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31. 发明申请

US20060234324A1 Method for enhancing activity of luciferase with fluorescence activity 有权
标题翻译：荧光素酶活性增强荧光活性的方法
公开(公告)号：US20060234324A1
公开(公告)日：2006-10-19
申请号：US11393233
申请日：2006-03-30
申请人： Satoshi Inouye , Satoko Sasaki
发明人： Satoshi Inouye , Satoko Sasaki
IPC分类号： C12Q1/66 , C07D487/14 , C07D487/04
CPC分类号： A61K49/0021 , A61K49/0013
摘要： Methods for enhancing luminescence of a luciferase (BFP-aq) with fluorescence activity derived from a calcium-binding photoprotein are provided. To a luciferase solution with fluorescence activity that contains an apoprotein, a calcium-binding photoprotein, which is constituted such that a coelenteramide or an analog thereof is coordinated inside, a coelenterazine that is the luminescent substrate of the luciferase or an analog thereof and a compound (e.g., imidazole etc.) having the function of removing an —NH-proton of the pyrazine ring of the imidazopyrazine skeleton in the coelenterazine or the analog thereof are added.
摘要翻译：提供了具有衍生自钙结合光蛋白的荧光活性的萤光素酶（BFP-aq）发光的方法。对于具有荧光活性的荧光素酶溶液，其包含脱辅基蛋白，钙结合光蛋白，其构成为使肠内酰胺或其类似物在内部配位，作为萤光素酶或其类似物的发光底物的肠腔菌素和化合物（例如，咪唑等），其具有除去肠腔素中咪唑并吡嗪骨架的吡嗪环的-NH-质子或其类似物的功能。

32. 发明授权

US5578476A Flavin reductase gene 失效
标题翻译：黄素还原酶基因
公开(公告)号：US5578476A
公开(公告)日：1996-11-26
申请号：US518223
申请日：1995-08-23
申请人： Shuhei Zenno , Satoshi Inouye , Kaoru Saigo
发明人： Shuhei Zenno , Satoshi Inouye , Kaoru Saigo
IPC分类号： C12N1/21 , C12N9/02 , C12N15/09 , C12N15/53 , C12R1/01 , C12R1/19 , C07H21/04 , C12N15/63
CPC分类号： C12N9/0036
摘要： Nucleic acids coding for the flavin reductase of a luminescent bacterium, Xenorhabdus luminescens, vectors comprising such nucleic acids, host cells into which such vectors have been introduced, recombinant methods for producing substantially purified recombinant flavin reductase, and recombinantly produced flavin reductase and equivalent proteins produced in accordance with such methods are provided.
摘要翻译：编码发光细菌的黄素还原酶的核酸，包含这种核酸的载体，其中引入了这些载体的宿主细胞，用于生产基本上纯化的重组黄素还原酶的重组方法和重组产生的黄素还原酶和产生的等同蛋白质的核酸根据这些方法提供。

33. 发明授权

US5288623A Process for secretory production of a calcium-binding protein 失效
标题翻译：钙结合蛋白的分泌生产方法
公开(公告)号：US5288623A
公开(公告)日：1994-02-22
申请号：US912582
申请日：1992-07-13
申请人： Shuhei Zenno , Satoshi Inouye
发明人： Shuhei Zenno , Satoshi Inouye
IPC分类号： C07K14/31 , C07K14/435 , C12N9/38 , C12N15/70 , C12N15/62
CPC分类号： C07K14/31 , C07K14/43504 , C12N15/70 , C12N9/2471 , C12Y302/01023 , C07K2319/02
摘要： A process for secretory production of a calcium-binding protein very useful for producing heterogenic proteins in Escherichia coli according to gene recombinant technique is provided, which process comprises using Escherichia coli and using a secretory expression system with the promoter of lipoprotein and the gene of signal peptide of outer membrane protein A, according to recombinant DNA technique.
摘要翻译：提供了一种根据基因重组技术在大肠杆菌中非常有用于生产异源蛋白质的钙结合蛋白的分泌生产方法，该方法包括使用大肠杆菌，并使用分泌表达系统与脂蛋白启动子和信号基因根据重组DNA技术，外膜蛋白A的肽。

34. 发明授权

US5139937A Process for producing apoaequorin 失效
标题翻译：生产APOAEQUORIN的方法
公开(公告)号：US5139937A
公开(公告)日：1992-08-18
申请号：US272900
申请日：1988-11-18
申请人： Satoshi Inouye , Shigeyuki Aoyama , Yoshiyuki Sakaki
发明人： Satoshi Inouye , Shigeyuki Aoyama , Yoshiyuki Sakaki
IPC分类号： C12N15/09 , C07K14/245 , C07K14/435 , C12P21/00 , C12P21/02 , C12R1/19
CPC分类号： C07K14/245 , C07K14/43595 , C07K2319/02
摘要： A process for producing the protein part of Ca.sup.+2 -binding photoprotein aequorin (apoaequorin) according to a recombinant DNA technique, and a process for purifying the resulting apoaequorin are provided, which production process comprises cultivating a strain having an expression vector piP-HE outside the bacterial bodies transformed into Escherichia coli, followed by separating the resulting culture solution into the bacterial bodies and a culture filtrate, and recovering the culture filtrate, and which purification process comprises adding an acid to the culture filtrate so as to give a pH of 4.7 or less, followed by recovering the resulting white precipitates, dissolving the white precipitates in a buffer solution, reducing the solution, subjecting the resulting apoaequorin fraction to adsorption treatment according to anion exchange chromatography and subjecting the resulting apoaequorin to gel filtration.

