Skip to main content
NCBI home page
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1985 Mar;49(3):577–581. doi: 10.1128/aem.49.3.577-581.1985

Studies on transfection and transformation of protoplasts of Bacillus larvae, Bacillus subtilis, and Bacillus popilliae.

N Bakhiet, D P Stahly
PMCID: PMC373552  PMID: 3922299

Abstract

Protoplasts of Bacillus larvae NRRL b-3555 and Bacillus subtilis RM125 (restrictionless, modificationless mutant) were transfected with DNA from the B. larvae bacteriophage PBL1c in the presence of polyethylene glycol. B. subtilis 168 and Bacillus popilliae NRRL B-2309M protoplasts could not be transfected with PBL1c DNA. Protoplasts of B larvae NRRL B-3555 were transformed with plasmids pC194 and pHV33 in the presence of polyethylene glycol. The frequency of transformation was much higher when the plasmids were isolated from B. larvae NRRL B-3555 transformants than when they were isolated from B. subtilis 168. These results indicate that the restriction-modification systems found in B. larvae NRRL B-3555 and B. subtilis 168 may be different. Conditions for protoplast formation and cell wall regeneration were developed for B. popilliae NRRL B-2309S. However, no transformation occurred with plasmids pC194 and pHV33 (isolated from B. subtilis 168).

Full text

PDF
577

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Allcock E. R., Reid S. J., Jones D. T., Woods D. R. Clostridium acetobutylicum Protoplast Formation and Regeneration. Appl Environ Microbiol. 1982 Mar;43(3):719–721. doi: 10.1128/aem.43.3.719-721.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chang S., Cohen S. N. High frequency transformation of Bacillus subtilis protoplasts by plasmid DNA. Mol Gen Genet. 1979 Jan 5;168(1):111–115. doi: 10.1007/BF00267940. [DOI] [PubMed] [Google Scholar]
  3. Costilow R. N., Coulter W. H. Physiological studies of an oligosporogenous strain of Bacillus popilliae. Appl Microbiol. 1971 Dec;22(6):1076–1084. doi: 10.1128/am.22.6.1076-1084.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dingman D. W., Bakhiet N., Field C. C., Stahly D. P. Isolation of two bacteriophages from Bacillus larvae, PBL1 and PBL0.5, and partial characterization of PBL1. J Gen Virol. 1984 Jun;65(Pt 6):1101–1105. doi: 10.1099/0022-1317-65-6-1101. [DOI] [PubMed] [Google Scholar]
  5. Dingman D. W., Stahly D. P. Medium Promoting Sporulation of Bacillus larvae and Metabolism of Medium Components. Appl Environ Microbiol. 1983 Oct;46(4):860–869. doi: 10.1128/aem.46.4.860-869.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dingman D. W., Stahly D. P. Protection of Bacillus larvae from Oxygen Toxicity with Emphasis on the Role of Catalase. Appl Environ Microbiol. 1984 Jun;47(6):1228–1237. doi: 10.1128/aem.47.6.1228-1237.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Ehrlich S. D. DNA cloning in Bacillus subtilis. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1433–1436. doi: 10.1073/pnas.75.3.1433. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Ehrlich S. D. Replication and expression of plasmids from Staphylococcus aureus in Bacillus subtilis. Proc Natl Acad Sci U S A. 1977 Apr;74(4):1680–1682. doi: 10.1073/pnas.74.4.1680. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fodor K., Alföldi L. Fusion of protoplasts of Bacillus megaterium. Proc Natl Acad Sci U S A. 1976 Jun;73(6):2147–2150. doi: 10.1073/pnas.73.6.2147. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gochnauer T. A. Some properties of a bacteriophage from Bacillus larvae. J Invertebr Pathol. 1970 Mar;15(2):149–156. doi: 10.1016/0022-2011(70)90228-4. [DOI] [PubMed] [Google Scholar]
  11. Goldfarb D. S., Rodriguez R. L., Doi R. H. Translational block to expression of the Escherichia coli Tn9-derived chloramphenicol-resistance gene in Bacillus subtilis. Proc Natl Acad Sci U S A. 1982 Oct;79(19):5886–5890. doi: 10.1073/pnas.79.19.5886. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gray O., Chang S. Molecular cloning and expression of Bacillus licheniformis beta-lactamase gene in Escherichia coli and Bacillus subtilis. J Bacteriol. 1981 Jan;145(1):422–428. doi: 10.1128/jb.145.1.422-428.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ito J., Kawamura F., Yanofsky S. Analysis of phi 29 and phi 15 genomes by bacterial restriction endonucleases, EcoR1 and Hpal. Virology. 1976 Mar;70(1):37–51. doi: 10.1016/0042-6822(76)90234-8. [DOI] [PubMed] [Google Scholar]
  14. Jentsch S., Günthert U., Trautner T. A. DNA methyltransferases affecting the sequence 5'CCGG. Nucleic Acids Res. 1981 Jun 25;9(12):2753–2759. doi: 10.1093/nar/9.12.2753. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kreft J., Bernhard K., Goebel W. Recombinant plasmids capable to replication in B. subtilis and E. coli. Mol Gen Genet. 1978 Jun 1;162(1):59–67. doi: 10.1007/BF00333851. [DOI] [PubMed] [Google Scholar]
  16. Landman O. E., Ryter A., Fréhel C. Gelatin-induced reversion of protoplasts of Bacillus subtilis to the bacillary form: electron-microscopic and physical study. J Bacteriol. 1968 Dec;96(6):2154–2170. doi: 10.1128/jb.96.6.2154-2170.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. McLaughlin J. R., Murray C. L., Rabinowitz J. C. Unique features in the ribosome binding site sequence of the gram-positive Staphylococcus aureus beta-lactamase gene. J Biol Chem. 1981 Nov 10;256(21):11283–11291. [PubMed] [Google Scholar]
  18. Okanishi M., Suzuki K., Umezawa H. Formation and reversion of Streptomycete protoplasts: cultural condition and morphological study. J Gen Microbiol. 1974 Feb;80(2):389–400. doi: 10.1099/00221287-80-2-389. [DOI] [PubMed] [Google Scholar]
  19. Primrose S. B., Ehrlich S. D. Isolation of plasmid deletion Mutants and study of their instability. Plasmid. 1981 Sep;6(2):193–201. doi: 10.1016/0147-619x(81)90066-4. [DOI] [PubMed] [Google Scholar]
  20. Stahl M. L., Pattee P. A. Computer-assisted chromosome mapping by protoplast fusion in Staphylococcus aureus. J Bacteriol. 1983 Apr;154(1):395–405. doi: 10.1128/jb.154.1.395-405.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Uozumi T., Hoshino T., Miwa K., Horinouchi S., Beppu T., Arima K. Restriction and modification in Bacillus species: genetic transformation of bacteria with DNA from different species, part I. Mol Gen Genet. 1977 Mar 28;152(1):65–69. doi: 10.1007/BF00264941. [DOI] [PubMed] [Google Scholar]
  22. Williams J. A., Yeggy J. P., Field C. C., Markovetz A. J. Resistance plasmids in Pseudomonas cepacia 4G9. J Bacteriol. 1979 Dec;140(3):1017–1022. doi: 10.1128/jb.140.3.1017-1022.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES