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Recombination between repeats in Escherichia coli by a recA-independent, proximity-sensitive mechanism
Authors:S T Lovett  T J Gluckman  P J Simon  V A Sutera and P T Drapkin
Institution:(1) Department of Biology and Rosenstiel Basic Medical Sciences Center, Brandeis University, 415 South St., 02254-9110 Waltham, MA, USA;(2) Present address: Northwestern University Medical School, 60611 Chicago, IL, USA;(3) Present address: Department of Anatomy and Cell Biology, Columbia University, College of Physicians and Surgeons, 630 W 168th St., 10032-2704 New York, NY, USA
Abstract:We have examined the influence of proximity on the efficiency of recombination between repeated DNA sequences in Escherichia coli. Our experiments have employed a plasmid-based assay to detect deletions between direct repeats of 100 bp. The rate of deletion of the juxtaposed direct repeats was reasonably high at 6 × 10–5 per cell. A comparison of recA+ and recA mutant strains showed that these deletion events are primarily the result of recA-independent recombination at these homologous sequences. Random restriction fragments of yeast or E. coli genomic DNA were used to separate the two repeats. Deletion rates decreased over two orders of magnitude with increasing separation of up to 7 kb. There was a surprisingly strong effect of even short sequence separations, with insertions of a few hundred base pairs exhibiting 10-fold reductions of deletion rates. No effect of recA on the efficiency of deletion was observed at any distance between repeats.
Keywords:Deletion  recA-independent recombination  Sister-strand exchange  Direct repeats  Escherichia coli genetic recombination
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