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A self-cloning system forActinomadura verrucosospora , a producer of the angucyclic antibiotic pradimicin A (PRM A), has been developed. The system is based on reproducible and reliable protoplasting and regeneration conditions forA. verrucosospora and a novel plasmid vector that consists of a replicon from a newly foundActinomadura plasmid and a selectable marker cloned from theActinomadura strain. The system has an efficiency of more than 105 CFU/microgram of DNA. Using this system, we have cloned and identified the polyketide synthase (PKS) genes essential for PRM A biosynthesis fromA. verrucosospora . Nucleotide sequence analysis of the 3.5-kbSal I-Sph I fragment showed that ketosynthase subunits (open reading frame 1 [ORF1] and ORF2) of the essential PKS genes have strong similarities (59 to 89%) to those for angucyclic antibiotic biosynthesis.

applied and environmental microbiology

Development of a Self-Cloning System for <i>Actinomadura verrucosospora</i> and Identification of Polyketide Synthase Genes Essential for Production of the Angucyclic Antibiotic Pradimicin

Development of a Self-Cloning System for Actinomadura verrucosospora and Identification of Polyketide Synthase Genes Essential for Production of the Angucyclic Antibiotic Pradimicin