利用PCR技术从Bacillus sp.nov SK006中扩增得到纤维蛋白溶解酶(Fibrinolitic enzymes,FE)基因的DNA片段,将其连接到pMD-T载体上,经测序后克隆至载体PET-22b(+),转化至E.coli BL21(DE3)中,得到的重组菌能高效表达FE。诱导表达后,SDS-PAGE显示,重组FE分子量约为28 ku。

A DNA encoding fibrinolytic enzyme(FE) was amplified from Bacillus sp.nov SK006 using PCR.Sequencing showed open reading frame is composed of 828 bp encoding a putative protein of 276 amino acid residues.The cDNA was cloned into an expression vector pET-22b(+) and expressed in Escherichia coli BL21(DE3).The recombinant protein was purified to electrophoretical homogeneity by affinity chromatography using Chelating Sepharose Fast Flow resin.The purified recombinant FE had an approximate molecular weight of 28 ku by SDS-PAGE analysis.