牛分枝杆菌外膜蛋白基因ompA的原核表达及免疫活性分析.pdfVIP

2009, 17(4) :43- 46 Chinese Journal of Animal Infectious iseases omp A 1 2 1 1 1 1 1 1 1 亓英芳 , 黄 瑛 , 杜艳芬 , 刘慧芳 , 赫明雷 , 司微 , 倪洪波 , 王春来 , 刘思国 (1. , 150001; 2. ,831100) : 为验证牛分枝杆菌外膜蛋白A 是否具有免疫活性, 以牛分枝杆菌Vallee 株基因组 NA 为模板扩增牛 分枝杆菌外膜蛋白基因omp A , 采用 NA 组技术将此基因片段连于表达载体 pGEX6p1, 构建 组质粒 pGEX6p1ompA, 并将其转化到大肠杆菌中BL21( E3) 中, IPTG 诱导(终浓度为 01mmol/ L) , 表达产物进行 S SPAGE 分析用glutathione sepharose 4B 纯化蛋白和Western blot 分析该蛋白的免疫学活性结果表明: pGEX6p1ompA 以可溶形式表达, 蛋白分子量大小为60 k a, 与预计大小相符, 纯化的蛋白具有良好的免疫学 活性 : 牛分枝杆菌;外膜蛋白ompA; 原核表达; 纯化; Western blot : S858.232.618:Q753 : A : 2009 ANALYSIS OF IMMUNOCOMPETENCE AND EXPRESSION OF OUTER MEMBRANE PROTEIN OMPA GENE OF M YCOBACTERI UM BOVI S 1 2 1 1 QI Yingfang , HUANG Ying , U Yanfen , LIU Huifang , 1 1 1 1 1 HE Minglei , SI Wei , NI Hongbo , WANG Chunlai , LIU Siguo (1State K ey L aboratory of Veterinary Biotechnology , H arbin Veterinary R esearch I nstitute, CAA S, H arbin 15 1, China; 2A nimal Center f or Disease Control and Prev ention of X inj iang , Changj i8311 , China) Abstract T he outer membrane protein A gene (omp A ) of Mycobacterium bovis Vallee strain was amplified by PCR and inserted into the p GEX 6p1 to construct the recombinant plasmid p GEX 6p1omp A . The recombinant plasmid was then transformed into Escherichia coli BL21 ( E3) . Protein expression was induced by IPTG with a final concentration of 0. 1 mmol/ L. T he expressed protein was purified by glutathione Sepharose 4B. The outer membrane protein A had a m