CN109734788B - 飞蝗粘蛋白Mucin17、其编码基因及其应用 - Google Patents

Abstract

本发明公开了一种飞蝗粘蛋白Mucin17、其编码基因及其应用。本发明首先从飞蝗中克隆了飞蝗粘蛋白基因Mucin17，并对飞蝗粘蛋白基因Mucin17设计引物，合成了用于干扰飞蝗粘蛋白基因Mucin17的dsRNA，且运用注射法将dsRNA导入飞蝗对飞蝗粘蛋白基因Mucin17进行RNAi。结果表明：有效沉默飞蝗粘蛋白基因Mucin17后可导致飞蝗死亡以及影响飞蝗翅和后足的发育。本发明为更深入理解粘蛋白在飞蝗发育中的作用机制，同时为基于RNA干扰的害虫防治提供新的靶标，具有理论意义和广阔的应用前景。

Description

飞蝗粘蛋白Mucin17、其编码基因及其应用

技术领域

本发明涉及生物技术领域，具体涉及飞蝗粘蛋白Mucin17、其编码基因及其应用。

背景技术

飞蝗是一种洲际性的农业害虫，主要分布于亚、欧、非、澳四大洲，我国大部分地区都有分布。其主要威胁玉米、谷子等禾本科农作物，在饥饿情况下，还取食大豆、白菜及向日葵等。飞蝗具有跳跃性以及功能强大的前后翅，具有远距离、大规模迁移的生理功能，使其活动空间从平面向立体扩展，行动半径大大提高，从而在觅食、求偶、避敌和扩大分布效率等方面都得到大幅度提高。因此，蝗灾一旦爆发，将会对我国的粮食安全和农业增收造成非常严重的影响。目前对其防治主要依靠化学防治，然而长期施用化学杀虫剂导致一系列问题，如环境污染，抗药性产生和对非靶标生物的危害等。因此，开发新型、可替代化学防治的方法变得极其紧迫。

粘蛋白(Mucin)是一种相对分子质量较高的糖蛋白家族，是人类等哺乳动物上皮细胞产生的粘液中的主要成分，广泛地分布于消化道，对消化道粘膜起保护和润滑作用。目前已发现21种粘蛋白家族成员，可分为膜结合型和分泌型两个亚家族。正常生理状况下Mucin起着润滑、保护、屏障等作用，参与细胞黏附、信号转导、细胞-细胞间的相互作用，异常表达的 Mucin参与调控肿瘤细胞的分化、增殖、浸润和转移以及化疗药物抵抗等过程。在昆虫中，肠道粘蛋白(Intestinal mucin)主要构成围食膜的成分，如烟草天蛾、棉铃虫等。在脊椎动物和无脊椎动物中，Mucin蛋白是一类高度O连接糖基化蛋白，在组织完整性、分泌及分泌小泡形成、生物发育等过程具有重要作用，可作为害虫防治的潜在靶标。

RNA干扰(RNAi)是一种由双链RNA分子引起的特异性转录后基因沉默的现象，自2006 年获得诺贝尔奖以来，RNAi技术一直跻身于科技前沿。RNAi不仅是研究基因功能的有力工具，同时在害虫防治方面也具有极大潜力。通过RNA干扰进行害虫防治具有如下特点：1)杀虫具有专一性，对非靶标生物无杀伤作用；2)RNA在自然界极易降解，无残留；3)对环境无毒无害，相对安全。因此，国内外学者将其称为第四代杀虫剂。基于RNA干扰进行害虫防治的基础技术是筛选靶标序列获得具有高致死作用的dsRNA。

发明内容

针对飞蝗迁飞性和跳跃性的特点，本发明基于RNA干扰技术，选取粘蛋白基因为对象，利用其重要生物学功能来进行害虫防治，安全高效，是一种环境友好型、对人类无毒无害、特异性的新型分子靶标，在害虫防治领域具有广阔的应用前景。

本发明提供的粘蛋白来源于飞蝗，其名称为Mucin17。所述Mucin17的基因序列如SEQ ID NO：1所示。该基因序列是基于飞蝗整虫转录组数据库，搜索获得的片段通过BioEdit软件结合飞蝗基因组序列对其进行拼接，然后经PCR扩增、克隆测序获得。

Mucin17基因编码的氨基酸序列是如SEQ ID NO：2所示的蛋白质。

本发明提供的一种飞蝗粘蛋白基因dsRNA，其核苷酸序列是SEQ ID NO：3，是根据SEQ ID NO：1通过primer premier 5.0软件设计含有T7启动子的上游引物SEQ ID NO：4和下游引物SEQ ID NO：5，通过PCR扩增获得SEQ ID NO：3，其产物含有T7启动子。产物经纯化后按照T7RiboMAX^TMExpress RNAi System(Promega)试剂盒说明体外转录合成dsRNA。

SEQ ID NO：3合成的dsRNA在飞蝗防治中的应用：通过微量进样器将SEQ ID NO：3合成的dsRNA注射进入飞蝗体腔内。结果表明：和对照相比，SEQ ID NO：3合成的dsRNA可以导致飞蝗蜕皮前及蜕皮中致死(致死率分别为33.3％和8.3％)、翅皱缩不能展开和后足发育畸形(表型率为50％)，最终导致死亡。

附图说明

图1：飞蝗粘蛋白基因Mucin17序列获得。A：Mucin17引物设计与拼接示意图；B：部分测序峰图；C：Mucin17基因翻译后氨基酸序列与结构示意图。

图2：飞蝗注射SEQ ID NO：3合成的dsRNA 24小时后，飞蝗粘蛋白基因Mucin17的mRNA沉默情况，β-actin做为内参基因，其中**，P<0.01。

图3：5龄飞蝗注射SEQ ID NO：3合成的dsRNA，飞蝗蜕皮前及蜕皮中致死(致死率分别为 33.3％和8.3％)、翅皱缩不能展开和后足发育畸形(表型率为50％)，最终导致死亡。A：注射 dsGFP的对照；B-D：注射SEQ ID NO：3合成的dsRNA，即dsMucin17；E-F：对照和异常翅的表型图；G-H：对照和异常足的表型图。

