红光熊蜂线粒体CoI基因的克隆与特性分析  

Cloning and characterization of cytochrome C oxidase subunitⅠfrom Bombus ignites

作　　者：王丹丹[1] 卢建璋 王星[1] WANG Dan-dan;LU Jian-zhang;WANG Xing(College of Agriculture,Eastern Liaoning University,Dandong 118003,China;Zhejiang University-University of Edinburgh Institute,Haining 314499,China)

机构地区：[1]辽东学院农学院,辽宁丹东118003 [2]浙江大学爱丁堡大学联合学院,浙江海宁314499

出　　处：《东北师大学报（自然科学版）》2021年第4期77-83,共7页Journal of Northeast Normal University(Natural Science Edition)

基　　金：中央财政林业科技推广示范资金资助项目(辽[2019]06号);辽宁省科学事业公益研究项目(2013002012).

摘　　要：通过对NCBI数据库的搜查,获得熊蜂CoI基因部分序列;设计引物,采用RT-PCR和RACE技术克隆了红光熊蜂CoI基因cDNA全长序列.采用DNAMAN 8.0,MEGA 7.0,Protparam和Automatic Modeling Mode等软件对该cDNA序列、基本理化性质、3-D结构、跨膜结构域及其系统进化关系进行了分析.结果表明:红光熊蜂CoI全长cDNA序列含1050 bp,编码349个氨基酸,对应的蛋白质相对分子质量为38350,预测其分子式为C_(1789)H_(2724)N_(422)O_(463)S_(25),理论等电点(pI)5.79,偏酸性,半衰期为30 h;蛋白质平均亲水系数0.706,不稳定性指数39.98,脂溶性指数110.92.该蛋白质具8个α-螺旋、8个跨膜结构域,为稳定的疏水性蛋白质.系统发育显示红光熊蜂CoI氨基酸序列与同属地熊蜂的进化关系较近,表明红光熊蜂线粒体CoI基因序列保守性较高.The objective of this study is to clone the full-length cDNA of cytochrome C oxidase subunitⅠ(CoI)from Bombus ignites,then analyze the sequence.The CoI-specific primers were designed based on the known sequence of CoI obtained by the NCBI database search.The full cDNA of B.ignites CoI gene was synthesized by RT-PCR and RACE techniques.The sequence of CoI gene,physicochemical property,three-dimensional structure,trans-membrane domain and phylogenetic were analyzed by DNAMAN 8.0,MEGA 7.0,Protparam and Automatic Modeling Mode software,respectively.The results showed that the ORF of CoI consisted of 1050 bp,which encoded a protein with 349 amino acid residues,a predicted formula of C_(1789)H_(2724)N_(422)O_(463)S_(25),and a molecular weight of 38350.The other characters of this protein included isoelectric point(pI)of 5.79,half-life period of 30 h,the unstable factor of 39.98,the total average hydrophilic coefficient of 0.706,and the fat solubility index of 110.92.Additionally,the protein has 8α-helix and 8 transmembrane domains,which indicated that the CoI protein is a stable hydrophobic protein.The phylogenetic tree demonstrated that B.ignites had the closest evolutionary relationship with B.terrestris at the amino acid level,which suggested that the mitochondrion CoI sequence in B.ignites was highly conservative in phylogeny.

关 键 词：红光熊蜂 线粒体 COI基因 克隆 RACE技术 

分 类 号：Q962[生物学—昆虫学]