摘要目的 构建新型呼肠病毒主要抗原蛋白σ1蛋白的真核表达质粒,研究其在真核细胞内的表达.方法 将S1基因克隆入真核表达载体pCAGGS/MCS,构建真核表达质粒pC-S并转染Vero细胞.通过SDS-PAGE和Western-Blot试验,对转染后24,48及72 h的细胞内蛋白表达进行研究.结果 酶切分析表明重组质粒构建成功.SDS-PAGE和Western-Blot的检测结果一致表明,转染后S1基因可在Vero细胞内表达且72 h的细胞内蛋白表达量最高.结论 通过构建重组真核表达质粒,可使S1基因在真核细胞内高效表达,为进一步研究新型呼肠病毒与宿主受体的相互作用打下基础.

abstractsObjective To construct the recombinant plasmid containing S1 gene of new type of reovirus,and to study the expression of protein σ1 in Vero cells.Methods The recombinant plasmid,named pC-S,was constructed by cloning S1 gene into vector pCAGGS/MCS.Then Vero cells were transfected with pC-S and collected at 24,48,72 h post transfection followed by SDS-PAGE and Western-Blot assay.Results Results Both SDS-PAGE and Western-Blot assay indicated that σ1 protein could be expressed well and the highest expression level was 72 h post transfection.Conclusions σ1 protein could be expressed well in Vero cells by transfected with recombinant plasmid containing S1 gene,and could give some implications for subsequent research on virus-host interactions.