具体实施方式

实施例1：飞蝗粘蛋白基因序列获得

1、飞蝗粘蛋白基因序列获得

基于本实验室获得的飞蝗整虫转录组数据库(GenBank登录号:GEZB00000000)，采用生物信息学方法对飞蝗粘蛋白基因(Mucin)进行搜索，获得飞蝗粘蛋白基因Mucin17部分序列。利用NCBI网站对序列进行Blast分析，同时结合飞蝗基因组序列(GenBank登录号:AVCP000000000)获得其编码序列，利用BioEdit软件进行拼接及比对后，获得飞蝗粘蛋白基因Mucin17全长开放阅读框序列。根据获得的飞蝗粘蛋白基因Mucin17全长开放阅读框序列，采用primer premier 5.0软件设计4对上、下游引物序列且每队引物之间与共同区域衔接，引物设计位置、长度及序列如图1所示(图1中的A)，引物送往生工生物工程(上海)有限公司合成。

选取生长健康，大小一致的飞蝗成虫(本实验室养虫室室内饲养)，解剖翅组织，并冷冻于液氮中。每3头一个生物学重复，设置3个生物学重复，依照Trizol试剂盒(TaKaRa公司) 提取翅组织总RNA。采用M-MLV反转录酶(TaKaRa公司)将所提总RNA反转录成第一链cDNA，并以此作为模板，结合设计的上下游引物(图1中A)，通过PCR扩增仪分别扩增获得Mucin17 基因片段，将得到的产物经Gel Extraction Kit(Sigma)试剂盒进行纯化、连接pEASY-T3 载体(北京全式金生物技术有限公司)并转化到大肠杆菌Trans T1感受态(北京全式金生物技术有限公司)中，挑取阳性克隆送往上海英潍捷基生物有限公司进行测序、拼接(测序峰图如图1中的B所示)，获得其全长开放阅读框序列，其序列为SEQ ID NO：1。利用ExPASy-Translate tool(https://web.expasy.org/translate/)将飞蝗粘蛋白基因Mucin17 序列翻译成氨基酸序列，其序列为SEQ ID NO：2。该基因编码2650个氨基酸，SMART网站 (http://smart.embl-heidelberg.de/)预测其含有两个表皮生长因子结构域(Epidermal growth factor domain，EGF)、一个钙结合类表皮生长因子结构域(Calcium-binding EGF-like domain，EGF_CA)、一个精子蛋白，肠激酶和糖蛋白结构域(Spermprotein,Enterokinase and Agrin domain，SEA)和一个跨膜结构域(Transmembranedomain，TM)(图1中的C)。

2、飞蝗粘蛋白基因特异性dsRNA合成

1)飞蝗粘蛋白基因dsRNA引物的设计

基于飞蝗粘蛋白基因Mucin17开放阅读框序列(SEQ ID NO：1)，采用primerpremier 5.0 软件设计dsRNA引物，其上下游引物序列分别为SEQ ID NO：4和SEQ ID NO：5。上下游引物均携带T7启动子序列。所有引物均由生工生物工程(上海)有限公司合成。

2)飞蝗粘蛋白基因dsRNA的合成

以上述获得的cDNA为模板，SEQ ID NO：4和SEQ ID NO：5作为上下游引物，进行PCR扩增。将扩增的PCR产物，经Gel Extraction Kit(sigma)试剂盒纯化后按照T7RiboMAX^TMExpress RNAi System(Promega)试剂盒说明体外转录合成dsRNA。使用NaNoDrop 2000(Thermo scientific)进行定量，使其终浓度达到2μg/μl。保存于-20℃低温冰箱备用。

实施例2：飞蝗粘蛋白基因dsRNA致死飞蝗实验

1、飞蝗粘蛋白基因dsRNA注射

选取生长健康、大小一致的30头5龄第2天若虫进行实验。将5μl(10μg)SEQ ID NO：3的dsRNA用25μl规格微量注射器顺着血流方法轻轻注射进入若虫侧腹部的二、三腹节之间。同时选取30头若虫设立为对照组。注射相同体积和浓度的dsGFP至对照组体内。分别将注射后飞蝗置于30℃恒温生化培养箱中饲养(光照：黑暗时间＝14h:10h，温度30±2℃，湿度60％)，每天饲喂新鲜小麦幼苗和麦麸。

2、飞蝗粘蛋白基因沉默检测

收集注射dsGFP和dsRNA 24h后的若虫各9头，解剖翅芽组织进行总RNA提取，并反转录成第一链cDNA，每组设置3个生物学重复，每个生物学重复3头若虫，采用Real-timePCR 方法分别检测目的基因(Mucin17)和管家基因(β-actin)的相对表达量，计算其沉默效率。结果表明，与对照组比较，注射dsRNA后，处理组飞蝗粘蛋白基因表达量极显著降低(图2)。

3、注射dsRNA后表型观察

5龄若虫经注射dsRNA后，注射dsGFP对照组6天后开始蜕皮并全部成功蜕至成虫，蜕至成虫后虫体形态活力正常，并于半天后开始正常进食。注射飞蝗粘蛋白基因dsRNA的处理组若虫共30头，与对照组若虫相比，飞蝗蜕皮前及蜕皮中致死(致死率分别为33.3％和8.3％)、翅皱缩不能展开和后足发育畸形(表型率为50％)，最终导致死亡(图3)。

序列表

<110> 中国农业科学院植物保护研究所

<120> 飞蝗粘蛋白Mucin17、其编码基因及其应用

<141> 2019-02-25

<160> 5

<170> SIPOSequenceListing 1.0

<210> 1

<211> 8290

<212> DNA

<213> 未知(Unknown)

