Colloque

Cloning and characterization of a polyketide synthase gene from Streptomyces venezuelae

Roy Mosher

Eswar Ramalingam

Janice Doull

Leo Vining

Ashish Paradkar

Membre a labase

Roy Mosher

Résumé du colloque

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 S. coelicolor A3(2) hybridized to a 2.4-kb BamHI fragment directly adjacent to the actI-hybridizing region. When the actI-hybridizing region of S. venezuelae DNA was inserted into pIJ702, and introduced into an actI mutant of S. coelicolor A3(2), actinorhodin production was restored, but the low frequency suggested repair of the actI mutation by homologous recombination. Isolation of an intact polyketide synthase gene and subcloning of the act-hybridizing regions is in progress.

Contexte

Section :

Biologie des actinomycètes

Thème du colloque :

Biologie des actinomycètes

Hôte : Université de Sherbrooke

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