摘要
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48h后的表达量较高;其中S1蛋白分子量约为90kDa,S2蛋白分子量约为60kDa。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.
出处
《上海交通大学学报（农业科学版）》
2017年第5期69-73,共5页
Journal of Shanghai Jiaotong University(Agricultural Science)
基金
国家重点研发计划(2016YFD0500100)
国家自然科学基金(31472211)
上海市自然基金(14ZR1421400)
关键词
猪流行性腹泻病毒
S基因
真核表达
porcine epidemic diarrhea virus
S gene
eukaryotic expression