<400> 1

gcacagctgg aggcgctgcg cagctccctg cgggcgacgg ccaccgcgga cgcgggggca 60

gacgcgcagg gctccggcgt ggtcgccacg gcgactctgc cgggcgggca gcaggtgcag 120

gtgatggcga tggaccaggc gcccgccgtg cagcccacca agacggcgga gccgccggcc 180

gtcgctccca ccgtcggtcc ggacgcgtcg cagatcgcgc cctcgatgga tacactgcag 240

ccggctaatg tggagaacat cgttacgggg tcaaccatca tcttcttcga cgaggaagac 300

ggtgtcgaca gtgactcggg gcccacggcg gccgccactc cggcactgga ctcctcgaca 360

gtcgccgccc tcagtgattc gcgcaccacc gtcgtgaatg ccggggcggc caccgtcatc 420

ttcccagacc cagagccgac cactgccgag ccggctgact ctcaacagga gagcaccaca 480

gtcagcacgg atgacgacga gtacggtacc ggcgacgaag aaaacggcgg aatcagccag 540

acaccgacga gtgacgcggg cccagcagca tcgacgtccg tggtgaccgt caccgcatca 600

ccagtcacca ccacggtcgc cgtacccgta accaagaagc ctgtggtagg acccggaggc 660

agcgagatac aggtcagcga cttgttgagc ttgggctcac ttggcatcaa cggtctcaat 720

gcattgggtc cagtcttcaa tgcgatggcc ggtctcattc aaggcaacct acagcagcaa 780

ccggatgcgg ccaagaggcg gaacgactct gtcaacaccg gaaacagagc cccggagctg 840

gtgcctggtg accgcgagga ctacccgcac cccgtgtacc ggccgcagcc gcaggcggac 900

accgagccac cgccgaggtc gcccatctac atcccggtgg gcggactcgc cgcagccgat 960

cacgacatcg aagctgcaga gagccagaac tacgaggcgc gcattaaaat caatgccctg 1020

ccagggtccg gcaaatgggg cgggagtgca gagtcggtgg gcggcgtggt gatcggccga 1080

ccgactctgg agaaaccgct cggcgacggc atccccatca gcccaggcga ggtcatcacc 1140

gccaacagcg acgtcatcgt gggcaagccc gccgtcgcgg gcccccgccc tcccaaaatc 1200

actgccgccg gacacgagca cgaccacgcc ctcgaccacg gcggcatcgc cgtgggcatg 1260

cggccgccgc cgccacaccg gaagccgtgg ctgcaccacc accagcaacc acccatgagg 1320

gacgagttcc tgggaccacc gccaccacca cctcctccgg gcagccacgt gggcggtgag 1380

gtccagggcc ggcctccggg ctccgccaac cactaccgcc tccaccacga taggccgcac 1440

cccggcctca atctcaacca caaccacaac cagaacaact tcaacaacaa ccagaagccg 1500

ccagcactca agctgcgtcc tcacgtggta gaaggtcttc cctacaagct ggagtccgtg 1560

gtgacgggct cgcaggcggt gctggtggcg gagggcggtg ccggtgaggt ggtgcacggg 1620

tccccggtgc cgcaccggca cgcggtgctg ccagcggggg cggacgcggc ctctctgctg 1680

gtcgacatcc ggccctcgca ggtggccaac gtgctcatcc ccaggggcag ccacaccgcc 1740

cacgtcttca gcgcacccgc gccctcgccg ctcgtcgccg acgacgtctt ccgcgacgac 1800

tcgcccttcc ccgttccgga ggcggctgcg gacttcgtgg gtctggacgc ggcgccgcag 1860

ctggagcacg gcgtgggcac cgtcatcgag gttcaccccg gggactcccc cacagagacc 1920

ggcggcgtcg tggccgggct gcagggcggc gacataaacg tcaacatcgc tccgggacag 1980

ggcatcagcg tggacgtggg ccccggcggc tcgatggtgc acgccaggcc ggactccgtc 2040

cccgggcccg gcaaggaggt gcaccgcatc cagccgagtg gcgtcgtgct cgacgctccg 2100

aggccgcagc tccagccgca gccacaccgc ccggagcagc ccctcgccca cccaccgctt 2160

ccccaccgcc cggagtcgcc cctcgcccac ccgccgccgc cacctccgcc ccctccgcct 2220

ccgtcgtcgt cgcagcagtc tccgtcgagg aggccgcccg tcacactgta ccgcccctcc 2280

acaggaactg gaggttccac ggcgggaggg aggccgcacc agtacaggtt tcctgtaccg 2340

gggcgtcctc tgcgaccgtc ggacttcatg acgccaccgc cgccgcctcc gcacctccca 2400

ctacgaccgc cggcgccgaa accgactccc ggagtgcacc agcacctccg cccccagcag 2460

ccaggtgggc cacttgtcat cccgcttaga cccgacgcgg ggcggccccc gcggcccggg 2520

aacgaccact tcgtccatgg ccacgaccac gagcacgacc tccacatcca cggtgacgac 2580

cacgaattgc acgtccacgg gctcgaggca gagcgcgacc acacgacgag gttgcacccg 2640

gtgaggatcg gcggcggcgg agtcgtcgtc aggcccggcc ccgggcctgt tgatgtgttt 2700

gtccggcccg acggagagct cgtctccgtg gaggacgacc tgggtgtcgc cggttcctcc 2760

