Summary
A recombinant λ phage carrying the recA gene of Rhizobium phaseoli was isolated from a R. phaseoli genomic library by complementation of the Fec− phenotype of the recombinant phage in Escherichia coli. When expressed in E. coli, the cloned recA gene was shown to restore resistance to both UV irradiation and the DNA alkylating agent methyl methanesulphonate (MMS). The R. phaseoli recA gene also promoted homologous recombination in E. coli. The cloned recA gene was only weakly inducible in E. coli recA strains by DNA damaging agents. The DNA sequence of the R. phaseoli recA gene was determined and compared with published recA sequences. No LexA-binding site was detected in the R. phaseoli recA upstream region.
Article PDF
Avoid common mistakes on your manuscript.
References
Ball TK, Wasmuth CR, Braunagel SC, Benedik MJ (1990) Expression of Serratia marcescens extracellular proteins requires recA. J Bacteriol 172:342–349
Better M, Helinski DR (1983) Isolation and characterization of the recA gene of Rhizobium meliloti. J Bacteriol 155:311–316
Buikema W (1985) Ph. D. thesis, Harvard University, Cambridge, Mass
Chen EY, Seeburg PH (1985) Supercoil sequencing: a fast and simple method for sequencing DNA. DNA 4:154–170
Clark AJ, Margulies AD (1965) Isolation and characterization of recombination-deficient mutants of Escherichia coli. Proc Natl Acad Sci USA 53:451–459
Flores M, Gonzalez V, Brom S, Martinez E, Piñero D, Romero D, Davila G, Palacios R (1987) Reiterated DNA sequences in Rhizobium and Agrobacterium spp. J Bacteriol 169:5782–5788
Flores M, Gonzalez V, Pardo MA, Leija A, Martinez E, Romero D, Piñero D, Davila G, Palacios R (1988) Genomic instability in Rhizobium phaseoli. J Bacteriol 170:1191–1196
Frischauf AM, Lehrach H, Povstka A, Murray NM (1983) Lambda replacement vectors carrying polylinker sequences. J Mol Biol 170:827–842
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY
Miles CA, Mountain A, Sastry GRK (1986) Cloning and characterization of the recA gene of Agrobacterium tumefaciens C58. Mol Gen Genet 204:161–165
Morett E, Moreno S, Espin G (1988) Transcription analysis of the three nih H genes of Rhizobium phaseoli with gene fusions. Mol Gen Genet 213:499–504
Owtthrim GW, Coleman JR (1989) Regulation of expression and nucleotide sequence of the Anabaena variabilis recA gene. J Bacteriol 171:5713–5719
Quinto C, de la Vega H, Flores M, Fernández L, Ballado T, Soberon G, Palacios R (1982) Reiteration of nitrogen fixation gene sequences in Rhizobium phaseoli. Nature 299: 724–726
Resnick D, Nelson DR (1988) Cloning and characterization of the Aeromonas caviae recA gene and construction of an A. caviae recA mutant. J Bacteriol 170:48–55
Sancar A, Stachelek C, Koningsberg W, Rupp WD (1980) Sequences of the recA gene and protein. Proc Natl Acad Sci USA 77:2611–2615
Sano Y, Kageyama M (1987) The sequence and function of the recA gene and its protein in Pseudomonas aeruginosa PAO. Mol Gen Genet 208:412–419
Sherry LK, McNamee KP, McEntee K (1984) Cloning and characterization of recA genes from Proteus vulgaris, Eruinia carotovora, Shigella flescneri and Escherichia coli B. J Bacteriol 160:153–160
Simon R, Priefer U, Pühler A (1983) A broad host range mobilization system for in vivo genetic engineering transposon mutagenesis in gram negative bacteria. Biotechnology 1:784–791
Singer JT (1989) Molecular cloning of the recA analog from the marine fish pathogen Vibrio anguillarum 775. J Bacteriol 171:6367–6371
Soberón-Chávez G, Nájera R, Olivera H, Segovia L (1986) Genetic rearrangements of a Rhizobium phaseoli symbiotic plasmid. J Bacteriol 167:487–491
Vieira J, Messing J (1982) The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with universal primers. Gene 19:259–268
Walker JE, Saraste M, Runswick MJ, Gay NJ (1982) Distantly related sequences in the α- and β-subunits of ATP synthase, myosin, kinases and other ATP-requiring enzymes and a common nucleotide binding fold. EMBO J 1:945–951
Author information
Authors and Affiliations
Additional information
Communicated by J. Schell
Rights and permissions
About this article
Cite this article
Michiels, J., Vande Broek, A. & Vanderleyden, J. Molecular cloning and nucleotide sequence of the Rhizobium phaseoli recA gene. Molec. Gen. Genet. 228, 486–490 (1991). https://doi.org/10.1007/BF00260644
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00260644