Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1979 Aug;139(2):597–607. doi: 10.1128/jb.139.2.597-607.1979

Isolation and characterization of replication-deficient mutants of ColE1 plasmids.

T Hashimoto-Gotoh, J Inselburg
PMCID: PMC216909  PMID: 378979

Abstract

Replication-defective mutants of plasmid ColE1 were isolated from a chimeric plasmid formed by ligating a temperature-sensitive replication derivative of pSC101, pHSG1, with a ColE1-Tn3-containing plasmid. The replication-defective ColE1 mutants isolated were all spontaneous deletion mutants that had lost the ColE1 replication origin and regions adjacent to it. The extent of a deletion was determined by analyzing restriction endonuclease-generated deoxyribonucleic acid fragments of the ColE1 plasmid component of the chimeras by both agarose and polyacrylamide gel electrophoresis. None of the chimeras containing the replication-defective ColE1 mutants was able to replicate in the presence of chloramphenicol. The expression of ColE1 incompatibility was either markedly reduced or not detectable in the replication mutants isolated.

Full text

PDF
597

Images in this article

Selected References

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

  1. Adler H. I., Fisher W. D., Cohen A., Hardigree A. A. MINIATURE escherichia coli CELLS DEFICIENT IN DNA. Proc Natl Acad Sci U S A. 1967 Feb;57(2):321–326. doi: 10.1073/pnas.57.2.321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bolivar F., Betlach M. C., Heyneker H. L., Shine J., Rodriguez R. L., Boyer H. W. Origin of replication of pBR345 plasmid DNA. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5265–5269. doi: 10.1073/pnas.74.12.5265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cabello F., Timmis K., Cohen S. N. Replication control in a composite plasmid constructed by in vitro linkage of two distinct replicons. Nature. 1976 Jan 29;259(5541):285–290. doi: 10.1038/259285a0. [DOI] [PubMed] [Google Scholar]
  4. Clewell D. B. Nature of Col E 1 plasmid replication in Escherichia coli in the presence of the chloramphenicol. J Bacteriol. 1972 May;110(2):667–676. doi: 10.1128/jb.110.2.667-676.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cohen S. N., Chang A. C., Boyer H. W., Helling R. B. Construction of biologically functional bacterial plasmids in vitro. Proc Natl Acad Sci U S A. 1973 Nov;70(11):3240–3244. doi: 10.1073/pnas.70.11.3240. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hashimoto-Gotoh T., Inselburg J. ColE1 plasmid incompatibility: localization and analysis of mutations affecting incompatibility. J Bacteriol. 1979 Aug;139(2):608–619. doi: 10.1128/jb.139.2.608-619.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hashimoto-Gotoh T., Sekiguchi M. Mutations of temperature sensitivity in R plasmid pSC101. J Bacteriol. 1977 Aug;131(2):405–412. doi: 10.1128/jb.131.2.405-412.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hashimoto T., Sekiguchi M. Isolation of temperature-sensitive mutants of R plasmid by in vitro mutagenesis with hydroxylamine. J Bacteriol. 1976 Sep;127(3):1561–1563. doi: 10.1128/jb.127.3.1561-1563.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Inselburg J. ColE1 plasmid mutants affecting growth of an Escherichia coli recB recC sbcB mutant. J Bacteriol. 1978 Jan;133(1):433–436. doi: 10.1128/jb.133.1.433-436.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Inselburg J. Isolation, mapping, and examination of effects of TnA insertions in ColE1 plasmids. J Bacteriol. 1977 Jan;129(1):482–491. doi: 10.1128/jb.129.1.482-491.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Inselburg J. Segregation into and replication of plasmid deoxyribonucleic acid in chromosomeless segregants of Escherichia coli. J Bacteriol. 1970 Jun;102(3):642–647. doi: 10.1128/jb.102.3.642-647.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Inselburg J., Ware P. Isolation and genetic analysis of deletion mutants of colicin E1 plasmids carrying a TnA insertion. J Bacteriol. 1977 Oct;132(1):321–331. doi: 10.1128/jb.132.1.321-331.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ishii K., Hashimoto-Gotoh T., Matsubara K. Random replication and random assortment model for plasmid incompatibility in bacteria. Plasmid. 1978 Sep;1(4):435–445. doi: 10.1016/0147-619x(78)90002-1. [DOI] [PubMed] [Google Scholar]
  14. Kopecko D. J., Cohen S. N. Site specific recA--independent recombination between bacterial plasmids: involvement of palindromes at the recombinational loci. Proc Natl Acad Sci U S A. 1975 Apr;72(4):1373–1377. doi: 10.1073/pnas.72.4.1373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Nisen P. D., Kopecko D. J., Chou J., Cohen S. N. Site-specific DNA deletions occurring adjacent to the termini of a transposable ampicillin resistance element (Tn3). J Mol Biol. 1977 Dec 25;117(4):975–978. doi: 10.1016/s0022-2836(77)80008-9. [DOI] [PubMed] [Google Scholar]
  16. Novick R. P., Hoppensteadt F. C. On plasmid incompatibility. Plasmid. 1978 Sep;1(4):421–434. doi: 10.1016/0147-619x(78)90001-x. [DOI] [PubMed] [Google Scholar]
  17. Oka A., Takanami M. Cleavage map of colicin E1 plasmid. Nature. 1976 Nov 11;264(5582):193–196. doi: 10.1038/264193a0. [DOI] [PubMed] [Google Scholar]
  18. Timmis K., Cabello F., Cohen S. N. Utilization of two distinct modes of replication by a hybrid plasmid constructed in vitro from separate replicons. Proc Natl Acad Sci U S A. 1974 Nov;71(11):4556–4560. doi: 10.1073/pnas.71.11.4556. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Tomizawa J. I., Ohmori H., Bird R. E. Origin of replication of colicin E1 plasmid DNA. Proc Natl Acad Sci U S A. 1977 May;74(5):1865–1869. doi: 10.1073/pnas.74.5.1865. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES