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朱砂叶螨烟碱型乙酰胆碱受体α4亚基基因的克隆与序列特征分析 预览

Cloning and sequence characteristics analysis of nicotinic acetylcholine receptor α4 subunit gene fromTetranychus cinnabarinus
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摘要 烟碱型乙酰胆碱受体(nAChR)是一种对神经递质乙酰胆碱起快速反应的激动剂门控离子通道,也是新烟碱类杀虫剂的作用靶标。此研究利用同源克隆结合RACE技术,从朱砂叶螨敏感品系中克隆了一条nAChR基因,全长为2 023bp,开放阅读框长1 635bp,编码544个氨基酸。预测分子量(Mr)为62.20kDa,等电点(pI)为5.00。根据同源性及系统发育分析,将其命名为Tcα4,提交至NCBI数据库,获得GenBank登陆号为KP694228。Tcα4具有nAChRα亚基典型结构特征:N端胞外结构域和C端跨膜结构域。Tcα4与二斑叶螨nAChRα4亚基基因序列同源性高达98.71%,与二斑叶螨其他nAChR亚基基因同源性仅在20.07%~43.16%之间,与其他昆虫nAChRα4亚基基因同源性在45.96%~56.07%之间。系统发育分析结果表明朱砂叶螨与二斑叶螨和肩突硬蜱nAChRα4亚基基因在同一分支,与其他昆虫nAChRα4亚基基因则较远。朱砂叶螨nAChRα4亚基基因的克隆为朱砂叶螨其他nAChR亚基基因的研究及以nAChR为作用靶标的新型杀螨剂的研究提供了一种新途径,奠定了基础。 Nicotinic acetylcholine receptor(nAChR)was the gated ion channel response to acetylcholine and one of the most important insecticide targets.In this study,the nAChRα4subunit gene of Tetranychus cinnabarinus(Boisduval)was cloned by RT-PCR and RACE.The cDNA had a length of 2 023 bp,and its Open Reading Frame(ORF)had a length of 1 635 bp encoded a protein of 544 amino acid residues.Molecular weight(Mr)of the protein is 62.20 kDa,and the isoelectric point(pI)is5.00.It was named as Tcα4(GenBank accession:No.KP694228)according to its amino acid homology to those of T.urticae and other insects.The nAChRα4subunit of T.cinnabarinus had rypical character of nAChR units with a N-terminal transmembrane domain and a C-terminal extracellular domain.Tcα4had a homology of 98.71%to T.urticae nAChRα4,and only20.07%~43.16% homology to other nAChRs of T.urticae;and had a range of 45.96%~56.07% homology to other insectsnAChRα4subunits.Phylogenetic tree analysis showed that Tcα4,T.urticae and Ixodes scapularis nAChRα4were classified in the same cluster,and were far from the others.The cloning and character analysis of Tcα4lay a solid foundation for the study of other nAChR subunits fromT.cinnabarinus and development of new biological miticide targeted to nAChR.
作者 高品 王帅 管学琼 卜春亚 师光禄 GAO Pin;WANG Shuai;GUAN Xueqiong;BU Chunya;SHI Guanglu;Plant Science and Technology College,Beijing University of Agriculture;Key Laboratory of Urban Agriculture(North)of Ministry of Agriculture,China,Beijing University of Agriculture;College of Biological Science and Engineering,Beijing University of Agriculture
出处 《北京农学院学报》 2015年第4期1-8,共8页 Journal of Beijing Agricultural College
基金 国家自然科学基金资助项目(31200483) 北京市自然科学基金资助项目(6122005) 教育部科学技术重点研究项目(212001) 北京市教育委员会科技计划面上项目(KM201210020001) 北京市教育委员会平台项目(KCT2014001)
关键词 朱砂叶螨 烟碱型乙酰胆碱受体 α4亚基 基因克隆 序列分析 Tetranychus cinnabarinus nicotinic acetylcholine receptor α4subunit gene cloning sequence analysis
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参考文献17

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