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A stringently controlled expression system for analysing lateral gene transfer between bacteria

S Jaenecke1, V de Lorenzo, K N Timmis

  • 1Department of Microbiology, GBF-National Research Centre for Biotechnology, Braunschweig, Germany.

Molecular Microbiology
|July 1, 1996
PubMed
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We developed a novel genetic circuit to detect and quantify horizontal gene transfer in microbial communities without needing specific selection. This system enables non-disruptive, real-time monitoring of gene flow in natural environments.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Environmental Science

Background:

  • Microbial communities exhibit remarkable adaptability driven by lateral gene transfer.
  • Understanding gene transfer in natural ecosystems is crucial but challenging.
  • Existing methods often require selective pressure for recipient organisms.

Purpose of the Study:

  • To develop a novel genetic circuit for detecting and quantifying horizontal gene transfer (HGT).
  • To enable HGT monitoring in natural environments without selection for recipient-specific phenotypes.
  • To facilitate non-disruptive, real-time assessment of gene flow.

Main Methods:

  • Engineered a reporter gene cassette (lacZ) controlled by a synthetic regulatory element.
  • Utilized a dual-repression system (lambda phage Cl-EK117 and IS 10 RNA-OUT) to ensure expression only upon transfer.

Related Experiment Videos

  • Validated the system in Pseudomonas putida to detect conjugational transfer frequencies as low as 10^-6.

    • Main Results:

    • Successfully detected and quantified HGT using the engineered genetic circuit.
    • Achieved detection of low-frequency conjugational transfer events (10^-6).
    • Demonstrated the system's effectiveness in a relevant bacterial host (Pseudomonas putida).

    Conclusions:

    • The developed genetic circuit provides a powerful tool for studying microbial gene flow.
    • Its modular design and broad host range facilitate environmental monitoring.
    • Enables real-time, in situ, and non-disruptive tracking of HGT in microbial communities.