重组PEDV-S干酪乳杆菌的构建及其表达

Cloning and Expression of PEDV-S on Lactobacillus casei

猪流行性腹泻病毒(PEDV)可引起猪的呕吐、腹泻及脱水。通过将其S1片段逆转录连接到干酪乳杆菌表面表达载体p LA上,使其表达蛋白,为进一步研究其特性及免疫原性奠定基础。采用TRIzol的方法从PEDV中提取RNA为模板,通过RT-PCR技术获得该病毒的S1片段,然后将该基因连接到载体p LA中并表达。重组菌经过培养表达之后,利用Western blot检测融合蛋白大小约为81 k Da;重组蛋白可被兔源的抗Pgs A血清和鼠源的抗PEDV血清所识别。表明重组干酪乳杆菌成功的表达了融合蛋白。

Porcine epidemic diarrhea（PEDV） can cause vomit,diarrhea and dehydration. The gene fragments encoding PEDV S1 protein was cloned into Lactobacillus casei surface expression vector p LA to express the protein,which laid the foundation for further study of characteristics and immunogenicity. By TRIzol method of extracting RNA from PEDV template,it got the virus by RT-PCR technique of S1 fragments,and then connected the gene to carry and express in the p LA. After expressing recombinant bacteria culture,the results showed that the molecular weight of about 81 k Da was detected. According to the result of Western-blot,the expressed protein possessed the antigenic specificity,which could be recognized by Rabbit anti-Pgs A and Mouse anti-PEDV serum.The recombinant strains were successfully induced to express protein.