recA Protein, E. coli

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Last modified: 2008-07-15 14:57:15

recA Protein is a multi-functional DNA-binding protein encoded by E.coli that plays integral roles in both homologous recombination and post-replicative DNA repair mechanisms. recA Protein is commonly used to study the molecular mechanisms involved in homologous recombination.

In vitro, recA Protein catalyses homologous recombination through a multiple-step ATP-dependent pathway. Initially, the protein binds preferentially to single-stranded DNA forming a nucleoprotein filament. The filament complex binds to naked duplex DNA and searches for regions of homology. Once a region of homology is found, strand displacement and exchange begins.¹ The various steps in this process have been used as a basis for research applications for recA Protein outside the realm of recombination studies.

Storage Buffer:

50% glycerol containing 50mM Tris-HCl, pH 7.5, 0.1M NaCl, 0.1mM EDTA, 1mM DTT, and 0.1% Triton® X-100.

Quality Control:

recA Protein is free of detectable RNase and DNA exo- and endonuclease activities. Physical purity is >95% as determined by SDS-PAGE.

Last modified: 2008-05-14 16:46:46

Applications

• Site-directed mutagenesis through displacement loop structures²
• Targeted site-specific cleavage of small and large DNA^3-5
• Enrichment of target sequences from libraries or other DNA pools^6,7
• Visualisation of DNA for electron microscopy⁸

Last modified: 2008-05-14 16:46:46

Protocols for: rec A Protein

(catalogue number RC44200 / RC441MG)

Please note: all protocols off site are the responsibility of the products supplier

Last modified: 2008-05-14 16:46:46

References

1. Radding, C.M. (1991) J. Biol. Chem. 266, 5355.
2. Shortle, D. et al. (1980) Proc. Natl. Acad. Sci. USA 77, 5375.
3. Ferrin, L.J. and Camerini-Otero, R.D. (1991) Science 254, 1494.
4. Koob, M. et al. (1986) Nucl. Acids Res. 20, 5831.
5. Ferrin, L.J. and Camerini-Otero, R.D. (1994) Nature Genet. 6, 379.
6. Honigberg, S.M. et al. (1986) Proc. Natl. Acad. Sci. USA 83, 9586.
7. Rigas, B. et al. (1986) Proc. Natl. Acad. Sci. USA 83, 9591.
8. Wasserman, S.A. and Cozzarelli, N.R. (1985) Proc. Natl. Acad. Sci. USA 82, 1079.

Last modified: 2008-05-14 16:46:46