Green Fluorescent Protein: Research Tool: Difference between revisions
New page: ==Use in the Laboratory== Green fluorescent protein has had great success as a marker protein in a variety of biological systems due to its inherent stability, which only adds to its many... |
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===Use of GFP in FRET=== | ===Use of GFP in FRET=== | ||
Förster resonance energy transfer (FRET) is a laboratory technique used to detect the proximity of two biological molecules by the fusion of fluorescent proteins with each of the proteins of interest. When the two fluorophores come within a certain proximity (typcially <100Å) in the proper orientation, the excited fluorophore (donor) emits energy that excites the second longer-wavelength fluorophore (acceptor) suc that it also fluoresces. Results can then be seen by the ratio of the donor and acceptor emission intensities.<ref>For a more extensive description and development of this topic, go to [http://en.wikipedia.org/wiki/F%C3%B6rster_resonance_energy_transfer Wikipedia]</ref> FRET is noninvasive and is therefore safe for using within live cells<ref name="Pollock" />. | |||
While the wild-type GFP protein has not been particularly useful as a sensor in FRET, several mutants of GFP have been manufactured to create proteins with distinct fluorescence spectra. FRET has been done between two GFP molecules or a single GFP molecule and a secondary fluorophore <ref name="Pollock" />. | While the wild-type GFP protein has not been particularly useful as a sensor in FRET, several mutants of GFP have been manufactured to create proteins with distinct fluorescence spectra. FRET has been done between two GFP molecules or a single GFP molecule and a secondary fluorophore <ref name="Pollock" />. | ||