表达猪繁殖与呼吸综合征病毒GP3基因的重组伪狂犬病病毒的构建

猪繁殖与呼吸综合征 (porcinereproductiveandrespiratorysyndrome ,PRRS)是引起怀孕母猪早产、流产、死胎及仔猪呼吸系统疾病的一种新发现的病毒性传染病[1] .该病毒的基因组为单股正链RNA ,约15kb ,含有 8个开放阅读框架 (ORFs) ,ORF1编码病毒非结构蛋白 (依赖RNA的RNA聚合酶 ) ,ORF2 ORF7编码病毒的结构蛋白 .其中ORF3含有 2 6 5个氨基酸 ,编码的GP3蛋白为高度糖基化的结构蛋白 ,有 7个糖基化位点 ,具有免疫原性[2 ,3 ] .目前 ,用于预防PRRS的疫苗主要是弱毒苗和灭活苗 ,虽然都有一定的免疫效果 ,但由于PRRS抗体依赖性…

Construction of the Recombinant Pseudorabies Virus Expressing the GP3 Gene of Porcine Reproductive and Respiratory Syndrome Virus

Based on the nucleotide sequence of porcine reproductive and respiratory syndrome virus (PRRSV) CH 1a strain, a pair of primers was designed. The GP3 gene of PRRSV HB 1 strain was cloned by RT PCR. The sequences analysis showed that the GP3 genes of HB 1 and CH 1a strain had 91% and 89% homology, at the levels of the nucleotide and amino acid, respectively. The GP3 gene was digested by Bam HⅠ and Eco RⅠ, and the fragment was inserted into the same sites of the universal vector pPgG uni. A recombinant virus transfer vector pPgG GP3 expressing PRRSV GP3 gene was constructed. The transfer vector pPgG GP3 digested by Kpn Ⅰ was co transfected PK 15 cells with the PRV TK -/gG -/LacZ + genomic DNA digested by Eco RⅠ using liposome method. The recombinant virus was purified by the phage test and PCR amplification. The expression of GP3 gene was indicated by Western blot analysis using anti sera. A recombinant PRV virus expressing PRRSV GP3 gene was obtained. The recombinant virus is very useful in the research of the genetic engineering vaccine against pseudorabies virus and PRRS virus.