AMPLIFICATION METHOD OF SURFACTIN BIOSYNTHESIS OPERON, RE-CONSTRUCTION PLASMID AND TRANSFORMED STRAIN CONTAINING OPERON

摘要

A method for isolating a surfacting biosynthesis operon is provided to obtain the operon encoding surfactin at a time and massively produce biosurfactant having excellent surface activity. A method for isolating surfactin biosynthesis operon(srfA operon) comprises: a step of preparing a PCR reactant containing 0.7-2.5 mM of Mg^2+, 0.5-3 U of rTth polymerase and 0.005-0.03 U vent polymerase based on the concentration of 50 mul of PCR reactant; and a step of performing long and accurate PCR(LA-PCR) using PCR reactant and isolating the surfactin biosynthesis operon from Bacillus subtilis C9. An isolated surfactin biosynthesis operon is denoted by sequence number 3(SEQ ID NO:3). A recombinant vector pSRF536 comprises the isolated surfactin biosynthesis operon. A transformed recombinant E.coli(KCTC 11248BP) having surface activity is produced by transforming with the recombinant vector pSRF536.