禽流感病毒M_(2e)基因与鸡IgG Fc基因在大肠杆菌中的融合表达和纯化  

Expression and Purification of Fusion Protein of Avain Influenza Virus Matrix 2 protein Ectodomain Segments(M_(2e)) and Avain IgG Fc in E.coli BL21

作　　者：蔡家利[1] 余华[2] 胡仁建[1] 于丽[2] 刘海军[1] 李静远[1] 叶芬[2] 

机构地区：[1]重庆理工大学,重庆400050 [2]西南大学动物科技学院,重庆400715

出　　处：《重庆理工大学学报（自然科学）》2010年第1期38-41,共4页Journal of Chongqing University of Technology：Natural Science

摘　　要：将含禽流感病毒M2蛋白膜外区(M2e)且偶联了鸡IgG Fc片段的重组质粒pET32a(+)的大肠杆菌,用终浓度为0.5 mmol/LIPTG在37℃下诱导表达,获得了包涵体表达的目的蛋白。Western-blotting试验表明,M2e能与兔抗禽流感H5阳性血清发生反应,且在47kD处出现一条特异性条带,具有良好的免疫学活性.表达产物通过His-Ni2+柱亲和层析后,得到了较好的纯化效果。本实验为进一步研究M2蛋白的生物学活性、检测方法及通用疫苗的开发奠定了基础。The purpose protein of inclusion body expression is obtained by conducting inducible expression on plasmid pErlB2a of E. coli BL21, which contains avian influenza virus matrix2 protein eetodomain segments （ M2e ） and couples avian IgG Fc segment, under the condition of IPTG of 0. 5mmol/L at 37℃. Western-blotting demonstrates that M2e, which can react with the rabbit anti-AIV H5 positive serum, and produces a special band in the 47kD area, and possesses good antigenicity. The fusion protein is purified by His-Ni2 ＋ affinity chromatography and good purification result is obtained. This paper lays a foundation for the further research of M2 protein immunological characteristics and the development of universal vaccines and detection methods.

关 键 词：禽流感病毒 M2e蛋白 pET32a(+)原核表达系统 纯化 

分 类 号：R392.1[医药卫生—免疫学]