美洲型与欧洲型PRRSV特异N蛋白抗原表位基因表达及其产物的纯化  

Expression of Specific N Protein Epitope Genes of European and American-Porcine reproductive and respiratory syndrome virus and Purification of recombinant Protein

作　　者：丛晓燕[1] 李俊[2] 吴家强[2] 张祯涛[3] 赵鸿雁[1] 曹帅[1] 王文志[2] 王金宝[2] 

机构地区：[1]莱阳农学院动物科技学院,山东青岛266109 [2]山东省农科院畜牧兽医研究所,山东济南250100 [3]山东省畜牧兽医总站,山东济南250100

出　　处：《动物医学进展》2006年第11期76-80,共5页Progress In Veterinary Medicine

摘　　要：对原核中以包涵体形式高效表达的美洲型与欧洲型PRRSV特异N蛋白抗原表位区段进行了变性、复性与纯化研究。将美洲型PRRSV重组质粒pGEX-6P-1-N转化入E.coliBL21(DE3)细胞中,成功表达了重组蛋白GST-N,超声裂解复性后目的蛋白经GSTrap FF纯化试剂盒纯化后,纯度可达90%以上。将欧洲型PRRSV重组质粒PQE30-N转化入E.coliM15细胞中表达了重组蛋白His-N。超声裂解后目的蛋白经HisTrapHP蛋白纯化试剂盒纯化后,纯度可达95%以上。Western blot结果表明,两种蛋白分别被美洲型和欧洲型PRRSV阳性血清识别。纯化出的大量美洲型与欧洲型PRRSV特异N蛋白抗原表位区段,为PRRSV的鉴别诊断提供了抗原基础。A US PRRSV recombinant plasmid pGEX-6P-1-N was constructed and transformed into the E. coli BL21（DE3）. The recombinant fusion protein GST-N was highly expressed in E. coli. The purity of fusion proteins affinity-purified using GSTrap FF Kit was above 95%. A EU PRRSV recombinant plasmid PQE30-N was constructed and transformed into the E. coli M15. The recombinant fusion protein HIS-N was highly expressed in E. coll. The purity of fusion proteins affinity-purified using HisTrapHP Kit was above 95%. Western-blot results showed that the both fusion proteins could react with the antibodies against US and EU PRRSV, respectively. The purified antigens allows the simultaneous detection and differentiation of antibodies against US and EU PRRSV.

关 键 词：猪繁殖与呼吸综合征病毒 美洲型 欧洲型 N蛋白抗原多肽 原核表达 纯化 

分 类 号：S852.659.6[农业科学—基础兽医学]