Abstract
We report a pH-insensitive and photostable ultramarine fluorescent protein, Sirius, with an emission peak at 424 nm, the shortest emission wavelength among fluorescent proteins reported to date. The pH-insensitivity of Sirius allowed prolonged visualization of biological events in an acidic environment. Two fluorescence resonance energy transfer (FRET) pairs, Sirius-mseCFP and Sapphire-DsRed, allowed dual-FRET imaging with single-wavelength excitation, enabling detection of Ca(2+) concentration and caspase-3 activation in the same apoptotic cells.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Substitution
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Animals
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Apoptosis
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Calcium Signaling
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Caspase 3 / metabolism
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Color
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Dictyostelium / microbiology
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Escherichia coli / genetics
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Escherichia coli / metabolism
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Fluorescence Resonance Energy Transfer
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Green Fluorescent Proteins / chemistry
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Green Fluorescent Proteins / genetics
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HeLa Cells
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Humans
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Hydrogen-Ion Concentration
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Luminescent Proteins / chemistry*
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Luminescent Proteins / genetics
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Molecular Sequence Data
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Mutagenesis
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Phagocytosis
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Photobleaching
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Protein Stability
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Recombinant Fusion Proteins / chemistry
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / metabolism
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Spectrometry, Fluorescence
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Spectrophotometry, Ultraviolet
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Transfection
Substances
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Cyan Fluorescent Protein
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Luminescent Proteins
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Recombinant Fusion Proteins
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blue fluorescent protein, Aequorea victoria
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fluorescent protein 583
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green fluorescent protein, Aequorea victoria
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Green Fluorescent Proteins
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Caspase 3