中文摘要：

目的 克隆人肝细胞生长因子受体（MET） 基因，构建不同结构域（1-507、1-587、553-970、553-1057、1018-1390）的原核表达载体，获得其原核表达产物，并纯化相应蛋白质，为基于MET 不同结构域的小分子抑制剂筛选提供依据。方法 采用PCR 技术从人肝细胞癌细胞系Hep G2 cDNA 文库中扩增出人MET 基因（1-507、1-587、553-970、553-1057、1018-1390），将其克隆到pGEX-4T-2 载体中，在大肠埃希菌BL21 中表达后，对原核表达产物进行纯化，以SDS-PAGE和Western blot 鉴定表达与纯化产物。结果 从人肝细胞癌细胞系Hep G2 cDNA 文库中扩增获得分别约1 524 bp、1 764 bp、1 257 bp、1 518 bp 和1 119 bp 的DNA 片段，并成功构建在pGEX-4T-2 载体上，经测序与目的序列完全一致；在BL21中诱导表达出相对分子质量约为82 kU、91 kU、72 kU、82 kU、67 kU 的目的蛋白，经纯化后获得了纯度较高的重组蛋白GST-c-MET。结论 成功获得了MET 不同结构域的重组蛋白GST-c-MET（1-507、1-587、553-970、553-1057、1018-1390），为后续研究MET 小分子抑制剂筛选奠定了实验基础。

英文摘要：

Objective To construct the prokaryotic expression vectors of human MET （1-507, 1-587, 553-970, 553-1057, 1018-1390） and obtain their purified proteins, and provide basis for screening small molecule inhibitors to human MET. Methods Human MET （1-507, 1-587, 553-970, 553-1057, 1018-1390） coding regions were amplified from human HCC Hep G2 cDNA library, and they were inserted into prokaryotic expression vector pGEX-4T-2. The recombinant plasmids pGEX-4T-2- MET were transformed into E.coli BL21. The expressed products were purified and identified by SDS-PAGE and Western blot analysis. Results The DNA fragments of 1 524, 1 764, 1 257, 1 518 and 1 119 bp were successfully amplified by PCR from human HCC Hep G2 cDNA library, and they were cloned into pGEX-4T-2 and identified by sequencing. The recombinant proteins with size 82, 91, 72, 82, 67 kU were successfully induced and purified, and highly purified recombination protein GST-c-MET was obtained. Conclusion The recombinant proteins of GST-c -MET （1-507, 1-587, 553-970, 553-1057, 1018-1390） are obtained successfully, which lay a foundation for further research on MET and small molecule inhibitors.