Establishment and optimization of the conjugal transfer system of fostriecin producer Streptomyces pulveraceus

Abstract:

[Objective] Establishment and optimization of the gene transfer system of Streptomyces pulveraceus which produces fostriecin based on normal conjugal transfer method. [Methods] Intergeneric genetic transfer system was based upon integrative plasmid pSET152 from donor E. coli ET12567/pUZ8002 to Streptomyces pulveraceus. [Results] The maximum transformation efficiency of Streptomyces pulveraceus by conjugation was obtained when MS medium containing 15% glycine was used, with the spores heat-shocked at 50 °C for 10 min before mixing with E. coli, using a final 20 mg/L concentration of apramycin for overlay after incubation of 18 h. Simultaneity, pSET153::gfp was constructed from pSET152 by combining the constitutive ermE+ promoter with green fluorescent protein gene (gfp). Green fluorescent protein gene was expressed. [Conclusion] In summary, intergeneric genetic transfer system was demonstrated and optimized, and glycin can increase the efficiency of exconjugates.