Background of the Invention:
Some cultured coral appear to produce a unique bright red fluorescence when exposed to stressors such as invading parasites including foraminifers, algae, and fungi. The tissue bordering these invading organisms slowly lifts off the skeleton, becomes disorganized histologically and appears bright pink to the naked eye. When excited with green light it brilliantly fluoresces red.
This fluorescence was first observed in live coral specimen using a dipping fluorescent microscope equipped with a hyperspectral camera. The excitation range (the light that will induce fluorescence) appeared specific to the green wavelengths and fluoresced in an area of red (gave off red light) that had not been observed in native coral red fluorescent proteins.
In addition the emitted light was 10-100 fold brighter than the native green fluorescence or the chlorophyll fluorescence of the specimen. The invention described herein is the process of sorting through a DNA library (tens of thousands of sequences) in order to find the specific sequence for this very bright protein, akin to finding a needle in a haystack, so that we could produce enough of the protein to characterize it biochemically and prove its uniqueness and desirability due to its extreme brightness.
There are numerous commercial applications for fluorescent proteins, many already described in the literature and in use daily; wherein this protein will substitute for fluorescent proteins of lower QY and EQ. The brightness of this RFP will lend itself to studies in live cell culture and small animals where the protein DNA sequence may be linked to host DNA to be co-expressed providing a reporter of the host gene expression…the fluorescent emission of this protein should out compete natural tissue auto-fluorescence giving an excellent ability to detect previously undetectable processes.
In addition, a bright far red shifted fluorescent protein can serve as a valuable color addition for multi-color labeling experiments in which a bright RFP can be paired with and visualized along green and yellow. The brightness of this FP will allow more accurate comparison of gene expression between tprRFP labelled expression products and bright green or yellow labelled expression products and the further red shifted emission of tprRFP can decrease occurrence of cross-talk and bleed through in these imaging experiments utilizing multiple colors for labeling.
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