1887

Abstract

We have attempted to undertake genetic analysis in using the technique of protoplast fusion that has been successfully applied in and Efficient production of protoplasts, fusion and regeneration techniques have been established. However, variability in numbers and types of recombinants using two-, three-, and four-factor crosses was observed throughout these studies. No linkages were detected, even between loci known to be linked by cotransduction with bacteriophage MP 13. These results were similar to those reported by Alföldi and coworkers using strain 216, even though the experimental design was significantly changed. During initial subculturing, segregants were observed in a 1 : 2 : 2 ratio of noncomplementing diploids : parental-l:parental-2. The ratio changed dramatically after seven subcultures. Double recombinants appeared after nine subcultures. These results corroborate those reported in and suggest that there is a locus-inactivation phenomenon present in which is not evident in or Until the mechanism is elucidated, protoplast fusion should not be used for chromosomal mapping in However, it can be used to transfer plasmids among the bacilli at a frequency of 10‒10 per regenerated protoplast.

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1985-04-01
2021-10-24
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References

  1. Baltz R. H. 1978; Genetic recombination in Streptomyces fradiae by protoplast fusion and cell regeneration. Journal of General Microbiology 107:93–102
    [Google Scholar]
  2. Birnboim H. C., Doly J. 1979; A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Research 7:1513–1523
    [Google Scholar]
  3. Bohin J. P., Khalifa K. B., Guillen N., Schaeffer P., Hirschbein L. 1982; Phenotypic expression in vivo and transforming activity in vitro: two related functions of folded bacterial chromosomes. Molecular and General Genetics 185:65–68
    [Google Scholar]
  4. Callahan J. P., Crawford I. P., Hess G. F., Vary P. S. 1983; Cotransductional mapping of the trp-his region of Bacillus megaterium . Journal of Bacteriology 154:1455–1458
    [Google Scholar]
  5. Chang S., Cohen S. N. 1979; High frequency transformation of Bacillus subtilis protoplasts by plasmid DNA. Molecular and General Genetics 168:111–115
    [Google Scholar]
  6. Dancer B. N. 1980; Transfer of plasmids among bacilli. Journal of General Microbiology 121:263–266
    [Google Scholar]
  7. Dancer B. N., Mandelstam J. 1981; Complementation of sporulation mutants in fused protoplasts of Bacillus subtilis . Journal of General Microbiology 123:17–26
    [Google Scholar]
  8. Fodor K., Alföldi L. 1976; Fusion of protoplasts of Bacillus megaterium . Proceedings of the National Academy of Sciences of the United States of America 732147–2150
    [Google Scholar]
  9. Fodor K., Alföldi L. 1979; Polyethylene-glycol induced fusion of bacterial protoplasts. Molecular and General Genetics 168:55–59
    [Google Scholar]
  10. Fodor K., Hadlaczky G., Alföldi L. 1975; Reversion of Bacillus megaterium protoplasts to the bacillary form. Journal of Bacteriology 121:390–391
    [Google Scholar]
  11. Fodor K., Lippai-Csanady L., Alföldi L. 1983; A search for biparentals in Bacillus megaterium . Experientia (Supplementum) 45:328–329
    [Google Scholar]
  12. Frehel C, Lheritier A., Sanchez-Rivas C., Schaeffer P. 1979; Electron microscopic study of Bacillus subtilis protoplast fusion. Journal of Bacteriology 137:1354–1361
    [Google Scholar]
  13. 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]
  14. Gabor M. H., Hotchkiss R. D. 1982; Analysis of randomly picked genetic recombinants from Bacillus subtilis protoplast fusion. In Genetic Exchange: a Celebration and a New Generation283–292 Streips U. N., Goodgal S. H., Guild W. R., Wilson G. A. New York: Marcel Dekker;
    [Google Scholar]
  15. Garbe J. C, Hess G. F., Franzen M. A., Vary P. S. 1984; Genetics of leucine biosynthesis in Bacillus megaterium QM B1551. Journal of Bacteriology 157:454–459
