Mutants of Streptomyces roseosporus that express enhanced recombination within partially homologous genes

• Published in: Microbiology (Society for General Microbiology) 1996-10, Vol.142 (10), p.2803-2813
• Main Authors: Hosted, Thomas J, Baltz, Richard H
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Bacterial Proteins - genetics; Bacteriology; Bacteriophages - genetics; Base Sequence; Biological and medical sciences; Chromosomes, Bacterial - genetics; Cloning, Molecular; Crossing Over, Genetic; DNA, Bacterial - genetics; DNA-Binding Proteins; Fundamental and applied biological sciences. Psychology; Genes, Bacterial - genetics; Genetics; Glucokinase - genetics; Glutamate-Ammonia Ligase - genetics; Glutamate-Ammonia Ligase - metabolism; Microbiology; Molecular Sequence Data; Mutagenesis; Mutation; Recombinant Fusion Proteins - biosynthesis; Recombination, Genetic; Sequence Homology, Amino Acid; Sequence Homology, Nucleic Acid; Sigma Factor; Streptomyces - enzymology; Streptomyces - genetics
• ISSN: 1350-0872; 1465-2080
• DOI: 10.1099/13500872-142-10-2803

Lilly Research Laboratories, A Division of Eli Lilly and Company, Indianapolis, IN 46285, USA 1 Author for correspondence: Richard H. Baltz. Tel: [phone]. Fax: [phone]. e-mail: [e-mail] 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 10 2 - to 10 4 -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