解放军医学院学报2017，Vol.38Issue(7)：657-661,5.DOI:10.3969/j.issn.2095-5227.2017.07.016
人肝细胞生长因子受体原核载体的构建表达及纯化
Prokaryotic expression and purification of human MET
摘要
Abstract
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.关键词
肝细胞生长因子受体/原核表达/纯化Key words
c-MET/prokaryotic expression/purification分类
生物科学引用本文复制引用
邱栾,赵珂,董小明,高瑞,张飞翔,张欣悦,詹轶群,李长燕,李建雄..人肝细胞生长因子受体原核载体的构建表达及纯化[J].解放军医学院学报,2017,38(7):657-661,5.基金项目
国家重点研发计划(2016YFC0105700) （2016YFC0105700）
国家自然科学基金(81273543 （）
81001042) （）
北京市科技新星计划(Z141107001814122)Supported by Research and Development Program of China (2016YFC0105700) （Z141107001814122）
the National Natural Science Foundation of China (81273543 （）
81001042) （）
Beijing Science and Technology New Star Program (Z141107001814122) （Z141107001814122）
