山东省淡色库蚊核糖体DNA第二内转录间隔区基因多态性分析

Genetic polymorphism analysis on rNDA-ITS2 from different geographical strains of Culex pipiens pallens in Shandong province

目的探讨山东省不同地区淡色库蚊核糖体DNA第二内转录间隔区(rDNA-ITS2)基因序列特征,分析其遗传变异。方法在山东省的济南、济宁、青岛、聊城、临沂、淄博市以及江苏省南京市采集淡色库蚊成蚊,并取实验室选育的抗氯氰菊酯与抗敌敌畏品系成蚊,提取蚊虫DNA,PCR扩增淡色库蚊的r DNA-ITS2,PCR产物纯化后连接至p MD19-T载体进行基因测序,生物信息学分析r DNA-ITS2基因多态性。结果 PCR特异扩增山东省6个不同地理株及南京市和实验室2个不同品系的淡色库蚊r DNA-ITS2区域,获得460 bp左右的扩增片段。测序结果显示淡色库蚊r DNA-ITS2出现36个变异位点,表现出基因的多态性,其r DNA-ITS2基因单核苷酸存在转换、颠换和缺失。结论山东省不同地理区域和不同抗性的淡色库蚊在r DNA-ITS2区出现遗传多态性,可能与蚊虫生态环境有一定的关系。

Objective To explore gene sequence characteristics of rNDA-ITS2 in different populations of Culex pipiens pallens from Shandong province and analyze the genetic variation of these mosquitoes.Methods Adult-female mosquitoeswere collected from different cities： Jinan, Jining, Qingdao, Liaocheng, Linyi, Zibo from Shandong and a sample from Nanjing. The pyrethroid-resistant and dichlorvos-resistant strains were reared in the laboratory. Single mosquito genomic DNA was extracted from Cx. pipiens pallens of these different populations and r NDA- ITS2 of Cx. pipiens pallens were specifically amplified by PCR. The PCR products were cloned into the p MD-19 T simple vectors and sequenced. Genetic polymorphisms of them were analyzed by biological information systems.Results The obtained r DNA-ITS2 sequences were approximate 460 bp in length, with 36 mutation sites. Culex pipiens pallens from different geographical environment showed gene polymorphisms in sequence analysis of r NDA-ITS2. There were transition,transversion and deletion in the mono nucleotide of r NDA-ITS2 gene.Conclusion Genetic polymorphisms presented on r NDA-ITS2 of Cx. pipiens pallens from different geographical regions in Shandong province and different resistance strains in the lab were probablyassociated with the ecological environment.