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J Bacteriol. 1991 May; 173(10): 3035–3040.
PMCID: PMC207895

Molecular cloning and characterization of the recA gene of Rhizobium phaseoli and construction of recA mutants.


The Rhizobium phaseoli recA gene has been cloned by interspecific complementation of the Fec phenotype of bacteriophage lambda. The cloned gene restored the recombination proficiency and conferred resistance to DNA-damaging agents (methyl methanesulfonate and nitrofurantoin) to an Escherichia coli recA mutant. The direction of transcription and the localization of the recA gene were determined by mutagenesis with phage MudIIPR13 and heterologous hybridization with an E. coli recA probe. An R. phaseoli recA::Spcr mutation was introduced in two R. phaseoli strains by homogenotization. The R. phaseoli recA mutants were more sensitive to DNA-damaging agents and exhibited a 100-fold reduction in recombination frequency as compared with their parental strains. A deletion of the symbiotic plasmid abolishing nodulation was found at high frequency (10(-2)) in R. phaseoli CNF42. This event was recA dependent. In R. phaseoli CFN285, two events of symbiotic instability were found at high frequency (10(-3]: one was a deletion in the symbiotic plasmid, and the other was the loss of whole symbiotic plasmid. In the CFN285 recA::Spcr mutant, only the loss of the symbiotic plasmid was observed.

