Abstract:

Fibrinolytic enzyme gene of Gloydius saxatilis was cloned, then analysis the gene sequence and function; construction and indentification of eukaryotic expression vector of FLE gene. Total RNA was extracted from venom cells from Gloydius saxatilis, FLE gene was amplified by RT-PCR. Then it was connected to pMD18-T vector by TA cloning. At last, sequencing and analysis the gene sequence. The FLE gene was inserted into the vector pEGFP-C1. This recombinant plasmid pEGFP-FLE was identified by digestion of endonuclease, PCR and sequencing. FLE gene was cloned successfully. Its length was 774 bp including whole open reading frame (ORF).The recombinant vector was constructed succeed. Gene sequence of FLE of Gloydius saxatilis was cloned successfully, the sequence was coding 257 animo acids and deduced its molecular weight was 30 KDa. The experiment will provide help for future research in eukaryotic expression.