猪流行性腹泻病毒SHpd/2012株S1和S2基因片段的真核表达
Eukaryotic Expression of S1 and S2 Gene Fragments from SHpd/2012 Strain of Porcine Epidemic Diarrhea Virus

为探究猪流行性腹泻病毒(PEDV)的新毒株SHpd/2012在感染细胞时与细胞的相互作用机制,根据已得到的序列设计引物,克隆了S1和S2基因片段,构建了pCAGGS-S1和pCAGGS-S2真核表达质粒。间接免疫荧光和Western-blot实验表明,转染后S1和S2都能在Vero细胞内表达,且转染48 h后的表达量较高;其中S1蛋白分子量约为90 kDa,S2蛋白分子量约为60 kDa。流行毒株SHpd/2012的S1与S2两肽段的成功表达,为之后进一步研究PEDV与Vero细胞的相互作用奠定了基础。
S1 and S2,fragments of S gene in SHpd/2012 strain of porcine epidemic diarrhea virus (PEDV),were cloned and their eukaryotic expression plasmids,namely pCAGGS-S1 and pCAGGS-S2,were constructed to investigate the mechanism of the interaction between SHpd/2012 strain of porcine epidemic diarrhea virus and Vero cells.Indirect immunofluorescence and Western-blot assay results showed that S1 and S2 could be expressed well in Vero cells and the amount of peptides were high 48 h after transfection.The molecular weights of S1 and S2 peptide were 90 and 60 kDa,respectively.The successful expression of the S1 and S2 proteins of the new strain SHpd/2012 laid the foundation for the further study of the interaction between PEDV and Vero cells.