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Journal of Bacteriology logoLink to Journal of Bacteriology
. 1977 Apr;130(1):167–172. doi: 10.1128/jb.130.1.167-172.1977

Method for the isolation of Escherichia coli mutants with enhanced recombination between chromosomal duplications.

E B Konrad
PMCID: PMC235189  PMID: 323226

Abstract

A method is described for the isolation of Escherichia coli mutants that show increased recombination between a pair of chromosomal duplications. These "hyper-rec" mutants display a variety of secondary phenotypes. I have isolated a large number of hyper-rec mutants and found them useful in screening for mutants that accumulate labeled DNA fragments after short pulses with [3H]thymidine. The mutants so recovered include ones that are defective in deoxyribonucleic acid ligase, deoxyribonucleic acid polymerase I and its associated 5' yields 3' exonuclease, and a group of mutants, dnaS, that accumulate abnormally short Okazaki fragments. Evidence is presented that suggests that the lac-att80 segment of the chromosome cannot be inverted.

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Selected References

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

  1. BECKWITH J. R. A DELETION ANALYSIS OF THE LAC OPERATOR REGION IN ESCHERICHIA COLI. J Mol Biol. 1964 Mar;8:427–430. doi: 10.1016/s0022-2836(64)80206-0. [DOI] [PubMed] [Google Scholar]
  2. Bachmann B. J., Low K. B., Taylor A. L. Recalibrated linkage map of Escherichia coli K-12. Bacteriol Rev. 1976 Mar;40(1):116–167. doi: 10.1128/br.40.1.116-167.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. De Lucia P., Cairns J. Isolation of an E. coli strain with a mutation affecting DNA polymerase. Nature. 1969 Dec 20;224(5225):1164–1166. doi: 10.1038/2241164a0. [DOI] [PubMed] [Google Scholar]
  4. Gottesman M. M., Hicks M. L., Gellert M. Genetics and function of DNA ligase in Escherichia coli. J Mol Biol. 1973 Jul 15;77(4):531–547. doi: 10.1016/0022-2836(73)90221-0. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. JACOB F., MONOD J. Genetic regulatory mechanisms in the synthesis of proteins. J Mol Biol. 1961 Jun;3:318–356. doi: 10.1016/s0022-2836(61)80072-7. [DOI] [PubMed] [Google Scholar]
  7. Konrad E. B., Lehman I. R. A conditional lethal mutant of Escherichia coli K12 defective in the 5' leads to 3' exonuclease associated with DNA polymerase I. Proc Natl Acad Sci U S A. 1974 May;71(5):2048–2051. doi: 10.1073/pnas.71.5.2048. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Konrad E. B., Lehman I. R. Novel mutants of Escherichia coli that accumulate very small DNA replicative intermediates. Proc Natl Acad Sci U S A. 1975 Jun;72(6):2150–2154. doi: 10.1073/pnas.72.6.2150. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Konrad E. B., Modrich P., Lehman I. R. Genetic and enzymatic characterization of a conditional lethal mutant of Escherichia coli K12 with a temperature-sensitive DNA ligase. J Mol Biol. 1973 Jul 15;77(4):519–529. doi: 10.1016/0022-2836(73)90220-9. [DOI] [PubMed] [Google Scholar]
  10. Kuempel P. L., Veomett G. E. A possible function of DNA polymerase in chromosome replication. Biochem Biophys Res Commun. 1970 Nov 25;41(4):973–980. doi: 10.1016/0006-291x(70)90180-4. [DOI] [PubMed] [Google Scholar]
  11. Marinus M. G., Morris N. R. Biological function for 6-methyladenine residues in the DNA of Escherichia coli K12. J Mol Biol. 1974 May 15;85(2):309–322. doi: 10.1016/0022-2836(74)90366-0. [DOI] [PubMed] [Google Scholar]
  12. Modrich P., Lehman I. R. Enzymatic joining of polynucleotides. IX. A simple and rapid assay of polynucleotide joining (ligase) activity by measurement of circle formation from linear deoxyadenylate-deoxythymidylate copolymer. J Biol Chem. 1970 Jul 25;245(14):3626–3631. [PubMed] [Google Scholar]
  13. Okazaki R., Arisawa M., Sugino A. Slow joining of newly replicated DNA chains in DNA polymerase I-deficient Escherichia coli mutants. Proc Natl Acad Sci U S A. 1971 Dec;68(12):2954–2957. doi: 10.1073/pnas.68.12.2954. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Olivera R. M., Bonhoeffer E. Replication of Escherichia coli requires DNA polymerase I. Nature. 1974 Aug 9;250(5466):513–514. doi: 10.1038/250513a0. [DOI] [PubMed] [Google Scholar]
  15. Pauling C., Hamm L. Properties of a temperature-sensitive radiation-sensitive mutant of Escherichia coli. Proc Natl Acad Sci U S A. 1968 Aug;60(4):1495–1502. doi: 10.1073/pnas.60.4.1495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Pauling C., Hamm L. Properties of a temperature-sensitive, radiation-sensitive mutant of Escherichia coli. II. DNA replication. Proc Natl Acad Sci U S A. 1969 Dec;64(4):1195–1202. doi: 10.1073/pnas.64.4.1195. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Siegel E. C. Ultraviolet-sensitive mutator strain of Escherichia coli K-12. J Bacteriol. 1973 Jan;113(1):145–160. doi: 10.1128/jb.113.1.145-160.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Tye B. K., Nyman P. O., Lehman I. R., Hochhauser S., Weiss B. Transient accumulation of Okazaki fragments as a result of uracil incorporation into nascent DNA. Proc Natl Acad Sci U S A. 1977 Jan;74(1):154–157. doi: 10.1073/pnas.74.1.154. [DOI] [PMC free article] [PubMed] [Google Scholar]

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