ST14 is one of the type II transmembrane serine proteases that correlates with the process of tumor metastasis. The C-terminal catalytic region (900 bp) of ST14 was cloned into the expression vector pGEX-4T-2 and the ...ST14 is one of the type II transmembrane serine proteases that correlates with the process of tumor metastasis. The C-terminal catalytic region (900 bp) of ST14 was cloned into the expression vector pGEX-4T-2 and the positive plasmid pGEX-4T-2-ST14 was transformed into E. coli BL21, then cultured and induced with IPTG. The chaperonin GroEL was found to be tightly associated with the fusion protein and co-purified with it by regular GST affinity chromatography. A method for the removal of contaminating GroEL from GST-ST14 fusion protein was described, the purity of product was 96.2%. Enzyme activity assay indicated that this fusion protein had serine protease activity.展开更多
文摘ST14 is one of the type II transmembrane serine proteases that correlates with the process of tumor metastasis. The C-terminal catalytic region (900 bp) of ST14 was cloned into the expression vector pGEX-4T-2 and the positive plasmid pGEX-4T-2-ST14 was transformed into E. coli BL21, then cultured and induced with IPTG. The chaperonin GroEL was found to be tightly associated with the fusion protein and co-purified with it by regular GST affinity chromatography. A method for the removal of contaminating GroEL from GST-ST14 fusion protein was described, the purity of product was 96.2%. Enzyme activity assay indicated that this fusion protein had serine protease activity.
