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Formation of recombinant lacZ + DNA in conjugational crosses with a recB mutant of Escherichia coli K12 depends on recF, recJ, and recO

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Summary

Conjugational recombination in Escherichia coli was investigated by monitoring synthesis of the lacZ + product, β-galactosidase, in crosses between lacZ mutants. We report here that mutation of recB and any combination of recF, recJ, or recO reduces enzyme production by a factor of between 10- and 25-fold whereas mutation of only one of these genes or any combination of recF, recJ, or recO has no more than a 2-fold effect. Mutation of recN has no effect either alone or in combination with the other mutations. We suggest that the products of recF, recJ, and recO may provide an efficient alternative to the RecBCD enzyme for the initiation of recombination in conjugational crosses but that RecBCD activity is needed in this case to produce a viable recombinant product.

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References

  • Amundsen SK, Taylor AF, Chaudhury AM, Smith GR (1986) recD: The gene for an essential third subunit of exonuclease V. Proc Natl Acad Sci USA 83:5558–5562

    Google Scholar 

  • Bachmann BJ (1972) Pedigrees of some mutant strains of Escherichia coli K12. Bacteriol Rev 36:525–557

    Google Scholar 

  • Bachmann BJ (1983) Linkage map of Escherichia coli K12, edition 7. Microbiol Rev 47:180–230

    Google Scholar 

  • Birge EA, Low KB (1974) Detection of transcribable recombination products following conjugation in Rec+, RecB, and RecC strains of Escherichia coli K12. J Mol Biol 83:447–457

    Google Scholar 

  • Bochner BR, Huang HC, Schieven GL, Ames BN (1980) Positive selection for loss of tetracycline resistance. J Bacteriol 143:926–933

    Google Scholar 

  • Clark AJ, Margulies AD (1965) Isolation and characterisation of recombination-deficient mutants of Escherichia coli K12. Proc Natl Acad Sci USA 53:451–459

    Google Scholar 

  • Cohen A, Laban A (1983) Plasmidic recombination in Escherichia coli K-12: the role of the recF gene product. Mol Gen Genet 189:471–474

    Google Scholar 

  • Emmerson PT, Howard-Flanders P (1967) Recombination-deficient mutants of Escherichia coli K-12 that map between thyA and argA. Genetics 60:19–30

    Google Scholar 

  • Hall JD, Howard-Flanders P (1972) Recombinant F′ factors from Escherichia coli K-12 strains carrying recB or recC. J Bacteriol 110:578–584

    Google Scholar 

  • Hickson ID, Robson CN, Atkinson KE, Hutton L, Emmerson PT (1985) Reconstitution of RecBC DNAse activity from purified Escherichia coli RecB and RecC proteins. J Biol Chem 260:1224–1229

    Google Scholar 

  • Horii ZI, Clark AJ (1973) Genetic analysis of the RecF pathway to genetic recombination in Escherichia coli K12: isolation and characterisation of mutants. J Mol Biol 80:327–344

    Google Scholar 

  • Howard-Flanders P, Theriot L (1966) Mutants of Escherichia coli K-12 defective in DNA repair and genetic recombination. Genetics 53:1137–1150

    Google Scholar 

  • Kolodner R, Fishel RA, Howard M (1985) Genetic recombination of bacterial plasmid DNA: Effect of RecF pathway mutations on plasmid recombination in Escherichia coli. J Bacteriol 163:1060–1066

    Google Scholar 

  • Kushner SR, Nagaishi H, Clark AJ (1972) Indirect suppression of recB and recC mutations by exonuclease I deficiency. Proc Natl Acad Sci USA 69:1366–1370

    Google Scholar 

  • Lloyd RG, Buckman C (1985) Identification and genetic analysis of sbcC mutations in commonly used recBC sbcB strains of Escherichia coli K-12. J Bacteriol 164:836–844

    Google Scholar 

  • Lloyd RG, Low B (1976) Some genetic consequences of changes in the level of recA gene function in Escherichia coli K12. Genetics 84:675–695

