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

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作  者:赵玉婷 谢洋洋[1] 周书亭 丁思嘉[1] 蔡雨豪 周艳君[3] 杨志彪[1] 

机构地区:[1]上海交通大学农业与生物学院,上海200240 [2]上海海洋大学水产与生命学院,上海201306 [3]中国农业科学院上海兽医研究所,上海200240

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)

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.

关 键 词:猪流行性腹泻病毒 S基因 真核表达 

分 类 号:S852.65[农业科学—基础兽医学]

 

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