CAJ | 학술논문

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.
목적 구건신형호장병독주요항원단백σ1단백적진핵표체질립,연구기재진핵세포내적표체.방법 장S1기인극륭입진핵표체재체pCAGGS/MCS,구건진핵표체질립pC-S병전염Vero세포.통과SDS-PAGE화Western-Blot시험,대전염후24,48급72 h적세포내단백표체진행연구.결과 매절분석표명중조질립구건성공.SDS-PAGE화Western-Blot적검측결과일치표명,전염후S1기인가재Vero세포내표체차72 h적세포내단백표체량최고.결론 통과구건중조진핵표체질립,가사S1기인재진핵세포내고효표체,위진일보연구신형호장병독여숙주수체적상호작용타하기출.
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.