc tttgtaagaa ctgattgttt ttgttggttg aaagaaaaaa tgggtagaag 7500
tgttaagtag tctacatggt tgctggtcaa gatccatgat ctagttcatc tgcataaatt 7560
aagcggttcc tatatggaaa cctgttctga aggtttgcat gtttgctgga gaaagaaaaa 7620
ctagcttgcg ggagaaagtc gatcatcaga aaactgaaga attctgttct gaaggtcaat 7680
atctttttga gttaaaaatc ctgttatttt ttgccgaagc tcgagatcag ccgcatcttc 7740
gtttcttctc ttgctagttc atccatgatc tcttgttata ttaaaaataa aaatcaaaac 7800
cattaacttt gttgaatgtt tctccagccc cgcgtaaact atatacacat tgcatgaact 7860
atacacaaaa aactgcccct tgaaagttac aactctaaaa catcaaggca aaacagagca 7920
ccgagcacga agagcagaaa acagagtaac cgatgttcca gcagcctgca agtgacgcca 7980
gttgacgcct gctgcaagag atcaattgga agaaactttc gccattgcag ctgccacgca 8040
aatgtcttca gtaacttgag cattactttc ttctgaaaag tcccctcaat ggcgtctgtt 8100
tctccttttc tgacaaaata aaggttacaa aaccagtgag atcatcaaca aatgtctatt 8160
agcttcacca gatcggaact gcggttcact tttgagtatc cattgtggct gcattaccca 8220
tcgacttaaa agtactccct ccgttccaaa atagatgact caactttgta caaagttagt 8280
ataaagttgg gtcatctatt ttggaacgga gggaatagat gggaacactt ttttttccgg 8340
aaacagtttt ttttcttcat ttgagcatac tgattttttt tttcaatttt tgttgatagt 8400
ataagaagga attaacactt caagttgtgg gatcggagta ccttgtatgt ggtcatctgc 8460
aggggtggca ttagcacgga gtggaggttc ggcaactcat ggtctttgac ctgggactgg 8520
tttgctccct ttgtcaagcc atcgtatgat ctccccggca gtaggtctgt ctccaggcat 8580
tgactttaca caatgtaggc cgatatcaat gcacattttt acttgcgctt ggaatttatc 8640
ctccagtgag ccatacaacg ttgctatttg tgaatcttgc ctccaattgt tctcgaccta 8700
catgatacat cagagccaag atttaaagac aaaataatgc cacctattat ataatcctaa 8760
gtcctaagag tatacaactt atagattcac aaaaaactta atgaatactc cctccattcc 8820
acaatgtagt gcctatagat ttttatgaaa gtcaaacttt gtcaactttg accaagtgta 8880
tagagaaaac tgtctacatc tacaatacca aatatgtaca ttctgaaaat ataacttatg 8940
atgtattcaa tgatgtgctt ttggtattct agatgtattt tcttttcttt ataaacatgg 9000
tcaaagtttg taatgtttga cttttgcaaa aaatctatag gcactacatt atggaacgga 9060
gggagtatat tgtatctcta aaagagatga tagtaatact agccatcgga tggcgtgaga 9120
tggatggtaa tattctagta tatctcacct ttcaagaggt aacagtaaca ctagccattg 9180
gatgacgcga aatggacggt tcgtattaat tgggaccatc ttacaacttc tcatctcttt 9240
actatgctca atatacttca ttaattatcc tacgcccaac gggagtaaaa ttataaaaaa 9300
tcatgtgaat ccttattttt ttgcaatcta ctgtagtact tacatgtaca tatatgcagc 9360
acaaactatg tgaaacatat agactcaaat gaagaataca tattagcttt tttatgtaca 9420
cacgtgtata agtgatgagg taatgaaaat acgtacacac agaggacaaa acattttctt 9480
ttttatgtac acacgtagat aaaagaggaa ttttccttgt tattttccct taatcaacaa 9540
aagcattgca tgatcggaag attgaataag ttttctcagt cttaataagt atgtcatggt 9600
tgcccttgat ctcttgatgt gttgcaatct ctagaatcat aagacccaag ctgtatatgt 9660
ccaccgtagc tagatatttg aggtcagtac ccttggagtg gaag 9704
<210> 5
<211> 41
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 5
aattcgagct cggtacccgg gttgttggcc aatcgttatc a 41
<210> 6
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 6
agacccctag tttcctgatg ata 23
<210> 7
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 7
tatcatcagg aaactagggg tct 23
<210> 8
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 8
gcatatattc atagcagagc agttg 25
<210> 9
<211> 25
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 9
caactgctct gctatgaata tatgc 25
<210> 10
<211> 23
<212> DNA
<213> 人工序列(artificial sequence)
<400> 10
ctgtgatgcg tggtatgtag ttg 23
<210> 11
<211> 23
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 11
caactacata ccacgcatca cag 23
<210> 12
<211> 42
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 12
tgcaggtcga ctctagagga tcgtccttca ttgatgtgtt cc 42
<210> 13
<211> 46
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 13
cgaattcgag ctcggtaccc ggggcaattg ttgctgtaat gtggta 46
<210> 14
<211> 46
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 14
aaacgacggc cagtgccaag ctcttccact ccaagggtac tgacct 46
<210> 15
<211> 41
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 15
tctagaggat ccccgggtac catgggcgga tacgagttcc a 41
<210> 16
<211> 43
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 16
ttcgagctct ctagaactag tttacttctt ccatccgaag tgt 43
Claims (8)
1.一种蛋白质在调控小麦对白粉病抗性中的应用，所述蛋白质的氨基酸序列如序列表中序列3所示。
2.一种蛋白质在增强或降低小麦对白粉病抗性中的应用，所述蛋白质的氨基酸序列如序列表中序列3所示。
3.编码权利要求1中所述蛋白质的核酸分子在培育对白粉病的抗性改变的小麦中的应用。
4.编码权利要求1中所述蛋白质的核酸分子在培育对白粉病的抗性增加或降低的转基因小麦中的应用。
5.根据权利要求3或4所述的应用，其特征在于：所述核酸分子为如下(B1)或(B2)：
(B1)序列表中序列1所示的DNA分子；
(B2)序列表中序列2所示的DNA分子。
6.一种制备转基因小麦的方法，其特征在于，包括如下步骤：在出发小麦中导入权利要求4或5中 所述核酸分子，得到对白粉病的抗性增强的转基因小麦。
7.一种制备转基因小麦的方法，其特征在于，包括如下步骤：抑制出发小麦中权利要求4或5中 所述核酸分子的表达，得到对白粉病的抗性降低的转基因小麦。
8.一种小麦育种方法，其特征在于，包括如下步骤：增加目的小麦中权利要求1中所述蛋白质的含量，使得目的小麦对白粉病的抗性增强；或，降低目的小麦中权利要求1所述蛋白质的含量，使得目的小麦对白粉病的抗性降低。
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