华北农学报 ›› 2015, Vol. 30 ›› Issue (1): 137-141. doi: 10.7668/hbnxb.2015.01.022

所属专题: 畜牧 生物技术

• 论文 • 上一篇    下一篇

猪伪狂犬病病毒河南分离株gE全基因的克隆与序列分析

高晓云1, 顾阳1, 潘鑫龙1, 郭小参2, 崔保安1, 陈红英1   

  1. 1. 河南农业大学 畜牧兽医工程学院, 河南 郑州 450002;
    2. 普莱克生物工程股份有限公司, 河南 洛阳 471000
  • 收稿日期:2014-11-10 出版日期:2015-02-28
  • 通讯作者: 陈红英(1965-),女,四川仁寿人,教授,博士,主要从事分子免疫学和分子病毒学研究。
  • 作者简介:高晓云(1988-),女,河南新县人,在读硕士,主要从事分子免疫学和分子病毒学研究。
  • 基金资助:
    河南省重大科技专项(111100110300)

Cloning and Sequence Analysis of Complete Genome of gE of Pseudorabies Virus Isolates from Henan

GAO Xiao-yun1, GU Yang1, PAN Xin-long1, GUO Xiao-can2, CUI Bao-an1, CHEN Hong-ying1   

  1. 1. College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China;
    2. Pulike Biological Engineering, INC., Luoyang 471000, China
  • Received:2014-11-10 Published:2015-02-28

摘要: 为了确定河南部分地区按照正常免疫程序接种猪伪狂犬病病毒(PRV)基因缺失苗的猪场是否发生了PRV野毒感染,了解新流行株的主要毒力基因gE是否发生变异,利用PCR方法,对南阳、周口、巩义、济源、漯河、原阳等地采集的疑似PRV感染的病料进行检测。对PCR检测为阳性的病料接种猪睾丸细胞,传至6代,分离出9株PRV,克隆其gE全基因并进行序列分析。结果表明,9株PRV均能扩增出1 862 bp的gE基因,且彼此之间同源性为99.9% ~100%,与其他PRV毒株同源性为97.5% ~99.6%。9株分离株处于一大分支上,且均含有2个氨基酸插入,在448位和510位均发生氨基酸置换。结果表明,分离株均为PRV野毒株,且gE基因有不同程度的变化,与河南省伪狂犬病的重新流行有密切联系。

关键词: 猪伪狂犬病毒, gE基因, 克隆, 序列分析

Abstract: The purpose of this study was to determine whether Pseudorabies virus (PRV) infection caused by wild strain occurred at the farms where PRV gene deletion vaccine was inoculated under normal immune procedure in some areas of Henan Province and whether variation had occurred in gE gene which was one of the most important virulence genes of PRV.Samples from swine with suspicious PRV infection were collected from Nanyang, Zhoukou and other regions of Henan and detected by PCR.PRV-positive samples were inoculated into swine testis cells and sub-cultured six times.The complete genomes of gE of 9 PRV strains were cloned, sequenced and analyzed.The result showed that the length of gE genomes of the 9 strains were 1 862 bp.The homologies between the 9 strains were 99.9% -100% and the homologies with other strains were 97.5% -99.6%.Phylogenetic tree showed the 9 PRV strains belonged to a relatively independent sub-branch, and contained 2 amino acid insertions.The replacement of amino acid occurred at the locus of 448 and the locus of 510.It indicated that all of the isolated PRV strains were wild isolates, which were related with the new prevalence.The variation had occurred in gE gene of these isolates.

Key words: Pseudorabies virus, gE gene, Cloning, Sequence analysis

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引用本文

高晓云, 顾阳, 潘鑫龙, 郭小参, 崔保安, 陈红英. 猪伪狂犬病病毒河南分离株gE全基因的克隆与序列分析[J]. 华北农学报, 2015, 30(1): 137-141. doi: 10.7668/hbnxb.2015.01.022.

GAO Xiao-yun, GU Yang, PAN Xin-long, GUO Xiao-can, CUI Bao-an, CHEN Hong-ying. Cloning and Sequence Analysis of Complete Genome of gE of Pseudorabies Virus Isolates from Henan[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2015, 30(1): 137-141. doi: 10.7668/hbnxb.2015.01.022.

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