| Description | RecA is essential for homologous recombination, reactions involving DNA repair and UV-induced mutagenesis in E coli (1,2,3). RecA binds to single stranded DNA, resulting in the polymerization of RecA into a nucleoprotein complex. This complex can align with complementary double stranded DNA, resulting in RecA catalysis of DNA strand exchange. RecA DNA binding is stimulated by ATP hydrolysis or non-hydrolyzable ATP analogs. The RecA-ATP-single stranded DNA complex also can function as a coprotease in the proteolytic cleavage of LexA, UmuD and certain bacteriophage proteins. In a healthy cell, LexA represses the expression of genes encoding DNA repair proteins (SOS genes). Upon injury of DNA, LexA catalyzes its own digestion, thereby allowing synthesis of necessary SOS proteins. However, LexA can only induce self-catalysis when activated by a ssDNA-RecA filament. A single filament will bind and activate several LexA proteins, each of which then cleaves other bound proteins. Thus, ssDNA-RecA, a product of DNA injury, stimulates DNA repair. |
| Applications | - Visualization of DNA structures with electron microscopy (3)
- D-loop mutagenesis (4)
- Screening libraries using RecA-coated probes (5,6)
- Cleavage of DNA at any single predetermined site (7,8,9)
- RecA mediated affinity capture for full length cDNA cloning (10, 11)
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| Source | An E. coli strain that carries the E. coli recA gene |
| Components | RecA: 2 mg/ml in 10 mM Tris-HCl, 1 mM DTT, 0.1 mM EDTA, 50% glycerol, pH 7.5 @ 25°C |
| Quality Control | The absence of exo- and endodeoxyribonucleases, non-specific DNase and RNase is confirmed by appropriate quality test. |
| Storage Condition | -20 °C |
References | West, S.C. (1992). Ann. Rev. Biochem.. 61, 603-640.Radding, C.M. (1991). J. Biol. Chem.. 266, 5355-5358.Wasserman, S.A. and Cozzarelli, N.R. (1985). Proc. Natl. Acad. Sci. USA. 82, 1079-1083.Shortle, D. et al. (1980). Proc. Natl. Acad. Sci. USA. 77, 5375-5379.Honigberg, S.M. et al. (1986). Proc. Natl. Acad. Sci. USA. 83, 9586-9590.Rigas, B. et al. (1986). Proc. Natl. Acad. Sci. USA. 83, 9591-9595.Ferrin, L.J. and Camerini-Otero, R.D. (1991). Science. 254, 1494-1497.Koob, M. et al. (1992). Nucl. Acids Res.. 20, 5831-5836.Koob, M. (1992). R. Wu(Ed.), Methods in Enzymology. 216, 321-329. San Diego: Academic Press.Zhumabayeva, B. et al. (1990). Biotechniques. 27, 834-845.Zhumabayeva, B. et al. (2001). Biotechniques. 30, 512-520. |