agacctctac cgcgcccacc gtcactctcg ccgaagcctt cgcgtagacc acctgtaacc 2820

gacctctttg caatctggcc gggtgacagg tacaatggca gcaggccaca gcacggtgcc 2880

gggcctccgc ccccgaggca gggggtcggg tctcgtcccg gagaccgaga gcgggagcag 2940

gagcggccgc cccagcagcc gaccctgcag cacggcttcc acgaggtgcg gccggtggcc 3000

gagccgggag acgtgcccac agttccaggt tttgtgaatg gccagccaaa accaccacca 3060

gacttccaca gccagtctca ccggcagcgt ccacccccca tcacggaggc cgacaacaac 3120

atcatcgtcg gacacgtgcc gcacttcaac gagttacaca gtccggttcc tccggacgac 3180

ttgcggccgc acgtagttgc cgtgggcgag cctacgaaca aggtggacga gcccccggga 3240

aatgtcgatc accatcagct ccatcaccat cacgtggctg gcgaggtctc cagcccggag 3300

atcggagtcg tccgtccggg tgaacgtcct ctgcagagtg gcatacgttt ccctggtgct 3360

ggtgatcatt ttgaagttga agaggacaca cagtcagttg agaaaagtga tccagacagc 3420

actggagcag catctggtac tgacggaagc ccagacccac cagcacatgg ctcagacagt 3480

gcagacagtg acgaagacag attggggtcc attttgcacg gtgggactca aactgaacgg 3540

cctctcgtga cactgcctcc ggacaacagc cccggtcacg accacgtcca caatacgaac 3600

actggcactg cgacacgtga gccggaacac agggacgaga cgccgtctcg aaacgagata 3660

cctattctcg gagtcgggaa agagaggcct ccacatctgc acgtaggagt ctccggctgg 3720

cagatcatcg gttctcagcc ggaccgtcct ggcactccac aagacggggc cagtaacggt 3780

ccagacgtcg ggagtgcaac tcccggccgg ggtcccccgc accgggtagc cggatatcgc 3840

cctccgggtt ggcacaccgg aggatcgggc cacctgatag tgactgtcgg cgacgagagt 3900

ggaccggtcc gcgaccgccc gtctggagac gactctgacg tccactacgg gcggccgcac 3960

gtctctcccg gcgggacgcc ggaggacgcg aggccccact ggtccggcga ggccccgaga 4020

ccgccgatcg agactcccgt cgtagatcgg gagacgtcgg gctacggcgt ggcagatttc 4080

gggcaggacg tgcctccgtt caaggccatg ggcgaggacg tttccgccgt ggtggactcg 4140

gtgctgaggc ctccgtcgct gaacctgccg ccggcaactg cggtacgtcc cacgcagcca 4200

cctccacctc cgccgccgcc gcccccgagg gtgacagtgc tcacggacca ccggcacaac 4260

aacaagccgc accggcagcc ggtcatcgtg ctgggacggc cggccgagtt tacgggggat 4320

gtcgcgaccc accggccgac gcagaggccc agcatgacag aggagggcag accgggagac 4380

ggggagccgc cggcagacat gacggaggtg gacaagccgc ccccgcctcc accacgcccc 4440

gccgtcgcag acctggagac ggcggcgagc acggcacgcc ccttcaagct gcggccacag 4500

gacgactcgg tgatggggct ggctccgccg ccgccccgtg tgacggtggt gggtgtcggc 4560

aatggaggcg gaccgccccc gctgcaagga gattccaccc accggccggt cctcatcgga 4620

ggtggcagga agccaccagt gcagacactg ccacccccac cgtccgagtt cgagcagccg 4680

cccccacccc tggtgaccac ccccaggcgc ccgcccttcg tagtgacgcc tcccaccccc 4740

gcatacagct ctgcccccac aacaaaaccg aagccctcga cttccacctc cgtcgatgac 4800

accaaagacg aactggaaga tgaggatcag gagctggcgg aaacatcctc cggcagcaaa 4860

ccgcccgcgc ccccgctgcc gcctctgtcg ccctctgtaa cgtcggcagc cgctcccgag 4920

ctggtagacg tgacgtaccg gccgcccgtc tacaccacgc agcggcctcg gccggtcacg 4980

cctctggatg ccaccaggaa gccgccgcag gtggtttcgc ccagccgcac ggaggcggac 5040

atgctggaga cggtgagtgt gggtcggccc gtgtcgcagc cactgccgcc cccaccgccg 5100

cctccgccac cgactcgtga ggcgccacca cgtcctcctt ccgtcgccct ggcacccaca 5160

cctgtcaggg atgccgcctc caccgaggtg aaggcgtctc cgcccccgcc acctgcggcc 5220

gccgcgacac cgcccgcaac actggaccgt acgtggacga cgttggcggt ggacctggag 5280

gggtcgcaga ctcgactgcc gaaccgacgc cggcccaccg cgacaccggc cgtcgctccc 5340

acctctgcga cgtcttcgga cacctcctcg aacaccggca aagtgacatc tgtggtggtg 5400

acatcgggtt ccaagacggt gctgttcgtg aagcctacgg ccgcccgact gccgacgcag 5460

acgacgaagt ctactcctac gccgcgaccg aggccgagac cccgtcccac caccgagacc 5520

ggaggactgt ttgacgactt cgacgacgtc gcggacgagg acgacgagga ggaatcggag 5580

