【摘要】 为重组表达猪繁殖与呼吸综合征病毒(PRRSV)M蛋白,本研究将RT-PCR获得的PRRSV CH-1a株M蛋白基因,克隆于pMD18-T载体中,经测序鉴定正确后,亚克隆至牛痘病毒重组转移质粒pSC11中,构建了转移重组质粒pSC11-PRRSV-M。将pSC11-PRRSV-M在脂质体的介导下转染WR株牛痘病毒感染的TK-143细胞,在含有X-gal的琼脂培养基上通过蓝斑筛选含有PRRSV M基因的重组病毒rWR-PRRSV-M。Western blot与IFA检测表明,重组病毒成功表达了PRRSV M蛋白,而且所表达的蛋白保持了良好的免疫原性。动物实验表明,rWR-PRRSV-M所表达的PRRSV M蛋白在免疫小鼠体内诱生了抗PRRSV的抗体。rWR-PRRSV-M的构建,为进一步探讨PRRSV M蛋白免疫原性提供了基础数据,也为PRRS重组活载体疫苗的研究奠定了基础。更多还原

【Abstract】 In this study,the M gene of Porcine reproductive and respiratory syndrome virus(PRRSV) CH-1a strain was amplified from the PRRSV genome by RT-PCR and cloned into the plasmid pSC11.The recombinant plasmid pSC11-PRRSV-M was transfected into TK-143 cells infected with vaccinia virus WR strain.Recombinants vaccinia virus of rWR-PRRSV-M was obtained by selecting blue plaques on the TK-143 cells overlaid with agar containing X-gal,and confirmed by PCR.The M gene expressed in TK-143 cells were identified by SDS-PAGE and confirmed by immunofluorescence assay(IFA).The rWR-PRRSV-M recombinant virus could induce specific humoral immune responses against PRRSV in the immunized mice.更多还原