中国兽医科学2009，Vol.39Issue(9)：796-802,7.
牛结核分枝杆菌CFP10-ESAT6融合蛋白的原核表达及其在牛结核病检测中的应用
Prokaryotic expression of recombinant CFP10-ESAT6 fusion protein from Mycobacterium tuberculosis and its application to detection of bovine tuberculosis
摘要
Abstract
The gene encoding protein CFP10-ESAT6 was amplified from Mycobacterium tuberculosis strain AN5 chromosomal DNA by recombinant PCR and cloned into the expression vector pET-32a to gen-erate the recombinant plasmid pET-CFP10/ESAT6. After being identified by BamH 1 +HindⅢ digestion and sequencing,the recombinant expression plasmid was transformed into Escherichia coli BL21(DE3). The fused protein rHIS-CFP10/ESAT6 was expressed with induction by IPTG. SDS-PAGE analysis showed that the fusion protein rHIS-CFP10/ESAT6 was 42 ku in molecular weight,and made up 40% of whole bacterial proteins. The soluble protein was purified with HIS affinity chromatography column. West-ern-blotting analysis indicated that the purified recombinant protein possessed good immunological activity, and could induce to generate high level of IFN-γ in whole blood,and the IFN-γ response to rHIS-CFP10/ ESAT-6 was better than that to PPDa and PPDb. The results showed that the recombinant fusion protein rHIS-CFP10/ESAT6 provided a basis for the diagnosis of tuberculosis.关键词
牛结核病/融合蛋白/CFP-10/ESAT-6Key words
bovine tuberculosis/fusion protein/CFP-10/ESAT-6分类
农业科技引用本文复制引用
徐贤坤,熊毅,龙剑明,刘棋,曾宪垠..牛结核分枝杆菌CFP10-ESAT6融合蛋白的原核表达及其在牛结核病检测中的应用[J].中国兽医科学,2009,39(9):796-802,7.基金项目
广西科技厅科技攻关项目(桂科攻0719004-3F) （桂科攻0719004-3F）
国家农业公益性行业科研专项(200803026) （200803026）
广西科技创新能力建设项目(08D5-01D) （08D5-01D）
