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1、主絢鐮蛆2010,36(4)：49—54PznntProtection小菜蛾GIuCI受體亞基eDNA基因克隆和序列分析梁延坡，吳青君，劉長仲，張友軍。，王少麗，徐寶云(1．甘肅農(nóng)業(yè)大學(xué)草業(yè)學(xué)院，蘭州730070；2．中國農(nóng)業(yè)科學(xué)院蔬菜花卉研究所，北京100081)摘要利用RT-PCR和RACE技術(shù)對小菜蛾[Plutellaxylostella(L)]4齡幼蟲谷氨酸門控的氯離子通道(GluC1)受體eDNA全長進(jìn)行了克隆和序列分析。通過設(shè)計簡并性上、下游引物擴(kuò)增出小菜蛾GluC1受體亞基基因192bp的cDNA片段，根據(jù)得到的片段序列設(shè)計3和5特異性引
2、物采用RACE技術(shù)進(jìn)行3和5末端的克隆，獲得了小菜蛾GluC1受體的cDNA的完整序列，C-enBank登錄號為GQ221939。該基因全長2144bp，其開放閱讀框(ORF)1344bp，編碼447個氨基酸。相似性分析表明：它與果蠅和赤擬谷盜的相似性均為73，與埃及伊蚊的相似性為75％，與東亞飛蝗的相似性為79；氨基酸分析后顯示它具有GluC1d亞基的典型特征，表明克隆的cDNA序列是小菜蛾GluCla亞基基因的序列。關(guān)鍵詞小菜蛾；谷氨酸門控的氯離子通道受體；反轉(zhuǎn)錄一多聚酶鏈?zhǔn)椒磻?yīng)；RACE；序列分析中圖分類號：Q965．9；S481．1文獻(xiàn)標(biāo)識碼：
3、ADOI：10．3969／j．issn．0529—1542．2010．04．010CloningandsequenceanalysisofcDNAofGluC1receptoralphagenefromthediamondbackmoth，tellaxylostellaLiangYanpo“，WuQingjun2，LiuChangzhong1，ZhangYoujunz，WangShaoliz，XuBaoyunz(1．CollegeofPrataculture，GansuAgriculturalUniversity，Lanzhou730070，China
4、；2．InstituteofVegetablesandFlowers，ChineseAcademyofAgriculturalSciences，Beijing100081，China)AbstractOnefull-lengthcDNAsequenceoftheGluC1receptoralphageneofthefourthinstarsofthediamond-backmoth，Plutellaxylostella(L．)wasamplified，cloned，sequencedandanalyzedbyusingRT-PCRandRACEmeth
5、od．Onefragmentof192bpcDNAoftheGluC1receptoralphagenewasobtainedbyusingdegeneratefor—wardandreverseprimers．Thenthe5一and3-endspecificprimersweredesignedaccordingtothefragmentsequenceforthe5一and3RACE．A2144bpfull-lengthcDNAofGluC1receptorwasobtained．ThisgenewasdepositedinGenBank(acc
6、essionno．GQ221939)．Ithasanopenreadingframeof1344nucleotides，encoding447aminoacidsresidues．SimilarityanalysisshowedthattheclonedeDNAsequencehad73％identitywiththepublishedsequencesofDrosophilamelanogasterandTriboliumcastaneum，75％and79％identitieswiththoseofAedesaegyptiandLocustamig
7、ratoria．TypicalfeaturesofGluC1subunitsasinothermembersoftheCys—looplig-and-gatedionchannelsuperfamilywerefoundthroughproteinanalysis．TheresultsindicatedthatthecDNAsequencewasGluClareceptorgene．KeywordsPlutellaxylostella；GluC1receptor；RT—PCR；RACE；sequenceanalysis谷氨酸是脊椎動物和無脊椎動物神經(jīng)系
8、統(tǒng)內(nèi)重內(nèi)谷氨酸既是興奮性的神經(jīng)遞質(zhì)。卜。又是抑制性的要的神經(jīng)遞質(zhì)，在脊椎動物體內(nèi)主要通過門控陽
