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Microbiology 142 (1996), 2803-2813; DOI  10.1099/13500872-142-10-2803
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Mutants of Streptomyces roseosporus that express enhanced recombination within partially homologous genes

Thomas J. Hosted and Richard H. Baltz1

Lilly Research Laboratories, A Division of Eli Lilly and Company, Indianapolis, IN 46285, USA

1Author for correspondence: Richard H. Baltz. Tel: +1 317 276 5179. Fax: +1 317 276 9086. e-mail: baltz_richard_h@lilly.com

ABSTRACT

Streptomyces roseosporus mutants that express enhanced recombination between partially homologous (homeologous) sequences were isolated by selection for recombination between the bacteriophage øC31 derivative KC570 containing the Streptomyces coelicolor glucose kinase (glk) gene and the S. roseosporus chromosome. The frequencies of homeologous recombination in the ehr mutants were determined by measuring the chromosomal insertion frequencies of plasmids containing S. coelicolor glnA or whiG genes. S. roseosporus ehr mutants showed 102- to 104-fold increases in homeologous recombination relative to Ehr+ strains, but no increase in homologous recombination. Southern hybridization analysis revealed single unique sites for the insertion of each of the plasmids, and the crossovers occurred in frame and in proper translational register, yielding functional chimeric glnA and whiG genes.

Keywords: Streptomyces roseosporus, homeologous recombination, transposition mutagenesis, glutamine synthetase, sporulation sigma factor




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