Naturally Fluorescent Field Mouse, Mus booduga (Gray, 1837) and Common House Gecko, Hemidactylus frenatus (Schlegel, 1836) as Model Organisms for Biomedical Studies
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Aequorea victoria
The unique and stunning spectroscopic properties of Green Fluorescent Protein (GFP) from the jellyfish Aequorea victoria, not to mention of its remarkable structural stability, have made it one of the most widely studied and used molecular tool in medicine, biochemistry, and cell biology. Its high fluorescent quantum yield is due to its chromophore, structure responsible of emitting green visible light when excited at 395 nm. Although it is noteworthy that there is enormous available information of the wonderful luminescent properties of GFP, the fact is that there are features and properties unexplored yet, particulary about its capabilities as molecular reporter in several biological processes. In this work, we used recombinant DNA technology to express the protein in bacteria; prepared the bacterial system both in liquid and solid media, and assembled an experimental set to expose those media to a laser beam; thereby we excited the protein chromophore and used emission spectroscopy in order to observe variations in fluorescence when the bacterial system is exposed to different temperatures.
Aequorea victoria
Chromophore
Quantum yield
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Aequorea victoria
Bioreporter
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Aequorea victoria
Wild type
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Aequorea victoria
Fluorescent protein
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The green fluorescent protein (GFP) of the jelly fish Aequorea victoria is a spontaneously fluorescent protein that can be incorporated into other proteins by genetic fusion. One of our approaches is to use two GFPs of different colors to permit fluorescence resonance energy transfer (FRET), which is highly sensitive to the relative orientation and distance between the two fluorophores and alters the ratio of their emission intensities. For example, cameleons are genetically-encoded fluorescent indicators for calcium. I describe here potentially unique features of GFP-based FRET, which would allow us to make probes for many intracellular signaling events that are currently assayed by grinding millions of cells.
Aequorea victoria
Fluorescent protein
Bimolecular fluorescence complementation
Yellow fluorescent protein
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Fluorescent protein
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Reporters based on the green fluorescent protein (GFP) from the jellyfish Aequorea victoria and GFP-like proteins from other marine organisms provide valuable tools to monitor gene transfer and expression noninvasively in living cells. Stable cell lines were generated from the Sp2/0-Ag14 hybridoma that express up to three spectral enhanced versions of GFP, the enhanced cyan fluorescent protein (ECFP), the enhanced green fluorescent protein (EGFP), and the enhanced yellow fluorescent protein (EYFP), and/or a variant of the Discosoma coral red fluorescent protein (DsRed). The panel of lines was used to demonstrate a flow cytometric procedure for simultaneous analysis of all four fluorescent proteins that utilizes dual-laser excitation at 488 nm and 407 nm. Additional schemes for simultaneous detection of two, three or four of these fluorescent proteins are also presented.
Aequorea victoria
Fluorescent protein
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Light is produced from several bioluminescent species through the action of green fluorescent proteins (GFPs) (reviewed in ref. 1). One such GFP from the jellyfish Aequorea victoria fluoresces following the transfer of energy from the Ca2+-activated photoprotein aequorin(2,3). This energy transfer mechanism is radiationless, and proceeds via the interaction of these two proteins in photogenic cells located at the base of the jellyfish. Expression of GFP in vivo does not alter the spectral properties of the protein (4). Full-length GFP appears to be required for fluorescence, however the chromophore responsible for light absorption is located within a hexapeptide at positions 64–69 (5). This region of GFP contains a Ser65-dehydro Tyr66-Gly67 cyclic tripeptide which functions as the minimal chromophore.
Aequorea victoria
Chromophore
Photoprotein
Aequorin
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Green fluorescent protein (GFP) is a sort of protein produced by jellyfish (Aequorea victoria) and sea pansy (Renilla reniformis) which absorbs blue light and emits green fluorescence without exogenous substrates or co-factors, and the fluorescence can be easily detected with fluorescent microscope or fluorescence-activated cell sorter (FACS).Green fluorescent protein gene is an important new reporter gene that can express in living cells without other external substrate and no specific in species, tissues or cells at present.With features of sensitivity in detection, simplicity in manipulation, and reliability in result, GFP is an important reporter gene of practical. Being an effective selective marker, GFP has a great potential application in fields of cell, developmental and molecular biology.
Aequorea victoria
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Aequorea victoria
Fluorescence Correlation Spectroscopy
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