非洲猪瘟病毒K205R蛋白基因原核表达的研究  

Prokaryotic Expression of K205R Gene of African Swine Fever Virus

作　　者：龚振华[1] 莫正芬 李葳[1] 李玉清[1] 刘开成[1] 史红[1] 刘国庆[1] 王君玮[1] 蒋正军[1] 王志亮[1] 

机构地区：[1]中国动物卫生与流行病学中心,山东青岛266032 [2]湖南桃江武潭镇中心,湖南桃江413407

出　　处：《中国动物检疫》2010年第12期35-36,共2页China Animal Health Inspection

基　　金：公益性行业科技专项-非洲猪瘟等重大外来动物疫病防控技术研究支持(200903037)

摘　　要：本研究合成非洲猪瘟的K205R基因,将K205R基因克隆至pET-30c(+)质粒,构建重组原核表达质粒pET30C-K205R。该重组质粒的阅读框架正确,表达的融合蛋白的N端和C端均带6×组氨酸。pET30C-K205R-BL21菌株经培养和诱导表达后,可生产K205R可溶性蛋白,K205R可溶性蛋白可用Ni柱纯化。纯化表达K205R蛋白的ELISA试验证明,表达的K205R蛋白与猪阳性非洲猪瘟血清具有较好的特异性反应。K205R gene of AFSV was synthesized and cloned to the plasmid pET30C to construct the recombinant prokaryotic expressed plasmid pET30C-K205R with correct reading frame and with 6x histamine both on N and C terminals of the expressed fusion protein. The recombinant plasmid pET30C-K205R was then transformed into E.coli DH5c~ and then into BL21 and the pET30C-K205R-BL21 strain was obtained and expressed by induction with IPTG, resulting in soluable protein K205R. ELISA with the purified K205R protein showed that the expressed K205R protein demonstrated very good specific reaction with positive ASFV serum.

关 键 词：非洲猪瘟病毒 K205R蛋白 原核表达 

分 类 号：S852.659.1[农业科学—基础兽医学]