摘要
利用DNAStar对牛结核分枝杆菌的3个主要抗原mpb70、mpb83、esat-6进行预测分析,将预测的抗原指数高的抗原表位经全基因合成后,克隆到表达载体pET28a(+)中,构建重组质粒pET28a-mpb-70-83-esat-6,转化入大肠杆菌BL21(DE3)中诱导表达,表达产物经SDS-PAGE鉴定,结果检测到相对分子质量约为21 000的重组蛋白。经NI-NTA纯化重组蛋白后进行Western-blot鉴定,结果显示该重组蛋白可被牛结核分枝杆菌的多克隆抗体识别,表明该蛋白具有良好的反应原性。
The three major antigens of Mycobacterium tuberculosis,mpb70,mpb83 and esat-6 were analyzed by DNAStar.The epitope which is in a high antigenic index was synthesised and cloned into the pET-28a（＋） expression vector.Constructed a recombinant plasmid called pET28a-mpb-70-83-esat-6.The recombinant plasmid was transformed into E.coli BL21 competent cells and then induced by 1 mmol/L IPTG.SDS-PAGE analysis revealed that a band of approximately 21 000 in molecular weight was expressed from the induced E.coli BL21 component cells.Using the NI-NTA to purify the recombination protein,Western-blot analysis displayed that the protein could be discriminated by Mycobacterium tuberculosis polyclonal antibody,indicating that the protein has a good reactionogenicity.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2011年第1期55-58,共4页
Chinese Journal of Veterinary Science
基金
国家"十一五"科技支撑计划资助项目(2006BAD04A10-4)
河北省重大技术创新专项计划资助项目(07227146Z-4)
