Protoplast Fusion in : Conditions for Efficient Genetic Recombination and Cell Regeneration Free

Abstract

Protoplasts from four different species of regenerated cells efficiently in hypertonic soft agar medium overlaid on partially dehydrated regeneration medium. The efficiencies of regeneration were strongly dependent upon the incubation temperatures for cell growth and for protoplast regeneration. Cell growth temperatures (before protoplast formation) required for efficient protoplast regeneration varied from species to species, and did not necessarily correlate with the optimum temperatures for protoplast regeneration. Under the best conditions, protoplasts from all four species were able to regenerate viable cells at nearly 100% efficiency and also formed confluent lawns of mycelia when plated in high concentrations. The temperatures for cell growth and protoplast regeneration also affected the frequencies of genetic recombinants obtained by protoplast fusion in and highest recombinant frequencies were obtained under conditions which favoured efficient protoplast regeneration. With the modified procedure described, maximum frequencies of genetic recombinants were obtained by treating parental protoplasts with 40 to 60% polyethylene glycol 1000.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-127-1-137
1981-11-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/micro/127/1/mic-127-1-137.html?itemId=/content/journal/micro/10.1099/00221287-127-1-137&mimeType=html&fmt=ahah

References

  1. Baltz R. H. 1978; Genetic recombination in Streptomvces fradiae by protoplast fusion and cell regeneration. Journal of General Microbiology 107:93–102
    [Google Scholar]
  2. Baltz R. H. 1980; Genetic recombination by protoplast fusion in Streptomyces. Developments in Industrial Microbiology 21:43–54
    [Google Scholar]
  3. Baltz R. H., Matsushima P. 1980; Applications of protoplast fusion, site directed mutagenesis and gene amplification to antibiotic yield improvement in Streptomyces. . In Genetics and Physiology of Actinomycetes: Proceedings of the Fourth Conference of Project III, Genetics of Microorganisms, of the US/USSR Joint Working Group on the Production of Substances by Microbial Means. pp. 124–148 Bradley S. G. Edited by Washington: American Society for Microbiology.;
    [Google Scholar]
  4. Bibb M. J., Ward J. M., Hopwood D. A. 1978; Transformation of plasmid DNA into Streptomyces at high frequency. Nature; London: 274398–400
    [Google Scholar]
  5. Bibb M. J., Ward J. M., Hopwood D. A. 1980a; The development of a cloning system for Streptomyces. Developments in Industrial Microbiology 21:55–64
    [Google Scholar]
  6. Bibb M. J., Schottel J. L., Cohen S. N. 1980b; A DNA cloning system for interspecies gene transfer in antibiotic-producing Streptomyces. Nature; London: 284526–531
    [Google Scholar]
  7. Chater K. F. 1980; Actinophage DNA. Developments in Industrial Microbiology 21:65–74
    [Google Scholar]
  8. Chater K. F., Wilde L. C. 1980; Streptomyces albus G mutants defective in the Sal GI restriction-modification system. Journal of General Microbiology 116:323–334
    [Google Scholar]
  9. Coetzee J. N., Sirgel F. A., Lecatsas G. 1979; Genetic recombination in fused spheroplasts of Providence alcalifaciens. Journal of General Microbiology 114:313–322
    [Google Scholar]
  10. Fodor K., Alföldi L. 1976; Fusion of protoplasts of Bacillus megaterium. Proceedings of the National Academy of Sciences of the United States of America 73:2147–2150
    [Google Scholar]
  11. Godfrey O., Ford L., Huber M. L. B. 1978; Interspecies matings of Streptomyces fradiae with Streptomyces bikiniensis mediated by conventional and protoplast fusion techniques. Canadian Journal of Microbiology 24:994–997
    [Google Scholar]
  12. Hopwood D. A., Wright H. M. 1978; Bacterial protoplast fusion: recombination in fused protoplasts of Streptomyces coelicolor. Molecular and General Genetics 162:307–317
    [Google Scholar]
  13. Hopwood D. A., Wright H. M. 1979; Factors affecting recombinant frequency in protoplast fusions of Streptomyces coelicolor. Journal of General Microbiology 111:137–143
    [Google Scholar]
  14. Hopwood D. A., Wright H. M., Bibb M. J., Cohen S. N. 1977; Genetic recombination through protoplast fusion in Streptomyces. Nature; London: 268171–174
    [Google Scholar]
  15. Kaneko H., Sakaguchi K. 1979; Fusion of protoplasts and genetic recombination of Brevibacterium flavum. Agricultural and Biological Chemistry 43:1007–1013
    [Google Scholar]
  16. Ochi K., Hitchcock 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]
  17. Oh Y. K., Speth J. L., Nash C. H. 1980; Protoplast fusion with Streptosporangium viridogriseum. Developments in Industrial Microbiology 21:219–226
    [Google Scholar]
  18. 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]
  19. Queener S. W., Baltz R. H. 1979; Genetics of industrial microorganisms. Annual Reports on Fermentation Processes 3:5–45
    [Google Scholar]
  20. Schaeffer P., Cami B., Hotchkiss R. D. 1976; Fusion of bacterial protoplasts. Proceedings of the National Academy of Sciences of the United States of America 73:2151–2155
    [Google Scholar]
  21. Seno E. T., Pieper R. L., Huber F. M. 1977; Terminal stages in the biosynthesis of tylosin. Antimicrobial Agents and Chemotherapy 11:455–461
    [Google Scholar]
  22. Suarez J. E., Chater K. F. 1980a; Polyethylene glycol-assisted transfection of Streptomyces protoplasts. Journal of Bacteriology 142:8–14
    [Google Scholar]
  23. Suarez J. E., Chater K. F. 1980b; DNA cloning in Streptomyces: a bifunctional replicon comprising pBR322 inserted into a Streptomyces phage. Nature; London: 286527–529
    [Google Scholar]
  24. Thompson C. J., Ward J. M., Hopwood D. A. 1980; DNA cloning in Streptomyces: resistance genes from antibiotic-producing species. Nature; London: 286527
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-127-1-137
Loading
/content/journal/micro/10.1099/00221287-127-1-137
Loading

Data & Media loading...

Most cited Most Cited RSS feed