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Related Experiment Videos

Conjugative plasmid transfer from Enterococcus faecalis to Escherichia coli.

P Trieu-Cuot1, C Carlier, P Courvalin

  • 1Centre National de la Recherche Scientifique Unité Associée 271, Institut Pasteur, Paris, France.

Journal of Bacteriology
|September 1, 1988
PubMed
Summary

Genetic transfer between bacteria was studied. A special plasmid enabled gene transfer from gram-positive Enterococcus faecalis to gram-negative Escherichia coli, showing conjugation is possible across these bacterial types.

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Area of Science:

  • Microbiology
  • Genetics
  • Molecular Biology

Background:

  • Bacterial conjugation is a key mechanism for horizontal gene transfer.
  • Understanding inter-kingdom transfer (Gram-positive to Gram-negative) is crucial for microbial genetics.
  • Shuttle vectors are essential tools for studying gene transfer between different bacterial species.

Purpose of the Study:

  • To investigate the feasibility of genetic information transfer via conjugation from Gram-positive to Gram-negative bacteria.
  • To characterize the transfer efficiency of a novel chimeric shuttle vector.
  • To demonstrate inter-kingdom bacterial conjugation.

Main Methods:

  • Construction of a chimeric shuttle plasmid (pAT191) combining elements of pBR322 and pAM beta 1.
  • Introduction of the shuttle vector into Enterococcus faecalis (Gram-positive).

Related Experiment Videos

  • Conjugation experiments between Enterococcus faecalis (donor) and Escherichia coli (recipient, Gram-negative).
  • Quantification of transfer frequency using colony forming units.

    • Main Results:

    • The chimeric shuttle vector pAT191, containing streptococcal plasmid pAM beta 1 tra functions, was successfully constructed.
    • Conjugative transfer of pAT191 from Enterococcus faecalis to Escherichia coli was achieved.
    • The average transfer frequency was determined to be 5 x 10(-9) per donor colony formed.

    Conclusions:

    • Genetic information can be transferred from Gram-positive (Enterococcus faecalis) to Gram-negative (Escherichia coli) bacteria via conjugation.
    • The developed shuttle vector (pAT191) is capable of mediating inter-kingdom bacterial conjugation.
    • This study validates the potential of shuttle vectors for facilitating gene transfer across diverse bacterial domains.