RecA Protein, E. coli

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Applications

• Site-directed mutagenesis through displacement loop structures.¹
• Targeted site-specific cleavage of small and large DNA.^2-4
• Enrichment of target sequences from libraries or other DNA pools.^5,6
• Visualization of DNA for electron microscopy.⁷
• Cloning or other strategies involving use of RecA Protein and a site-specific oligonucleotide to block endonuclease cleavage at the complementary site in a target DNA molecule.

RecA Protein is a multifunctional DNA-binding protein produced by E.coli that plays an integral role in both homologous recombination and postreplicative DNA repair mechanisms. RecA Protein is commonly used to study the molecular mechanisms involved in homologous recombination.

In vitro, RecA Protein helps promote homologous recombination through a multiple-step ATP-dependent process. Initially, the protein binds preferentially to ssDNA 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.⁸ Many of the research applications for RecA derive from this recombination process.

Storage Buffer: 50% glycerol containing 50 mM Tris-HCl (pH 7.5), 0.1 M NaCl, 0.1 mM EDTA, 1 mM 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.

References

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