[Construction of fusion recombinant plasmid coding for RGD peptide and urokinase B chain, its expression in Escherichia coli and preliminary characterization of its biological activity].
A synthetic RGD (Arg-Gly-Asp) peptide-coding sequence was fused with urokinase B-chain cDNA and then cloned into the prokaryotic expression vector pBV220. The fused gene was expressed in E. coli DH5 alpha under the control of PRPL promoter by 42 degrees C induction. The expression level of the fusion protein was over 9.2% of the total bacterial proteins as a or of inactive inclusion body. The purified fusion protein was obtained with similar antigenicity as urokinase shown by Western blotting. Its in vitro fibrinolysis and anti-platelet aggregation activity was also evaluated by bioassay.
Zhu
F X
FX
Beijing Institute of Radiation Medicine, Beijing 100850.
Xing
G C
GC
He
F C
FC
D015153:Blotting, Western; D004926:Escherichia coli / Q000235:genetics*; D009842:Oligopeptides / Q000235:genetics*; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000494:pharmacology; D014568:Urokinase-Type Plasminogen Activator / Q000235:genetics* / Q000494:pharmacology
D009842:Oligopeptides; D011993:Recombinant Fusion Proteins; C047981:arginyl-glycyl-aspartic acid; D014568:Urokinase-Type Plasminogen Activator