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Related Experiment Videos

Precise excision of bacteriophage Mu DNA.

C Abbes1, G Sezonov, D Joseleau-Petit

  • 1Institut Jacques Monod, Centre National de la Recherche Scientifique (CNRS), Université Paris 6, Université Paris 7, France.

Canadian Journal of Microbiology
|September 29, 2001
PubMed
Summary
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The temperate bacteriophage Mu can precisely excise from bacterial DNA, restoring gene function. However, this precise excision is linked to a loss of the ability to form colonies.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Bacteriophage Research

Background:

  • The temperate bacteriophage Mu is a transposable element known for random integration into bacterial DNA, causing mutations.
  • Previous observations suggested that Mu might not precisely excise, unlike other transposable elements, as Mu-induced mutants typically do not revert.

Purpose of the Study:

  • To investigate whether the temperate bacteriophage Mu can precisely excise from bacterial DNA.
  • To determine the consequences of Mu's precise excision on bacterial viability and gene function.

Main Methods:

  • Utilized a lacZ::Muc62(Ts) strain, which is unable to form Lac+ colonies due to Mu prophage integration.
  • Cloned a functional lacZ+ gene in vivo using a mini-Mu plasmid under conditions of prophage induction.

Related Experiment Videos

  • Analyzed recovered lac+ plasmids to assess the integrity of the bacterial DNA sequence at the integration site.

    • Main Results:

    • Recovery of lacZ+ genes demonstrated that the wild-type sequence was restored at the Mu integration site.
    • This indicates that precise excision of the Mu prophage does occur.
    • Despite the restoration of the lacZ+ gene, the bacterial strains consistently lost colony-forming ability.

    Conclusions:

    • Precise excision of the temperate bacteriophage Mu from bacterial DNA is possible.
    • Precise excision events mediated by Mu are strongly associated with a loss of bacterial colony-forming ability.
    • This finding challenges the previous assumption of Mu's inability to undergo precise excision and highlights a novel consequence of this process.