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

doi:10.15933/j.cnki.1004-3268.2021.06.015

Journal of Henan Agricultural Sciences ›› 2021, Vol. 50 ›› Issue (6): 125-133.DOI: 10.15933/j.cnki.1004-3268.2021.06.015

• Animal Husbandry and Veterinary Medicine • Previous Articles Next Articles

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

HUANG Zhengyang¹,CHEN Yujing¹,HUANG Ke¹,LENG Chaoliang^1,2,3,LIU Yangkun^1,2,3,YAO Lunguang^1,2,3,LI Na^1,2,3

(1.School of Life Science and Agricultural Engineering,Nanyang Normal University,Nanyang 473061,China;2.Henan Provincial Engineering and Technology Center of Health Products for Livestock and Poultry,Nanyang 473061,China;3.Henan Provincial Engineering and Technology Center of Animal Disease Diagnosis and Integrated Control,Nanyang 473061,China)

Received:2020-11-26 Published:2021-06-15 Online:2021-06-15

猪A组轮状病毒HN2019-01株的分离鉴定及VP6基因序列分析

黄正阳¹，陈宇婧¹，黄克¹，冷超粮^1,2,3，刘阳坤^1,2,3，姚伦广^1,2,3，李娜^1,2,3

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)

通讯作者: 李娜（1983-），女，河南南阳人，讲师，博士，主要从事基因工程疫苗与动物疾病诊断研究。E-mail:lina330077@163.com
作者简介:黄正阳（1995-），男，河南南阳人，在读硕士研究生，研究方向：基因工程疫苗与动物疾病诊断。E-mail:15090135771@163.com
基金资助:
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）

Abstract

Abstract: 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.

Key words: Rotavirus(RV), RVA, Virus isolation, VP6 gene, Sequence analysis

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。

关键词: 轮状病毒, 组轮状病毒, 病毒分离, VP6基因, 序列分析

CLC Number:

S855.3

HUANG Zhengyang, CHEN Yujing, HUANG Ke, LENG Chaoliang, LIU Yangkun, YAO Lunguang, LI Na. Isolation and VP6 Gene Sequence Analysis of Porcine Group A Rotavirus HN2019-01 Strain[J]. Journal of Henan Agricultural Sciences, 2021, 50(6): 125-133.

黄正阳, 陈宇婧, 黄克, 冷超粮, 刘阳坤, 姚伦广, 李娜. 猪A组轮状病毒HN2019-01株的分离鉴定及VP6基因序列分析[J]. 河南农业科学, 2021, 50(6): 125-133.

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

猪A组轮状病毒HN2019-01株的分离鉴定及VP6基因序列分析

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