Journal
JOURNAL OF MOLECULAR MICROBIOLOGY AND BIOTECHNOLOGY
Volume 19, Issue 3, Pages 117-122Publisher
KARGER
DOI: 10.1159/000321497
Keywords
Site-specific recombination; Flp recombinase; Coliphage HK022; Integrase; Recombinase-mediated cassette exchange; Atomic force microscopy
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Funding
- Leon Reich scholarship fund
- BARD [MB-8713-08]
- United States-Israel Binational Agricultural Research and Development Fund
- US National Institutes of Health [RO1-GM085848]
- NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES [R01GM085848] Funding Source: NIH RePORTER
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A comparison between the efficiency of recombinase-mediated cassette exchange (RMCE) reactions catalyzed in Escherichia coli by the site-specific recombinases Flp of yeast and Int of coliphage HK022 has revealed that an Flp-catalyzed RMCE reaction is more efficient than an Int-HK022 catalyzed reaction. In contrast, an RMCE reaction with 1 pair of frt sites and 1 pair of att sites catalyzed in the presence of both recombinases is very inefficient. However, the same reaction catalyzed by each recombinase individually supplied in a sequential order is very efficient, regardless of the order. Atomic force microscopy images of Flp with its DNA substrates show that only 1 pair of recombination sites forms a synaptic complex with the recombinase. The results suggest that the RMCE reaction is sequential. Copyright (C) 2010 S. Karger AG, Basel
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