Prokaryotic expression and puri fication of human MET

Objective To construct the prokaryotic expression vectors of human MET (1-507, 1-587, 553-970, 553-1057, 1018-1390) and obtain their puri fied 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 ampli fied 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 ampli fied 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 puri fied, and highly puri fied 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.