表达高致病性猪蓝耳病病毒GP5重组伪狂犬病病毒的构建及鉴定  

Construction of recombinant pseudorabies virus expressing GP5 of highly pathogenic PRRSV HuN4 strain

作　　者：陈瑾[1] 田志军[1] 彭金美[1] 王瑜[2] 周艳君[1] 安同庆[1] 童光志[1,3] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/猪传染病研究室,黑龙江哈尔滨150001 [2]华中农业大学动物医学学院,湖北武汉430075 [3]中国农业科学院上海兽医研究所,上海200232

出　　处：《中国预防兽医学报》2009年第1期20-23,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家“973”计划(2005CB523200);十一五国家科技支撑计划(2006BAD06A04/18)

摘　　要：将PCR扩增的高致病性猪蓝耳病病毒(HP-PRRSV)HuN4株GP5基因与含绿色荧光蛋白(EGFP)基因的表达盒插入到伪狂犬病毒通用转移载体pgG-uni中,构建了转移载体pgG-HPGP5-EGFP。通过磷酸钙法将转移载体pgG-HPGP5-EGFP和PRV Bartha-K61株的基因组共转染Vero细胞,经过8轮荧光蚀斑筛选和PCR鉴定获得了稳定表达EGFP的重组病毒,命名为rPRV-HPGP5。IFA和western blot结果证实HP-PRRSV HuN4株GP5在重组病毒中得到了有效表达。该重组病毒的获得为高致病性猪蓝耳病基因工程疫苗的研究奠定了基础。In this study, the GP5 gent of highly pathogenic porcine reproductive and respiratory syndrome virus （HP-PRRSV） HuN4 strain and EGFP gene were inserted in to the pgG-uni downstream of pseudorabies virus （PRV） gG promoter to construct the transfer vector pgG-HPGP5-EGFP. The purified genomic DNA of Bartha-K61 strain and pgG-HPGP5-EGFP were co-transfected into Vero cells by calcium phosphate procedure. A recombinant virus stably expressing EGFP gene was obtained after eight cycles of fluorescence plague purification and PCR identification. Western blot and IFA assays confirmed that HP-PRRSV GP5 protein had been expressed in recombinant virus. The potential of the recombinant virus as a genetically engineering vaccine against HP-PRRS was discussed.

关 键 词：高致病性蓝耳病病毒 HUN4 GP5 伪狂犬病病毒 重组 

分 类 号：S852.659.6[农业科学—基础兽医学] S858.28[农业科学—兽医学]