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In a galactose-isoleucine medium at 37°C Streptomyces venezuelae ISP5230 produces a group of related pigment antibiotics. The structures of these products place them in the family of polyketide secondary metabolites (S. Ayers, personal communication). A library of S. venezuelae ISP5230 genomic DNA was prepared in a lambda replacement vector and probed with the actI gene for polyketide synthase from Streptomyces coelicolor A3(2). A clone, possessing a 9.3-kb insert of S. venezuelae DNA, hybridized strongly to actI. The actI-hybridizing region was localized to a 1.8-kb SstI fragment at one end of the insert. The actIII gene encoding the polyketide reductase of …