小鼠MIF在大肠杆菌中的表达及纯化  

Expression and purification of murine MIF in E.col

作　　者：侯桂华[1] 于敏[1] 李璐娜[1] 

机构地区：[1]山东大学医学院实验核医学研究所

出　　处：《山东大学学报（医学版）》2003年第5期469-471,共3页Journal of Shandong University:Health Sciences

基　　金：山东省卫生厅立项课题(2001CA1CAB3)

摘　　要：目的:获得足够量的高纯度小鼠巨噬细胞移动抑制因子(MIF)重组蛋白。方法:酶切重组克隆质粒pMD18-MIF,将片段插入原核表达载体pQE31,构建相应的重组表达质粒pQE31-MIF,将此质粒转化大肠杆菌M15,得到转化子。经IPTG诱导表达后进行菌体蛋白的Westernblotting免疫印迹分析,并用Ni-NTA凝胶进行纯化。结果:MIF基因重组体构建成功,克隆的目的基因片段在M15中产生的融合表达产物在Westernblotting检测中具有与抗MIF单克隆抗体发生特异性结合反应的特性。得到了纯化的目的蛋白。结论:MIF基因片段可在大肠杆菌中有效表达,表达产物可以与抗MIF单克隆抗体发生特异性结合反应,并得到纯化的MIF。Objective:To obtain enough and high purified MIF recombinant protein.Methods:Digesting recombinant cloning vector pMD18-MIF to obtain MIF,then inserting this fragment into the expressing vector pQE31to construct a recombinant expressing plasmid,which was transformed into the E.coli M15.IPTG induced transformant was analyzed with Western blotting and purified with Ni-NTA agarose.Results:Induced expressing product was shown to have antigenic reactivity to the anti-MIF McAb so that high purity MIF protein was obtained.Conclusion:The MIF gene was expressed effec-tively in M15and high purity MIF protein obtained may provide a basis for further research.

关 键 词：巨噬细胞游走抑制因子 小鼠 基因表达 大肠杆菌 

分 类 号：R392[医药卫生—免疫学]