桔小实蝇V-ATPaseG亚基基因的克隆及组织表达特异性分析  

Cloning and tissue-specific expression analysis of V-ATPase G subunit gene in Bactrocera dorsalis(Hendel)(Diptera:Tephritidae)

作　　者：胡黎明[1] 申建梅[1] 宾淑英[1] 林进添[1] 

机构地区：[1]仲恺农业工程学院农学院,广州510225

出　　处：《昆虫学报》2011年第12期1452-1458,共7页Acta Entomologica Sinica

基　　金：国家自然科学基金项目(30971923)

摘　　要：空泡型ATP酶(vacuolar-type H+-ATPase,V-ATPase)作为质子泵几乎在所有的真核生物细胞中发挥重要作用。本研究利用RT-PCR和RACE技术获得了桔小实蝇Bactrocera dorsalis(Hendel)V-ATPase G亚基序列全长,命名为BdorATPG。测序结果表明,BdorATPG阅读框全长354bp,编码117个氨基酸。氨基酸序列比对表明,BdorATPG的N端序列与其他物种的ATPG亚基对应区域具有较高的序列一致性。BdorATPG与拟暗果蝇Drosophila pseudoobscura ATPG亚基的氨基酸序列一致性最高,为88.9%。三维结构模建结果表明,BdorATPG N端(第1～59位氨基酸)序列为α-螺旋结构,亲水性和疏水性氨基酸在螺旋两侧呈对称分布。BdorATPG在不同组织中的荧光定量PCR分析表明,BdorATPG在各组织中都有表达,其中在触角中的表达量最高;在雄虫生殖节中的表达量是雌虫中的6.04倍。结果提示BdorATPG可能在雄虫生殖生理过程中发挥重要作用。Vacuolar-type H＋-ATPase（V1V0-ATPase,V-ATPase）,a type of proton pump,functions in almost every eukaryotic cell.In this study,a V-ATPase G subunit gene was cloned from Bactrocera dorsalis and named as BdorATPG.The open reading frame of BdorATPG is 354 bp in length,encoding 117 amino acid residues.Homology analysis showed that BdorATPG has high similarity with corresponding regions of V-ATPase G subunits from other dipteran insects,and shares 88.9% amino acid identity with the V-ATPase G subunit from Drosophila pseudoobscura.Three-dimensional structure modeling results showed that the N terminal（1-59 amino acids） sequence of BdorATPG is α-helix structure,and the hydrophilic and hydrophobic residues are almost symmetrically distributed on both sides of the helix.Real-time PCR analysis revealed that BdorATPG mRNA was expressed in all tissues,while the highest expression level was detected in the antenna.The BdorATPG mRNA level in male genital segments was about 6.04-fold as high as that in female genital segments,suggesting that V-ATPase plays an especially important role in male reproductive physiological processes.

关 键 词：桔小实蝇 V-ATPASE G亚基 基因克隆 荧光定量 组织特异性表达 

分 类 号：Q966[生物学—昆虫学]