GFP Flashcards
what is fluorescence?
emission of light by substance that has absorbed light/radiation
what is immunofluorescence?
method to localize molecules on fixes cells or tissues
where does the original green fluorescent protein come from?
a jellyfish
what creates the green luminescence?
aequorin and GFP convert Ca2+ luminescent signals
what does GFP do?
absorbs blue light and emits green light
what is special about GFP?
doesn’t need prosthetic group, can be entirely genetically encoded
what allows direct visualization of protein?
fusing GFP to coding sequence
what directs GFP to particular cell comparetments?
peptide locations signals being added to it
what do mutations do to GFP?
changes the absorption and emission colours (can also change maturation time, brightness stability, etc)
what are some things we can do with fluorescent proteins?
track movement of proteins/cells in vivo
visualize intracellular Ca2+ concentrations (monitor activity of neurons in vivo)
what is photobleaching?
photochemical alteration of fluorophore (makes it unable to fluoresce)
what are photoactivatable fluorescent proteins?
fluorescent proteins that display changes in spectral properties after specific light
what is fluorescence resonance energy transfer (FRET)?
technique to gauge the distance between two chromophores (in proximity = FRET)
what is the MS2/MCP system?
allows tracking RNA molecules
