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. 1985 Apr;162(1):29–34. doi: 10.1128/jb.162.1.29-34.1985

General recombination in Escherichia coli K-12: in vivo role of RecBC enzyme.

S D Yancey, R D Porter
PMCID: PMC218947  PMID: 3884592

Abstract

Heterozygous lacZ- merodiploids of Escherichia coli K-12 have been used to study the role of the RecBC enzyme in general recombination. The transcribable intermediate assay detects the product of early steps in recombination without requiring the formation of viable recombinant colonies. Recombination is initiated by infection with lambda precA+. We have found that transcribable intermediate formation in crosses between F42 lac and chromosomal lac is dependent on F fertility functions and an active RecBC enzyme. Thus, the products of the recB and recC genes are required in early steps of recombination between these two substrates. Introduction of the F42 lac donor DNA by conjugation immediately after infection with lambda precA+ abolishes the requirement for an active RecBC enzyme.

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Selected References

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

  1. Birge E. A., Low K. B. Detection of transcribable recombination products following conjugation in rec+, reCB- and recC-strains of Escherichia coli K12. J Mol Biol. 1974 Mar 15;83(4):447–457. doi: 10.1016/0022-2836(74)90506-3. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Capaldo F. N., Barbour S. D. Isolation of the nonviable cells produced during normal growth of recombination-deficient strains of Escherichia coli K-12. J Bacteriol. 1973 Sep;115(3):928–936. doi: 10.1128/jb.115.3.928-936.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Capaldo F. N., Ramsey G., Barbour S. D. Analysis of the growth of recombination-deficient strains of Escherichia coli K-12. J Bacteriol. 1974 Apr;118(1):242–249. doi: 10.1128/jb.118.1.242-249.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chaconas G., de Bruijn F. J., Casadaban M. J., Lupski J. R., Kwoh T. J., Harshey R. M., DuBow M. S., Bukhari A. I. In vitro and in vivo manipulations of bacteriophage Mu DNA: cloning of Mu ends and construction of mini-Mu's carrying selectable markers. Gene. 1981 Jan-Feb;13(1):37–46. doi: 10.1016/0378-1119(81)90041-x. [DOI] [PubMed] [Google Scholar]
  6. Chaudhury A. M., Smith G. R. Escherichia coli recBC deletion mutants. J Bacteriol. 1984 Nov;160(2):788–791. doi: 10.1128/jb.160.2.788-791.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Csonka L. N., Clark A. J. Deletions generated by the transposon Tn10 in the srl recA region of the Escherichia coli K-12 chromosome. Genetics. 1979 Oct;93(2):321–343. doi: 10.1093/genetics/93.2.321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dagert M., Ehrlich S. D. Prolonged incubation in calcium chloride improves the competence of Escherichia coli cells. Gene. 1979 May;6(1):23–28. doi: 10.1016/0378-1119(79)90082-9. [DOI] [PubMed] [Google Scholar]
  9. Dykstra C. C., Prasher D., Kushner S. R. Physical and biochemical analysis of the cloned recB and recC genes of Escherichia coli K-12. J Bacteriol. 1984 Jan;157(1):21–27. doi: 10.1128/jb.157.1.21-27.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Eckhardt T. A rapid method for the identification of plasmid desoxyribonucleic acid in bacteria. Plasmid. 1978 Sep;1(4):584–588. doi: 10.1016/0147-619x(78)90016-1. [DOI] [PubMed] [Google Scholar]
  11. Goldmark P. J., Linn S. Purification and properties of the recBC DNase of Escherichia coli K-12. J Biol Chem. 1972 Mar 25;247(6):1849–1860. [PubMed] [Google Scholar]
  12. Hickson I. D., Emmerson P. T. Identification of the Escherichia coli recB and recC gene products. Nature. 1981 Dec 10;294(5841):578–580. doi: 10.1038/294578a0. [DOI] [PubMed] [Google Scholar]
