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. 1984 Jan;157(1):152–157. doi: 10.1128/jb.157.1.152-157.1984

Isolation and characterization of Bacillus subtilis mutants altered in competence.

R Fani, G Mastromei, M Polsinelli, G Venema
PMCID: PMC215145  PMID: 6317647

Abstract

We isolated and characterized four Bacillus subtilis competence-deficient mutants. The mutants were obtained by nitrosoguanidine mutagenesis and by screening for mutants unable to be transformed both on solid and in liquid medium. Most of the mutants obtained in this way were tested for their sensitivity to the DNA-damaging agents methyl methanesulfonate, mitomycin C, and UV light. Among the mutants which did not show an increased sensitivity to these agents, four were chosen for further characterization. Data were obtained which indicate that the mutants are reduced in chromosomal and plasmid transformation and in transfection, whereas they are not altered in transduction and in protoplast transformation. Transformation experiments carried out by mixing a culture of a mutant with a culture of a wild-type strain gave some complementation for competence with one of the strains. The mutants were also characterized for their capacity to bind, take up, and break down transforming DNA; furthermore, the four competence mutations were mapped, and the results indicate that they belong to four different genes.

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Selected References

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

  1. Akrigg A., Ayad S. R., Barker G. R. The nature of a competence-inducing factor in Bacillus subtilis. Biochem Biophys Res Commun. 1967 Sep 27;28(6):1062–1067. doi: 10.1016/0006-291x(67)90090-3. [DOI] [PubMed] [Google Scholar]
  2. Biswal N., Kleinschmidt A. K., Spatz H. C., Trautner T. A. Physical properties of the DNA of bacteriophage SP50. Mol Gen Genet. 1967;100(1):39–55. doi: 10.1007/BF00425774. [DOI] [PubMed] [Google Scholar]
  3. Buitenwerf J., Venema G. Transformation in bacillus subtilis: fate of transforming DNA in transformation deficient mutants. Mol Gen Genet. 1977 Mar 7;151(2):203–213. doi: 10.1007/BF00338696. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. DAVIS B. D., MINGIOLI E. S. Mutants of Escherichia coli requiring methionine or vitamin B12. J Bacteriol. 1950 Jul;60(1):17–28. doi: 10.1128/jb.60.1.17-28.1950. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Dedonder R. A., Lepesant J. A., Lepesant-Kejzlarová J., Billault A., Steinmetz M., Kunst F. Construction of a kit of reference strains for rapid genetic mapping in Bacillus subtilis 168. Appl Environ Microbiol. 1977 Apr;33(4):989–993. doi: 10.1128/aem.33.4.989-993.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dooley D. C., Hadden C. T., Nester E. W. Macromolecular synthesis in Bacillus subtilis during development of the competent state. J Bacteriol. 1971 Nov;108(2):668–679. doi: 10.1128/jb.108.2.668-679.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dubnau D., Cirigliano C. Fate of transforming DNA following uptake by competent Bacillus subtilis. Formation and properties of products isolated from transformed cells which are derived entirely from donor DNA. J Mol Biol. 1972 Feb 28;64(1):9–29. doi: 10.1016/0022-2836(72)90318-x. [DOI] [PubMed] [Google Scholar]
  9. Fein J. E., Rogers H. J. Autolytic enzyme-deficient mutants of Bacillus subtilis 168. J Bacteriol. 1976 Sep;127(3):1427–1442. doi: 10.1128/jb.127.3.1427-1442.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gryczan T. J., Contente S., Dubnau D. Characterization of Staphylococcus aureus plasmids introduced by transformation into Bacillus subtilis. J Bacteriol. 1978 Apr;134(1):318–329. doi: 10.1128/jb.134.1.318-329.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hoch J. A., Barat M., Anagnostopoulos C. Transformation and transduction in recombination-defective mutants of Bacillus subtilis. J Bacteriol. 1967 Jun;93(6):1925–1937. doi: 10.1128/jb.93.6.1925-1937.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Joenje H., Venema G. Different nuclease activities in competent and noncompetent Bacillus subtilis. J Bacteriol. 1975 Apr;122(1):25–33. doi: 10.1128/jb.122.1.25-33.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lepesant-Kejzlarová J., Lepesant J. A., Walle J., Billault A., Dedonder R. Revision of the linkage map of Bacillus subtilis 168: indications for circularity of the chromosome. J Bacteriol. 1975 Mar;121(3):823–834. doi: 10.1128/jb.121.3.823-834.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lepesant J. A., Kunst F., Lepesant-Kejzlarová J., Dedonder R. Chromosomal location of mutations affecting sucrose metabolism in Bacillus subtilis Marburg. Mol Gen Genet. 1972;118(2):135–160. doi: 10.1007/BF00267084. [DOI] [PubMed] [Google Scholar]
  15. Mulder J. A., Venema G. Isolation and partial characterization of Bacillus subtilis mutants impaired in DNA entry. J Bacteriol. 1982 Apr;150(1):260–268. doi: 10.1128/jb.150.1.260-268.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. NESTER E. W., LEDERBERG J. Linkage of genetic units of Bacillus subtilis in DNA transformation. Proc Natl Acad Sci U S A. 1961 Jan 15;47:52–55. doi: 10.1073/pnas.47.1.52. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Riva S., Polsinelli M., Falaschi A. A new phage of Bacillus subtilis with infectious DNA having separable strands. J Mol Biol. 1968 Jul 28;35(2):347–356. doi: 10.1016/s0022-2836(68)80029-4. [DOI] [PubMed] [Google Scholar]
  18. Smith H. O., Danner D. B., Deich R. A. Genetic transformation. Annu Rev Biochem. 1981;50:41–68. doi: 10.1146/annurev.bi.50.070181.000353. [DOI] [PubMed] [Google Scholar]
  19. Smith H., de Vos W., Bron S. Transformation in Bacillus subtilis: properties of DNA-binding-deficient mutants. J Bacteriol. 1983 Jan;153(1):12–20. doi: 10.1128/jb.153.1.12-20.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Spizizen J. TRANSFORMATION OF BIOCHEMICALLY DEFICIENT STRAINS OF BACILLUS SUBTILIS BY DEOXYRIBONUCLEATE. Proc Natl Acad Sci U S A. 1958 Oct 15;44(10):1072–1078. doi: 10.1073/pnas.44.10.1072. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Streips U. N., Young F. E. Transformation in Bacillus subtilis using excreted DNA. Mol Gen Genet. 1974;133(1):47–55. doi: 10.1007/BF00268676. [DOI] [PubMed] [Google Scholar]
  22. Venema G. Bacterial transformation. Adv Microb Physiol. 1979;19:245–331. doi: 10.1016/s0065-2911(08)60200-3. [DOI] [PubMed] [Google Scholar]
  23. Yamagishi H., Takahashi I. Transducing particles of PBS 1. Virology. 1968 Dec;36(4):639–645. doi: 10.1016/0042-6822(68)90194-3. [DOI] [PubMed] [Google Scholar]
  24. de Vos W. M., Venema G. Fate of plasmid DNA in transformation of Bacillus subtilis protoplasts. Mol Gen Genet. 1981;182(1):39–43. doi: 10.1007/BF00422764. [DOI] [PubMed] [Google Scholar]

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