淡色库蚊抗药性相关新基因——视蛋白基因的克隆与分析  

Cloning of a novel deltamethrin-resistance associated opsin gene from Culex pipiens pallens

作　　者：孙艳[1] 杨明夏[1] 马磊[1] 李秀兰[1] 公茂庆[1] 胡小邦[1] 顾燕[1] 朱昌亮[1] 

机构地区：[1]南京医科大学病原生物学系,江苏南京210019

出　　处：《南京医科大学学报（自然科学版）》2005年第5期321-323,共3页Journal of Nanjing Medical University(Natural Sciences)

基　　金：国家自然科学基金资助项目(30170835)

摘　　要：目的:获取淡色库蚊抗药性相关视蛋白基因(NYD鄄OP)全长cDNA序列。方法:根据抑制性差减杂交(SSH)结合cDNA芯片分离获得的淡色库蚊抗药性相关视蛋白基因的289bp片段,设计上、下游引物,分别以cDNA文库和反转录二链产物为模板,采用快速扩增cDNA末端法(rapidamplificationofcDNAend,RACE)扩增视蛋白基因5'、3'端,经对位拼接获得全长序列,并用相应的软件进行分析。结果:获得6个淡色库蚊视蛋白基因全长序列,开放阅读框分别为1116bp(GeneBank/NCBIAY297441,AY297442)及1119bp(GeneBank/NCBIAY297443,AY297444,AY299337,AY299338),以上序列经推导分别编码371及372个氨基酸。进行蛋白质序列分析,与烟草天蛾视蛋白的同源性是77%。结论:获得6个淡色库蚊抗药性相关视蛋白基因全长cDNA序列。Objective:To clone a novel deltamethrin-resistance associated opsin gene from Culex pipiens pallens. Methods:The left and right gene-specific primers were designed on the basis of the nucleotide sequences of the 289 bp known from the combination of suppression substractive hybridization(SSH) and gene expression profiling by cDNA arrays and used in 5’ and 3’ RACE to amplify opsin cDNA ends. The templates of 5’ and 3’ RACE were Plasmid cDNA library and the second-strand cDNA fragment amplified by RT-PCR, respectively. The 5’ and 3’ cDNAs of opsin obtained were sequenced, spliced and made bioinformatics analysis using softwares. Results:The six full-length sequences of opsin gene were obtained, named NYD-OP1,OP2,OP3,OP4,OP5 and OP6 respectively. There was an open reading frame of 1 116 bp in the sequence of NYD-OP1-2 cDNA(GenBank/NCBI AY297441,AY297442) and 1 119 bp in that of NYD-OP3-6(Genebank/NCBI AY297443,AY297444,AY299337,AY299338) respectively. Conclusion:The cDNAs of opsin gene were successfully cloned from Culex pipiens pallens.

关 键 词：淡色库蚊 抗药性 克隆 视蛋白基因 

分 类 号：R381.1[医药卫生—医学寄生虫学]