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  • Barcak GJ, Tomb JF, Laufer CS, Smith HO. Two Haemophilus influenzae Rd genes that complement the recA-like mutation rec-1. J Bacteriol. 1989 May;171(5):2451–2457. [PMC free article] [PubMed]
  • Better M, Helinski DR. Isolation and characterization of the recA gene of Rhizobium meliloti. J Bacteriol. 1983 Jul;155(1):311–316. [PMC free article] [PubMed]
  • Bolivar F, Rodriguez RL, Greene PJ, Betlach MC, Heyneker HL, Boyer HW, Crosa JH, Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed]
  • Boyer HW, Roulland-Dussoix D. A complementation analysis of the restriction and modification of DNA in Escherichia coli. J Mol Biol. 1969 May 14;41(3):459–472. [PubMed]
  • Cohen G. Electron microscopy study of early lytic replication forms of bacteriophage P1 DNA. Virology. 1983 Nov;131(1):159–170. [PubMed]
  • Cox MM, Lehman IR. Enzymes of general recombination. Annu Rev Biochem. 1987;56:229–262. [PubMed]
  • Ditta G, Stanfield S, Corbin D, Helinski DR. Broad host range DNA cloning system for gram-negative bacteria: construction of a gene bank of Rhizobium meliloti. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7347–7351. [PMC free article] [PubMed]
  • Eckhardt T. A rapid method for the identification of plasmid desoxyribonucleic acid in bacteria. Plasmid. 1978 Sep;1(4):584–588. [PubMed]
  • Eitner G, Adler B, Lanzov VA, Hofemeister J. Interspecies recA protein substitution in Escherichia coli and Proteus mirabilis. Mol Gen Genet. 1982;185(3):481–486. [PubMed]
  • Farrand SK, O'Morchoe SP, McCutchan J. Construction of an Agrobacterium tumefaciens C58 recA mutant. J Bacteriol. 1989 Oct;171(10):5314–5321. [PMC free article] [PubMed]
  • Flores M, González V, Brom S, Martínez E, Piñero D, Romero D, Dávila G, Palacios R. Reiterated DNA sequences in Rhizobium and Agrobacterium spp. J Bacteriol. 1987 Dec;169(12):5782–5788. [PMC free article] [PubMed]
  • Flores M, González V, Pardo MA, Leija A, Martínez E, Romero D, Piñero D, Dávila G, Palacios R. Genomic instability in Rhizobium phaseoli. J Bacteriol. 1988 Mar;170(3):1191–1196. [PMC free article] [PubMed]
  • Gay P, Le Coq D, Steinmetz M, Berkelman T, Kado CI. Positive selection procedure for entrapment of insertion sequence elements in gram-negative bacteria. J Bacteriol. 1985 Nov;164(2):918–921. [PMC free article] [PubMed]
  • Girard ML, Flores M, Brom S, Romero D, Palacios R, Dávila G. Structural complexity of the symbiotic plasmid of Rhizobium leguminosarum bv. phaseoli. J Bacteriol. 1991 Apr;173(8):2411–2419. [PMC free article] [PubMed]
  • Goldberg I, Mekalanos JJ. Cloning of the Vibrio cholerae recA gene and construction of a Vibrio cholerae recA mutant. J Bacteriol. 1986 Mar;165(3):715–722. [PMC free article] [PubMed]
  • Goodman HJ, Parker JR, Southern JA, Woods DR. Cloning and expression in Escherichia coli of a recA-like gene from Bacteroides fragilis. Gene. 1987;58(2-3):265–271. [PubMed]
  • Grunstein M, Hogness DS. Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene. Proc Natl Acad Sci U S A. 1975 Oct;72(10):3961–3965. [PMC free article] [PubMed]
  • Horii T, Ogawa T, Ogawa H. Organization of the recA gene of Escherichia coli. Proc Natl Acad Sci U S A. 1980 Jan;77(1):313–317. [PMC free article] [PubMed]
  • Karn J, Brenner S, Barnett L, Cesareni G. Novel bacteriophage lambda cloning vector. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5172–5176. [PMC free article] [PubMed]
  • Kawashima H, Horii T, Ogawa T, Ogawa H. Functional domains of Escherichia coli recA protein deduced from the mutational sites in the gene. Mol Gen Genet. 1984;193(2):288–292. [PubMed]
  • Keener SL, McNamee KP, McEntee K. Cloning and characterization of recA genes froM Proteus vulgaris, Erwinia carotovora, Shigella flexneri, and Escherichia coli B/r. J Bacteriol. 1984 Oct;160(1):153–160. [PMC free article] [PubMed]
  • Knight KL, Hess RM, McEntee K. Conservation of an ATP-binding domain among RecA proteins from Proteus vulgaris, Erwinia carotovora, Shigella flexneri, and Escherichia coli K-12 and B/r. J Bacteriol. 1988 Jun;170(6):2427–2432. [PMC free article] [PubMed]
  • Kokjohn TA, Miller RV. Molecular cloning and characterization of the recA gene of Pseudomonas aeruginosa PAO. J Bacteriol. 1985 Aug;163(2):568–572. [PMC free article] [PubMed]
  • Koomey JM, Falkow S. Cloning of the recA gene of Neisseria gonorrhoeae and construction of gonococcal recA mutants. J Bacteriol. 1987 Feb;169(2):790–795. [PMC free article] [PubMed]
  • McEvoy JL, Murata H, Chatterjee AK. Molecular cloning and characterization of an Erwinia carotovora subsp. carotovora pectin lyase gene that responds to DNA-damaging agents. J Bacteriol. 1990 Jun;172(6):3284–3289. [PMC free article] [PubMed]
  • Noel KD, Sanchez A, Fernandez L, Leemans J, Cevallos MA. Rhizobium phaseoli symbiotic mutants with transposon Tn5 insertions. J Bacteriol. 1984 Apr;158(1):148–155. [PMC free article] [PubMed]
  • Owttrim GW, Coleman JR. Molecular cloning of a recA-like gene from the cyanobacterium Anabaena variabilis. J Bacteriol. 1987 May;169(5):1824–1829. [PMC free article] [PubMed]
  • Prentki P, Krisch HM. In vitro insertional mutagenesis with a selectable DNA fragment. Gene. 1984 Sep;29(3):303–313. [PubMed]
  • Quinto C, De La Vega H, Flores M, Leemans J, Cevallos MA, Pardo MA, Azpiroz R, De Lourdes Girard M, Calva E, Palacios R. Nitrogenase reductase: A functional multigene family in Rhizobium phaseoli. Proc Natl Acad Sci U S A. 1985 Feb;82(4):1170–1174. [PMC free article] [PubMed]
  • Ratet P, Schell J, de Bruijn FJ. Mini-Mulac transposons with broad-host-range origins of conjugal transfer and replication designed for gene regulation studies in Rhizobiaceae. Gene. 1988;63(1):41–52. [PubMed]
  • Resnick D, Nelson DR. Cloning and characterization of the Aeromonas caviae recA gene and construction of an A. caviae recA mutant. J Bacteriol. 1988 Jan;170(1):48–55. [PMC free article] [PubMed]
  • Rigby PW, 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. [PubMed]
  • Romero D, Brom S, Martínez-Salazar J, Girard ML, Palacios R, Dávila G. Amplification and deletion of a nod-nif region in the symbiotic plasmid of Rhizobium phaseoli. J Bacteriol. 1991 Apr;173(8):2435–2441. [PMC free article] [PubMed]
  • Sapienza C, Rose MR, Doolittle WF. High-frequency genomic rearrangements involving archaebacterial repeat sequence elements. Nature. 1982 Sep 9;299(5879):182–185. [PubMed]
  • Smith GR. Homologous recombination in procaryotes. Microbiol Rev. 1988 Mar;52(1):1–28. [PMC free article] [PubMed]
  • Soberón-Chávez G, Nájera R, Olivera H, Segovia L. Genetic rearrangements of a Rhizobium phaseoli symbiotic plasmid. J Bacteriol. 1986 Aug;167(2):487–491. [PMC free article] [PubMed]
  • Southern EM. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. [PubMed]
  • Szybalski EH, Szybalski W. A comprehensive molecular map of bacteriophage lambda. Gene. 1979 Nov;7(3-4):217–270. [PubMed]
  • Walker GC. Mutagenesis and inducible responses to deoxyribonucleic acid damage in Escherichia coli. Microbiol Rev. 1984 Mar;48(1):60–93. [PMC free article] [PubMed]
  • Xu BW, Paszty C, Lurquin PF. A plasmid-based method to quantitate homologous recombination frequencies in gram-negative bacteria. Biotechniques. 1988 Sep;6(8):752–760. [PubMed]

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