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猪流行性腹泻是由猪流行性腹泻病毒(porcine epidemic diarrhea virus,PEDV)引起的一种可致仔猪严重腹泻的高度接触性肠道传染病。S蛋白在病毒感染和入侵宿主细胞的过程中具有重要作用。为了研究S蛋白的特点,本研究扩增了PEDV新流行毒株SHpd/2012的纤突蛋白S基因分段片段S11、S12、S13、S21和S22,将得到的目的片段导入原核表达载体pGEX-6P-1中,并转化到大肠杆菌BL21(DE3)感受态细胞中进行表达、鉴定,获得了正确的PEDV S的分段蛋白S11(24-277aa)、S12(272-571aa)、S13(560-797aa)、S21(793-1109aa)及S22(1102-1384aa),为后续单克隆抗体的制备和致病机制的研究奠定了基础。
Swine porcine epidemic diarrhea is a highly contagious enteropathy caused by porcine epidemic diarrhea virus (PEDV) that causes severe diarrhea in piglets. S protein plays an important role in viral infection and invasion of host cells. In order to study the characteristics of S protein, we amplified S11, S12, S12, S13, S21 and S22 fragments of the spike protein S gene of the new epidemic strain SHPd / 2012 in PEDV. The resulting fragment was introduced into prokaryotic expression vector pGEX- 6P-1 and transformed into E. coli BL21 (DE3) competent cells for expression and identification. The correct PEDV S fragment proteins S11 (24-277aa), S12 (272-571aa), S13 (560 -797aa), S21 (793-1109aa) and S22 (1102-1384aa), which laid the foundation for the preparation of the next monoclonal antibody and the study of pathogenic mechanism.