Abstract: To further explore the relationship of gene between the full peptide and the mature peptide,we designed and synthesized a pair of primers according to DNA sequence(GenBank FJ463037)of B.pseudomycoides 34 kDa fibrinolytic enzyme published and fibrinolytic enzyme active site.We proformed PCR from the recombinant plasmid pGEX-BpFE and obtained fibrinolytic enzyme mature peptide gene-G(951 bp encoding,317 amino acid)to construct pET-28a-G vector.The recombinant plasmid of pET-28a-G was transformed into host bacterium of E.coli BL21 to construct pET-28 a-G/BL2 1 engineering strains successfully.SDS-PAGE analysis was performed after pET-28a-G/ BL21 being induced by IPTG(1 mmol/L)to detect intracellular expression of the fusion protein.Fusion protein had an apparent molecular weight of about 40 kDa,which was consistent with the predicted results.Fibrin-Plate method showed that the intracellular protein had activity,and end products after nickel affinity chromatography purified retained fibrinolie activity.As the result,we successfully obtained the recombinant bacteria in which 34 kDa fibrinolytic enzyme mature peptide gene expression with activity.

Key words: B．pseudomycoides, Fibrinolytic enzyme, Mature peptide, Expression, Purification

CLC Number: 

• Q786