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Characterization of site-specific recombination by the integrase MJ1 from enterococcal bacteriophage phiFC1

Mi-Ok Park1, Ki-Hong Lim, Tae-Hyung Kim

  • 1School of Life Sciences and Biotechnology, Korea University, Seoul 136-701, Korea.

Journal of Microbiology and Biotechnology
|December 7, 2007
PubMed

Insights

Bacteriophage phiFC1 integrase (MJ1) efficiently catalyzes site-specific recombination between phage (attP) and bacterial (attB) attachment sites. This integrase functions in vitro without additional factors and recognizes specific DNA sequences, suggesting a simple mechanism.

Area of Science:

  • Molecular Biology
  • Microbiology
  • Biochemistry

Background:

  • Bacteriophage phiFC1 integrase (MJ1) mediates site-specific recombination between phage (attP) and host (attB) attachment sites.
  • MJ1 integrase has demonstrated activity in both its natural host, Enterococcus faecalis, and the non-host bacterium Escherichia coli.

Purpose of the Study:

  • To investigate the biochemical properties of the MJ1 integrase.
  • To elucidate the mechanism by which MJ1 integrase performs site-specific recombination.

Main Methods:

  • In vitro recombination assays were performed to assess MJ1 integrase activity.
  • Electrophoretic mobility shift assays (EMSA) and DNase I footprinting were used to study MJ1 integrase-DNA interactions.

Main Results:

  • MJ1 integrase catalyzed attP-attB recombination in vitro without requiring additional host factors.
  • MJ1 integrase demonstrated efficient binding to attP and attB sites.
  • DNase I footprinting identified specific, relatively short DNA sequences (approx. 50 bp) recognized by MJ1 integrase, including an overlapping region in attB and attP.

Conclusions:

  • MJ1 integrase directly catalyzes integrative recombination between attP and attB.
  • The recombination mechanism mediated by MJ1 integrase appears to be simple and unidirectional, similar to serine integrases.

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