    [Google Scholar]
  16. Guillen N., Gabor M. H., Hotchkiss R. D., Hirschbein L. 1982; Isolation and characterization of the nucleoid of non-complementing diploids from protoplast fusion in Bacillus subtilis . Molecular and General Genetics 185:69–74
    [Google Scholar]
  17. Guillen N., Sanchez-Rivas C., Hirschbein L. 1983; Absence of functional RNA encoded by a silent chromosome in noncomplementary diploids obtaiNcd from protoplast fusion in Bacillus subtilis . Molecular and General Genetics 191:81–85
    [Google Scholar]
  18. Hopwood D. A. 1981; Genetic studies with bacterial protoplasts. Annual Review of Microbiology 35:237–272
    [Google Scholar]
  19. Hopwood D. A., Wright H. M. 1978; Bacterial protoplast fusion: recombination in fused proto-plasts of Streptomyces coelicolor . Molecular and General Genetics 162:307–317
    [Google Scholar]
  20. Hopwood D. A., Wright H. M., Ward J. M. 1979; Applications of protoplasts in Streptomyces genetics . In Protoplasts - Applications in Microbial Genetics5–11 Peberdy J. F. Nottingham: Nottingham University Press;
    [Google Scholar]
  21. Hotchkiss R. D., Gabor M. H. 1980; Biparental products of bacterial protoplast fusion showing unequal parental chromosome expression. Proceedings of the National Academy of Sciences of the United States of America 113553–3557
    [Google Scholar]
  22. Kieselburg M. K., Weickert M., Vary P. S. 1984; Analysis of resident and transformant plasmids in Bacillus megaterium . Bio / Technology 2:254–259
    [Google Scholar]
  23. Lammi C. J., Vary J. C. 1972; Deoxyribonucleic acid synthesis during outgrowth of Bacillus megaterium QMB1551 spores. In Spores V277–282 Halvorson H. O., Hanson R. S., Campbell L. L. Washington DC: American Society for Microbiology;
    [Google Scholar]
  24. Luchansky J. B., Pattee P. A. 1984; Isolation of transposon Tn55l insertions near chromosomal markers of interest in Staphylococcus aureus. Journal of Bacteriology 159:894–899
    [Google Scholar]
  25. Sanchez-Rivas C. 1982; Direct selection of complementing diploids from PEG-induced fusion of Bacillus subtilis protoplasts. Molecular and General Genetics 185:329–333
    [Google Scholar]
  26. Sanchez-Rivas C., Garro A. J. 1979; Bacterial fusion assayed by a prophage complementation test. Journal of Bacteriology 137:1340–1345
    [Google Scholar]
  27. Sanchez-Rivas C, Levi-Meyrueis C, Lazard-Monier F., Schaeffer P. 1982; Diploid state of phenotypically recombinant progeny arising after protoplast fusion in Bacillus subtilis . Molecular and General Genetics 188:272–278
    [Google Scholar]
  28. Schaeffer P., Cami B., Hotchkiss R. D. 1976; Fusion of bacterial protoplasts. Proceedings of the National Academy of Sciences of the United States of American2151–2155
    [Google Scholar]
  29. Shay L., Vary J. C. 1978; Biochemical studies of glucose initiated germination in Bacillus megaterium. Biochimica et biophysica acta 538:284–292
    [Google Scholar]
  30. Stahl M. L., Pattee P. A. 1983; Computer-assisted chromosomal mapping by protoplast fusion in Staphylococcus aureus . Journal of Bacteriology 154:395–405
    [Google Scholar]
  31. Vary J. C. 1972; Spore germination of Bacillus megaterium QM B1551 mutants. Journal of Bacteriology 112:640–642
    [Google Scholar]
  32. Vary P. S. 1979; Transduction in Bacillus megaterium . Biochemical and Biophysical Research Communications 88:1119–1124
    [Google Scholar]
  33. Vary P. S., Garbe J. C, Franzen M., Frampton E. W. 1982; MP13, a generalized transducing bacteriophage for Bacillus megaterium . Journal of Bacteriology 149:1112–1119
    [Google Scholar]
  34. Wyrick P. B., Rogers H. J. 1973; Isolation and characterization of cell wall-defective variants of Bacillus subtilis and Bacillus licheniformis . Journal of Bacteriology 116:456–465
    [Google Scholar]
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