Description: RecA Protein (E. coli) is necessary for genetic recombination, reactions involving DNA repair and UV-induced mutagenesis. RecA promotes the autodigestion of the LexA repressor, UmuD protein and lambda repressor. Cleavage of LexA derepresses more than 20 genes (1). In vitro studies indicate that in the presence of ATP, RecA promotes the strand exchange of single-strand DNA fragments with homologous duplex DNA. The reaction has three distinct steps: (i) RecA polymerizes on the single-strand DNA, (ii) the nucleoprotein filament binds the duplex DNA and searches for a homologous region, (iii) the strands are exchanged (2).
Source: An E. coli strain ER2502 that carries an overexpressed RecA gene from E. coli.
Application: - 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)
Supplied in: 10 mM Tris-HCl 1 mM DTT 0.1 mM EDTA 50% Glycerol pH 7.5 @ 25°C
Supplied with: 10x RecA Reaction Buffer
10x RecA Reaction Buffer: 700 mM Tris-HCl 100 mM MgCl2 50 mM DTT pH 7.6 @ 25°C
Unit Definition: Sold by mass of pure protein determined at OD280 (A280 = 0.516 at 1 mg/mL, 1cm).
Recommended Storage Condition: -20°C
References: 1. West, S.C. (1992) Ann. Rev. Biochem., 61, 603-640. 2. Zhumabayeva, B. et al. (1990) Biotechniques, 27, 834-845. 3. Zhumabayeva, B. et al. (2001) Biotechniques, 30, 512-520. 4. Radding, C.M. (1991) J. Biol. Chem., 266, 5355-5358. 5. Wasserman, S.A. and Cozzarelli, N.R. (1985) Proc. Natl. Acad. Sci. USA, 82, 1079-1083. 6. Shortle, D. et al. (1980) Proc. Natl. Acad. Sci. USA, 77, 5375-5379. 7. Honigberg, S.M. et al. (1986) Proc. Natl. Acad. Sci. USA, 83, 9586-9590. 8. Rigas, B. et al. (1986) Proc. Natl. Acad. Sci. USA, 83, 9591-9595. 9. Ferrin, L.J. and Camerini-Otero, R.D. (1991) Science, 254, 1494-1497. 10. Koob, M. et al. (1992) Nucl. Acids Res., 20, 5831-5836. 11. Koob, M. (1992) R. Wu (Eds.), Methods in Enzymology, 216, pp. 321-329. San Diego: Academic Press.
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