RecA Protein, E. coli

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.