    Google Scholar 

  • Lloyd RG, Thomas A (1984) A molecular model for conjugational recombination in Escherichia coli K12. Mol Gen Genet 197:328–336

    Google Scholar 

  • Lloyd RG, Low B, Godson GN, Birge EA (1974) Isolation and characterisation of an Escherichia coli K12 mutant with a temperature-sensitive RecA phenotype. J Bacteriol 120:407–415

    Google Scholar 

  • Lloyd RG, Picksley SM, Prescott C (1983) Inducible expression of a gene specific to the RecF pathway for recombination in Escherichia coli K12. Mol Gen Genet 190:162–167

    Google Scholar 

  • Lloyd RG, Benson FE, Shurvinton CE (1984) Effect of ruv mutations on recombination and DNA repair in Escherichia coli. Mol Gen Genet 194:303–309

    Google Scholar 

  • Lovett SJ, Clark AJ (1983) Genetic analysis of regulation of the RecF pathway of recombination in Escherichia coli K-12. J Bacteriol 153:1471–1478

    Google Scholar 

  • Lovett SJ, Clark AJ (1984) Genetic analysis of the recJ gene of Escherichia coli K-12. J Bacteriol 157:190–196

    Google Scholar 

  • Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, New York

    Google Scholar 

  • Nakayama H, Nakayama K, Nakayama R, Irino N, Nakayama Y, Hanawalt PC (1984) Isolation and genetic characterization of a thymineless death-resistant mutant of Escherichia coli K12: Identification of a new mutation (recQ1) that blocks the RecF recombination pathway. Mol Gen Genet 195:474–480

    Google Scholar 

  • Picksley SM, Attfield PV, Lloyd RG (1984a) Repair of DNA double-strand breaks in Escherichia coli K12 requires a functional recN product. Mol Gen Genet 195:267–274

    Google Scholar 

  • Picksley SM, Lloyd RG, Buckman C (1984b) Genetic analysis and regulation of inducible recombination in Escherichia coli K12. Cold Spring Harbor Symp Quant Biol XLIX:469–474

    Google Scholar 

  • Ponticelli AS, Schultz DW, Taylor AF, Smith GR (1985) Chidependent DNA strand cleavage by RecBC enzyme. Cell 41:145–151

    Google Scholar 

  • Porter RD, Welliver RA, Witkowski TA (1982) Specialized transduction with λplac5:dependence of recB. J Bacteriol 150:1485–1488

    Google Scholar 

  • Radding CM (1982) Homologous pairing and strand exchange in genetic recombination. Annu Rev Genet 16:405–437

    Google Scholar 

  • Seifert HS, Porter RD (1984) Enhanced recombination between λplac5 and F42lac: Identification of cis- and trans-acting factors. Proc Natl Acad Sci USA 81:7500–7504

    Google Scholar 

  • Taylor A, Smith GR (1980) Unwinding and rewinding of DNA by the RecBC enzyme. Cell 22:447–457

    Google Scholar 

  • Taylor A, Smith GR (1985) Substrate specificity of the DNA unwinding activity of the RecBC enzyme of Escherichia coli. J Mol Biol 185:431–443

    Google Scholar 

  • Taylor AF, Schultz DW, Ponticelli AS, Smith GR (1985) RecBC enzyme nicking at Chi sites during DNA unwinding: Location and orientation-dependence of the cutting. Cell 41:153–163

    Google Scholar 

  • Templin A, Kushner SR, Clark AJ (1972) Genetic analysis of mutations indirectly suppressing recB and recC mutations. Genetics 72:205–215

    Google Scholar 

  • Walker GC (1984) Mutagenesis and inducible responses to deoxyribonucleic acid damage in Escherichia coli. Microbiol Rev 48:60–93

    Google Scholar 

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Lloyd, R.G., Evans, N.P. & Buckman, C. Formation of recombinant lacZ + DNA in conjugational crosses with a recB mutant of Escherichia coli K12 depends on recF, recJ, and recO . Mol Gen Genet 209, 135–141 (1987). https://doi.org/10.1007/BF00329848

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  • DOI: https://doi.org/10.1007/BF00329848

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