High efficiency of a sequential recombinase-mediated cassette exchange reaction in escherichia coli

Natalia Malchin*, Tatiana Molotsky, Ilya Borovok, Yuri Voziyanov, Alexander B. Kotlyar, Ezra Yagil, Mikhail Kolot

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

6 Scopus citations

Abstract

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.

Original languageEnglish
Pages (from-to)117-122
Number of pages6
JournalJournal of Molecular Microbiology and Biotechnology
Volume19
Issue number3
DOIs
StatePublished - Dec 2010

Funding

FundersFunder number
National Institute of General Medical SciencesR01GM085848

    Keywords

    • Atomic force microscopy
    • Coliphage HK022
    • Flp recombinase
    • Integrase
    • Recombinase-mediated cassette exchange
    • Site-specific recombination

    Fingerprint

    Dive into the research topics of 'High efficiency of a sequential recombinase-mediated cassette exchange reaction in escherichia coli'. Together they form a unique fingerprint.

    Cite this