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Related Experiment Videos

Fluorescence-based sensing system for copper using genetically engineered living yeast cells.

Ranjit S Shetty1, Sapna K Deo, Yue Liu

  • 1Department of Chemistry, University of Kentucky, Lexington, KY 40506-0055, USA.

Biotechnology and Bioengineering
|October 30, 2004
PubMed
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A novel whole-cell biosensor detects copper ions using genetically engineered yeast. This system quantifies copper by measuring green fluorescent protein (GFP) production, offering a sensitive and selective analytical method.

Area of Science:

  • Biotechnology
  • Environmental Science
  • Analytical Chemistry

Background:

  • Developing sensitive and selective methods for detecting heavy metal ions is crucial for environmental monitoring and human health.
  • Whole-cell biosensors offer a promising platform for environmental analysis due to their inherent biological recognition capabilities.

Purpose of the Study:

  • To develop and validate a whole-cell optical sensing system for the detection of copper ions.
  • To utilize Saccharomyces cerevisiae engineered with a GFP reporter system for copper quantification.

Main Methods:

  • Engineering Saccharomyces cerevisiae with plasmid pYEX-GFPuv to express the green fluorescent protein (GFPuv) reporter gene under the control of a copper-inducible promoter.
  • Utilizing the Ace1 transcriptional activator protein to regulate GFPuv expression in response to copper ion presence.

Related Experiment Videos

  • Quantifying copper concentration by measuring GFPuv fluorescence intensity via optical detection at specific excitation and emission wavelengths.

    • Main Results:

    • The developed biosensor system demonstrated the ability to detect copper ions at concentrations as low as 5 x 10(-7) M.
    • The system exhibited selectivity for copper ions, with minimal interference from other metal ions, except for silver.
    • Successful application of the sensing system across various analytical platforms and in real-world sample analyses was shown.

    Conclusions:

    • A robust and sensitive whole-cell biosensor for copper detection has been successfully established.
    • The system leverages a genetically modified yeast strain and optical detection for accurate copper quantification.
    • This approach holds potential for diverse environmental and analytical applications requiring copper ion monitoring.