山东省不同蚊种28S rDNA D3区序列特征分析  

Sequences analysis of D3 domain of 28S ribosomal DNA of different species of mosquitoes in Shandong Province

作　　者：郭秀霞 王海防 王文倩 张崇星 刘宏美 刘丽娟 黄晓丹 程鹏 公茂庆 GUO Xiuxia;WANG Haifang;WANG Wenqian;ZHANG Chongxing;LIU Hongmei;LIU Lijuan;HUANG Xiaodan;CHENG Peng;GONG Maoqing(Shandong First Medical University&Shandong Academy of Medical Sciences,Shandong Institute of Parasitic Diseases,Jining,Shandong 272033,China)

机构地区：[1]山东第一医科大学(山东省医学科学院),山东省寄生虫病防治研究所,山东济宁272033

出　　处：《中华卫生杀虫药械》2024年第5期455-459,共5页Chinese Journal of Hygienic Insecticides and Equipments

基　　金：山东省医学科学院医药卫生科技创新工程;山东省自然科学基金(ZR2021MH122);山东省医药卫生科技发展计划(202101050203,202312070627,202302081244)。

摘　　要：目的分析山东地区常见9种蚊虫核糖体DNA 28S第3结构域(28S rDNA D3)基因序列特征,为蚊虫种类分子鉴定提供依据。方法在山东济宁北湖地区采集蚊虫并形态学鉴定,提取不同蚊种基因组DNA,PCR扩增其28S rDNA D3基因片段并进行测序分析,基因序列在GenBank中进行同源性比对,ClustalX 1.81、DNAMAN、DNASTAR和Mega6软件分析序列特征并构建系统进化树探讨系统发生关系。结果成功获得淡色库蚊、三带喙库蚊、二带喙库蚊、白纹伊蚊、刺扰伊蚊、中华按蚊、骚扰阿蚊、常型曼蚊及黄色轲蚊的28S rDNA D3基因,其长度为361~422 bp,GC含量为52.9%~57.4%,包含保守位点297个,96个位点为变异位点,平均发生碱基转换15个,颠换17个,转换与颠换之比为0.90,平均遗传距离为0.073,所有蚊虫的28S rDNA D3基因分子鉴定与形态学鉴定基本吻合。构建系统发育树显示,昆虫纲不同目物种分别为不同的分支,库蚊属和伊蚊属各为一支,常型曼蚊和黄色轲蚊聚为一支,中华按蚊为独立的分支。结论28S rDNA D3基因在蚊科的属和种的分类阶元鉴定中具有一定的意义。Objective To explore gene characteristics of D3 domain of 28S ribosomal DNA in nine common mosquito species of Shandong Province,so as to provide a basis for the molecular identification of the mosquitoes.Methods Mosquitoes were collected from Jining City and identified in the laboratory.Single mosquito genomic DNA was extracted from different species and 28S rDNA D3 was specifically amplified by PCR and sequenced.Gene sequences were blasted in GenBank,and the gene traits were analyzed by ClustalX 1.81,DNAMAN,DNASTAR and Mega6.The phylogenetic tree was constructed.Results The 28S rDNA D3 gene fragments of Culex pipiens pallens,Culex tritaeniorhynchus,Culex bitaeniorhynchus,Aedes albopictus,Aedes vexans,Anopheles sinensis,Armigeres obturbans,Mansonia uniformis,Coquillettidia ochracea were amplified,from 361 bp to 422 bp,GC contents were 52.9%to 57.4%.There were 297 conserved sites,96 variable sites,15 transitionsal pairs,17 transversional pairs,transition:transversion bias was 0.90,and the mean genetic distance was 0.073.The molecular identification of the species based on 28S rDNA D3 was consistent with the traditional morphological identification.The phylogenetic tree showed that the mosquitoes of different orders were in different branches.The genus Culex and genus Aedes belong to different branches.Mansonia uniformis and Coquillettidia ochraces were grouped together,and Anopheles sinensis was an independent branch.Conclusion 28S rDNA D3 can serve as an useful classification and identification molecular marker for different mosquito species and genus.

关 键 词：蚊虫 核糖体DNA 28S第3结构域 序列分析 

分 类 号：R384.1[医药卫生—医学寄生虫学]