Expression, purification and characterization of recombinant plasminogen activator from Gloydius brevicaudus venom in Escherichia coli

• The PA gene was cloned from the venom gland of Gloydius brevicaudus using RT-PCR.
• The MW of rGBV-PA expressed in E. coli was 27.9 kDa.
• rGBV-PA can reduce the plasma ELT in rabbits and prevent thrombosis in rats.

The plasminogen activator (PA) in snake venom, a serine protease, can convert plasminogen to active plasmin, indirectly causing the degradation of fibrin. It is difficult to purify sufficient snake venom PA (SV-PA) for clinical applications due to the low SV-PA content in venom. The gene encoding PA was obtained from the venom gland of Gloydius brevicaudus using RT-PCR with primers designed according to the N-terminal amino acids of G. brevicaudus venom PA (GBV-PA), was cloned into the prokaryotic expression vector pET-42a, and recombinant GBV-PA (rGBV-PA) was expressed via Isopropyl-β-d-1-Thiogalactopyranoside (IPTG) induction. Like human tissue PA, the purified renatured rGBV-PA could significantly reduce the rabbit plasma euglobulin lysis time (ELT) and prevent thrombus formation in the inferior vena cava of rats. Within the dosage range, the dosage and effects were positively correlated.