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LIGAND FLUORESCENT SENSOR PROTEIN AND USE THEREOF UPDATE

外国特許コード F170009106
整理番号 (S2016-0169-N0)
掲載日 2017年6月15日
出願国 世界知的所有権機関(WIPO)
国際出願番号 2016JP085902
国際公開番号 WO 2017094885
国際出願日 平成28年12月2日(2016.12.2)
国際公開日 平成29年6月8日(2017.6.8)
優先権データ
  • 特願2015-237524 (2015.12.4) JP
  • 201662340533 (2016.5.24) US
発明の名称 (英語) LIGAND FLUORESCENT SENSOR PROTEIN AND USE THEREOF UPDATE
発明の概要(英語) The present invention involves a ligand fluorescent sensor protein that specifically responds to ligands, leading to a change in fluorescence properties. The ligand fluorescent sensor protein includes a first fluorescent protein domain, an N terminal-side linker, a ligand-bonding domain, a C terminal-side linker, and a second fluorescent protein domain. The fluorescent protein used in the ligand fluorescent sensor protein has a β-barrel structure. The first fluorescent protein domain includes a β1–β3 β-sheet region from the N-terminal of the fluorescent protein, an α-helix region continuing therefrom, and a β4–β6 β-sheet region. The second fluorescent protein domain includes a β7–β11 β-sheet region of the same fluorescent protein as the first fluorescent protein domain. The N-terminal-side linker and the C-terminal-side linker are each independently a polypeptide comprising one or a plurality of amino acids.
従来技術、競合技術の概要(英語) BACKGROUND ART
Fluorescence vital imaging is, observed the inside of the living tissue or organ, and the molecular weight of live cells where the kinetics of new studies as a method of real-time analysis. Fluorescence imaging and optical equipment in recent years by technological innovation, various tissue or organ, a wide variety of biological phenomenon by applying fluorescent biological imaging, and live cells to the position of the motions of molecules become.
Fluorescence vital in imaging, a molecule (ligand) existing within the cell receptor that binds to the specific use, the movement of the living tissue or organ to which a ligand can be detected. Specifically, as a receptor, for example, in vivo antibody that binds to a molecule that binds specifically to the antibody present in the organism or antigen, or a specific substrate for the enzyme to a particular enzyme on the substrate, or ATP, cAMP, or cGMP binding domain of the molecule in the living body such as a fluorescent sensor proteins can be used in the like, the concentration of the ligand of a cell or within a time-varying distribution can be analyzed to visualize (space-time dynamics).
Protein is a fluorescent sensor, a single type, the ratio-type, and FRET (fluorescence resonance energy transfer) is largely classified into a type. A single type, each of the fluorescent sensor protein that is specified for the fluorescence intensity at a single wavelength changes depending on the concentration of the ligand type. The ratio-type is, for each identified protein 2 each fluorescent sensor at the fluorescence wavelength of one of the relative intensity ratio changes depending on the concentration of the ligand type. FRET type, different fluorophore 2 protein fragment of a fusion protein comprising a fragment of one when the excited state, depending on the concentration of the other fragment of the ligand is neared to the distance, the other of the excited state energy from the fragment to the fragment of a non-radiative transition Forster Resonance Energy (Forster resonance energy transfer) using the phenomenon that the, the concentration of the ligand to the former fragment fluorescence intensity and relative intensity of the fluorescence intensity of the latter fragment type ratio changes. For example, up to now, a single type (for example, see non-patent document 1), FRET-type (for example, see non-patent document 2), and the ratio-type (for example, non-patent document 3, reference 4) of the fluorescent sensor protein ATP have been reported.
  • 出願人(英語)
  • ※2012年7月以前掲載分については米国以外のすべての指定国
  • THE UNIVERSITY OF TOKYO
  • WASEDA UNIVERSITY
  • 発明者(英語)
  • Tsuboi Takashi
  • Tetsuya Kitaguchi
  • Satoshi Arai
  • UEDA HIROSHI
国際特許分類(IPC)
指定国 National States: AE AG AL AM AO AT AU AZ BA BB BG BH BN BR BW BY BZ CA CH CL CN CO CR CU CZ DE DJ DK DM DO DZ EC EE EG ES FI GB GD GE GH GM GT HN HR HU ID IL IN IR IS JP KE KG KN KP KR KW KZ LA LC LK LR LS LU LY MA MD ME MG MK MN MW MX MY MZ NA NG NI NO NZ OM PA PE PG PH PL PT QA RO RS RU RW SA SC SD SE SG SK SL SM ST SV SY TH TJ TM TN TR TT TZ UA UG US UZ VC VN ZA ZM ZW
ARIPO: BW GH GM KE LR LS MW MZ NA RW SD SL SZ TZ UG ZM ZW
EAPO: AM AZ BY KG KZ RU TJ TM
EPO: AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR
OAPI: BF BJ CF CG CI CM GA GN GQ GW KM ML MR NE SN ST TD TG

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