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Novel polypeptide exhibiting fluorescent properties and use thereof 実績あり

外国特許コード F190009896
整理番号 08118-EP
掲載日 2019年8月26日
出願国 欧州特許庁(EPO)
出願番号 14757001
公報番号 2963115
公報番号 2963115
出願日 平成26年2月28日(2014.2.28)
公報発行日 平成28年1月6日(2016.1.6)
公報発行日 令和2年4月1日(2020.4.1)
国際出願番号 JP2014055160
国際公開番号 WO2014133158
国際出願日 平成26年2月28日(2014.2.28)
国際公開日 平成26年9月4日(2014.9.4)
優先権データ
  • 特願2013-040097 (2013.2.28) JP
  • 2014JP55160 (2014.2.28) WO
発明の名称 (英語) Novel polypeptide exhibiting fluorescent properties and use thereof 実績あり
発明の概要(英語) In order to provide a novel fluorescent protein and use thereof, the polypeptide according to the present invention has fluorescent properties in the presence of bilirubin and includes (1) the amino acid sequence of SEQ ID NO: 1, (2) an amino acid sequence having, for example, substitution of 1 to 21 amino acids in the amino acid sequence of SEQ ID NO: 1, (3) an amino acid sequence having 85% or more sequence identity to the amino acid sequence of SEQ ID NO: 1, or (4) the amino acid sequence encoded by a polynucleotide that hybridizes with a polynucleotide consisting of a sequence complementary to the polynucleotide encoding the polypeptide according to the amino acid sequence in (1) under a stringent condition.
従来技術、競合技術の概要(英語) Background Art
Fluorescent proteins, such as green fluorescent protein (GFP), are indispensable as a tool for visualizing cells, tissue, biological individuals, and so on.
Most of fluorescent proteins have been isolated from invertebrates such as coral, sea anemones, and arthropods. However, for example, Non Patent Literatures 1 and 2 and Patent Literatures 1 and 2 report that a vertebrate, Japanese eel (Anguilla japonica), has a fluorescent protein.
Citation List
Patent Literature
Patent Literature 1: JP Patent Publication (Kokai) No. 2007-254371 A (Date of publication: October 4, 2007)
Patent Literature 2: JP Patent Publication (Kokai) No. 2008-141988 A (Date of publication: June 26, 2008)

Non Patent Literature
Non Patent Literature 1: Masao HONDA, Jinsuke KISHINO, Miyuki IMAMURA, Seiichi HAYASHI: Abstracts of the Fiscal Year Heisei 16 meeting of the Japanese Society of Fisheries Science (April 2, 2004, p. 203, 1101)
Non Patent Literature 2: Hayashi et al., Fisheries Science, 75, 1461-1469, 2009
特許請求の範囲(英語) [claim1]
1. A polypeptide-bilirubin complex constituted by bringing bilirubin into contact with a polypeptide or a fusion polypeptide in a form free from bilirubin, wherein the polypeptide is selected from the group consisting of the polypeptides of the following (1) to (3) having fluorescent properties in the presence of bilirubin, and the fusion polypeptide comprises the polypeptide selected from the group consisting of the polypeptides of the following (1) to (3) having fluorescent properties in the presence of bilirubin and another polypeptide:
(1) a polypeptide including the amino acid sequence of SEQ ID NO: 1;
(2) a polypeptide including an amino acid sequence having substitution, deletion, insertion, and/or addition of 1 to 21 amino acids in the amino acid sequence of SEQ ID NO: 1; and
(3) a polypeptide having 85% or more sequence identity to the amino acid sequence of SEQ ID NO: 1.

[claim2]
2. An in vitro method for detecting bilirubin in a biological or non-biological sample, the method comprising:
a contact step of bringing a polypeptide or a fusion polypeptide into contact with the biological or non-biological sample; and
a detection step of detecting fluorescence emitted by the polypeptide or the fusion polypeptide, after the contact step, wherein the polypeptide is selected from the group consisting of the polypeptides of the following (1) to (3) having fluorescent properties in the presence of bilirubin, and the fusion polypeptide comprises the polypeptide selected from the group consisting of the polypeptides of the following (1) to (3) having fluorescent properties in the presence of bilirubin and another polypeptide:
(1) a polypeptide including the amino acid sequence of SEQ ID NO: 1;
(2) a polypeptide including an amino acid sequence having substitution, deletion, insertion, and/or addition of 1 to 21 amino acids in the amino acid sequence of SEQ ID NO: 1; and
(3) a polypeptide having 85% or more sequence identity to the amino acid sequence of SEQ ID NO: 1.

[claim3]
3. The method according to claim 2, wherein
the biological or non-biological sample is a sample derived from blood or urine collected from a living body.

[claim4]
4. An in vitro method for inspecting the presence or absence of a predisposing factor or onset of liver disease or hemolytic disease in a biological or non-biological sample, the method comprising:
a contact step of bringing a polypeptide or a fusion polypeptide into contact with the biological or non-biological sample;
a detection step of detecting fluorescence emitted by the polypeptide or the fusion polypeptide, after the contact step, wherein the polypeptide is selected from the group consisting of the polypeptides of the following (1) to (3) having fluorescent properties in the presence of bilirubin, and the fusion polypeptide comprises the polypeptide selected from the group consisting of the polypeptides of the following (1) to (3) having fluorescent properties in the presence of bilirubin and another polypeptide:
(1) a polypeptide including the amino acid sequence of SEQ ID NO: 1;
(2) a polypeptide including an amino acid sequence having substitution, deletion, insertion, and/or addition of 1 to 21 amino acids in the amino acid sequence of SEQ ID NO: 1; and
(3) a polypeptide having 85% or more sequence identity to the amino acid sequence of SEQ ID NO: 1;
and
a testing step of inspecting the presence or absence of the predisposing factor or onset of liver disease or hemolytic disease based on the detection result in the detection step.

[claim5]
5. The method according to claim 4, wherein the biological or non-biological sample is a sample derived from blood or urine collected from a living body.

[claim6]
6. A bilirubin detecting kit comprising at least one selected from a polypeptide, a polynucleotide encoding the polypeptide, a recombinant vector comprising the polynucleotide, a transformant comprising the polynucleotide or the recombinant vector, and a fusion polypeptide; and at least one selected from a reagent and/or a tool for detecting bilirubin, a reagent and/or a tool for preparing a sample for detection, a manual of the detection kit, a sample for a control in detection, and reference data for analyzing a detection result,
wherein the polypeptide is selected from the group consisting of the polypeptides of the following (1) to (3) having fluorescent properties in the presence of bilirubin, and the fusion polypeptide comprises the polypeptide selected from the group consisting of the polypeptides of the following (1) to (3) having fluorescent properties in the presence of bilirubin and another polypeptide:
(1) a polypeptide including the amino acid sequence of SEQ ID NO: 1;
(2) a polypeptide including an amino acid sequence having substitution, deletion, insertion, and/or addition of 1 to 21 amino acids in the amino acid sequence of SEQ ID NO: 1; and
(3) a polypeptide having 85% or more sequence identity to the amino acid sequence of SEQ ID NO: 1.
  • 出願人(英語)
  • RIKEN
  • 発明者(英語)
  • MIYAWAKI, Atsushi
  • KUMAGAI, Akiko
国際特許分類(IPC)
指定国 Contracting States: AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR

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