欧洲型PRRSV N蛋白特异性抗原表位基因串联表达及其产物的纯化  

Tandem expression and purification of the specific antigen epitope of N protein of EU-PRRSV

作　　者：李冰[1,2] 唐峰[2] 卢赫[2] 冯方周 丁壮[1] 

机构地区：[1]吉林大学动物医学院,吉林长春130062 [2]辽宁医学院畜牧兽医学院,辽宁锦州121001

出　　处：《中国兽医学报》2015年第1期1-4,35,共5页Chinese Journal of Veterinary Science

基　　金：辽宁省教育厅科研项目(L2014327)

摘　　要：采用SOE-PCR方法将编码欧洲型猪繁殖与呼吸综合征病毒(PRRSV)N蛋白的aa2~12和aa40~46核苷酸片段进行重复串联,获得编码双表位重复串联基因NE,构建表达载体pGEX-4T-1-NE,将其转入BL21(DE3)pLysS菌中,获得融合蛋白。Western-blot检测结果表明,该融合蛋白可被欧洲型PRRSV阳性血清识别,而不能被美洲型PRRSV阳性血清、猪瘟病毒阳性血清和伪狂犬病病毒阳性血清识别。纯化所得的融合蛋白,为建立检测欧洲型PRRSV提供了抗原基础。The epitope areas of EU-PRRSV N aa2-12 and aa40-46 were amplified and fused repeatedly by gene SOEing PCR technique.The new genes(NE)encoding the mulitepitope peptide were generated.Expression vector pGEX-4T-1-NE was constructed,turned into BL21(DE3)pLysS cells and inducted under the IPTG and obtained the fusion protein.Detection of expressed protein by western-blot showed that the prokaryotic expression of the fusion protein can be identified by the positive serum samples of EU-PRRS,except NA-PRRS,swine fever and porcine pseudorabies.The result indicated that the purification of fusion protein provided an antigen basis for detecting Europe type PRRSV.

关 键 词：欧洲型PRRSV N蛋白 特异性抗原表位 原核表达 纯化 

分 类 号：S852.65[农业科学—基础兽医学]