Imaging fluorescence resonance energy transfer between two green fluorescent proteins in living yeast.
We show that fluorescence resonance energy transfer between two mutants of the green fluorescent protein (GFP) can be monitored by imaging microscopy in living yeast. This work is based on the constitutive expression of a GFP-containing fusion protein and the inducible expression of the tobacco etch virus (TEV) protease. In the fusion protein, the P4.3 GFP mutant is linked to the YS65T GFP mutant by a spacer bearing the TEV protease-specific cleavage site.
Sagot
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Laboratoire de Biologie Cellulaire de la Levure, Institut de Biochimie et Génétique Cellulaires, CNRS UPR 9026, Bordeaux, France.
Bonneu
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Balguerie
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Aigle
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D010450:Endopeptidases / Q000235:genetics; D004735:Energy Transfer; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism*; D008856:Microscopy, Fluorescence / Q000379:methods*; D009154:Mutation; D017800:Potyvirus / Q000201:enzymology; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D013050:Spectrometry, Fluorescence / Q000379:methods
D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D010450:Endopeptidases; C099488:TEV protease