Abstract: Objective: Fibrinolytic enzyme gene of Gloydius intermedius was cloned,then analysis the gene sequence and function. Methods:Total RNA was extracted from venom cells from Gloydius intermedius,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. Results:FLE gene was successfully cloned.Its length was 777bp including whole open reading frame(ORF). Conclusion:Gene sequence of FLE of Gloydius intermedius was cloned successfully and deduced the animo acid sequence.The ORF codes 258 animo acids,including lots of hydrophobic animo acids.19 animo acids is a secretory signal peptide.Based on the animo acid sequences of other fibrinolytic enzyme,the His65、Asp110 and Ser204 constitute the catalytic resduces and the frame is high identity;the whole sequence includes 12 cys what matching 6 disulfide bridges are essential to the tertiary structure and stability of FLE .Cloning the gene of FLE successfully affords basis for the research of genetic character of this gene and prokaryotic expression or eukaryotic expression.