TOP > 外国特許検索 > Probe for detection and quantification of nitric oxide, and method for detecting and quantifying nitric oxide using the same

Probe for detection and quantification of nitric oxide, and method for detecting and quantifying nitric oxide using the same 実績あり

外国特許コード F120006716
整理番号 A161-18US2
掲載日 2012年5月28日
出願国 アメリカ合衆国
出願番号 201113103317
公報番号 20110312014
公報番号 8748117
出願日 平成23年5月9日(2011.5.9)
公報発行日 平成23年12月22日(2011.12.22)
公報発行日 平成26年6月10日(2014.6.10)
国際出願番号 JP2005012722
国際公開番号 WO2006004217
国際出願日 平成17年7月5日(2005.7.5)
国際公開日 平成18年1月12日(2006.1.12)
優先権データ
  • 特願2004-198239 (2004.7.5) JP
  • 11/631,422 (2007.2.26) US
  • 2005JP012722 (2005.7.5) WO
発明の名称 (英語) Probe for detection and quantification of nitric oxide, and method for detecting and quantifying nitric oxide using the same 実績あり
発明の概要(英語) Probe 1 for detection and quantification of nitric oxide, which comprises two subunits 21 and 22 of soluble guanylate cyclase 2 and cGMP-visualization probes 3 respectively connected with each subunit, wherein the cGMP-visualization probe generates signal upon recognizing guanosine 3′,5′-cyclic monophospate.
特許請求の範囲(英語) [claim1]
1. A method for detecting and quantifying nitric oxide in vitro in a cell from an animal cell line or cultured somatic cells, which comprises: co-expressing in the cell a first hybrid protein consisting of an alpha -subunit of soluble guanylate cyclase and a first cGMP-visualization probe, and a second hybrid protein consisting of a beta -subunit of soluble guanylate cyclase and a second cGMP-visualization probe, and forming a dimer of the first hybrid protein and the second hybrid protein;
contacting the dimer of the first hybrid protein and the second hybrid protein with guanosine 5'-triphosphate in the cell;
detecting a signal from the first cGMP-visualization probe and the second cGMP-visualization probe, wherein the signal indicates the presence of nitric oxide in the cell; and
quantifying the amount of nitric oxide in the cell by comparison of the signal change with a calibration curve between nitric oxide concentrations and the signal changes,
wherein the first cGMP visualization probe and second cGMP-visualization probe each comprise:
a polypeptide that binds specifically to cyclic guanosine monophosphate (cGMP), wherein the polypeptide is cGMP-dependent protein kinase Ialpha in which amino acids 1-47 are deleted, and
two chromophores, cyan fluorescent protein and yellow fluorescent protein, one of which is linked to the N-terminal of the polypeptide and the other of which is linked to the C-terminal of the polypeptide,
and wherein the method is capable of detecting a 1 nanomolar concentration of nitric oxide in the cell.
  • 発明者/出願人(英語)
  • UMEZAWA YOSHIO
  • SATO MORITOSHI
  • JAPAN SCIENCE AND TECHNOLOGY AGENCY
国際特許分類(IPC)
米国特許分類/主・副
  • 435/7.6
参考情報 (研究プロジェクト等) CREST Endocrine Disrupters AREA
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