大肠杆菌NrfA蛋白表达、纯化及多克隆抗体的制备Expression,Purification of E.coli NrfA Protein and Preparation of Polyclonal Antibody Against NrfA
何婷婷;龚钢明;高然;
摘要(Abstract)：
[目的]克隆大肠杆菌NrfA基因,构建pET-28a(+)-NrfA表达载体,制备相应的多克隆抗体并对其进行鉴定。[方法]以大肠杆菌基因组DNA为模板,PCR扩增得到NrfA基因编码区,构建pET-28a(+)-NrfA表达载体;经IPTG诱导表达并纯化重组蛋白;再免疫新西兰雄兔,制备多克隆抗体;用ELISA方法检测抗体的效价,Western Blotting检测抗体的特异性。[结果]构建的表达载体pET-28a(+)-NrfA在大肠杆菌中诱导后可高效表达NrfA蛋白;免疫获得的多克隆抗体用ELISA检测,其效价为1∶204 900;经Western Blotting分析,抗体的特异性较好。[结论]成功克隆大肠杆菌的NrfA基因,并构建了其表达载体,制备的NrfA多克隆抗体具有较高的效价和良好的特异性,为研究细菌有关NrfA奠定了基础。
关键词(KeyWords)： NrfA基因;原核表达;多克隆抗体
基金项目(Foundation):
作者(Author): 何婷婷;龚钢明;高然;
Email:
DOI: 10.13989/j.cnki.0517-6611.2012.10.138
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