35. 发明授权

US09678067B2 Method for extending light-emitting time of calcium-binding photoprotein solution 有权
公开(公告)号：US09678067B2
公开(公告)日：2017-06-13
申请号：US11411715
申请日：2006-04-26
申请人： Satoshi Inouye , Satoko Sasaki
发明人： Satoshi Inouye , Satoko Sasaki
IPC分类号： G01N33/533 , G01N33/58
CPC分类号： G01N33/533 , G01N33/582 , G01N2333/43595
摘要： Methods for extending light-emitting time of a solution of a calcium-binding photoprotein that instantaneously emits light by binding to calcium ions are provided. In the light-emitting reaction system of a solution of a calcium-binding photoprotein, a light-emitting reaction is performed in the presence of an anion capable of binding to the calcium ion or the cation that can be substituted for the calcium ion and/or a cation that can bind to the calcium-binding site of the calcium-binding photoprotein with a lower affinity than the calcium ion or the cation that can be substituted for the calcium ion without activating the calcium-binding photoprotein.

36. 发明授权

US08710195B2 Fluorescent proteins 有权
标题翻译：荧光蛋白
公开(公告)号：US08710195B2
公开(公告)日：2014-04-29
申请号：US10567857
申请日：2004-08-12
申请人： Satoshi Inouye
发明人： Satoshi Inouye
IPC分类号： C07K1/13 , G01N33/533 , C07D241/00
CPC分类号： C07K14/43595 , G01N21/76 , G01N33/84
摘要： A fluorescent protein (bFP) having chemiluminescence activity is a complex composed of the apoprotein of a calcium-binding photoprotein, coelenteramid or an analog thereof, and calcium ions or divalent or trivalent ions that can be substituted for the calcium ions. In the complex, the ratio of the number of molecules of the apoprotein to that of the coelenteramid is 1:1 and the ratio of the number of molecules of the apoprotein to that of the divalent or trivalent ions is 1:1 to 1:4. The fluorescent protein is used as a marker because it catalyzes luminescence of coelenterazine and has fluorescence capability. Removal of calcium ions etc. from this fluorescent protein (bFP) having luminescence activity provides a novel fluorescent protein (gFP). Mixing this gFP with the coelenterazine provides a calcium-binding photoprotein, which emit light instantaneously, enabling use as a marker.
摘要翻译：具有化学发光活性的荧光蛋白（bFP）是由钙结合光蛋白，coelenteramid或其类似物的脱辅基蛋白和可以代替钙离子的钙离子或二价或三价离子组成的复合物。在络合物中，脱辅基蛋白的分子数与肠腔蛋白的分子数之比为1:1，脱辅基蛋白的分子数与二价或三价离子的分子数之比为1:1至1:4 。荧光蛋白被用作标记物，因为它催化coelenterazine的发光并具有荧光能力。从具有发光活性的荧光蛋白（bFP）中去除钙离子等提供新的荧光蛋白（gFP）。将这种gFP与coelenterazine混合提供钙结合的光蛋白，其瞬时发光，可用作标记物。

37. 发明申请

US20110313140A1 PROCESS FOR PRODUCING COELENTERAMIDE OR AN ANALOG THEREOF 有权
标题翻译：生产咖啡酰胺或其类似物的方法
公开(公告)号：US20110313140A1
公开(公告)日：2011-12-22
申请号：US13209773
申请日：2011-08-15
申请人： Satoshi Inouye
发明人： Satoshi Inouye
IPC分类号： C07D241/20 , C07K14/00
CPC分类号： C07D241/20 , C07K1/1077 , C07K14/43595
摘要： A process for producing coelenteramide or its analog in a high yield has been desired. The invention provides a process for producing di-O-methylcoelenteramide or its analog of formula (3) which comprises reacting O-methylcoelenteramine or its analog with 4-methoxyphenylacetyl halide or its analog. The invention also provides a process for producing coelenteramide or its analog, which comprises demethylation of di-O-methylcoelenteramide or its analog.
摘要翻译：希望以高产率生产肠酰胺胺或其类似物的方法。本发明提供了一种生产二-O-甲基苯胺酰胺或其式（3）的类似物的方法，其包括使O-甲基肉毒碱或其类似物与4-甲氧基苯基乙酰卤或其类似物反应。本发明还提供了一种用于制备肠酰胺胺或其类似物的方法，其包括二-O-甲基醋酸纤维素或其类似物的脱甲基化。

