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Related Experiment Videos

High expression of plasmid-encoded tetracycline resistance gene in E. coli causes a decrease in membrane-bound ATPase

M S Valenzuela1, K A Siddiqui, B L Sarkar

  • 1Division of Biomedical Sciences, Meharry Medical College, Nashville, Tennessee 37208, USA. valenz44@ccvax.mme.edu

Plasmid
|July 1, 1996
PubMed
Summary
This summary is machine-generated.

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Tetracycline resistance gene expression in E. coli reduces membrane-bound ATPase activity, impacting cell growth. This effect is linked to the intact tet gene and is more pronounced with tac promoter-controlled expression.

Area of Science:

  • Molecular Biology
  • Bacterial Physiology
  • Genetics

Background:

  • The tetracycline resistance gene (tet) on plasmid pBR322 has known pleiotropic effects beyond tetracycline efflux.
  • Previous studies indicated that expressing the tet gene on plasmid pKH47 inhibits Escherichia coli growth.
  • Understanding these pleiotropic effects is crucial for bacterial genetic engineering and physiology.

Purpose of the Study:

  • To investigate the impact of tet gene expression on membrane-bound ATPase activity in E. coli.
  • To determine if reduced ATPase activity affects bacterial growth, particularly in minimal media.
  • To compare the effect of tet gene expression from different plasmids and promoters.

Main Methods:

  • Culturing E. coli strains harboring various plasmids, including pKH47 and an unrelated plasmid with a tac promoter.

Related Experiment Videos

  • Measuring membrane-bound ATPase activity in bacterial cell extracts.
  • Assessing bacterial growth rates in minimal medium with succinate as the sole carbon source.

    • Main Results:

    • E. coli cells containing plasmid pKH47 exhibited reduced membrane-bound ATPase activity.
    • This reduction in ATPase activity was dependent on the presence of an intact tet gene.
    • Reduced ATPase activity correlated with impaired growth in succinate-based minimal medium, an effect amplified when tet gene expression was driven by the tac promoter.

    Conclusions:

    • Expression of the tetracycline resistance gene (tet) negatively impacts E. coli membrane-bound ATPase activity.
    • This impairment of ATPase activity contributes to growth inhibition, especially under nutrient-limited conditions.
    • The findings highlight an uncharacterized pleiotropic effect of tet gene expression relevant to bacterial physiology and plasmid biology.