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

刘飞, 郭秀霞, 程鹏, 王海防, 刘丽娟, 杨培培, 赵玉强, 王怀位, 公茂庆

doi:10.11853/j.issn.1003.4692.2015.06.003

中国媒介生物学及控制杂志 >

2015 , Vol. 26 >Issue 6: 550 - 554

DOI: https://doi.org/10.11853/j.issn.1003.4692.2015.06.003

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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

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1 山东省医学科学院, 山东省寄生虫病防治研究所, 山东省医药卫生媒介生物学重点实验室, 山东济宁 272033;
2 济南大学山东省医学科学院医学与生命科学学院, 济南 250012;
3 济宁市第一人民医院, 山东济宁 272033

刘飞,女,在读硕士,检验技师,从事病原生物学研究,Email:707137516@qq.com

收稿日期: 2015-08-21

网络出版日期: 2015-12-20

基金资助

国家自然科学基金(81271877,81471985)

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

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1 Shandong Institute of Parasitic Diseases, Shandong Academy of Medical Sciences, Department of Medical Entomology, Shandong Province Key Laboratory of Health and Vector Biology, Jining 272033, Shandong Province, China;
2 School of Medicine and Life Sciences, University of Jinan, Shandong Academy of Medical Sciences, Jinan 250012, Shandong Province, China;
3 Shandong Jining No. 1 People's Hospital, Jining 272033, Shandong Province, China

Received date: 2015-08-21

Online published: 2015-12-20

Supported by

Supported by the National Natural Science Foundation of China(No. 81271877, 81471985)

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摘要

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rDNA-ITS2,PCR产物纯化后连接至pMD19-T载体进行基因测序,生物信息学分析rDNA-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rDNA-ITS2区域,获得460 bp左右的扩增片段。测序结果显示淡色库蚊rDNA-ITS2出现36个变异位点,表现出基因的多态性,其rDNA-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rDNA-ITS2区出现遗传多态性,可能与蚊虫生态环境有一定的关系。

关键词： 淡色库蚊; 核糖体DNA第二内转录间隔区; 基因多态性

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刘飞, 郭秀霞, 程鹏, 王海防, 刘丽娟, 杨培培, 赵玉强, 王怀位, 公茂庆 . 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[J]. 中国媒介生物学及控制杂志, 2015 , 26(6) : 550 -554 . DOI: 10.11853/j.issn.1003.4692.2015.06.003

Abstract

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 mosquitoes were 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 rNDA-ITS2 of Cx. pipiens pallens were specifically amplified by PCR. The PCR products were cloned into the pMD-19T simple vectors and sequenced. Genetic polymorphisms of them were analyzed by biological information systems. Results The obtained rDNA-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 rNDA-ITS2. There were transition,transversion and deletion in the mono nucleotide of rNDA-ITS2 gene. Conclusion Genetic polymorphisms presented on rNDA-ITS2 of Cx. pipiens pallens from different geographical regions in Shandong province and different resistance strains in the lab were probably associated with the ecological environment.

Key words： Culex pipiens pallens; rDNA-ITS2; Genetic polymorphism

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