Abstract

A Streptomyces coelicolor gene, called spaA, homologous to the stationary phase regulatory gene rspA of Escherichia coli [Huisman and Kolter (1994) Science 265, 537–539], was cloned using the Streptomyces ambofaciens rspA homologue spa2 [Schneider et al. (1993) J. Gen. Microbiol. 139, 2559–2567] as a probe. Considerable differences in sequence and in genetic context were detected between spa2 of S. ambofaciens and spaA of S. coelicolor. A cloned internal fragment of spaA was used to direct integration of a phage vector into the spaA gene. The disruption caused delayed antibiotic production (undecylprodigiosin and actinorhodin) and led on further incubation to increased actinorhodin production at high, but not low, cell density. This phenotype was apparent only on the nutritionally poorest of three media tested. The attempted use of an integrating plasmid-based system for gene replacement of spaA gave rise to extensive deletions of adjacent chromosomal DNA.

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