acgggcacgg ccggcgggtc gcggggcaag gtggacctgg gcgtcgctag ctccaaccag 5640

gagaaggtgg tggtcggctc ggcgcccgcc tccaccctct acgtcaccca cacgcacaca 5700

ctcacagtaa cgactacgga gacgcaggtg gtgagcacag ccggccggcc gccggtgacg 5760

cgcactgtgg tgctcaccaa gacgcagacg tcaacagtcg tggacacagt caccgagacc 5820

cagacgctgc tccggccgac cagcgtctat gccaccgtca ccaccacggt gagtgccatt 5880

cggcccttca cagagggggt gtctagcacc agcaccgtct ccacggtgac gacacctgcg 5940

cccaccaccg gctcgcagcc tccgccgccc ccgccacctg cggggacggg tgaccgaccc 6000

ggagaccgca cggctggcgc gggagacacc ttctttgtgg tggtgaccga cccgcaccac 6060

aagttgccgc ctcccagcgg agcacccctg ccacccggca tcatcgagta cgaggtggac 6120

aaggaggacg agcaggaacc gcggcaggac ggcggcggcg caggtggcaa cgtgctgctg 6180

acggcgggcg tgggcggcag cggtgcggcg gcgacctcgg cctgccggcc cgagtgcagc 6240

gcggcgcgca acgaggtgtg ccagcgcgcc gcggacggcc acccacgttg cctctgcagg 6300

ccgggcttcg ccaggatctt cccagaccgg ccctgcaagc cgacatacac atatacgcta 6360

cggctagtgc tgtcgcgcca tggcgaccgc cccatcacct tctcgcccgg cctgggagat 6420

acaggcagtg acaccttcgc tgagacgtct cgcgctgccg ccgaggggct cgaccggctg 6480

gtgatgcagt ccgacctgcg cgacgtcttc cacggtgtct ctgttgaggc gttcctgccg 6540

cccgacgaag gcagcaccgc cggcgttatc gtcaccttcc aagtcaggct gtcagacaat 6600

accgacgagg agcgactgaa ggaggtattc cgaaagtcac tcagggctac caactacagc 6660

ctgggtggca cggatgtctt tgccgacaag gagcgcatcc agcacattga ggcagaagac 6720

ttcgacgagt gcgcgagccg gcagtaccac gactgttcgg agaacgcgca ctgcttcaac 6780

ctgcgcggca cgtacacgtg cagctgccgc gagggcttcg ccgacctctc cgagaacccg 6840

ctgttcccgg gcaggctctg ctcagcggag ctgatcgggt gtgagcggtg ccacttccac 6900

ggcacgtgcg gcgacgacca ccaccaggtg cagtgcgagt gcttccagtg gtacgccggg 6960

gacacctgcc acatcaacct caaggcgctg ctgatcgcac tggtgacgat cggctgcgta 7020

ctggtggcgc ttctgctggt gtgcgtgctg atgacgtgcg cgcgtggagg cagtcgcggt 7080

gggcgcaccc ggggacggtc gcggcgcgag gggccgttgg agcggcgcgc catgatccac 7140

gactcggcca gcgaggccag cggcgaccag acgctgccca ggagccagac tccggcagcg 7200

gtgttcggca agaaggcgcg accggtgtcg gcgccgacgg cgacggtggt agccgtggtt 7260

cccccgcctc cagcgggctt cgcccccacg cccgtaccgg tgccccctcc actgcagccg 7320

cccccggagc agcgcgaccg ctcgctgacg gtgatgatcc cgcgcgccaa gtaccggccg 7380

tcgcccgtgg cgcccccgca ccagccactg cccacgctgg tctccatgtc gacgttcggc 7440

gatgccgaga aacgcgccat cgctcacgag agcaagctgc tcgccctgct ggagggagcc 7500

tccaagcagg atggccagct acagtcccat catcatcatc atcaacacca gcagcagaca 7560

tcaaaacgaa aacctagcaa cacaacgagc caagcaacag aggcgtccac tcccaggaag 7620

aattcatctt cgaggaaacc ttcgaatgga aacattccac caccagcacc gggagccctg 7680

gtctcggcgg gcttcgaggt gtcggcgact gtcggcaagc ctgcgacgga ctccgacacg 7740

gaggcatcgg cagtcgtcca caacggttcc acgttgcgga cgacagacag tgtacaaatc 7800

accgttgaag agatgtcgga gccatcggta aaggctgggt ctgctgtgga aacgctgacc 7860

gtgtccgaag ctcgatcttg tgacgagaca acgatacatc caccgacaaa gtcgctccac 7920

agcaattaca gctcaaaaca ctcatcaaac aacctcaaca atgatgaggg ccacacaatg 7980

gctgagagag acatcggctc tacgttcgtg atgccacagt cgcacttgta cacaccagac 8040

aggggaagtg acatttcaaa tttcgactca ctgtgagtaa agccttcagt gcaactgcag 8100

aagtttgcta catcacttgc actgcgaggc tttcgggagt ggagatgcga cacttctgaa 8160

cggagctatc atgtgtcgac ttgaaacgaa ggaaattgtg aaaagaatgt gactgtaaca 8220

ggctcaattt caaagacata ttgagtgagt tattgtattt aatataatgg aggactgaat 8280

actgttaact 8290

<210> 2

<211> 2650

<212> PRT

<213> 未知(Unknown)

