title: Amplification of the entire kanamycin biosynthetic gene cluster during empirical strain improvement of Streptomyces kanamyceticus.
abstract: Streptomyces kanamyceticus 12-6 is a derivative of the wild-type strain developed for industrial kanamycin (Km) production. Southern analysis and DNA sequencing revealed amplification of a large genomic segment including the entire Km biosynthetic gene cluster in the chromosome of strain 12-6. At 145 kb, the amplifiable unit of DNA (AUD) is the largest AUD reported in Streptomyces. Striking repetitive DNA sequences belonging to the clustered regularly interspaced short palindromic repeats family were found in the AUD and may play a role in its amplification. Strain 12-6 contains a mixture of different chromosomes with varying numbers of AUDs, sometimes exceeding 36 copies and producing an amplified region >5.7 Mb. The level of Km production depended on the copy number of the Km biosynthetic gene cluster, suggesting that DNA amplification occurred during strain improvement as a consequence of selection for increased Km resistance. Amplification of DNA segments including entire antibiotic biosynthetic gene clusters might be a common mechanism leading to increased antibiotic production in industrial strains.
authors.affiliation: Department of Molecular Microbiology, John Innes Centre, Colney, Norwich NR4 7UH, United Kingdom.
authors.forename: Koji
authors.initials: K
authors.lastname: Yanai
authors.forename: Takeshi
authors.initials: T
authors.lastname: Murakami
authors.forename: Mervyn
authors.initials: M
authors.lastname: Bibb
mesh_terms: D001483:Base Sequence; D002876:Chromosomes, Bacterial / Q000235:genetics; D005784:Gene Amplification / Q000235:genetics*; D017353:Gene Deletion; D018628:Gene Dosage; D015964:Gene Expression Regulation, Bacterial; D007612:Kanamycin / Q000096:biosynthesis*; D008969:Molecular Sequence Data; D005810:Multigene Family / Q000235:genetics*; D009154:Mutation / Q000235:genetics; D013302:Streptomyces / Q000145:classification* / Q000235:genetics*
chemical_list: D007612:Kanamycin