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公开(公告)号：US5744320A
公开(公告)日：1998-04-28
申请号：US472546
申请日：1995-06-07
CPC分类号： G01N33/581 , C12Q1/66 , Y10S435/963 , Y10S435/975
摘要： The present invention relates to single and dual-reporter luminescence assays utilizing general and specific reagents to quench enzyme-mediated reactions. In one embodiment of the invention, a reagent is added to the assay which non-specifically quenches enzyme-mediated luminescent reactions. In another embodiment of the invention, a reagent is added to the assay which simultaneously quenches one enzyme-mediated luminescent reaction while activating another distinct enzyme-mediated luminescent reaction. An assay kit containing specific quench reagents, and the reagents themselves are also disclosed.
摘要翻译： 本发明涉及使用一般和特异性试剂来淬灭酶介导的反应的单和双重报道发光测定。 在本发明的一个实施方案中，将试剂加入到非特异性淬灭酶介导的发光反应的测定中。 在本发明的另一个实施方案中，将试剂加入到测定中，同时淬灭一种酶介导的发光反应，同时激活另一种不同的酶介导的发光反应。 还公开了包含特定淬灭试剂的测定试剂盒和试剂本身。
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公开(公告)号：US5670356A
公开(公告)日：1997-09-23
申请号：US354240
申请日：1994-12-12
申请人： Bruce A. Sherf , Keith V. Wood
发明人： Bruce A. Sherf , Keith V. Wood
CPC分类号： C12N9/0069
摘要： A modified form of beetle luciferase, which has been engineered for improved genetic reporting, is disclosed. The modified form contains one or more new features. Chief among these is removal of the peroxisomal translocation sequence to yield a cytoplasmic form of the enzyme. Other changes include removal of potentially interfering restriction sites and genetic regulatory sites from the gene, improvement of the codon usage for mammalian cells. The modified luciferase reporter enzyme is also devoid of potential N-glycosylation targets to minimize post-translational modification and remains in the cytoplasm of host cells to optimize substrate availability.
摘要翻译： 披露了改进的甲壳质萤光素酶的形式，其已被设计用于改进遗传报告。 修改后的表单包含一个或多个新功能。 其中主要是去除过氧化物酶体易位序列以产生酶的细胞质形式。 其他变化包括从基因中去除潜在的干扰性限制性位点和遗传调控位点，改善哺乳动物细胞的密码子使用。 修饰的萤光素酶报道酶也没有潜在的N-糖基化靶标以最小化翻译后修饰并保留在宿主细胞的细胞质中以优化底物可利用性。
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公开(公告)号：US5798263A
公开(公告)日：1998-08-25
申请号：US708533
申请日：1996-09-05
申请人： Keith V. Wood , Bruce A. Sherf
发明人： Keith V. Wood , Bruce A. Sherf
CPC分类号： G01N21/763
摘要： The presently disclosed invention is drawn to an automatic luminometer apparatus capable of measuring two distinct luminescent reactions from within a single, non-compartmentalized sample container. The present apparatus may be dimensioned, configured, and programmed to automatically perform dual-reporter luminescent assays using multi-well sample plates, such as 96-well microtiter plates.
摘要翻译： 本发明公开了一种自动发光计装置，其能够在单个非隔室样品容器内测量两种不同的发光反应。 本装置的尺寸，尺寸和尺寸可以使用多孔样品板（例如96孔微量滴定板）自动进行双重报道发光测定。
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4.发明授权Compositions comprising a dehalogenase substrate and a contrast agent and methods of use 有权包含脱卤酶底物和造影剂的组合物和使用方法公开(公告)号：US08921620B2
公开(公告)日：2014-12-30
申请号：US13450217
申请日：2012-04-18
申请人： Keith V. Wood , Dieter Klaubert , Georgyi V. Los , Robert F. Bulleit , Mark McDougall , Chad Zimprich
发明人： Keith V. Wood , Dieter Klaubert , Georgyi V. Los , Robert F. Bulleit , Mark McDougall , Chad Zimprich
CPC分类号： C12Q1/34 , C07C217/08 , C07C271/16 , C07D209/48 , C07D233/64 , C07D311/78 , C07D311/82 , C07D405/06 , C07D493/10 , C07D495/04 , C07F5/022 , C12N9/14 , C12N9/86 , C12Y308/01005 , G01N33/5005 , G01N33/58 , Y10S436/80
摘要： A mutant hydrolase optionally fused to a protein of interest is provided. The mutant hydrolase is capable of forming a bond with a substrate for the corresponding nonmutant (wild-type) hydrolase which is more stable than the bond formed between the wild-type hydrolase and the substrate. Substrates for hydrolases comprising one or more functional groups are also provided, as well as methods of using the mutant hydrolase and the substrates of the invention. Also provided is a fusion protein capable of forming a stable bond with a substrate and cells which express the fusion protein.
