广西猪瘟病毒E2基因克隆及序列分析  

Cloning and sequencing of classical swine fever virus E2 gene in Guangxi

作　　者：陆增荣[1,2] 张民秀[1] 肖雄[1] 王艺[1] 刘芳[1] 刘欢[1] 卢冰霞[1] 刘磊[1] 郭旋[1] 黎木兰[1] 黄福标[1] 黄伟坚[1] 

机构地区：[1]广西大学动物科学技术学院,南宁530005 [2]广西丽原生物股份有限公司,南宁530001

出　　处：《南方农业学报》2012年第4期528-531,共4页Journal of Southern Agriculture

基　　金：广西科学研究与技术开发计划项目(桂科攻1123007-3)

摘　　要：【目的】了解广西猪瘟病毒(CSFV)流行毒株的变异规律及其与疫苗株的基因差异,为正确选择猪瘟疫苗和防制广西猪瘟提供参考依据。【方法】以广西本地的阳性猪瘟病料为研究对象,应用RT-PCR扩增CSFV的E2基因,经克隆、测序后用DNASTAR软件对其序列进行比对分析,并绘制遗传进化树。【结果】从41份疑似猪瘟病料中共扩增出4个阳性样品的E2基因(1140bp),经序列比对分析发现,扩增获得的4株CSFV毒株(GXGG1、GXLC1、GXLC2和GXNN1)与兔化弱毒株(HCLV)、Shimen强毒株的核苷酸同源性为82.1%～82.3%和82.9%～83.4%、其推导氨基酸同源性为88.4%～89.2%和88.9%～90.0%,4株毒株间的E2基因核苷酸及其推导氨基酸同源性分别为90.8%～99.6%和94.2%～99.5%,且均属于基因群Ⅱ;E2基因推导的氨基酸表明,E2蛋白空间结构及抗原结构的氨基酸位点693Cys、737Cys、792Cys、818Cys、828Cys、856Cys、833Pro、834Thr、837Arg均未发生变异,但其单抗识别位点G713E、N729D、K734R发生变异。【结论】近年来广西CSFV流行毒株的变异未出现较大差异,与HCLV株、Shimen强毒株亲缘关系较远,但与Paderborn株、GXWZ02株亲缘关系较近。[ Objective ]Variations regulation of classical swine fever virus （CSFV） prevalent strains in Guangxi and its difference with vaccine strain was studied in order to select a swine fever vaccine for controlling this disease in Guangxi. [Method]The CSFV E2 gene in local positive diseased swine samples was amplified using RT-PCR, cloned into the pMD18-T vector and sequenced. The sequences were comparatively analyzed using DNASTART Software and heredity evolutionary tree was drawn. [Result]Four positive strains （GXGG1, GXLC1, GXLC2 and GXNN1 ） of E2 gene were amplified from 41 CSF samples. The length of E2 gene was 1140 bp. The homologies of nucleotide sequence of 4 strains were 82.1-82.3% and 82.9-83.4%, and the homologies of deduced amino acid sequence were 88.4-89.2% and 88.9-90.0% with HCLV and Shimen strain, respectively. The homologies of nucleotide and amino acid sequence of 4 strains were 90.8- 99.6% and 9g.2-99.5%, respectively. These strains were classified into 2 groups, and the 4 strains belonged to the genotype Ⅱ. No variation occurred at E2 protein structure and amino acid sites： 693Cys, 737Cys, 792Cys, 818Cys, 828Cys, 856Cys, 833Pro, 834Thr and 837Arg, however, the epitope（G713E, N729D, K734R） changed. [Conclusion]No big variation happened to common CSFV strains during recent years in Guangxi, and they were not found close to HCLV and Shimen. Nevertheless, CSFV strains shared high homology with Paderborn and GXWZ02.

关 键 词：猪瘟病毒 E2基因 序列分析 同源性 氨基酸变异 广西 

分 类 号：S858.28[农业科学—临床兽医学]