猪瘟病毒T细胞表位与猪细小病毒VP2融合基因在重组杆状病毒中的表达

将猪细小病毒分离株LJL12 VP2基因和猪瘟病毒多肽基因E290克隆至真核表达载体pMel Ba-cA,将该重组质粒与Bac-N-Blue DNA共转染昆虫细胞,并对表达产物进行分析,构建了表达猪瘟病毒T细胞表位和猪细小病毒VP2融合蛋白的重组杆状病毒。结果显示,获得了含VP2基因和E290基因的重组质粒pM-VP2-E290,昆虫细胞sf9的表达产物经SDS-PAGE、Western-blot检测后确定所表达的蛋白质分子质量大小约为67 ku,且具有天然蛋白的抗原特性。免疫电镜观察可见表达产物形成的病毒样颗粒。证实,成功构建了同时含有PPV VP2基因和CSFV特异性T细胞表位基因的重组杆状病毒,并在昆虫细胞中得到了高效表达。

Expression of fusion gene consisting of T cell antigen epitope gene of classical swine fever virus and VP2 protein gene of porcine parvovirus in recombinant baculovirus

To construct recombinant baculovirus expressing fusion gene of T cell antigen epitope gene of classical swine fever virus(CSFV) with VP2 protein gene of porcine parvovirus(PPV),the VP2 gene of PPV isolate LJL12 and E290 gene of CSFV were co-cloned into pMel BacA vector.Then,the recombinant plasmid and Bac-N-Blue DNA were co-transfected into sf9 cells.The expressed fusion protein was analyzed by SDS-PAGE and Western-blotting.The expressed product which was approximately 67 ku in size reacted strongly with the PPV-specific antibody.The virus-like particles formed by expressed product were observed using an immunoelectron microscopy.The results confirmed that the recombinant baculovirus containing T cell antigen epitope gene of CSFV and VP2 protein gene of PPV was constructed successfully and expressed efficiently in sf9 cells.