猪流行性腹泻病毒SHpd/2012株S蛋白的表达鉴定
Expression and Identification of S Protein of Porcine Epidemic Diarrhea Virus SHpd/2012

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。
PED (porcine epidemic diarrhea) is a highly contagious and infectious disease of piglets with diarrhea caused by PEDV (porcine epidemic diarrhea virus,PEDV).The disease can spread to a wide range,and has a high level of morbidity and mortality rate,which is the most serious and harmful type of viral diarrhea in the world,and causes huge economic losses to the pig industry and farmers.However,the pathogenic mechanism of new strains is unclear.In addition,due to the changes of host immune selection and virus evolution,S protein is prone to mutation.In order to study the characteristics and functions of S protein,we amplified five fragments (namely S11,S12,S13,S21 and S22) of the spike protein (S) gene of the new-found strain SHpd/2012 of PEDV,cloned into pGEX-6P-1 and then transformed into Escherichia coli BL21 (DE3) competent cells for expression.The PEDV’s S11(24-277aa),S12(272-571aa),S13(560-797aa),S21(793-1109aa) and S22(1102-1384aa) peptides were obtained and confirmed by Western blot.Our work provided the basis for the further study on preparation of monoclonal antibody and pathogenesis.