桔小实蝇takeout基因的克隆和序列分析及时空表达  

Cloning,sequence analysis and spatio-temporal expression of takeout in Bactrocera dorsalis(Hendel)

作　　者：胡黎明[1] 申建梅[1] 宾淑英[1] 廖泓之[1] 林进添[1] 

机构地区：[1]仲恺农业工程学院农学院,广东广州510225

出　　处：《南京农业大学学报》2012年第4期75-80,共6页Journal of Nanjing Agricultural University

基　　金：国家自然科学基金项目(30971923)

摘　　要：采用RT-PCR及RACE技术克隆了桔小实蝇(Bactrocera dorsalis(Hendel))takeout基因的cDNA全长序列,命名为BdorTO。测序结果表明:BdorTO开放阅读框全长747 bp,编码248个氨基酸,相对分子质量约为28.15×103,等电点为5.29。Signal P软件分析表明,该蛋白N端具21个氨基酸的信号肽,为分泌型蛋白。氨基酸序列比对表明,BdorTO蛋白与其他昆虫TO蛋白的序列一致性较低,其中与刺舌蝇(Glossina morsitans)的序列一致性最高,一致性值为48.6%。荧光定量PCR分析表明,BdorTO在触角中的表达量最高,推测BdorTO可能参与了桔小实蝇嗅觉感受过程;在雄虫生殖节中的表达量明显高于雌虫生殖节,提示BdorTO可能与桔小实蝇雄虫的生殖生理过程密切相关。Using RT-PCR and RACE methods,the takeout gene from Bactrocera dorsalis was cloned and named as BdorTO.The sequencing results showed that the open reading frame in BdorTO was 747 bp,encoding 248 amino acid residues.The protein molecular weight is about 28.15×103 and the isoelectric point is 5.29.Signal P analysis showed that BdorTO was a secreted protein with signal peptide containing 21 amino acid residues in the N-terminal.The homology analysis of amino acid sequence showed that the BdorTO had lower similarity with TO from other insects and shared the highest similarity（48.6%）with the TO sequence from Glossina morsitans.The tissue expression level of BdorTO was highset in antennae,which implied that BdorTO had incorporated in the olfactory sensation.The fact that the expression level of BdorTO in the male genital segments was significantly higher than that in the female suggests that BdorTO might be closely related to the reproduction physiology of the male B.dorsalis.

关 键 词：桔小实蝇 Takeout蛋白 基因克隆 荧光定量PCR 

分 类 号：Q965.9[生物学—昆虫学]