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Green fluorescent protein

M Chalfie1

  • 1Department of Biological Sciences, Columbia University, New York, NY 10027, USA.

Photochemistry and Photobiology
|October 1, 1995
PubMed
Summary
This summary is machine-generated.

Green fluorescent protein (GFP) from Aequorea victoria can be expressed in various cells, serving as a marker for gene expression and protein localization. Mutations allow for altered fluorescent properties, expanding its utility.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Background:

  • Bioluminescent coelenterates utilize green fluorescent protein (GFP) for light production via energy transfer.
  • Key GFPs studied are from Aequorea victoria and Renilla reniformis, sharing a common chromophore.
  • Differences in GFP fluorescence are attributed to variations in the protein environment surrounding the chromophore.

Purpose of the Study:

  • To investigate the potential of green fluorescent protein (GFP) as a reporter for gene expression and protein localization.
  • To explore the functional expression of Aequorea victoria GFP in heterologous systems.
  • To assess the feasibility of engineering GFP variants with modified fluorescent characteristics.

Main Methods:

  • Cloning of the Aequorea victoria gfp cDNA.

Related Experiment Videos

  • Heterologous expression of gfp in bacterial, plant, and animal cells.
  • Site-directed mutagenesis of the gfp cDNA to generate variants.

    • Main Results:

    • Expression of Aequorea victoria GFP in the absence of other Aequorea proteins yielded a fluorescent product.
    • Successful heterologous expression of GFP was demonstrated across diverse cell types, including mammalian cells.
    • Engineered GFP variants exhibited enhanced fluorescence intensity and altered spectral properties.

    Conclusions:

    • Green fluorescent protein (GFP) can function as an effective reporter for gene expression and protein localization in living and fixed cells.
    • The cloning and heterologous expression of gfp enable its use as a versatile molecular tool.
    • Mutational analysis of GFP allows for the development of novel variants with tailored fluorescent properties for various applications.