结核分枝杆菌Rv1400c多抗制备及反应原性分析  

Preparation of Polyclonal Antibody against the Recombinant Protein Rv1400c from Mycobacterium tuberculosis and Reactinogenicity Analysis of the Protein

作　　者：陶荣珊[1,2] 胡太超[1] 李金伟 刘思国[4] 王全凯[1,5] 

机构地区：[1]吉林农业大学动物科学技术学院,长春130118 [2]北海绿瑞贸易有限责任公司,北海536000 [3]东丰药业,东丰136300 [4]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/动物细菌病研究室,哈尔滨150001 [5]吉林省中韩动物科学研究院,长春130600

出　　处：《经济动物学报》2015年第1期16-19,24,共5页Journal of Economic Animal

基　　金：国家国际科技合作专项(2011DFA32900);国家自然科学基金项目(31100659;31272538);国家重点基础研究发展计划[973计划(2012CB518801)];长科技合(2011239)

摘　　要：将pET28b-Rv1400c重组表达质粒转化到大肠杆菌BL21(DE3)中,筛选出阳性菌株并增菌培养后IPTG诱导表达,离心后沉淀用SKL变性溶解,用亲和层吸树脂纯化Rv1400c蛋白,通过SDS-PAGE和Western-blot分析目的蛋白表达与纯化情况;利用Rv1400c蛋白免疫新西兰兔,制备多克隆抗体;Western-blot分析鹿结核阳性血清的反应情况。SDS-PAGE和Western-blot结果显示:Rv1400c以包涵体形式表达,蛋白分子质量为39 ku;目的蛋白免疫新西兰兔后,ELISA检测血清效价达到1∶51 200。纯化的Rv1400c重组抗原蛋白能与鹿的阳性血清发生体外结合反应。The recombinant plasmid pET-28b-Rv1400 c was transformed into E.coliBL21（DE3）for expression induced with IPTG（1mmol/ L）. The expressed product was purified by Ni-NTA af-finity chromatography. The recombinant protein Rv1400 c was analyzed by SDS-PAGE and Western-blot. The recombinant protein Rv1400 c was immunized the New Zealand rabbits to prepare polycio-nal antibody. The positive deer serum was analyzed by Western-blot. The result showed that theRv1400 c was successfully expressed in the form of inclusion body. The molecular weight of proteinwas 39 ku. The New Zealand rabbits immunized the purified recombinant protein could produce highspecific antibodies with the titer to 1∶51 200 and the antibodies expressed good specificity, and therecombinant protein could be combined by the positive deer serum in vitro.

关 键 词：结核分枝杆菌 Rv1400c 包涵体 复性 反应原性 

分 类 号：S852.618[农业科学—基础兽医学]