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Efficient transformation ofMicromonospora melanosporea protoplasts byStreptomyces plasmid

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Abstract

An efficient and relatively simple procedure forMicromonospora melanosporea protoplast preparation and transformation is described. Transformation ofM. melanosporea protoplast by theStreptomyces plasmid pIJ702 was optimized by altering parameters affecting the formation, regeneration, and transformation of protoplasts. Improvement of regeneration medium resulted in relatively quick growth of transformants (only 7 days). As a result of these experiments we describe a new transformation method that has routinely yielded 106 transformants/µg plasmid DNA.

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Literature Cited

  1. Bibb MJ, Ward JM, Hopwood DA (1978) Transformation of plasmid DNA intoStreptomyces at high frequency. Nature 274:398–400

    Article  PubMed  Google Scholar 

  2. Caso JL, Hardisson D, Suarez JE (1987) Transfection inMicromonospora spp. Appl Environ Microbiol 53:2544–2547

    PubMed  Google Scholar 

  3. Hasegawa M, Dairi T, Ohta T, Hashimoto E (1990) Novel gene cloning system ofMicromonospora which revealed the gene-organization for fortimicin biosynthesis. Abstract Book of the 6th International Symposium on Genetics of Industrial Microorganisms 1990:87

  4. Hopwood DA, Bibb MJ, Chater KF, Kieser T, Bruton CJ, Kieser HM, Lydiate DJ, Smith CP, Ward JM, Schrempf H (1985) Genetic manipulation of Streptomyces, a laboratory manual. Norwich, UK: The John Innes Foundation

    Google Scholar 

  5. Kelemen GH, Financsek I, Jarai M (1989) Efficient transformation ofMicromonospora purpurea with pIJ702 plasmid. J Antibiot 42:325–328

    PubMed  Google Scholar 

  6. Kieser T (1984) Factors affecting the isolation of ccc DNA fromStreptomyces lividans andEscherichia coli. Plasmid 12:19–36

    Article  PubMed  Google Scholar 

  7. Lampel JS, Strohl WR (1986) Transformation and transfection of anthracycline-producing streptomycetes. Appl Environ Microbiol 51:126–131

    PubMed  Google Scholar 

  8. MacNeil DJ (1987) Introduction of plasmid DNA intoStreptomyces lividans by electroporation. FEMS Microbial Lett 42:239–244

    Article  Google Scholar 

  9. Matkovic B, Piendl W, Bock A (1984) Ribosomal resistance as a wide-spread self-defence mechanism in aminoglycoside-producing Micromonospora species. FEMS Microbiol Lett 24:273–276

    Article  Google Scholar 

  10. Matsushima P, Baltz RH (1985) Efficient plasmid transformation ofStreptomyces ambofaciens andStreptomyces fradiae protoplasts. J Bacteriol 163:180–185

    PubMed  Google Scholar 

  11. Matsushima P, Baltz RH (1988) Genetic transformation ofMicromonospora rosaria byStreptomyces plasmid pIJ702. J Antibiot 41:583–585

    PubMed  Google Scholar 

  12. Matsushima P, Cox KL, Baltz RH (1987) Highly transformable mutants ofStreptomyces fradiae defective in several restriction systems. Mol Gen Genet 206:393–400

    Article  PubMed  Google Scholar 

  13. Matsushima P, McHenney MA, Baltz RH (1987) Efficient transformation ofAmycolatopsis orientalis (Nocardia orientalis) protoplasts byStreptomyces plasmids. J Bacteriol 169:2298–2300

    PubMed  Google Scholar 

  14. Pidcock KA, Montenecourt BS, Sands JA (1985) Genetic recombination and transformation in protoplasts ofThermomonospora fuska. Appl Environ Microbiol 50:693–695

    Google Scholar 

  15. Piendl W, Bock A (1982) Ribosomal resistance in the gentamicin producer organismMicromonospora purpurea. Antimicrob Agents Chemother 22:231–236

    PubMed  Google Scholar 

  16. Thompson CJ, Ward JM, Hopwood DA (1982) Cloning of antibiotic resistance and nutritional genes in streptomycetes. J Bacteriol 151:668–677

    PubMed  Google Scholar 

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Authors and Affiliations

  1. Institute of Molecular Genetics and Genetic Engineering, Vojvode Stepe 283, P.O. Box 794, 11001, Belgrade, Yugoslavia

    Milorad Kojic, Ljubisa Topisirovic &  Branka Vasiljevic

Authors
  1. Milorad Kojic
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  2. Ljubisa Topisirovic
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  3. Branka Vasiljevic
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Kojic, M., Topisirovic, L. & Vasiljevic, B. Efficient transformation ofMicromonospora melanosporea protoplasts byStreptomyces plasmid. Current Microbiology 23, 343–345 (1991). https://doi.org/10.1007/BF02104137

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  • DOI: https://doi.org/10.1007/BF02104137

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