机构地区: 黑龙江八一农垦大学动物科技学院
出 处: 《中国动物传染病学报》 2012年第6期36-39,共4页
摘 要: 从副结核分支杆菌K-10菌株基因组中扩增出516 bp的MAP1588C基因片段,插入原核表达载体pET-28b(+),转化至大肠杆菌BL21(DE3),在0.5 mmol/L的IPTG诱导下进行表达;利用融合蛋白的组氨酸标签进行亲和层析纯化重组蛋白;通过抗6 His标签抗体的Western blot验证了纯化蛋白产物;另外,副结核阳性牛血清可检测到该蛋白条带。结果表明,表达的重组蛋白MAP1588C的蛋白分子量为21 kDa,具有良好抗原性,有望将此抗原用于建立副结核的ELISA诊断方法。 The gene of MAP1588C was cloned from Mycobacterium paratuberculosis(MAP) strain K-10 and inserted into pET-28b(+) vector.The resulting recombinant plasmid p28B-MAP8 was transformed into E.coli BL21(DE3).Then,the product expressed was purified using Ni2+ affinity chromatography.The purified MAP1588C was approximately 21 kDa and reacted with anti-6His antibody and MAP cattle antiserum in Western blotting.These results indicated that the recombinant MAP1588C maintained good reactivity and could be used for development of an ELISA method for diagnosis of MAP.