普通大蓟马MuRhodopsin基因的全长克隆及生物信息学分析

Cloning and bioinformatics analysis of MuRhodopsin gene in Megalurothrips usitatus

为了解蓟马害虫视蛋白分子结构及特征,根据普通大蓟马(Megalurothrips usitatus)转录组信息克隆获得了视紫红质MuRhodopsin的基因cDNA全长,并利用DNAMan 9.0和SWISSMODEL等软件对视紫红质MuRhodopsin的基因进行序列同源性比对并构建基因系统进化树,分析理化性质及结合域特点,并对该蛋白进行结构预测。结果表明:该基因全长1140 bp,共编码380个氨基酸,相对分子质量为42.73 kDa,理论等电点为8.47。序列比对以及同源性分析结果表明,MuRhodopsin为横纹视紫红质(rhabdomeric opsins,r-opsins)与西花蓟马(Frankliniella occidentalis)及棕榈蓟马(Thrips palmi)具有高度同源性。结构域分析表明该蛋白具有7个跨膜结构域,包含了1个G蛋白偶联受体家族区域,隶属于G蛋白偶联受体家族;具有6个N-豆蔻酰化位点,3个N-糖基化位点,3个酪蛋白激酶Ⅱ磷酸化位点,4个蛋白激酶C磷酸化位点;其第322位氨基酸—赖氨酸(K)为与发色团(视黄醛)的重要结合位点,具有典型的视蛋白能够激活IP3/Ca2+信号通路引起光依赖的去极化反应。

It is of important basic data significance to understand the molecular structure and characteristics of opsin of Megalurothrips usitatus pest by cloning Rhodopsin gene of M.usitatus for further analysis of light environment differences.The full-length gene of the rhodopsin,MuRhodopsin,was cloned based on the transcriptome information of thrips.The sequence homology was compared using DNAMan 9.0 and SWISSMODEL software,and the phylogenetic tree was constructed.The full-length MuRhodopsin gene was 1140 bp,encoding 380 amino acids,with a relative molecular weight of 42.73 kDa and a theoretical isoelectric point of 8.47.Sequence alignment and homology analysis showed that MuRhodopsin was rhabdomeric opsins(ropsins)with high homology with that of Frankliniella occidentalis and Thrips palmi.The domain analysis showed that the protein had 7 transmembrane domains,including 1 G protein-coupled receptor family region,belonging to the G protein-coupled receptor family,and contained 6 N-cardamoylation sites,3 Nglycosylation sites,3 casein kinase II phosphorylation sites,and 4 protein kinase C phosphorylation sites.The 322nd amino acid lysine(K)of the protein is an important binding site to chromophore(retinal),which is a typical opsin that can activate the IP3/Ca2+signaling pathway and cause light-dependent depolarization reaction.The structure and function of Rhodopsin in M.usitatus were elucidated,providing a theoretical basis for further study of its visual pathway.