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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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Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been...
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Related Experiment Video

Updated: Dec 21, 2025

How to Quantify the Fraction of Photoactivated Fluorescent Proteins in Bulk and in Live Cells
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A photostable monomeric superfolder green fluorescent protein.

Fernando M Valbuena1, Ivy Fitzgerald2, Rita L Strack1

  • 1Department of Molecular Genetics and Cell Biology, The University of Chicago, Chicago, Illinois, USA.

Traffic (Copenhagen, Denmark)
|May 17, 2020
PubMed
Summary

Researchers developed msGFP2, a new green fluorescent protein (GFP) variant. This improved protein tagging tool offers enhanced photostability and minimizes disruption to partner proteins, making it ideal for general use.

Keywords:
GFPcytotoxicityfluorescent proteinmCherrymScarletmonomericphotobleachingsuperfolder

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Biotechnology

Background:

  • Green fluorescent protein (GFP) from Aequorea victoria is widely used for protein tagging.
  • Existing variants like EGFP and superfolder GFP have limitations, including photobleaching and functional interference.

Purpose of the Study:

  • To engineer a novel GFP variant, msGFP2, that combines monomeric superfolder properties with improved photostability and reduced functional interference.
  • To provide an optimal GFP derivative for general-purpose protein tagging applications.

Main Methods:

  • Engineering of msGFP2 by modifying N- and C-terminal peptides of a previously developed monomeric superfolder GFP (msGFP).
  • Comparative analysis of msGFP2's photostability, folding efficiency, and impact on partner protein function against EGFP and mEGFP.

Main Results:

  • msGFP2 demonstrates photostability comparable to EGFP, overcoming the faster photobleaching of earlier superfolder variants.
  • msGFP2 retains monomeric superfolder properties, ensuring efficient folding under various conditions.
  • msGFP2 exhibits reduced disruption of partner protein functions compared to EGFP and mEGFP.

Conclusions:

  • msGFP2 represents a significant advancement in GFP engineering for protein tagging.
  • Its enhanced photostability and minimal functional interference make it a superior choice for general protein labeling in biological research.