CAJ | 학술논문

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。
종부결핵분지간균K-10균주기인조중확증출516 bp적MAP1588C기인편단,삽입원핵표체재체pET-28b（＋）,전화지대장간균BL21（DE3）,재0.5 mmol/L적IPTG유도하진행표체;이용융합단백적조안산표첨진행친화층석순화중조단백;통과항6 His표첨항체적Western blot험증료순화단백산물;령외,부결핵양성우혈청가검측도해단백조대。결과표명,표체적중조단백MAP1588C적단백분자량위21 kDa,구유량호항원성,유망장차항원용우건립부결핵적ELISA진단방법。
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.