摘要翻译： 提供任选融合到感兴趣的蛋白质的突变体水解酶。 突变体水解酶能够与相应的非突变（野生型）水解酶的底物形成键，其比野生型水解酶和底物之间形成的键更稳定。 还提供了包含一个或多个官能团的水解酶的底物，以及使用本发明的突变体水解酶和底物的方法。 还提供能够与底物形成稳定结合的融合蛋白和表达融合蛋白的细胞。
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5.发明授权Oplophorus-derived luciferases, novel coelenterazine substrates, and methods of use 有权Oplophorus来源的萤光素酶，新型肠溶素底物和使用方法公开(公告)号：US08669103B2
公开(公告)日：2014-03-11
申请号：US13287986
申请日：2011-11-02
申请人： Brock Binkowski , Lance P. Encell , Mary Hall , Matthew B. Robers , Michael R. Slater , Keith V. Wood , Monika G. Wood
发明人： Brock Binkowski , Lance P. Encell , Mary Hall , Matthew B. Robers , Michael R. Slater , Keith V. Wood , Monika G. Wood
CPC分类号： C12Q1/66 , A01K2217/072 , C07D487/04 , C12N9/0004 , C12N9/0069 , C12N15/52 , C12N2009/02 , C12Y113/12005 , C12Y113/12007
摘要： An isolated polynucleotide encoding a modified luciferase polypeptide and substrates. The OgLuc variant polypeptide has at least 60% amino acid sequence identity to SEQ ID NO: 1 and at least one amino acid substitution at a position corresponding to an amino acid in SEQ ID NO: 1. The OgLuc variant polypeptide has at least one of enhanced luminescence, enhanced signal stability, and enhanced protein stability relative to the corresponding polypeptide of the wild-type Oplophorus luciferase.
摘要翻译： 编码修饰的萤光素酶多肽和底物的分离的多核苷酸。 OgLuc变体多肽与SEQ ID NO：1具有至少60％的氨基酸序列同一性，并且在对应于SEQ ID NO：1中的氨基酸的位置具有至少一个氨基酸取代.OgLuc变体多肽具有至少一个 相对于野生型Oplophorus萤光素酶的相应多肽，增强的发光，增强的信号稳定性和增强的蛋白质稳定性。
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公开(公告)号：US20130183760A1
公开(公告)日：2013-07-18
申请号：US13274643
申请日：2011-10-17
IPC分类号： C12N15/63
摘要： The invention provides vectors and methods for directional cloning.
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7.发明授权Luminogenic and fluorogenic compounds and methods to detect molecules or conditions 有权发光和荧光化合物以及检测分子或条件的方法公开(公告)号：US08476450B2
公开(公告)日：2013-07-02
申请号：US12217494
申请日：2008-07-03
申请人： James J. Cali , William Daily , Erika Hawkins , Dieter Klaubert , Jianquan Liu , Poncho Meisenheimer , Michael Scurria , John Shultz , James Unch , Michael P. Valley , Keith V. Wood , Wenhui Zhou
发明人： James J. Cali , William Daily , Erika Hawkins , Dieter Klaubert , Jianquan Liu , Poncho Meisenheimer , Michael Scurria , John Shultz , James Unch , Michael P. Valley , Keith V. Wood , Wenhui Zhou
IPC分类号： A61K31/428 , C07D417/04 , C07D417/14
CPC分类号： C07D277/66 , C07D277/68 , C07D311/16 , C07D417/04 , C07D417/14 , C07D493/10 , C07F9/65583 , C09B11/24 , C09B19/00 , C09B57/00 , C09B57/02 , C12Q1/66 , G01N2500/00
摘要： A method to detect the presence or amount of at least one molecule in a sample which employs a derivative of luciferin or a derivative of a fluorophore is provided. Compounds and compositions for carrying out the methods of the invention are also provided.
摘要翻译： 提供了检测采用荧光素衍生物或荧光团衍生物的样品中至少一种分子的存在或量的方法。 还提供了用于实施本发明方法的化合物和组合物。
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8.发明申请METHOD OF IMMOBILIZING A PROTEIN OR MOLECULE VIA A MUTANT DEHALOGENASE THAT IS BOUND TO AN IMMOBILIZED DEHALOGENASE SUBSTRATE AND LINKED DIRECTLY OR INDIRECTLY TO THE PROTEIN OR MOLECULE 有权通过对固定化的脱氧核糖核酸结合的突变性死亡来维持蛋白质或分子的方法，并直接或间接连接到蛋白质或分子公开(公告)号：US20120330001A1
公开(公告)日：2012-12-27
申请号：US13477593
申请日：2012-05-22
申请人： Aldis Darzins , Lance Encell , Tonny Johnson , Dieter Klaubert , Georgyi V. Los , Mark McDougall , Keith V. Wood , Monika G. Wood , Chad Zimprich
发明人： Aldis Darzins , Lance Encell , Tonny Johnson , Dieter Klaubert , Georgyi V. Los , Mark McDougall , Keith V. Wood , Monika G. Wood , Chad Zimprich
IPC分类号： C07C235/08 , C07C19/075 , C08B1/00 , C07C19/01
CPC分类号： C12N9/14 , B82Y5/00 , B82Y10/00 , B82Y30/00 , C07K2299/00 , C12N9/0069 , C12Q1/26 , C12Q1/34 , C12Q1/66 , C12Y113/12005 , C12Y308/01005
摘要： A mutant hydrolase optionally fused to a protein of interest is provided. The mutant hydrolase is capable of forming a bond with a substrate for the corresponding nonmutant (wild-type) hydrolase which is more stable than the bond formed between the wild-type hydrolase and the substrate and has at least two amino acid substitutions relative to the wild-type hydrolase. Substrates for hydrolases comprising one or more functional groups are also provided, as well as methods of using the mutant hydrolase and the substrates of the invention. Also provided is a fusion protein capable of forming a stable bond with a substrate and cells which express the fusion protein.
