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Related Experiment Videos

Structure and function of the shufflon in plasmid R64.

Atsuko Gyohda1, Nobuhisa Furuya, Akiko Ishiwa

  • 1Department of Biology, Tokyo Metropolitan University, Hachioji, Tokyo 192-0397, Japan.

Advances in Biophysics
|October 21, 2004
PubMed
Summary
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Site-specific recombination in plasmid R64

Area of Science:

  • Microbiology
  • Molecular Biology
  • Genetics

Background:

  • Site-specific recombination generates biological diversity in prokaryotes.
  • Plasmid R64 utilizes a unique shufflon system with seven recombination sites (sfx) and a recombinase (Rci).

Purpose of the Study:

  • To investigate the mechanism of shufflon multiple inversion system in plasmid R64.
  • To elucidate the role of Rci recombinase and sfx site asymmetry in DNA inversion.

Main Methods:

  • Overexpression and purification of Rci protein.
  • In vitro recombination assays using R64 shufflon system.
  • Analysis of sfx sequence elements and their impact on inversion frequency.

Main Results:

Related Experiment Videos

  • Shufflon inversions create diverse pilV genes, determining recipient specificity in bacterial matings.
  • Rci mediates recombination via a 5' protruding 7-bp staggered cut in asymmetric sfx sites.
  • Symmetric sfx sequences and altered Rci binding enhance inversion frequency.

    • Conclusions:

    • The asymmetry of sfx sites and Rci's binding properties regulate shufflon recombination.
    • The Rci C-terminal domain is crucial for specific binding to asymmetric sfx sites.
    • Shufflon system provides a model for understanding DNA inversion and biological diversity.