1887

Abstract

The regeneration of streptomycete protoplasts is a major step following genetic manipulations such as fusion and DNA-mediated transformation. Reports of studies on the regeneration of protoplasts from are limited and for this reason the experiments described in this paper were carried out. An investigation of protoplast formation and cytology was made to gain further insight into the loss of protoplast viability in osmotically stabilized support media. Protoplasts with the highest regeneration frequency were isolated from mycelium, grown in a two-stage culture system (without glycine), using lysozyme dissolved in a sucrose osmoticum containing 1% bovine serum albumin. The latter promoted improved protoplast viability. A systematic survey was made of the components of regeneration medium R5, previously used for , and other potentially advantageous components and conditions, in an attempt to raise the regeneration frequency of the protoplasts. An improved regeneration medium (R6) and protocol which supported higher and more consistent levels of regeneration of protoplasts resulted from these experiments. These improved procedures for protoplast isolation and regeneration proved to be suitable for other streptomycete species.

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1989-08-01
2022-01-16
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References

  1. Akamatsu T., Sekiguchi J. 1981; Studies on regeneration media for Bacillus subtilis protoplasts.. Agricultural and Biological Chemistry 45:2887–2894
    [Google Scholar]
  2. Araujo J.M., De, Andrade Lyra F.D., De, Kurylowicz W. 1984; Recombination in antibiotic producing Streptomyces by protoplast fusion.. Revista Microbiologica São Paulo 15:67–81
    [Google Scholar]
  3. Bailey C.R., Butler M.J., Normansell I.D., Rowlands R.T., Winstanley D.J. 1984; Cloning a Streptomyces clavuligerus genetic locus involved in clavulanic acid biosynthesis.. Biotechnology 2:808–811
    [Google Scholar]
  4. Baltz R.H. 1980; Genetic recombination by protoplast fusion in Streptomyces.. Developments in Industrial Microbiology 21:43–54
    [Google Scholar]
  5. Baltz R.H., Matsushima P. 1980; Applications of protoplast fusion, site directed mutagenesis and gene amplification to antibiotic yield improvement in Streptomyces. The Biology of the Actinomyces and Related Organisms 15:18–34
    [Google Scholar]
  6. Baltz R.H., Matsushima P. 1981; Protoplast fusion in Streptomyces: Conditions for efficient genetic recombination and cell regeneration. Journal of General Microbiology 127:137–146
    [Google Scholar]
  7. Bibb M.J., Ward J.M., Hopwood D.A. 1978; Transformation of plasmid DNA into Streptomyces at high frequency. Nature, London 274:398–400
    [Google Scholar]
  8. Calich G.V.L., Purchio A., Paula G.C. 1978; A new fluorescent viability test for fungal cells. Mycopathologia 66:175–177
    [Google Scholar]
  9. Chater K.F., Hopwood D.A., Kieser T., Thompson C.J. 1982; Gene cloning in Streptomyces. Current Topics in Microbiology and Immunology 96:69–95
    [Google Scholar]
  10. Gabor M.H., Hotchkiss R.D. 1979; Parameters governing bacterial regeneration and genetic recombination after fusion of Bacillus subtilis protoplasts. Journal of Bacteriology 137:1346–1353
    [Google Scholar]
  11. Higgens C.E., Kastner R.F. 1971; Streptomyces clavuligerus sp. nov, a β-lactam antibiotic producer. International Journal of Systematic Bacteriology 21:326–331
    [Google Scholar]
  12. Hitchcock M.J.M., Katz E. 1978; Actinomycin biosynthesis by protoplasts derived from Streptomyces parvulus.. Antimicrobial Agents and Chemotherapy 13:104–114
    [Google Scholar]
  13. Hopwood D.A. 1981a; Genetic studies of antibiotics and other secondary metabolites. Symposia of the Society for General Microbiology 31:187–218
