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. 1981 Feb;145(2):861–866. doi: 10.1128/jb.145.2.861-866.1981

Isolation and analysis of multicopy extragenic suppressors of dnaA mutations.

S J Projan, J A Wechsler
PMCID: PMC217190  PMID: 6257653

Abstract

Recombinant plasmids were constructed from restriction enzyme digests of Escherichia coli chromosomal deoxyribonucleic acid and pMB9 plasmid deoxyribonucleic acid and selected for correction of the dnaA phenotype. The three plasmids isolated, all retransformed dnaA cells, both recA+ and recA, such that all tetracycline-resistant transformants selected at permissive temperature simultaneously became temperature resistant. Restriction enzyme mapping of the plasmids showed all three to be different, and it was subsequently shown that none contained the dnaA+ gene. Though each of the three plasmids suppressed three different temperature-sensitive dnaA alleles, none corrected the phenotype of an unsuppressed dnaA amber allele. It was concluded, therefore, that each plasmid contained a unique extragenic suppressor of dnaA and that the suppression was observed because of the elevated gene dosage of the cloned material. The plasmids were unstable in the absence of selection.

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Selected References

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

  1. Abe M., Tomizawa J. Chromosome replication in Escherichia coli K12 mutant affected in the process of DNA initiation. Genetics. 1971 Sep;69(1):1–15. doi: 10.1093/genetics/69.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bachmann B. J., Low K. B. Linkage map of Escherichia coli K-12, edition 6. Microbiol Rev. 1980 Mar;44(1):1–56. doi: 10.1128/mr.44.1.1-56.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Beyersmann D., Messer W., Schlicht M. Mutants of Escherichia coli B-r defective in deoxyribonucleic acid initiation: dnaI, a new gene for replication. J Bacteriol. 1974 Jun;118(3):783–789. doi: 10.1128/jb.118.3.783-789.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Carl P. L. Escherichia coli mutants with temperature-sensitive synthesis of DNA. Mol Gen Genet. 1970;109(2):107–122. doi: 10.1007/BF00269647. [DOI] [PubMed] [Google Scholar]
  6. Clewell D. B., Helinski D. R. Supercoiled circular DNA-protein complex in Escherichia coli: purification and induced conversion to an opern circular DNA form. Proc Natl Acad Sci U S A. 1969 Apr;62(4):1159–1166. doi: 10.1073/pnas.62.4.1159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. Gotfried F., Wechsler J. A. Dominance of dnaA+ to dnaA in Escherichia coli. J Bacteriol. 1977 May;130(2):963–964. doi: 10.1128/jb.130.2.963-964.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hansen F. G., von Meyenburg K. Characterization of the dnaA, gyrB and other genes in the dnaA region of the Escherichia coli chromosome on specialized transducing phages lambda tna. Mol Gen Genet. 1979 Sep;175(2):135–144. doi: 10.1007/BF00425529. [DOI] [PubMed] [Google Scholar]
  10. Hirota Y., Ryter A., Jacob F. Thermosensitive mutants of E. coli affected in the processes of DNA synthesis and cellular division. Cold Spring Harb Symp Quant Biol. 1968;33:677–693. doi: 10.1101/sqb.1968.033.01.077. [DOI] [PubMed] [Google Scholar]
  11. Kohiyama M. DNA synthesis in temperature sensitive mutants of Escherichia coli. Cold Spring Harb Symp Quant Biol. 1968;33:317–324. doi: 10.1101/sqb.1968.033.01.036. [DOI] [PubMed] [Google Scholar]
  12. Lederberg E. M., Cohen S. N. Transformation of Salmonella typhimurium by plasmid deoxyribonucleic acid. J Bacteriol. 1974 Sep;119(3):1072–1074. doi: 10.1128/jb.119.3.1072-1074.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lindahl G., Hirota Y., Jacob F. On the process of cellular division in Escherichia coli: replication of the bacterial chromosome under control of prophage P2. Proc Natl Acad Sci U S A. 1971 Oct;68(10):2407–2411. doi: 10.1073/pnas.68.10.2407. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Miki T., Kimura M., Hiraga S., Nagata T., Yura T. Cloning and physical mapping of the dnaA region of the Escherichia coli chromosome. J Bacteriol. 1979 Dec;140(3):817–824. doi: 10.1128/jb.140.3.817-824.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  16. SAITO H., MIURA K. I. PREPARATION OF TRANSFORMING DEOXYRIBONUCLEIC ACID BY PHENOL TREATMENT. Biochim Biophys Acta. 1963 Aug 20;72:619–629. [PubMed] [Google Scholar]
  17. Schaller H., Otto B., Nüsslein V., Huf J., Herrmann R., Bonhoeffer F. Deoxyribonucleic acid replication in vitro. J Mol Biol. 1972 Jan 28;63(2):183–200. doi: 10.1016/0022-2836(72)90369-5. [DOI] [PubMed] [Google Scholar]
  18. Smith J. D., Barnett L., Brenner S., Russell R. L. More mutant tyrosine transfer ribonucleic acids. J Mol Biol. 1970 Nov 28;54(1):1–14. doi: 10.1016/0022-2836(70)90442-0. [DOI] [PubMed] [Google Scholar]
  19. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  20. Tanaka T., Weisblum B. Construction of a colicin E1-R factor composite plasmid in vitro: means for amplification of deoxyribonucleic acid. J Bacteriol. 1975 Jan;121(1):354–362. doi: 10.1128/jb.121.1.354-362.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Thomas M., Davis R. W. Studies on the cleavage of bacteriophage lambda DNA with EcoRI Restriction endonuclease. J Mol Biol. 1975 Jan 25;91(3):315–328. doi: 10.1016/0022-2836(75)90383-6. [DOI] [PubMed] [Google Scholar]
  22. Wechsler J. A., Gross J. D. Escherichia coli mutants temperature-sensitive for DNA synthesis. Mol Gen Genet. 1971;113(3):273–284. doi: 10.1007/BF00339547. [DOI] [PubMed] [Google Scholar]
  23. Wechsler J. A. dnaA alleles are recessive. J Bacteriol. 1980 Nov;144(2):856–857. doi: 10.1128/jb.144.2.856-857.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Wehr C. T., Waskell L., Glaser D. A. Characteristics of cold-sensitive mutants of Escherichia coli K-12 defective in deoxyribonucleic acid replication. J Bacteriol. 1975 Jan;121(1):99–107. doi: 10.1128/jb.121.1.99-107.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]

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