title: A gene cloning system for 'Streptomyces toyocaensis'.
abstract: We explored different methods of introducing DNA into 'Streptomyces toyocaensis' and Streptomyces virginiae to construct stable recombinant strains. Plasmid pIJ702 isolated from Streptomyces lividans transformed protoplasts of 'S. toyocaensis' at a frequency of 7 x 10(3) transformants (mu g DNA)-1. pIJ702 prepared from 'S. toyocaensis' transformed 'S. toyocaensis' protoplasts at a frequency of 1 center dot 5 x 10(5) (mu g DNA)-1, suggesting that 'S. toyocaensis' expresses restriction and modification. Plasmid pRHB126 was transduced by bacteriophage FP43 into 'S. toyocaensis' at a frequency of 1.2 x 10(-6) (p.f.u)-1. Plasmids pOJ436 and pRHB304 were introduced into 'S. toyocaensis' by conjugation from Escherichia coli S17-1 at frequencies of about 2 x 10(-4) and 1 x 10(-4) per recipient, respectively. Analysis of several exconjugants indicated that pOJ436 and pRHB304 inserted into a unique phiC31 attB site and that some of the insertions had minimal deleterious effects on glycopeptide A47934 production. The results indicate that 'S. toyocaensis' is a suitable host for gene cloning, whereas S. virginiae does not appear to be.
authors.affiliation: Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN 46285, USA.
authors.forename: Patti
authors.initials: P
authors.lastname: Matsushima
authors.affiliation: Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, IN 46285, USA.
authors.forename: Richard H
authors.initials: RH
authors.lastname: Baltz
mesh_terms: D000900:Anti-Bacterial Agents / Q000096:biosynthesis; D001435:Bacteriophages / Q000235:genetics; D002876:Chromosomes, Bacterial / Q000235:genetics; D003001:Cloning, Molecular / Q000379:methods*; D003227:Conjugation, Genetic; D004269:DNA, Bacterial / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D005822:Genetic Vectors; D010455:Peptides; D010957:Plasmids / Q000235:genetics; D011523:Protoplasts; D013302:Streptomyces / Q000235:genetics* / Q000378:metabolism; D014161:Transduction, Genetic; D014170:Transformation, Genetic; D016662:Virus Integration / Q000235:genetics
chemical_list: D000900:Anti-Bacterial Agents; D004269:DNA, Bacterial; D010455:Peptides