    [Google Scholar]
  14. Hopwood D.A. 1981b; Genetic studies with bacterial protoplasts. Annual Review of Microbiology 35:237–272
    [Google Scholar]
  15. Hopwood D.A., Bibb M.J., Chater K.F., Kieser T., Bruton C.J., Kieser H.M., Lydiate D.J., Smith C.P., Ward J.M., Schrempf H. 1985 Genetic Manipulation of Streptomyces. A Laboratory Manual Norwich: The John Innes Foundation;
    [Google Scholar]
  16. Hopwood D.A., Wright H.M., Bibb M.J., Cohen S.N. 1977; Genetic recombination through protoplast fusion in Streptomyces. Nature, London 268:171–174
    [Google Scholar]
  17. Illing G.T. 1987 Protoplast fusion and regeneration in Streptomyces clavuligerus PhD thesis University of Nottingham, Nottingham;
    [Google Scholar]
  18. Keller U., Poschmann S., Krengel U., Klein-Kauf H., Kraepelin G. 1983; Studies of protoplast fusion in Streptomyces chrysomallus.. Journal of General Microbiology 129:1725–1731
    [Google Scholar]
  19. Kim K.S., Ryu Y.D.D., Lee S.Y. 1983; Application of protoplast fusion technique to genetic recombination of Micromonospora rosaria.. Enzyme and Microbial Technology 5:273–280
    [Google Scholar]
  20. Liu C.J., Pogell B.M., Redshaw P.A. 1985; Regulation of conditional aerial mycelium mutants of Streptomyces. Journal of General Microbiology 131:1015–1021
    [Google Scholar]
  21. Matsushima P., Baltz R.H. 1985; Efficient plasmid transformation of Streptomyces ambofaciens and Streptomyces fradiae protoplasts. Journal of Bacteriology 163:180–185
    [Google Scholar]
  22. Mirdamadi-Tehrani J., Mitchell J.I., Williams S.T., Ritchie D.A. 1986; Factors affecting protoplast formation and regeneration by four species of Streptomyces. Letters in Applied Microbiology 3:27–30
    [Google Scholar]
  23. Ochi K., Hitchcock M.J.M., Katz E. 1979; High frequency fusion of Streptomyces parvulus or Streptomyces antibioticus protoplasts induced by polyethylene glycol. Journal of Bacteriology 139:984–992
    [Google Scholar]
  24. Ogawa H., Imai S., Satoh A., Kojima H. 1983; An improved method for preparation of Streptomyces and Micromonospora protoplasts. Journal of Antibiotics 36:184–186
    [Google Scholar]
  25. Oh Y.H., Speth J.L., Nash C.H. 1980; Protoplast fusion with Streptosporangium virido- griseum.. Developments in Industrial Microbiology 21:219–226
    [Google Scholar]
  26. Okanishi M., Suzuki K., Umezawa H. 1974; Formation and reversion of streptomycete protoplasts : cultural conditions and morphological study. Journal of General Microbiology 80:389–400
    [Google Scholar]
  27. Robinson M., Lewis E., Napier E. 1981; Occurrence of reiterated DNA sequences in strains of Streptomyces produced by an interspecific protoplast fusion. Molecular and General Genetics 182:336–360
    [Google Scholar]
  28. Rolinson G.N. 1984; The history and background of augmentin. In Augmentin: Clavulanate-potentiated Amoxycillin pp. 5–9 Croyden E. A. P., Michel M. F. Edited by London: Academic Press;
    [Google Scholar]
  29. Sagara Y., Fukui K., Ota F., Yoshida N., Kashiyama T., Fujimoto M. 1971; Rapid formation of protoplasts of Streptomyces griseoflavus and their fine structure. Japanese Journal of Microbiology 15:73–84
    [Google Scholar]
  30. Such T.M. 1981 Studies on protoplasts of Streptomyces MPhil thesis University of Nottingham, Nottingham;
    [Google Scholar]
  31. Thompson C.J., Ward J.M., Hopwood D.A. 1982; Cloning of antibiotic resistance and nutritional genes from antibiotic producing species. Journal of Bacteriology 151:668–677
    [Google Scholar]
  32. Zinder N.D., Arndt W.F. 1956; Production of protoplasts of Escherichia coli by lysozyme treatment. Proceedings of the National Academy of Sciences of the United States of America 42:586–590
    [Google Scholar]
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