RecA-mediated joint molecule formation between O-methylated RNA/DNA hairpins and single-stranded targets |
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Authors: | P. A. Havre and E. B. Kmiec |
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Affiliation: | (1) Department of Microbiology/Immunology, Kimmel Cancer Institute, Thomas Jefferson University, 233 S. 10th Street, Philadelphia, PA 19107, USA Fax: +1-215-923-1098; e-mail: kmiec@lac.jci.tju.edu, US |
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Abstract: | Chimeric oligonucleotides consisting of one DNA strand paired with an O-methylated RNA strand interrupted by six DNA residues have been used in gene targeting experiments. Here we demonstrate that these hairpins can form a heteroduplex (or joint molecule) with single-stranded DNA targets in a reaction mediated by the E. coli RecA protein. One end of the double hairpin may unwind to form a 14-base-RecA filament which initiates the reaction. Chimeric oligonucleotides containing only O-methylated RNA residues on one strand or truncated hairpins lacking this 14-base segment did not participate in RecA-driven heteroduplex formation under these reaction conditions. The results presented here represent a first step in studying one facet of a strategy which uses O-methylated RNA residues as participants in homologous pairing events. Received: 2 June 1997 / Accepted: 29 September 1997 |
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Keywords: | RecA Recombinase Strand transfer DNA repair |
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