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Related Experiment Videos

"Fluorescent timer": protein that changes color with time.

A Terskikh1, A Fradkov, G Ermakova

  • 1School of Medicine, Stanford University, Stanford, CA 94305, USA. Alexey.Terskikh@Stanford.edu

Science (New York, N.Y.)
|November 25, 2000
PubMed
Summary
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Researchers developed a novel fluorescent timer protein (E5) that shifts from green to red fluorescence over time. This tool enables precise measurement of gene expression dynamics in whole organisms, independent of protein levels.

Area of Science:

  • Molecular Biology
  • Developmental Biology
  • Biochemistry

Background:

  • Fluorescent proteins are vital tools for monitoring biological processes.
  • Existing fluorescent reporters often lack the ability to track dynamic changes in gene expression over time.
  • A method to measure promoter activation and down-regulation in real-time is needed.

Purpose of the Study:

  • To develop a novel fluorescent protein that acts as a timer for gene expression.
  • To validate the utility of this "fluorescent timer" in tracking promoter activity in vivo.
  • To demonstrate its application in different model organisms and developmental contexts.

Main Methods:

  • Generation and characterization of a red fluorescent protein mutant (drFP583 E5).
  • Assessment of fluorescence color conversion rate and its independence from protein concentration.

Related Experiment Videos

  • In vivo labeling experiments in Caenorhabditis elegans using a heat shock-dependent promoter.
  • In vivo labeling experiments in developing Xenopus embryos using the Otx-2 promoter.

    • Main Results:

    • The E5 mutant exhibits a time-dependent shift in fluorescence from green to red.
    • The rate of color conversion is constant and not influenced by protein concentration.
    • E5 successfully monitored temporal gene expression from a heat shock promoter in C. elegans.
    • E5 effectively tracked Otx-2 promoter activity during Xenopus development.

    Conclusions:

    • The E5 mutant serves as a reliable "fluorescent timer" for quantitative analysis of gene expression.
    • This tool allows for the monitoring of both promoter activation and down-regulation across different biological systems.
    • E5 provides a powerful new method for studying gene expression dynamics on a whole-organism scale.