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当前位置: 首页 > 文章 > 牛结核分枝杆菌重组多抗原表位基因的原核表达与鉴定 中国兽医学报 2011,31 (1) 55-58
Position: Home > Articles > Expression of recombinant epitope genes of Mycobacterium tuberculosis and identification of its expression product Chinese Journal of Veterinary Science 2011,31 (1) 55-58

牛结核分枝杆菌重组多抗原表位基因的原核表达与鉴定

作  者：
冯向辉;包永占;王建永;赵月兰;左玉柱;秦建华
单  位：
河北农业大学
关键词：
牛结核分枝杆菌;mpb70;mpb83;esat-6;原核表达
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。
译  名：
Expression of recombinant epitope genes of Mycobacterium tuberculosis and identification of its expression product
作  者：
FENG Xiang-hui1,BAO Yong-zhan1,WANG Jian-yong2,ZHAO Yue-lan2,ZUO Yu-zhu1,QIN Jian-hua1(1.College of Animal Science and Technology,Hebei Agricultural University,Baoding,Hebei 071000,China;2.College of Chinese Traditional Veterinary Science,Hebei Agricultural University,Dingzhou,Hebei 073000,China)
关键词：
Mycobacterium tuberculosis;mpb70,mpb83;esat-6;prokaryotic expression
摘  要：
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.

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