|発明の名称 （英語）||Fluorescent protein|
|発明の概要（英語）||The object of the present invention is to provide a novel fluorescent protein in which on and off of fluorescence thereof can be controlled by irradiation with lights of two different wavelengths. The present invention provides a fluorescent protein shown in the following (a) or (b); (a) a protein which has the amino acid sequence shown in SEQ ID NO: 3 or 7; or (b) a protein which has an amino acid sequence comprising a deletion, substitution, and/or addition of one or several amino acids with respect to the amino acid sequence shown in SEQ ID NO: 3 or 7, and which has fluorescence characteristics of exhibiting a photochromic effect.|
Green fluorescent protein (GFP) derived from a jelly fish, Aequorea victoria, has many uses in biological systems. Recently, various GFP mutants that are changed in color, improved in folding characteristic, enhanced in luminance, and modified in pH sensitivity have been prepared based on random mutagenesis techniques and semi-rational mutagenesis techniques. Using genetic manipulation technology, another protein is fused to a fluorescent protein such as GFP, and monitoring of its expression and transport are being carried out.
As one of the most frequently used GFP mutants, yellow fluorescent protein (YFP) is listed. YFP shows fluorescence with the longest wavelength among jellyfish Aequorea GFP mutants. The values of ε and Φ of most YFPs are 60,000 to 100,000 M－1cm－1 and 0.6 to 0.8, respectively (Tsien, R. Y. (1998). Ann. Rev. Biochem. 67, 509-544), and these values are comparable to those of common luminophores (fluorescein, rhodamine, etc.). Accordingly, improvement in absolute luminance of YFP is almost reaching the limit.
As other examples of the GFP mutants, there is cyan fluorescent protein (CFP), and enhanced cyan fluorescent protein (ECFP) is known. Further, red fluorescent protein (RFP) is also isolated from a Discoma sp., and DsRed is known. Thus, four typed of fluorescent proteins (green, yellow, cyan, and red) have been developed one after another, and the spectral range has been greatly extended.
1. A fluorescent protein shown in the following (a) or (b):
(a) a protein which has the amino acid sequence shown in SEQ ID NO: 3; or
(b) a protein which has
an amino acid sequence comprising a deletion, substitution, and/or addition of one to twenty amino acids with respect to the amino acid sequence shown in SEQ ID NO: 3,
an aspartic acid residue at position 172, a histidine residue at position 194, and a serine residue at position 205 with respect to SEQ ID NO: 3,
and which has fluorescence characteristics of exhibiting a photochromic effect.
2. A photochromic material comprising the fluorescent protein of claim 1.
3. A DNA encoding the fluorescent protein according to claim 1.
4. A recombinant vector having the DNA of claim 3.
5. An isolated host cell having the DNA of claim 3.
6. An isolated host cell having the recombinant vector of claim 4.
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