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Related Experiment Videos

Model suicide vector for containment of genetically engineered microorganisms.

A K Bej1, M H Perlin, R M Atlas

  • 1Department of Biology, University of Louisville, Kentucky 40292.

Applied and Environmental Microbiology
|October 1, 1988
PubMed
Summary

A novel suicide vector (pBAP19h) effectively eliminated genetically engineered microorganisms in soil. However, high selective pressure in vitro led to the emergence of resistant cells, compromising containment effectiveness.

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The Molecularly Crowded Cytoplasm of Bacterial Cells: Dividing Cells Contrasted with Viable but Non-culturable (VBNC) Bacterial Cells.

Current issues in molecular biology·2012

Area of Science:

  • Microbiology
  • Genetic Engineering
  • Biotechnology

Background:

  • Genetically engineered microorganisms (GEMs) require containment strategies.
  • Suicide vectors offer a method to control GEM populations.
  • The pBAP19h plasmid utilizes a hok gene for cell lethality.

Purpose of the Study:

  • To evaluate the efficacy of the pBAP19h suicide vector for GEM containment.
  • To assess the vector's performance in vitro and in soil microcosms.
  • To investigate potential failure mechanisms of the suicide vector system.

Main Methods:

  • Construction of the pBAP19h plasmid with a lac promoter-controlled hok gene and carbenicillin resistance.
  • In vitro experiments assessing cell elimination with and without carbenicillin.

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  • Soil microcosm experiments to evaluate GEM containment under selective pressure.

    • Main Results:

    • In vitro, hok induction eliminated vector-bearing cells without carbenicillin.
    • Carbenicillin presence led to the emergence of Hok-resistant cells, indicating system failure.
    • Soil microcosms showed 90-99% elimination of GEMs, but some Hok-resistant survivors with impaired growth were observed.

    Conclusions:

    • The pBAP19h suicide vector demonstrates efficacy in controlling GEMs in soil.
    • High selective pressure can lead to the evolution of resistance, compromising containment.
    • Mutations conferring Hok resistance may impact cell viability and growth, requiring further investigation.