Nurturing Life Sciences
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#Unit 13. Methods in Biology
Fluorescent probes are derived from a naturally occurring fluorescent protein, the green fluorescent protein (GFP) of the jellyfish Aequorea Victoria.
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When excited by absorption of a photon of light, GFP emits a photon (that is, it fluoresces) in the green region of the spectrum.
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The light-absorbing/ emitting center of GFP (its chromophore) comprises an oxidized form of the tripeptide –Ser65–Tyr66–Gly67
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This protein, from the bioluminescent jellyfish Aequorea victoria, fluoresces with a peak wavelength of 508 nm (green light) when irradiated by ultraviolet or blue light (optimally 400 nm).
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Cyclization between Ser and Gly is probably rapid, and the oxidation of the Tyr side chain (by O2) is probably the rate-limiting step of fluorophore generation.
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Genetic engineering has introduced site-specific mutations that enhance fluorescence intensity and shift the wavelength of the emitted light to different colors, thereby making it possible to simultaneously monitor the expression of two or more different genes.