38. 发明申请

US20110288280A1 COELENTERAMIDE ANALOGS 有权
标题翻译：椰油胺类似物
公开(公告)号：US20110288280A1
公开(公告)日：2011-11-24
申请号：US13146710
申请日：2010-02-08
申请人： Takamitsu Hosoya , Kohei Oka , Satoshi Inouye
发明人： Takamitsu Hosoya , Kohei Oka , Satoshi Inouye
IPC分类号： C07D241/20 , C07K2/00
CPC分类号： C07D241/20 , C07K14/43595
摘要： There has been a need for coelenteramide analogs or the like that produce fluorescent proteins which exhibit different fluorescent characteristics from those of the existing fluorescent proteins. Disclosed is a compound represented by general formula (1) (wherein R1 represents a substituted or unsubstituted aryl, a substituted or unsubstituted arylalkyl, a straight or branched alkyl which may optionally be substituted with an alicyclic group, an alicyclic group or a heterocyclic group; R2 represents hydrogen or —(SO2)R4; R3 represents hydrogen, hydroxyl, methoxy or acetoxy; R4 represents a substituted or unsubstituted aryl, a substituted or unsubstituted arylalkyl or a straight or branched alkyl which may optionally be substituted with an alicyclic group; and X1 represents —C(═S)— or —SO2—).
摘要翻译：需要产生与现有荧光蛋白不同的荧光特性的荧光蛋白质的coelenteramide类似物等。公开了由通式（1）表示的化合物（其中R 1表示取代或未取代的芳基，取代或未取代的芳烷基，可任选被脂环基，脂环基或杂环基取代的直链或支链烷基; R 2表示氢或 - （SO 2）R 4; R 3表示氢，羟基，甲氧基或乙酰氧基; R 4表示取代或未取代的芳基，取代或未取代的芳基烷基或可任选被脂环族基团取代的直链或支链烷基; X 1表示-C（= S） - 或-SO 2 - ）。

39. 发明授权

US07396655B2 Method for enhancing activity of luciferase with fluorescence activity 有权
标题翻译：荧光素酶活性增强荧光活性的方法
公开(公告)号：US07396655B2
公开(公告)日：2008-07-08
申请号：US11393233
申请日：2006-03-30
申请人： Satoshi Inouye , Satoko Sasaki
发明人： Satoshi Inouye , Satoko Sasaki
IPC分类号： C12Q1/66
CPC分类号： A61K49/0021 , A61K49/0013
摘要： Methods for enhancing luminescence of a luciferase (BFP-aq) with fluorescence activity derived from a calcium-binding photoprotein are provided. To a luciferase solution with fluorescence activity that contains an apoprotein, a calcium-binding photoprotein, which is constituted such that a coelenteramide or an analog thereof is coordinated inside, a coelenterazine that is the luminescent substrate of the luciferase or an analog thereof and a compound (e.g., imidazole etc.) having the function of removing an —NH-proton of the pyrazine ring of the imidazopyrazine skeleton in the coelenterazine or the analog thereof are added.
摘要翻译：提供了具有衍生自钙结合光蛋白的荧光活性的荧光素酶（BFP-aq）发光的方法。对于具有荧光活性的荧光素酶溶液，其包含脱辅基蛋白，钙结合光蛋白，其构成为使肠内酰胺或其类似物在内部配位，作为萤光素酶或其类似物的发光底物的肠腔菌素和化合物（例如，咪唑等），其具有除去肠腔素中咪唑并吡嗪骨架的吡嗪环的-NH-质子或其类似物的功能。

40. 发明申请

US20060246534A1 Method for extending light-emitting time of calcium-binding photoprotein solution 有权
标题翻译：延长钙结合光蛋白溶液发光时间的方法
公开(公告)号：US20060246534A1
公开(公告)日：2006-11-02
申请号：US11411715
申请日：2006-04-26
申请人： Satoshi Inouye , Satoko Sasaki
发明人： Satoshi Inouye , Satoko Sasaki
IPC分类号： C12Q1/26 , C07D487/14 , C07D487/02
CPC分类号： G01N33/533 , G01N33/582 , G01N2333/43595
摘要： Methods for extending light-emitting time of a solution of a calcium-binding photoprotein that instantaneously emits light by binding to calcium ions are provided. In the light-emitting reaction system of a solution of a calcium-binding photoprotein, a light-emitting reaction is performed in the presence of an anion capable of binding to the calcium ion or the cation that can be substituted for the calcium ion and/or a cation that can bind to the calcium-binding site of the calcium-binding photoprotein with a lower affinity than the calcium ion or the cation that can be substituted for the calcium ion without activating the calcium-binding photoprotein.
摘要翻译：提供了通过结合钙离子来瞬时发光的钙结合光蛋白溶液的发光时间延长的方法。在钙结合光蛋白溶液的发光反应体系中，在能够结合钙离子的阴离子或阳离子的存在下进行发光反应，该阳离子可以代替钙离子和/ 或可以以比钙离子或阳离子更低的亲和力结合钙结合光蛋白的钙结合位点的阳离子，而阳离子可以代替钙离子而不活化钙结合光蛋白。

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