目的：构建His-INMAP原核表达载体，表达、纯化蛋白，为研究INMAP体外磷酸化做准备。方法：利用GFP-INMAP进行PCR获得长度为1.1 kb的INMAP全长cDNA片段，把该片段构建到原核表达载体pET30a（+）中，转化大肠杆菌BL-21（DE3）菌株。菌株经IPTG诱导，产生His-INMAP包涵体蛋白。包涵体蛋白经过去垢剂Triton X-100洗涤、6 mol/L尿素溶解变性，使用Ni柱亲和层析对蛋白进行纯化，利用低浓度的尿素透析对蛋白进行复性。结果：菌落PCR鉴定及基因测序表明INMAP基因cDNA 片段被构建到表达载体pET30a（+）中；SDS-PAGE结果显示纯化的蛋白呈单一条带，与目的蛋白大小一致，表明蛋白纯化成功。结论：成功构建了His-INMAP原核表达载体，并经过诱导表达，纯化得到了纯度较高的INMAP蛋白。

Aim: Construting His-INMAP prokaryotic expression vector, expressing and purifying the protein product, in order to lay a base for the in vitro phosphorylation study of INMAP. Methods: PCR with GFP-INMAP primer to get full length (1.1kbp) cDNA fragment of INMAP for recombination within prokaryotic expression vector pET30a (+) and transforme it into E. coli BL-21 (DE3) strain. Induced by IPTG, His-INMAP fused proteins were produced by the bacteria within inclusion bodies. Inclusion body proteins, washed with detergent Triton X-100 and denatured with 6 mol / L urea solution, were purified by Ni2+ affinity chromatography, followed by renaturation reaction utilizing urea dialysis method. Results: Colony identification and gene sequencing showed that INMAP cDNA fragments were constructed into expression vector pET30a (+); SDS-PAGE showed that the purified protein showed a single band, consistent with the size of the target protein, so the protein purification was successful. Conclusion: The His-INMAP prokaryotic expression vector was successfully constructed, and after expression induction and purification, high purity of INMAP protein was obtained.