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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 investigate the sequence of rDNA-ITS2 gene of ribosomal DNA of Culex pipiens pallens in different areas of Shandong Province and analyze its genetic variation. Methods Adult mosquitoes of Culex pipiens pallens were collected from Jinan, Jining, Qingdao, Liaocheng, Linyi, Zibo, and Nanjing of Jiangsu Province. The mosquitoes were extracted from mosquitoes by PCR The PCR products of rDNA-ITS2 of Culex pipiens pallens were amplified and then ligated to p-MD19-T vector for gene sequencing. Bioinformatics analysis of r DNA-ITS2 gene polymorphism was performed. Results PCR amplification of rDNA-ITS2 region of 6 Culex pipiens pallens from 6 different geographical isolates in Shandong Province and 2 different strains in Nanjing and laboratories resulted in the amplification of 460 bp. Sequencing results showed that there were 36 mutation sites in DNA-ITS2 of Culex pipiens pallens, which showed polymorphism of the gene. The single nucleotide of rDNA-ITS2 gene was transposed, transposed and deleted. Conclusion Genetic diversity of Culex pipiens pallens from different geographical regions and different resistant areas in Shandong province was observed in rDNA-ITS2 region, which may be related to the ecological environment of mosquitoes.