  13. Karu A. E., MacKay V., Goldmark P. J., Linn S. The recBC deoxyribonuclease of Escherichia coli K-12. Substrate specificity and reaction intermediates. J Biol Chem. 1973 Jul 25;248(14):4874–4884. [PubMed] [Google Scholar]
  14. Low B. Rapid mapping of conditional and auxotrophic mutations in Escherichia coli K-12. J Bacteriol. 1973 Feb;113(2):798–812. doi: 10.1128/jb.113.2.798-812.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Manis J. J., Kline B. C. Restriction endonuclease mapping and mutagenesis of the F sex factor replication region. Mol Gen Genet. 1977 Apr 29;152(3):175–182. doi: 10.1007/BF00268815. [DOI] [PubMed] [Google Scholar]
  16. Muskavitch K. M., Linn S. A unified mechanism for the nuclease and unwinding activities of the recBC enzyme of Escherichia coli. J Biol Chem. 1982 Mar 10;257(5):2641–2648. [PubMed] [Google Scholar]
  17. Porter R. D. Enhanced recombination between F42lac and lambda plac5: dependence on F42lac fertility functions. Mol Gen Genet. 1981;184(3):355–358. doi: 10.1007/BF00352504. [DOI] [PubMed] [Google Scholar]
  18. Porter R. D., Lark M. W., Low K. B. Specialized transduction with lambda plac5: dependence on recA and on configuration of lac and att lambda. J Virol. 1981 May;38(2):497–503. doi: 10.1128/jvi.38.2.497-503.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Porter R. D., McLaughlin T., Low B. Transduction versus "conjuduction": evidence for multiple roles for exonuclease V in genetic recombination in Escherichia coli. Cold Spring Harb Symp Quant Biol. 1979;43(Pt 2):1043–1047. doi: 10.1101/sqb.1979.043.01.113. [DOI] [PubMed] [Google Scholar]
  20. Porter R. D., Welliver R. A., Witkowski T. A. Specialized transduction with lambda plac5: dependence on recB. J Bacteriol. 1982 Jun;150(3):1485–1488. doi: 10.1128/jb.150.3.1485-1488.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Rosamond J., Telander K. M., Linn S. Modulation of the action of the recBC enzyme of Escherichia coli K-12 by Ca2+. J Biol Chem. 1979 Sep 10;254(17):8646–8652. [PubMed] [Google Scholar]
  22. Seifert H. S., Porter R. D. Enhanced recombination between lambda plac5 and mini-F-lac: the tra regulon is required for recombination enhancement. Mol Gen Genet. 1984;193(2):269–274. doi: 10.1007/BF00330679. [DOI] [PubMed] [Google Scholar]
  23. Stahl F. W. Special sites in generalized recombination. Annu Rev Genet. 1979;13:7–24. doi: 10.1146/annurev.ge.13.120179.000255. [DOI] [PubMed] [Google Scholar]
  24. Stahl M. M., Kobayashi I., Stahl F. W., Huntington S. K. Activation of Chi, a recombinator, by the action of an endonuclease at a distant site. Proc Natl Acad Sci U S A. 1983 Apr;80(8):2310–2313. doi: 10.1073/pnas.80.8.2310. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Taylor A., Smith G. R. Unwinding and rewinding of DNA by the RecBC enzyme. Cell. 1980 Nov;22(2 Pt 2):447–457. doi: 10.1016/0092-8674(80)90355-4. [DOI] [PubMed] [Google Scholar]
  26. Willetts N. S., Mount D. W. Genetic analysis of recombination-deficient mutants of Escherichia coli K-12 carrying rec mutations cotransducible with thyA. J Bacteriol. 1969 Nov;100(2):923–934. doi: 10.1128/jb.100.2.923-934.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Willetts N. The transcriptional control of fertility in F-like plasmids. J Mol Biol. 1977 May 5;112(1):141–148. doi: 10.1016/s0022-2836(77)80161-7. [DOI] [PubMed] [Google Scholar]
  28. Willis D. K., Uhlin B. E., Amini K. S., Clark A. J. Physical mapping of the srl recA region of Escherichia coli: analysis of Tn10 generated insertions and deletions. Mol Gen Genet. 1981;183(3):497–504. doi: 10.1007/BF00268771. [DOI] [PubMed] [Google Scholar]
  29. Yancey S. D., Porter R. D. Negative complementation of recA protein by recA1 polypeptide: in vivo recombination requires a multimeric form of recA protein. Mol Gen Genet. 1984;193(1):53–57. doi: 10.1007/BF00327413. [DOI] [PubMed] [Google Scholar]

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