新型呼肠病毒S1基因真核表达质粒的构建及表达

目的构建新型呼肠病毒主要抗原蛋白盯1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基因克隆人真核表达载体pCAGGS／MCS,构建真核表达质粒pc—s并转染~ero细胞。通过SDS—PAGE和Western—Blot试验,对转染后24,48及72h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72h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。

Construction of recombinant plasmid expressing S1 gene of new type of reovirus

Objective 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.