Construction of conjugal transfer system of Streptomyces cinnamonensis and effect of PCR-mediated nsdA gene disruption on its secondary metabolism

Abstract

Intergeneric transfer of plasmid vectors pSET152 and pHL212 from donor Escherichia coli ET12567(pUZ8002) and S17-1 to Streptomyces cinnamonensis were demonstrated and optimized. The nsdA gene disruption structure was constructed by PCR-targeting system and then introduced into S. cinnamonensis BIB2005 through intergeneric conjugal transfer. After PCR analysis, the screened AprRKanS conjugant BIB309 was confirmed to be the nsdA mutant. Compared with the start strain, the yield of monensin of the nsdA mutant BIB309 increased 270 percent in the level of flask.