<400> 2

Met Ala Met Asp Gln Ala Pro Ala Val Gln Pro Thr Lys Thr Ala Glu

1 5 10 15

Pro Pro Ala Val Ala Pro Thr Val Gly Pro Asp Ala Ser Gln Ile Ala

20 25 30

Pro Ser Met Asp Thr Leu Gln Pro Ala Asn Val Glu Asn Ile Val Thr

35 40 45

Gly Ser Thr Ile Ile Phe Phe Asp Glu Glu Asp Gly Val Asp Ser Asp

50 55 60

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

65 70 75 80

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

85 90 95

Thr Val Ile Phe Pro Asp Pro Glu Pro Thr Thr Ala Glu Pro Ala Asp

100 105 110

Ser Gln Gln Glu Ser Thr Thr Val Ser Thr Asp Asp Asp Glu Tyr Gly

115 120 125

Thr Gly Asp Glu Glu Asn Gly Gly Ile Ser Gln Thr Pro Thr Ser Asp

130 135 140

Ala Gly Pro Ala Ala Ser Thr Ser Val Val Thr Val Thr Ala Ser Pro

145 150 155 160

Val Thr Thr Thr Val Ala Val Pro Val Thr Lys Lys Pro Val Val Gly

165 170 175

Pro Gly Gly Ser Glu Ile Gln Val Ser Asp Leu Leu Ser Leu Gly Ser

180 185 190

Leu Gly Ile Asn Gly Leu Asn Ala Leu Gly Pro Val Phe Asn Ala Met

195 200 205

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

210 215 220

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

225 230 235 240

Pro Gly Asp Arg Glu Asp Tyr Pro His Pro Val Tyr Arg Pro Gln Pro

245 250 255

Gln Ala Asp Thr Glu Pro Pro Pro Arg Ser Pro Ile Tyr Ile Pro Val

260 265 270

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

275 280 285

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

290 295 300

Trp Gly Gly Ser Ala Glu Ser Val Gly Gly Val Val Ile Gly Arg Pro

305 310 315 320

Thr Leu Glu Lys Pro Leu Gly Asp Gly Ile Pro Ile Ser Pro Gly Glu

325 330 335

Val Ile Thr Ala Asn Ser Asp Val Ile Val Gly Lys Pro Ala Val Ala

340 345 350

Gly Pro Arg Pro Pro Lys Ile Thr Ala Ala Gly His Glu His Asp His

355 360 365

Ala Leu Asp His Gly Gly Ile Ala Val Gly Met Arg Pro Pro Pro Pro

370 375 380

His Arg Lys Pro Trp Leu His His His Gln Gln Pro Pro Met Arg Asp

385 390 395 400

Glu Phe Leu Gly Pro Pro Pro Pro Pro Pro Pro Pro Gly Ser His Val

405 410 415

Gly Gly Glu Val Gln Gly Arg Pro Pro Gly Ser Ala Asn His Tyr Arg

420 425 430

Leu His His Asp Arg Pro His Pro Gly Leu Asn Leu Asn His Asn His

435 440 445

Asn Gln Asn Asn Phe Asn Asn Asn Gln Lys Pro Pro Ala Leu Lys Leu

450 455 460

Arg Pro His Val Val Glu Gly Leu Pro Tyr Lys Leu Glu Ser Val Val

465 470 475 480

Thr Gly Ser Gln Ala Val Leu Val Ala Glu Gly Gly Ala Gly Glu Val

485 490 495

Val His Gly Ser Pro Val Pro His Arg His Ala Val Leu Pro Ala Gly

500 505 510

Ala Asp Ala Ala Ser Leu Leu Val Asp Ile Arg Pro Ser Gln Val Ala

515 520 525

Asn Val Leu Ile Pro Arg Gly Ser His Thr Ala His Val Phe Ser Ala

530 535 540

Pro Ala Pro Ser Pro Leu Val Ala Asp Asp Val Phe Arg Asp Asp Ser

545 550 555 560

Pro Phe Pro Val Pro Glu Ala Ala Ala Asp Phe Val Gly Leu Asp Ala

565 570 575

Ala Pro Gln Leu Glu His Gly Val Gly Thr Val Ile Glu Val His Pro

580 585 590

Gly Asp Ser Pro Thr Glu Thr Gly Gly Val Val Ala Gly Leu Gln Gly

595 600 605

Gly Asp Ile Asn Val Asn Ile Ala Pro Gly Gln Gly Ile Ser Val Asp

610 615 620

Val Gly Pro Gly Gly Ser Met Val His Ala Arg Pro Asp Ser Val Pro

625 630 635 640

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

645 650 655

Asp Ala Pro Arg Pro Gln Leu Gln Pro Gln Pro His Arg Pro Glu Gln

660 665 670

Pro Leu Ala His Pro Pro Leu Pro His Arg Pro Glu Ser Pro Leu Ala

675 680 685

His Pro Pro Pro Pro Pro Pro Pro Pro Pro Pro Pro Ser Ser Ser Gln

690 695 700

Gln Ser Pro Ser Arg Arg Pro Pro Val Thr Leu Tyr Arg Pro Ser Thr

705 710 715 720

Gly Thr Gly Gly Ser Thr Ala Gly Gly Arg Pro His Gln Tyr Arg Phe

725 730 735

Pro Val Pro Gly Arg Pro Leu Arg Pro Ser Asp Phe Met Thr Pro Pro

740 745 750

Pro Pro Pro Pro His Leu Pro Leu Arg Pro Pro Ala Pro Lys Pro Thr

755 760 765

Pro Gly Val His Gln His Leu Arg Pro Gln Gln Pro Gly Gly Pro Leu

770 775 780

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

785 790 795 800

Asp His Phe Val His Gly His Asp His Glu His Asp Leu His Ile His

805 810 815

Gly Asp Asp His Glu Leu His Val His Gly Leu Glu Ala Glu Arg Asp

820 825 830

His Thr Thr Arg Leu His Pro Val Arg Ile Gly Gly Gly Gly Val Val

835 840 845

Val Arg Pro Gly Pro Gly Pro Val Asp Val Phe Val Arg Pro Asp Gly

850 855 860

Glu Leu Val Ser Val Glu Asp Asp Leu Gly Val Ala Gly Ser Ser Arg

865 870 875 880

Pro Leu Pro Arg Pro Pro Ser Leu Ser Pro Lys Pro Ser Arg Arg Pro

885 890 895

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

900 905 910

Ser Arg Pro Gln His Gly Ala Gly Pro Pro Pro Pro Arg Gln Gly Val

915 920 925

Gly Ser Arg Pro Gly Asp Arg Glu Arg Glu Gln Glu Arg Pro Pro Gln

930 935 940

Gln Pro Thr Leu Gln His Gly Phe His Glu Val Arg Pro Val Ala Glu

945 950 955 960

Pro Gly Asp Val Pro Thr Val Pro Gly Phe Val Asn Gly Gln Pro Lys

965 970 975

Pro Pro Pro Asp Phe His Ser Gln Ser His Arg Gln Arg Pro Pro Pro

