猪瘟病毒E2基因和牛疱疹病毒Ⅰ型UL49基因的共表达

为了探讨牛疱疹病毒Ⅰ型(BHV-1)UL49基因编码的膜蛋白VP22对猪瘟病毒E2基因表达水平的影响,构建了单独表达E2蛋白的pcDNA3.1-E2及融合表达E2-VP22的pcDNA3.1-E2-UL49。将pcDNA3.1-E2与pcDNA3.1-E2-UL49分别转染293 T细胞,间接免疫荧光试验结果显示,单独表达E2蛋白的细胞,其荧光主要固缩于核周,而融合表达E2-VP22蛋白的细胞在核周及细胞质内都有大量的荧光;Western-blot分析结果显示,VP22蛋白与E2蛋白都获得了表达,但融合了VP22蛋白的E2蛋白表达量要相对高一些(P<0.01),表明VP22蛋白可以促进E2蛋白的表达。

Co-expression of classical swine fever virus E2 gene with bovine herpesvirus 1 UL49 gene

In order to investigate the influence of VP22 protein on expression of E2 gene in vitro,two recombinant eukaryotic expression vectors,pcDNA3.1-E2 and pcDNA3.1-E2-UL49,were constructed by inserting E2 gene and E2-VP22 protein gene into pcDNA3.1( ),respectively.The expressed products in 293 T cells transfected with pcDNA3.1-E2 and pcDNA3.1-E2-UL49 were detected by immunofluorescent antibody test and Western-blotting.The cells containing pcDNA3.1E2 were IFA-positive to CSFV antisera and the expressed protein E2 was found around the nucleus,but the expressed fusion protein E2-VP22 was located both in the cytoplasm and around the nucleus.Western-blotting analysis indicated that both the protein E2 and the fusion protein E2-VP22 could react with CSFV specific antibodies and the E2-VP22 could also react with BHV-1 specific antibody.The expression level of the E2-VP22 protein gene was(higher) than that of the E2 gene(P<0.01).The results suggested that the VP22 protein had the function of protein transduction.