Ⅰ.Screening, Molecular Cloning, And Identifying Of Encoding Gene Of Fibrinolytic Activity Protein From Clamworm Ⅱ.Investigate The Biologic Activity Of Fibrinolytic Activity Protein From Clamworm And Its Distribution In Vivo Of Clamworm

Objective Screening ,molecular cloning , and identifying of encoding gene of fibrinolytic activity protein from cDNA library established from clamworm digestive tract epithelial cells by SMART(Switching Mechanism At 5' end of the RNA Transcript) technology . Methods Trizol reagent from clamworm digestive tract epithelial cells extracted methods Total RNA. The mRNA was purified and reverse transcripted into cDNA. The cDNA was linked into plasmid pDNR-Lib after digested by nucleate endonuclease Sfi I to construct cDNA library. Multiple clones were obtained and sequenced by performing PCRs by designed primers in Kit of SMART (Switching Mechanism At 5' end of the RNA Transcript). The corresponding proteins encoded by sequenced genes were functionally predicted by molecular biological software and simulated by their second structures and third ones. Results The encoding gene of fibrinolytic activity protein was obtained as well as the recombined Escherichia Coli containing the gene sequence. The simulated results showed similarity comparing with mammalian serine proteases and had the conserved catalytic amino acid residues. Conclusion Recombined Escherichia Coli containing aimed sequence was successfully constructed in prokaryotic expression system. The protein encoded by the gene sequence has possibility in becoming into one new pattern medicine, which shows fibrinolytic activity.