980 985 990

Ile Thr Glu Ala Asp Asn Asn Ile Ile Val Gly His Val Pro His Phe

995 1000 1005

Asn Glu Leu His Ser Pro Val Pro Pro Asp Asp Leu Arg Pro His Val

1010 1015 1020

Val Ala Val Gly Glu Pro Thr Asn Lys Val Asp Glu Pro Pro Gly Asn

1025 1030 1035 1040

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

1045 1050 1055

Ser Pro Glu Ile Gly Val Val Arg Pro Gly Glu Arg Pro Leu Gln Ser

1060 1065 1070

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

1075 1080 1085

Thr Gln Ser Val Glu Lys Ser Asp Pro Asp Ser Thr Gly Ala Ala Ser

1090 1095 1100

Gly Thr Asp Gly Ser Pro Asp Pro Pro Ala His Gly Ser Asp Ser Ala

1105 1110 1115 1120

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

1125 1130 1135

Thr Glu Arg Pro Leu Val Thr Leu Pro Pro Asp Asn Ser Pro Gly His

1140 1145 1150

Asp His Val His Asn Thr Asn Thr Gly Thr Ala Thr Arg Glu Pro Glu

1155 1160 1165

His Arg Asp Glu Thr Pro Ser Arg Asn Glu Ile Pro Ile Leu Gly Val

1170 1175 1180

Gly Lys Glu Arg Pro Pro His Leu His Val Gly Val Ser Gly Trp Gln

1185 1190 1195 1200

Ile Ile Gly Ser Gln Pro Asp Arg Pro Gly Thr Pro Gln Asp Gly Ala

1205 1210 1215

Ser Asn Gly Pro Asp Val Gly Ser Ala Thr Pro Gly Arg Gly Pro Pro

1220 1225 1230

His Arg Val Ala Gly Tyr Arg Pro Pro Gly Trp His Thr Gly Gly Ser

1235 1240 1245

Gly His Leu Ile Val Thr Val Gly Asp Glu Ser Gly Pro Val Arg Asp

1250 1255 1260

Arg Pro Ser Gly Asp Asp Ser Asp Val His Tyr Gly Arg Pro His Val

1265 1270 1275 1280

Ser Pro Gly Gly Thr Pro Glu Asp Ala Arg Pro His Trp Ser Gly Glu

1285 1290 1295

Ala Pro Arg Pro Pro Ile Glu Thr Pro Val Val Asp Arg Glu Thr Ser

1300 1305 1310

Gly Tyr Gly Val Ala Asp Phe Gly Gln Asp Val Pro Pro Phe Lys Ala

1315 1320 1325

Met Gly Glu Asp Val Ser Ala Val Val Asp Ser Val Leu Arg Pro Pro

1330 1335 1340

Ser Leu Asn Leu Pro Pro Ala Thr Ala Val Arg Pro Thr Gln Pro Pro

1345 1350 1355 1360

Pro Pro Pro Pro Pro Pro Pro Pro Arg Val Thr Val Leu Thr Asp His

1365 1370 1375

Arg His Asn Asn Lys Pro His Arg Gln Pro Val Ile Val Leu Gly Arg

1380 1385 1390

Pro Ala Glu Phe Thr Gly Asp Val Ala Thr His Arg Pro Thr Gln Arg

1395 1400 1405

Pro Ser Met Thr Glu Glu Gly Arg Pro Gly Asp Gly Glu Pro Pro Ala

1410 1415 1420

Asp Met Thr Glu Val Asp Lys Pro Pro Pro Pro Pro Pro Arg Pro Ala

1425 1430 1435 1440

Val Ala Asp Leu Glu Thr Ala Ala Ser Thr Ala Arg Pro Phe Lys Leu

1445 1450 1455

Arg Pro Gln Asp Asp Ser Val Met Gly Leu Ala Pro Pro Pro Pro Arg

1460 1465 1470

Val Thr Val Val Gly Val Gly Asn Gly Gly Gly Pro Pro Pro Leu Gln

1475 1480 1485

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

1490 1495 1500

Pro Val Gln Thr Leu Pro Pro Pro Pro Ser Glu Phe Glu Gln Pro Pro

1505 1510 1515 1520

Pro Pro Leu Val Thr Thr Pro Arg Arg Pro Pro Phe Val Val Thr Pro

1525 1530 1535

Pro Thr Pro Ala Tyr Ser Ser Ala Pro Thr Thr Lys Pro Lys Pro Ser

1540 1545 1550

Thr Ser Thr Ser Val Asp Asp Thr Lys Asp Glu Leu Glu Asp Glu Asp

1555 1560 1565

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

1570 1575 1580

Leu Pro Pro Leu Ser Pro Ser Val Thr Ser Ala Ala Ala Pro Glu Leu

1585 1590 1595 1600

Val Asp Val Thr Tyr Arg Pro Pro Val Tyr Thr Thr Gln Arg Pro Arg

1605 1610 1615

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

1620 1625 1630

Pro Ser Arg Thr Glu Ala Asp Met Leu Glu Thr Val Ser Val Gly Arg

1635 1640 1645

Pro Val Ser Gln Pro Leu Pro Pro Pro Pro Pro Pro Pro Pro Pro Thr

1650 1655 1660

Arg Glu Ala Pro Pro Arg Pro Pro Ser Val Ala Leu Ala Pro Thr Pro

1665 1670 1675 1680

Val Arg Asp Ala Ala Ser Thr Glu Val Lys Ala Ser Pro Pro Pro Pro

1685 1690 1695

Pro Ala Ala Ala Ala Thr Pro Pro Ala Thr Leu Asp Arg Thr Trp Thr

1700 1705 1710

Thr Leu Ala Val Asp Leu Glu Gly Ser Gln Thr Arg Leu Pro Asn Arg

1715 1720 1725

Arg Arg Pro Thr Ala Thr Pro Ala Val Ala Pro Thr Ser Ala Thr Ser

1730 1735 1740

Ser Asp Thr Ser Ser Asn Thr Gly Lys Val Thr Ser Val Val Val Thr

1745 1750 1755 1760

Ser Gly Ser Lys Thr Val Leu Phe Val Lys Pro Thr Ala Ala Arg Leu

1765 1770 1775

Pro Thr Gln Thr Thr Lys Ser Thr Pro Thr Pro Arg Pro Arg Pro Arg

1780 1785 1790

Pro Arg Pro Thr Thr Glu Thr Gly Gly Leu Phe Asp Asp Phe Asp Asp

1795 1800 1805

Val Ala Asp Glu Asp Asp Glu Glu Glu Ser Glu Thr Gly Thr Ala Gly

1810 1815 1820

Gly Ser Arg Gly Lys Val Asp Leu Gly Val Ala Ser Ser Asn Gln Glu

1825 1830 1835 1840

Lys Val Val Val Gly Ser Ala Pro Ala Ser Thr Leu Tyr Val Thr His

1845 1850 1855

Thr His Thr Leu Thr Val Thr Thr Thr Glu Thr Gln Val Val Ser Thr

1860 1865 1870

Ala Gly Arg Pro Pro Val Thr Arg Thr Val Val Leu Thr Lys Thr Gln

1875 1880 1885

Thr Ser Thr Val Val Asp Thr Val Thr Glu Thr Gln Thr Leu Leu Arg

1890 1895 1900

Pro Thr Ser Val Tyr Ala Thr Val Thr Thr Thr Val Ser Ala Ile Arg

1905 1910 1915 1920

Pro Phe Thr Glu Gly Val Ser Ser Thr Ser Thr Val Ser Thr Val Thr

