狂犬病毒GSQY-Dog-China-2013株全基因组序列测定和遗传进化分析  

Complete Genome Sequencing and Phylogenetic Analysis of Rabies Virus GSQY-Dog-China-2013 Strain

作　　者：邢广旭[1] 姜中毅 焦文强[1] 王方雨[1] 张改平[1] XING Guang-xu;JIANG Zhong-yi;JIAO Wen-qiang;WANG Fang-yu;ZHANG Gai-ping(Henan Provincial Key Laboratory of Animal Immunology,Henan Academy of Agricultural Sciences,Zhengzhou 450002,China;Gansu Provincial Center for Disease Control and Prevention,Lanzhou 730000,China)

机构地区：[1]河南省农业科学院动物免疫学重点实验室,河南郑州450002 [2]甘肃省疫病预防控制中心,甘肃兰州730000

出　　处：《中国兽医杂志》2023年第1期8-13,共6页Chinese Journal of Veterinary Medicine

基　　金：河南省农业科学院优秀青年科技基金(2020YQ25);河南省科技攻关项目(212102110376)。

摘　　要：为了全面分析我国狂犬病病毒遗传进化情况,本试验使用反转录PCR(RT-PCR)方法将狂犬病病毒GSQY-Dog-China-2013分为10个片段进行扩增,回收目的片段并与pMD20-T simple载体连接,转化JM109感受态细胞,挑取阳性克隆进行测序;使用DNASTAR软件中的Edit Seq通过拼接相邻片段之间的重叠序列获得GSQY-Dog-China-2013全基因组序列,构建系统进化树进行遗传进化分析。结果显示,GSQY-Dog-China-2013全长11924核苷酸(nt),包含5个开放阅读框,分别编码核蛋白(N)、磷蛋白(P)、基质蛋白(M)、糖蛋白(G)和转录大蛋白(L);基于全基因组序列的进化树分析显示,GSQY-Dog-China-2013株和国内分离株形成一个分支,在国内分离株中与SXYL15株和SXBJ15株形成一个分支,亲缘关系最接近,提示GSQY-Dog-China-2013可能源自我国陕西省。本试验系甘肃省首次报道狂犬病病毒全基因组序列,进一步丰富了我国狂犬病病毒遗传进化数据,为狂犬病全面防治以及疫苗研发提供了数据支持。In order to fully elucidate the evolution of rabies virus circulating in China,this study utilized reverse transcription PCR(RT-PCR)to amplify the divided 10 fragments of rabies virus GSQY-Dog-China-2013 strain.The PCR products were purified and ligated with pMD20-T simple vector and transformed into competent cell JM109,positive clones were selected for sequencing.The whole genome sequence of GSQY-Dog-China-2013 was obtained by using Edit Seq of DNASTAR software,and the phylogenetic tree was constructed to analyze the genetic evolution.The results showed that the full length of GSQY-Dog-China-2013 was 11924 nt,and comprised five open reading frames,encoding nucleocapsid protein(N),phosphoprotein(P),matrix protein(M),glycoprotein(G)and large RNA-dependent RNA polymerase(L).Phylogenetic analysis revealed that GSQY-Dog-China-2013 formed a branch with the rabies virus isolated from China,and had a very close relationship to SXYL15 and SXBJ15,suggesting that GSQY-Dog-China-2013 might have originated from Shaanxi Province.Thus,this study was the first to report the complete genome sequence of rabies virus in Gansu Province,which further enriches the genetic and evolutionary data of rabies virus in China,and provides data support for the comprehensive prevention and control of rabies and vaccine research and development.

关 键 词：狂犬病病毒 GSQY-Dog-China-2013 全基因组序列测定 遗传进化分析 

分 类 号：S831[农业科学—畜牧学]