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Fluorescent protein achieved

Foreign code F190009898
File No. 06267-US
Posted date Aug 26, 2019
Country United States of America
Application number 56104004
Gazette No. 20070072259
Gazette No. 7960530
Date of filing Jun 16, 2004
Gazette Date Mar 29, 2007
Gazette Date Jun 14, 2011
International application number JP2004008790
International publication number WO2004111236
Date of international filing Jun 16, 2004
Date of international publication Dec 23, 2004
Priority data
  • P2003-170330 (Jun 16, 2003) JP
  • 2004JP08790 (Jun 16, 2004) WO
Title Fluorescent protein achieved
Abstract An object of the present invention is to provide a novel fluorescent protein derived from favia favus. The present invention provides a fluorescent protein derived from favia favus having the following properties: (1) an excitation maximum wavelength is 507 nm; (2) a fluorescence maximum wavelength is 517 nm; (3) a molar absorption coefficient at 482 nm is 80,000; (4) a quantum yield is 0.68; and (5) pH sensitivity of the fluorescence maximum is stable at pH 5 to pH 11.
Outline of related art and contending technology BACKGROUND ART
Green fluorescent protein (GFP) derived from Aequorea victoria, a jellyfish, has many purposes in biological systems. Recently, various GFP mutants have been produced based on the random mutagenesis and semi-rational mutagenesis, wherein a color is changed, a folding property is improved, luminance is enhanced, or pH sensitivity is modified. Fluorescent proteins such as GFP are fused with other proteins by gene recombinant technique, and monitoring of the expression and transportation of the fusion proteins is carried out.
One of the most commonly used types of GFP mutant is Yellow fluorescent protein (YFP). Among Aequorea-derived GFP mutants, YFP exhibits the fluorescence with the longest wavelength. The values ε and Φ of the majority of YEPs 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). These values are comparable to those of the general fluorescent group (fluorescein, rhodamine, etc.). Accordingly, improvement of the absolute luminance of YFP is nearly approaching its limit.
In addition, cyan fluorescent protein (CFP) is another example of the GFP mutant. Of this type of protein, ECFP (enhanced cyan fluorescent protein) has been known. Moreover, red fluorescent protein (RFP) has been isolated from sea anemone (Discosoma sp.). Of this type of protein, DsRed has been known. Thus, 4 types of fluorescent proteins, that are, green fluorescent protein, yellow fluorescent protein, cyan fluorescent protein, and red fluorescent protein, have successfully been developed. The range of the spectrum has significantly been expanded.
Also, there are many cnidarians which emit fluorescence. Cloning of genes of fluorescent proteins from cnidarians has been attempted, but cloning of more genes is necessary in order to increase repertoire of fluorescent and biochemical properties.
Scope of claims [claim1]
1. An isolated DNA comprising any of the following nucleotide sequences:
(a) the nucleotide sequence shown in SEQ ID NO: 2, 13, 15, 17, 19, or 21; or
(b) the nucleotide sequence of SEQ ID NO: 2 with 1 to 60 nucleotide substitutions, and encoding a fluorescent protein.

2. A recombinant vector having the DNA of claim 1.

3. A transformant having the DNA of claim 1.
  • Inventor, and Inventor/Applicant
  • Miyawaki Atsushi
  • Tsutsui Hidekazu
  • Karasawa Satoshi
IPC(International Patent Classification)