Improved green fluorescence

The green fluorescent protein (GFP) from the jellyfish Aequorea victoria has attracted interest since the demonstration that heterologous expression of the cloned gene can generate striking green fluorescence. Despite the potential of recombinant GFP as a marker for gene expression or cell lineage o...

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Bibliographic Details
Published in:Nature (London) 1995-02, Vol.373 (6516), p.663-664
Main Authors: Heim, Roger, Cubitt, Andrew B, Tsien, Roger Y
Format: Article
Language:English
Subjects:
Aequorea victoria
Animals
Cnidaria
Green Fluorescent Proteins
Humanities and Social Sciences
Luminescent Proteins - chemistry
Luminescent Proteins - genetics
multidisciplinary
Pennatulacea
Point Mutation
Renilla reniformis
Science
Science (multidisciplinary)
scientific-correspondence
Scyphozoa
Serine - chemistry
Spectrum Analysis
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Summary:The green fluorescent protein (GFP) from the jellyfish Aequorea victoria has attracted interest since the demonstration that heterologous expression of the cloned gene can generate striking green fluorescence. Despite the potential of recombinant GFP as a marker for gene expression or cell lineage or as an in situ tag for fusion proteins, the wild-type protein from A. victoria has significant deficiencies. Its excitation spectrum shows peaks at both 396 and 475 nm. The longer-wavelength excitation peak has the advantages of greater photostability and better matching to stand fluorescein filter sets, but is relatively low in amplitude. A closely related protein from the sea pen Renilla reniformis has the same high quantum yield of emission (0.7-0.8), yet shows only one absorbance and excitation peak with an extinction coefficient per monomer more than 10 times that of the longer-wavelength peak of Aequorea GFP. We now report that simple point mutations in Aequorea GFP ameliorate its main problems and bring its spectra much closer to that of Renilla.
ISSN:0028-0836
1476-4687
DOI:10.1038/373663b0