小鼠巨噬细胞移动抑制因子(MIF)原核表达载体pQE31-MIF的构建

Construction of mice migration inhibitory factor (MIF) procaryotic expression vector pQE31-MIF

目的 :获得小鼠巨噬细胞移动抑制因子(MIF)全长基因克隆 ,并构建重组表达载体pQE3 1 MIF。方法 :用RT PCR技术 ,以特异性寡核苷酸为引物 ,从小鼠脾脏总RNA中扩增MIF基因 ,插入pMD1 8 T载体 ,用限制性核酸内切酶和DNA序列分析鉴定重组克隆。构建并鉴定原核表达载体pQE3 1 MIF。结果 :成功克隆了小鼠MIF基因 ,构建了重组质粒pMD1 8 MIF及重组原核表达质粒。序列分析显示获得的MIF基因cD NA序列与文献报导一致。结论 :成功构建了重组表达质粒pQE3 1 MIF 。

Objective: To clone mice macrophage migration inhibitory factor (MIF)cDNA and to construct mice MIF procaryotic exprssion vector. Methods: RT PCR was used to amplify the target gene with total RNA from mice spleen as template and specific oligo nucleotides as primers of MIF. The gene was inserted into pMD18 T and then sequenced.Then the procaryotic expression vector pQE31 MIF containing MIF cDNA was constructed and identified. Results:PCR amplified DNA fragment was cloned into pMD18 T and clonal recombinant of pMD18 MIF and procaryoticexpression vector pQE31 MIF was successfully constructed. Conclusion: Clonal recombinant of pMD18 MIF is obtained and MIF cDNA sequence is the same with that reported previously and procaryotic expression vector pQE31 MIF was successfully constructed.