1925 1930 1935

Thr Pro Ala Pro Thr Thr Gly Ser Gln Pro Pro Pro Pro Pro Pro Pro

1940 1945 1950

Ala Gly Thr Gly Asp Arg Pro Gly Asp Arg Thr Ala Gly Ala Gly Asp

1955 1960 1965

Thr Phe Phe Val Val Val Thr Asp Pro His His Lys Leu Pro Pro Pro

1970 1975 1980

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

1985 1990 1995 2000

Glu Asp Glu Gln Glu Pro Arg Gln Asp Gly Gly Gly Ala Gly Gly Asn

2005 2010 2015

Val Leu Leu Thr Ala Gly Val Gly Gly Ser Gly Ala Ala Ala Thr Ser

2020 2025 2030

Ala Cys Arg Pro Glu Cys Ser Ala Ala Arg Asn Glu Val Cys Gln Arg

2035 2040 2045

Ala Ala Asp Gly His Pro Arg Cys Leu Cys Arg Pro Gly Phe Ala Arg

2050 2055 2060

Ile Phe Pro Asp Arg Pro Cys Lys Pro Thr Tyr Thr Tyr Thr Leu Arg

2065 2070 2075 2080

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

2085 2090 2095

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

2100 2105 2110

Ala Glu Gly Leu Asp Arg Leu Val Met Gln Ser Asp Leu Arg Asp Val

2115 2120 2125

Phe His Gly Val Ser Val Glu Ala Phe Leu Pro Pro Asp Glu Gly Ser

2130 2135 2140

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

2145 2150 2155 2160

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

2165 2170 2175

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

2180 2185 2190

Gln His Ile Glu Ala Glu Asp Phe Asp Glu Cys Ala Ser Arg Gln Tyr

2195 2200 2205

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

2210 2215 2220

Thr Cys Ser Cys Arg Glu Gly Phe Ala Asp Leu Ser Glu Asn Pro Leu

2225 2230 2235 2240

Phe Pro Gly Arg Leu Cys Ser Ala Glu Leu Ile Gly Cys Glu Arg Cys

2245 2250 2255

His Phe His Gly Thr Cys Gly Asp Asp His His Gln Val Gln Cys Glu

2260 2265 2270

Cys Phe Gln Trp Tyr Ala Gly Asp Thr Cys His Ile Asn Leu Lys Ala

2275 2280 2285

Leu Leu Ile Ala Leu Val Thr Ile Gly Cys Val Leu Val Ala Leu Leu

2290 2295 2300

Leu Val Cys Val Leu Met Thr Cys Ala Arg Gly Gly Ser Arg Gly Gly

2305 2310 2315 2320

Arg Thr Arg Gly Arg Ser Arg Arg Glu Gly Pro Leu Glu Arg Arg Ala

2325 2330 2335

Met Ile His Asp Ser Ala Ser Glu Ala Ser Gly Asp Gln Thr Leu Pro

2340 2345 2350

Arg Ser Gln Thr Pro Ala Ala Val Phe Gly Lys Lys Ala Arg Pro Val

2355 2360 2365

Ser Ala Pro Thr Ala Thr Val Val Ala Val Val Pro Pro Pro Pro Ala

2370 2375 2380

Gly Phe Ala Pro Thr Pro Val Pro Val Pro Pro Pro Leu Gln Pro Pro

2385 2390 2395 2400

Pro Glu Gln Arg Asp Arg Ser Leu Thr Val Met Ile Pro Arg Ala Lys

2405 2410 2415

Tyr Arg Pro Ser Pro Val Ala Pro Pro His Gln Pro Leu Pro Thr Leu

2420 2425 2430

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

2435 2440 2445

Glu Ser Lys Leu Leu Ala Leu Leu Glu Gly Ala Ser Lys Gln Asp Gly

2450 2455 2460

Gln Leu Gln Ser His His His His His Gln His Gln Gln Gln Thr Ser

2465 2470 2475 2480

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

2485 2490 2495

Pro Arg Lys Asn Ser Ser Ser Arg Lys Pro Ser Asn Gly Asn Ile Pro

2500 2505 2510

Pro Pro Ala Pro Gly Ala Leu Val Ser Ala Gly Phe Glu Val Ser Ala

2515 2520 2525

Thr Val Gly Lys Pro Ala Thr Asp Ser Asp Thr Glu Ala Ser Ala Val

2530 2535 2540

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

2545 2550 2555 2560

Val Glu Glu Met Ser Glu Pro Ser Val Lys Ala Gly Ser Ala Val Glu

2565 2570 2575

Thr Leu Thr Val Ser Glu Ala Arg Ser Cys Asp Glu Thr Thr Ile His

2580 2585 2590

Pro Pro Thr Lys Ser Leu His Ser Asn Tyr Ser Ser Lys His Ser Ser

2595 2600 2605

Asn Asn Leu Asn Asn Asp Glu Gly His Thr Met Ala Glu Arg Asp Ile

2610 2615 2620

Gly Ser Thr Phe Val Met Pro Gln Ser His Leu Tyr Thr Pro Asp Arg

2625 2630 2635 2640

Gly Ser Asp Ile Ser Asn Phe Asp Ser Leu

2645 2650

<210> 3

<211> 430

<212> DNA

<213> 未知(Unknown)

<400> 3

cggcaaagga cacgggggga cacgggccaa gacgggcgcg gaagccacgg ccgcccgacg 60

ccgacgcaga cgacgaagca cccacgccgc gaccgaggcc gagaccccgc ccaccaccga 120

gaccggagga cggacgaccg acgacgcgcg gacgaggacg acgaggagga acggagacgg 180

gcacggccgg cgggcgcggg gcaaggggac cgggcgcgca gcccaaccag gagaaggggg 240

gcggccggcg cccgccccac cccacgcacc cacacgcaca caccacagaa cgacacggag 300

acgcaggggg agcacagccg gccggccgcc gggacgcgca cggggccacc aagacgcaga 360

cgcaacagcg ggacacagca ccgagaccca gacgcgcccg gccgaccagc gcagccaccg 420

caccaccacg 430

<210> 4

<211> 45

<212> DNA

<213> 未知(Unknown)

<400> 4

gcgtaatacg actcactata ggcggcaaag tgacatctgt ggtgg 45

<210> 5

<211> 43

<212> DNA

<213> 未知(Unknown)

<400> 5

gcgtaatacg actcactata ggcgtggtgg tgacggtggc ata 43

Claims (6)

1.一种飞蝗粘蛋白，其特征在于，其氨基酸序列如SEQ ID NO：2所示。

2.编码权利要求1所述飞蝗粘蛋白的基因。

3.权利要求2所述的基因，命名为Mucin17，其cDNA序列如SEQ ID NO：1所示。

4.一种飞蝗粘蛋白基因合成的dsRNA，以SEQ ID NO：3所示的核苷酸序列为模板，通过体外转录合成而得到。

5.权利要求4所述dsRNA的获得方法，步骤如下：根据权利要求3中的SEQ ID NO：1设计的上游引物SEQ ID NO：4和下游引物SEQ ID NO：5，通过PCR扩增获得SEQ ID NO：3，其产物含有T7启动子；产物经纯化后按照T7 RiboMAX^TMExpress RNAi System试剂盒说明体外转录合成dsRNA。

6.权利要求4所述的飞蝗粘蛋白基因合成的dsRNA在害虫防治中的应用。

Images