摘要翻译： 提供任选融合到感兴趣的蛋白质的突变体水解酶。 突变体水解酶能够与相应的非突变（野生型）水解酶的底物形成键，其比野生型水解酶和底物之间形成的键更稳定，并且相对于野生型水解酶具有至少两个氨基酸取代 野生型水解酶。 还提供了包含一个或多个官能团的水解酶的底物，以及使用本发明的突变体水解酶和底物的方法。 还提供能够与底物形成稳定结合的融合蛋白和表达融合蛋白的细胞。
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9.发明授权Compositions comprising a dehalogenase substrate and a fluorescent label and methods of use 有权包含脱卤酶底物和荧光标记的组合物和使用方法公开(公告)号：US08257939B2
公开(公告)日：2012-09-04
申请号：US12975020
申请日：2010-12-21
申请人： Keith V. Wood , Georgyi V. Los , Robert F. Bulleit , Dieter Klaubert , Mark McDougall , Chad Zimprich
发明人： Keith V. Wood , Georgyi V. Los , Robert F. Bulleit , Dieter Klaubert , Mark McDougall , Chad Zimprich
CPC分类号： C12Q1/34 , C07C217/08 , C07C271/16 , C07D209/48 , C07D233/64 , C07D311/78 , C07D311/82 , C07D405/06 , C07D493/10 , C07D495/04 , C07F5/022 , C12N9/14 , C12N9/86 , C12Y308/01005 , G01N33/5005 , G01N33/58 , Y10S436/80
摘要： A mutant hydrolase optionally fused to a protein of interest is provided. The mutant hydrolase is capable of forming a bond with a substrate for the corresponding nonmutant (wild-type) hydrolase which is more stable than the bond formed between the wild-type hydrolase and the substrate. Substrates for hydrolases comprising one or more functional groups are also provided, as well as methods of using the mutant hydrolase and the substrates of the invention. Also provided is a fusion protein capable of forming a stable bond with a substrate and cells which express the fusion protein.
摘要翻译： 提供任选融合到感兴趣的蛋白质的突变体水解酶。 突变体水解酶能够与相应的非突变（野生型）水解酶的底物形成键，其比野生型水解酶和底物之间形成的键更稳定。 还提供了包含一个或多个官能团的水解酶的底物，以及使用本发明的突变体水解酶和底物的方法。 还提供能够与底物形成稳定结合的融合蛋白和表达融合蛋白的细胞。
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10.发明申请OPLOPHORUS-DERIVED LUCIFERASES, NOVEL COELENTERAZINE SUBSTRATES, AND METHODS OF USE 有权欧莱酮衍生化石蜡，新型椰油基质及其使用方法公开(公告)号：US20120174242A1
公开(公告)日：2012-07-05
申请号：US13287986
申请日：2011-11-02
申请人： Brock Binkowski , Lance P. Encell , Mary Hall , Matthew B. Robers , Michael R. Slater , Keith V. Wood , Monika G. Wood
发明人： Brock Binkowski , Lance P. Encell , Mary Hall , Matthew B. Robers , Michael R. Slater , Keith V. Wood , Monika G. Wood
IPC分类号： C12N9/02 , C12N15/63 , A01K67/027 , C12N5/10 , C12N1/21 , C12N1/15 , C12N1/19 , C12N15/53 , C12Q1/66
CPC分类号： C12Q1/66 , A01K2217/072 , C07D487/04 , C12N9/0004 , C12N9/0069 , C12N15/52 , C12N2009/02 , C12Y113/12005 , C12Y113/12007
摘要： An isolated polynucleotide encoding a modified luciferase polypeptide and substrates. The OgLuc variant polypeptide has at least 60% amino acid sequence identity to SEQ ID NO: 1 and at least one amino acid substitution at a position corresponding to an amino acid in SEQ ID NO: 1. The OgLuc variant polypeptide has at least one of enhanced luminescence, enhanced signal stability, and enhanced protein stability relative to the corresponding polypeptide of the wild-type Oplophorus luciferase.
摘要翻译： 编码修饰的萤光素酶多肽和底物的分离的多核苷酸。 OgLuc变体多肽与SEQ ID NO：1具有至少60％的氨基酸序列同一性，并且在对应于SEQ ID NO：1中的氨基酸的位置具有至少一个氨基酸取代.OgLuc变体多肽具有至少一个 相对于野生型Oplophorus萤光素酶的相应多肽，增强的发光，增强的信号稳定性和增强的蛋白质稳定性。
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