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Highly transformable mutants of Streptomyces fradiae defective in several restriction systems

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Summary

Streptomyces fradiae JS85 is a mutant defective in tylosin production and an efficient recipient for conjugal transfer of tylosin genes. JS85 was mutagenized with N-methyl-N′-nitro-N-nitrosoguanidine (MNNG) and derivatives defective in restriction were isolated by sequential selection for increased transformability by several plasmid DNAs. From the number of mutation and selection cycles required to eliminate most restriction, it was estimated that wild type S. fradiae expressed at least five restriction systems. From the patterns of restriction enzyme digestion of chromosomal DNA observed in the series of mutants that became progressively less restricting, it was suggested that wild type S. fradiae normally expresses modification (and presumably restriction) systems similar or analogous to PstI, XhoI, ScaI and EcoRI. The least restricting mutant of S. fradiae was readily transformable by many plasmids, including a bifunctional cosmid vector containing a large insert of Streptomyces DNA.

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References

  • Baltz RH (1978) Genetic recombination in Streptomyces fradiae by protoplast fusion and cell regeneration. J Gen Microbiol 107:93–102

    Google Scholar 

  • Baltz RH (1980) Genetic recombination by protoplast fusion in Streptomyces. Dev Ind Microbiol 21:43–54

    Google Scholar 

  • Baltz RH (1982) Genetics and biochemistry of tylosin production: A model for genetic engineering in antibiotic-producing Streptomyces. In: Hollaender A (ed) Genetic engineering of microorganisms for chemicals. Plenum, New York, pp 431–444

    Google Scholar 

  • Baltz RH (1986a) Mutagenesis in Streptomyces spp. In: Demain AL, Solomon A (eds) Manual of industrial microbiology and biotechnology. American Society for Microbiology, Washington DC, pp 184–190

    Google Scholar 

  • Baltz RH (1986b) Mutation in Streptomyces. In: Day LE, Queener SE (eds) The bacteria, vol 9, Antibiotic-producing Streptomyces. Academic Press, New York, pp 61–94

    Google Scholar 

  • Baltz RH, Matsushima P (1981) Protoplast fusion in Streptomyces: conditions for efficient genetic recombination and cell regeneration. J Gen Microbiol 127:137–146

    Google Scholar 

  • Baltz RH, Matsushima P (1983) Advances in protoplast fusion and transformation in Streptomyces. Experientia [Suppl] 46:143–148

    Google Scholar 

  • Baltz RH, Seno ET (1981) Properties of Streptomyces fradiae mutants blocked in biosynthesis of the macrolide antibiotic tylosin. Antimicrob Agents Chemother 20:214–225

    Google Scholar 

  • Baltz, RH, Stonesifer J (1985) Phenotypic changes associated with loss of expression of tylosin biosynthesis and resistance genes in Streptomyces fradiae. J Antibiot (Tokyo) 38:1226–1236

    Google Scholar 

  • Baltz RH, Seno ET, Stonesifer J, Matsushima P, Wild GM (1981) Genetics and biochemistry of tylosin production by Streptomyces fradiae. In: Schlessinger D (ed) Microbiology 1981. American Society for Microbiology, Washington DC, pp 371–375

    Google Scholar 

  • Baltz, RH, Seno, ET, Stonesifer J, Matsushima P, Wild GM (1982) Genetics and biochemistry of tylosin production. In: Umezawa H, Demain AL, Hata T, Hutchinson CR (eds) Trends in antibiotic research. Japan Antibiotics Research Association, Tokyo, pp 65–72

    Google Scholar 

  • Baltz RH, Seno ET, Stonesifer J, Wild GM (1983) Biosynthesis of the macrolide antibiotic tylosin. A preferred pathway from tylactone to tylosin. J Antibiot (Tokyo) 36:131–141

    Google Scholar 

  • Bibb MJ, Hopwood DA (1981) Genetic studies of the fertility plasmid SCP2 and its SCP2* variants in Streptomyces coelicolor A3(2). J Gen Microbiol 126:427–442

    Google Scholar 

  • Chater KF, Wilde L (1980) Streptomyces albus G mutants defective in the SalGI restriction-modification system. J Gen Microbiol 116:323–334

    Google Scholar 

  • Cox KL, Baltz RH (1984) Restriction of bacteriophage plaque formation in Streptomyces spp. J Bacteriol 159:499–504

    Google Scholar 

  • Fishman SE, Hershberger CL (1983) Amplified DNA in Streptomyces fradiae. J Bacteriol 155:459–466

    Google Scholar 

  • Katz, E, Hopwood DA, Thompson CJ (1982) Cloning and expression of the tyrosinase gene from Streptomyces antibioticus in Streptomyces lividans. J Gen Microbiol 129:2703–2714

    Google Scholar 

  • Larson JL, Hershberger CL (1984) Shuttle vectors for cloning recombinant DNA in Escherichia coli and Streptomyces griseofuscus C581. J Bacteriol 157:314–317

    Google Scholar 

  • Matsushima P, Baltz, RH (1985) Efficient plasmid transformation of Streptomyces ambofaciens and Streptomyces fradiae protoplasts. J Bacteriol 163:180–185

    Google Scholar 

  • Rao RN, Richardson MA, Kuhstoss S (1987) Cosmid shuttle vectors for cloning and analysis of Streptomyces DNA. Methods Enzymol (in press)

  • Richardson M, Mabe JA, Beerman NE, Nakatsukasa WM, Fayerman, JT (1982) Development of cloning vehicles from the Streptomyces plasmid pFJ103. Gene 20:451–457

    Google Scholar 

  • Roberts JR (1984) Restriction and modification enzymes and their recognition sequences. Nucleic Acids Res 12:167–204

    Google Scholar 

  • Seno ET, Baltz RH (1981) Properties of S-adenosyl-L-methionine: macrocin O-methyltransferase in extracts of Streptomyces fradiae strains which produce normal or elevated levels of tylosin and in mutants blocked in specific O-methylations. Antimicrob Agents Chemother 20:370–377

    Google Scholar 

  • Seno ET, Baltz RH (1982) S-adenosyl-L-methionine: macrocin O-methyltransferase activities in a series of Streptomyces fradiae mutants that produce different levels of the macrolide antibiotic tylosin. Antimicrob Agents Chemother 21:758–763

    Google Scholar 

  • Stonesifer J, Baltz RH (1985) Mutagenic DNA repair in Streptomyces. Proc Natl Acad Sci USA 82:1180–1183

    Google Scholar 

  • Stonesifer J, Matsushima P, Baltz RH (1986) High frequency conjugal transfer of tylosin genes and amplifiable DNA in Streptomyces fradiae. Mol Gen Genet 202:348–355

    Google Scholar 

  • Thompson CJ, Ward JM, Hopwood DA (1980) DNA cloning in Streptomyces: resistance genes from antibiotic producing species. Nature 286:525–527

    Google Scholar 

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Communicated by H. Saedler

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Matsushima, P., Cox, K.L. & Baltz, R.H. Highly transformable mutants of Streptomyces fradiae defective in several restriction systems. Mol Gen Genet 206, 393–400 (1987). https://doi.org/10.1007/BF00428877

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  • DOI: https://doi.org/10.1007/BF00428877

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