A GENERAL-APPROACH FOR IDENTIFYING AND CLONING PEPTIDE SYNTHETASE GENES

A wide variety of bioactive peptides are synthesized nonribosomally by multienzyme complexes employing the thiotemplate mechanism. Based on the known and highly conserved structures of several genes encoding mu ltifunctional peptide synthetases, we developed a universal polymerase chain reaction (PCR) approach for amplifying, cloning and identifying parts of putative peptide synthetase genes. We showed, by cloning fra gments of peptide synthetase genes from the phaseolotoxin-producing st rain Pseudomonas syringae pv. phaseolica and from Bacillus licheniform is, which produces the branched peptide antibiotic bacitracin, that th is approach is applicable. It gives a new and potentially general acce ss to the biosynthetic genes of many nonribosomally synthesized peptid es.