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A DNA sequence encoding the Aequorea victoria GFP (Ser 2-Lys 238) was expressed, with a polyhistide tag at the N-terminus.
The green fluorescent protein (GFP) is a protein composed of 238 amino acid residues (26.9kDa) that exhibits bright green fluorescence when exposed to light in the blue to ultraviolet range. Although many other marine organisms have similar green fluorescent proteins, GFP traditionally refers to the protein first isolated from the jellyfish Aequorea victoria. The GFP from A. victoria has a major excitation peak at a wavelength of 395 nm and a minor one at 475 nm. Its emission peak is at 509 nm, which is in the lower green portion of the visible spectrum. The fluorescence quantum yield (QY) of GFP is 0.79. The GFP from the sea pansy (Renilla reniformis) has a single major excitation peak at 498 nm.
The recombinant Aequorea victoria GFP consisting of 253 amino acids and has a calculated molecular mass of 28.7KDa. It migrates as an 35 kDa band in SDS-PAGE under reducing conditions as predicted.
Predicted N terminal
> 90 % as determined by SDS-PAGE
Lyophilized from sterile PBS, pH 7.5
Store it under sterile conditions at -70oC. It is recommended that the protein be aliquoted for optimal storage. Avoid repeated freeze-thaw cycles.
A hardcopy of COA with reconstitution instruction is sent along with the products.
Aequorea victoria, also sometimes called the crystal jelly, is a bioluminescent hydrozoan jellyfish, or hydromedusa, that is found off the west coast of North America. This species is thought to be synonymous with Aequorea aequorea of Osamu Shimomura, the discoverer of green fluorescent protein (GFP). Shimomura together with Martin Chalfie and Roger Y. Tsien were awarded the 2008 Nobel Prize in Chemistry for the discovery and development of this protein as an important biological research tool. Originally the victoria species was supposed to designate the variant found in the Pacific, and the aequorea designation was used for specimens found in the Atlantic and Mediterranean. The species name used in GFP purification was later disputed by M.N. Arai and A. Brinckmann-Voss (1980), who decided to separate them on the basis of 40 specimens collected from around Vancouver Island. Osamu Shimomura notes that this species in general shows great variation: from 1961 to 1988 he collected around 1 million individuals in the waters surrounding the Friday Harbor Laboratories of University of Washington, and in many cases there were pronounced variations in the form of the jellyfish. In September 2009, Aequorea victoria was spotted in the Moray Firth, an unusual occurrence, as crystal jellies had never been seen or reported in British waters. The specimen is now on display in Macduff Marine Aquarium in Aberdeenshire, Scotland.
GFP ribbon diagram.
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