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Related Concept Videos

Reporter Genes02:11

Reporter Genes

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Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
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Highly Sensitive and Rapid Fluorescence Detection with a Portable FRET Analyzer
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A highly selective cell-based fluorescent biosensor for genistein detection.

Lucy Fang-I Chao1, Dany Liu1, Verena Siewers1

  • 1Department of Life Sciences, Division of Systems and Synthetic Biology, Chalmers University of Technology, Gothenburg SE-41296, Sweden.

Engineering Microbiology
|December 4, 2024
PubMed
Summary

Researchers developed an improved biosensor in yeast to detect genistein, a beneficial compound from legumes. This tool enhances screening for high-producing microbial strains, accelerating the development of genistein-producing cell factories.

Keywords:
BiosensorFlavonoidGenisteinIsoflavone

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Area of Science:

  • Biotechnology
  • Synthetic Biology
  • Microbial Engineering

Background:

  • Genistein, a legume-derived isoflavone, offers significant health benefits, driving interest in microbial production.
  • Developing efficient microbial cell factories for genistein requires effective screening methods for high-producing strains.

Purpose of the Study:

  • To engineer a sensitive and specific cell-based biosensor for genistein detection in *Saccharomyces cerevisiae*.
  • To facilitate high-throughput screening of microbial strains for enhanced genistein production.

Main Methods:

  • Repurposing the Gal4DBD-ERα-VP16 (GEV) transcriptional activator in yeast to create a genistein-responsive system.
  • Engineering the GEV sensor protein, its promoter (using *REV1*p), and the reporter promoter (removing Mig1 sites, adding Gal4p sites) for improved performance.
  • Replacing the VP16 transactivating domain with the VPR transactivator.

Main Results:

  • The engineered biosensor achieved a 20-fold increase in GFP signal intensity upon genistein induction compared to the uninduced state.
  • The biosensor demonstrated high specificity, responding only to genistein among eight tested flavonoids.
  • A linear response to genistein concentration and intracellular genistein levels was observed.

Conclusions:

  • The developed biosensor is a sensitive and specific tool for genistein detection.
  • This biosensor can significantly aid in the high-throughput screening of genistein-producing yeast cell factories.
  • Further optimization could accelerate the biotechnological production of genistein.