PRODUCTION OF A HYBRID PROTEIN CONSISTING OF THE N-TERMINAL FRAGMENT OF UROKINASE AND THE C-TERMINAL DOMAIN OF URINARY TRYPSIN-INHIBITOR INESCHERICHIA-COLI
D. Sugino et al., PRODUCTION OF A HYBRID PROTEIN CONSISTING OF THE N-TERMINAL FRAGMENT OF UROKINASE AND THE C-TERMINAL DOMAIN OF URINARY TRYPSIN-INHIBITOR INESCHERICHIA-COLI, Biotechnology and applied biochemistry, 27, 1998, pp. 145-152
We have constructed a hybrid protein (ATFHI) consisting of an N-termin
al fragment from urokinase (ATF) and HI-8, which is the C-terminal dom
ain of urinary trypsin inhibitor. The fusion genes for the hybrid prot
eins were engineered by PCR and cloned into expression plasmids, Under
the control of the toc promoter, fusion genes were efficiently expres
sed in Escherichia cell, The hybrid proteins, produced as inclusion bo
dies in E. coil, were refolded by a dialysis method and purified by io
n-exchange chromatography, ATFHI exhibited bifunctional activity relat
ed to antimetastatic effects: the urokinase receptor-binding activity
of ATF and the inhibitory activity of HI-8 on plasmin.