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Related Experiment Videos

DNA inversion on conjugative plasmid pVT745.

Jinbiao Chen1, Donald J Leblanc, Dominique M Galli

  • 1School of Dentistry, Department of Oral Biology, Indiana University, 1121 W. Michigan Street, Indianapolis, IN 46202, USA.

Journal of Bacteriology
|October 11, 2002
PubMed
Summary
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A novel site-specific recombinase, Inv, mediates DNA inversion in Actinobacillus actinomycetemcomitans plasmids. This inversion, crucial for plasmid stability, is regulated by host factors and can impact plasmid transfer efficiency.

Area of Science:

  • Molecular Biology
  • Microbiology
  • Genetics

Background:

  • Plasmid pVT745 from Actinobacillus actinomycetemcomitans facilitates inter-strain DNA transfer.
  • Plasmid derivatives can undergo structural rearrangements post-transfer, impacting genetic stability.

Purpose of the Study:

  • To investigate the mechanism behind a 9-kb DNA segment inversion observed in pVT745 derivatives.
  • To identify the genetic elements and host factors involved in this plasmid rearrangement.

Main Methods:

  • Plasmid transfer experiments between Actinobacillus actinomycetemcomitans strains.
  • Analysis of plasmid DNA structure in transconjugants.
  • Site-specific recombinase gene identification and functional analysis.

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Main Results:

  • A 9-kb DNA segment inversion in pVT745 derivatives was mediated by a novel site-specific recombinase, Inv.
  • The inversion process is RecA-independent and relies on two specific 22-bp recombination sites on the plasmid.
  • Efficient inversion requires a host factor present in most strains but absent or inactive in the original host, VT745.

Conclusions:

  • The Inv recombinase plays a role in plasmid DNA rearrangement, likely influencing plasmid stability and transfer.
  • A host-dependent factor regulates Inv activity, suggesting a complex interplay between plasmid and host.
  • Inactivation of the invertase gene significantly increases transconjugant numbers, indicating its role in post-mating plasmid loss.