桔小实蝇肌球蛋白轻链2基因的克隆及表达分析  

Cloning and expression profiling of myosin light chain 2 gene in Bactrocera dorsalis(Hendel)(Diptera:Tephritidae)

作　　者：申建梅[1] 胡黎明[1] 宾淑英[1] 林进添[1] 

机构地区：[1]仲恺农业工程学院,外来有害生物预警与控制研究所,广州510225

出　　处：《昆虫学报》2011年第5期508-514,共7页Acta Entomologica Sinica

基　　金：国家自然科学基金项目(30971923)

摘　　要：肌球蛋白轻链在生物的运动过程中具有重要作用,本研究利用RT-PCR和RACE技术获得了桔小实蝇Bactrocera dorsalis(Hendel)肌球蛋白轻链2(myosin light chain 2,MLC2)的cDNA全长序列,命名为BdorMLC2。测序结果表明,BdorMLC2开放阅读框全长669 bp,编码222个氨基酸残基。在线软件SMART分析显示,BdorMLC2具有2个Ca2+结合基序(EFh)结构域,可以结合Ca2+,属于肌钙蛋白C超家族成员。氨基酸序列比对表明,BdorMLC2与其他昆虫的肌球蛋白具有较高的序列一致性,其中与黑腹果蝇Drosophila melanogasterMLC2的序列一致性高达93.2%。BdorMLC2在不同组织和时期的荧光定量PCR分析表明,BdorMLC2在桔小实蝇雄虫胸部的含量最高,在前足、中足、后足和翅中也有较高的表达;BdorMLC2几乎在桔小实蝇发育的各个时期都有表达,刚羽化成虫的表达量显著高于其他发育阶段。结果提示,BdorMLC2可能与桔小实蝇的肌肉收缩运动密切相关。本研究为深入研究桔小实蝇肌球蛋白的功能提供了理论依据。Myosin light chain 2（MLC2） plays an important role in biological motility.Myosin light chain 2 gene was cloned from Bactrocera dorsalis and named as BdorMLC2.The open reading frame of BdorMLC2 is 669 bp in length encoding 222 amino acid residues.On-line SMART analysis revealed that BdorMLC2 has two EFh functional domains that can bind Ca2＋,and belongs to one of the members of the troponin C superfamily.Homology analysis of amino acid sequence showed that BdorMLC2 has very high similarity with MLC from other insects and shares 93.2% identity with the MLC sequence from Drosophila melanogaster.Tissue expression analysis revealed that BdorMLC2 transcript was observed clearly in thorax of male B.dorsalis,and highly expressed in head（antennae removed）,foreleg,middle legs,hind legs and wings.The temporal expression pattern further indicated that BdorMLC2 was expressed in different developmental stage.The highest expression level was found in newly-emerged adult compared to other developmental stages.The results suggest that BdorMLC2 might be closely related to muscle contraction of B.dorsalis.This study provides a basis for further researching the function of BdorMLC2.

关 键 词：桔小实蝇 肌球蛋白轻链2 基因克隆 表达谱分析 荧光定量PCR 

分 类 号：Q966[生物学—昆虫学]