猪A组轮状病毒HN2019-01株的分离鉴定及VP6基因序列分析  

Isolation and VP6 Gene Sequence Analysis of Porcine Group A Rotavirus HN2019-01 Strain

作　　者：黄正阳 陈宇婧 黄克 冷超粮[1,2,3] 刘阳坤 姚伦广 李娜[1,2,3] HUANG Zhengyang;CHEN Yujing;HUANG Ke;LENG Chaoliang;LIU Yangkun;YAO Lunguang;LI Na(School of Life Science and Agricultural Engineering,Nanyang Normal University,Nanyang 473061,China;Henan Provincial Engineering and Technology Center of Health Products for Livestock and Poultry,Nanyang 473061,China;Henan Provincial Engineering and Technology Center of Animal Disease Diagnosis and Integrated Control,Nanyang 473061,China)

机构地区：[1]南阳师范学院生命科学与农业工程学院,河南南阳473061 [2]河南省畜禽保健品工程技术研究中心,河南南阳473061 [3]河南省动物疫病诊断与综合防控工程技术研究中心,河南南阳473061

出　　处：《河南农业科学》2021年第6期125-133,共9页Journal of Henan Agricultural Sciences

基　　金：国家自然科学基金面上项目(31870917);国家自然科学基金项目(32002302);河南省高等学校重点科研项目(18A230011);河南省科技攻关项目(182102310077);河南省“疫苗工程”高校科技创新团队项目(20IRTSTHN024);南阳师范学院高层次人才启动专项(2018ZX011)。

摘　　要：为了解猪轮状病毒(RV)在河南省的流行情况及其分子遗传特征,利用反转录PCR方法,从河南某猪场腹泻样品中扩增得到RV特异性核酸片段,并将腹泻样品利用MA-104细胞进行RV的分离和传代,将分离得到的RV命名为RVA/Pig-wt/HN2019-01。对分离毒株VP6基因片段的序列测定及生物信息学分析结果表明,分离毒株VP6核苷酸序列与猪源毒株西南株-1C(SWU-1C)相似性最高,病毒分类上同属I5亚群。VP6蛋白全长397个氨基酸,存在4个N糖基化位点和18个潜在磷酸化位点,二级结构主要由α-螺旋组成,三级空间结构与牛轮状病毒RF株VP6蛋白高度相似,且在2个细胞毒性T淋巴细胞(CTL)抗原表位区域的氨基酸序列保守。To investigate the prevalence and molecular genetic characteristics of porcine rotaviruses(RV)in Henan,the positive diarrheal stool samples of RV from a pig farm were examined using reverse transcription-polymerase chain reaction(RT-PCR).A novel strain of RV,RVA/Pig-wt/HN2019-01,was isolated and passaged by MA-104 cells.VP6 gene amplification and bioinformatics analysis were performed to further characterize the RV isolated strain at the molecular level.As the result of VP6 gene amplification and sequence analysis,the RVA/Pig-wt/HN2019-01 showed the highest nucleotide identity with the previously reported porcine RV SWU-1C strain,which both were clustered to the I5 subgroup.The VP6 protein composed of 397 amino acids in length,possessing 4 N-glycosylation and 18 potential phosphorylation sites.Secondary structure of VP6 protein was found to be mainly alpha helix-based,and the spatial structure was highly similar to the VP6 of bovine rotavirus RF strain.Two cytotoxic T lymphocyte(CTL)epitopes regions of rotavirus VP6 were conserved among different strains.

关 键 词：轮状病毒 A组轮状病毒 病毒分离 VP6基因 序列分析 

分 类 号：S855.3[农业科学—临床兽医学]