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1. 发明公开

EP3508570A1 MUTANT PROTEASE BIOSENSORS WITH ENHANCED DETECTION CHARACTERISTICS 有权
公开(公告)号：EP3508570A1
公开(公告)日：2019-07-10
申请号：EP19155964.0
申请日：2011-05-11
申请人： Promega Corporation
发明人： Binkowski, Brock , Butler, Braeden L. , Encell, Lance P. , Fan, Frank , Hook, Brad , Otto, Paul , Vidugiris, Gediminas , Wigdal, Susan , Zimmerman, Kristopher
IPC分类号： C12N9/02 , C12Q1/66 , A61K49/00 , C12Q1/37 , A01K67/027 , C12N5/00 , C12N15/00
摘要： A polynucleotide encoding a biosensor polypeptide comprising a modified circularly-permuted thermostable luciferase and a linker linking the C-terminal portion of the thermostable luciferase to the N-terminal portion of the thermostable luciferase. The modified circularly-permuted thermostable luciferase is modified relative to an unmodified circularly-permuted thermostable luciferase. The linker contains a sensor region capable of interacting with a target molecule in a cell. The modified circularly-permuted thermostable luciferase has an enhanced response after interaction of the biosensor with the target molecule relative to the unmodified circularly-permuted thermostable luciferase in the presence of the target molecule. Alternatively, the modified circularly-permuted thermostable luciferase has an enhanced response after interaction of the biosensor with the target molecule relative to the modified circularly-permuted thermostable luciferase in the absence of the target molecule.

2. 发明授权

EP2327768B1 Permuted and nonpermuted luciferase biosensors 有权转让
标题翻译：改变顺序和nichtpermutierte荧光素酶生物传感器
公开(公告)号：EP2327768B1
公开(公告)日：2015-09-09
申请号：EP11155576.9
申请日：2007-04-02
申请人： Promega Corporation
发明人： Binkowski, Brock , Fan, Frank , Wigdal, Susan , Wood, Keith V. , Wood, Monika G.
IPC分类号： C12N9/02 , C12N15/53 , C12Q1/26 , C12Q1/66 , G01N33/573 , C12N5/10 , C12N1/15 , C12N1/21
CPC分类号： C12N9/0069 , C12N9/0004 , C12Q1/66 , C12Y113/12007 , G01N33/84

3. 发明公开

EP2327768A3 Permuted and nonpermuted luciferase biosensors 有权转让
标题翻译：改变顺序和nichtpermutierte荧光素酶生物传感器
公开(公告)号：EP2327768A3
公开(公告)日：2011-10-12
申请号：EP11155576.9
申请日：2007-04-02
申请人： PROMEGA CORPORATION
发明人： Binkowski, Brock , Fan, Frank , Wigdal, Susan , Wood, Keith V. , Wood, Monika G.
IPC分类号： C12N9/02 , C12N15/53 , C12Q1/26 , C12Q1/66 , G01N33/573 , C12N5/10 , C12N1/15 , C12N1/21
CPC分类号： C12N9/0069 , C12N9/0004 , C12Q1/66 , C12Y113/12007 , G01N33/84
摘要： A modified luciferase protein which is a sensor for molecules including cAMP, cGMP, calcium, chelators thereof, kinases, or phosphatases is provided. Also provided is a circularly permuted anthozoan luciferase protein and a decapod crustacean luciferase protein, optionally containing one or more heterologous amino acid sequences, at least one of which directly or indirectly interacts with a molecule of interest. Further provided is a modified anthozoan luciferase protein and a decapod crustacean luciferase protein containing an insertion of one or more heterologous amino acid sequences, at least one of which directly or indirectly interacts with a molecule of interest.

4. 发明公开

EP2327768A2 Permuted and nonpermuted luciferase biosensors 有权转让
标题翻译： Permutierte und nichtpermutierte萤光素酶-Biosensoren
公开(公告)号：EP2327768A2
公开(公告)日：2011-06-01
申请号：EP11155576.9
申请日：2007-04-02
申请人： PROMEGA CORPORATION
发明人： Binkowski, Brock , Fan, Frank , Wigdal, Susan , Wood, Keith V. , Wood, Monika G.
IPC分类号： C12N9/02 , C12N15/53 , C12Q1/26 , G01N33/573 , C12N5/10 , C12N1/15 , C12N1/21
CPC分类号： C12N9/0069 , C12N9/0004 , C12Q1/66 , C12Y113/12007 , G01N33/84
摘要： A modified luciferase protein which is a sensor for molecules including cAMP, cGMP, calcium, chelators thereof, kinases, or phosphatases is provided. Also provided is a circularly permuted anthozoan luciferase protein and a decapod crustacean luciferase protein, optionally containing one or more heterologous amino acid sequences, at least one of which directly or indirectly interacts with a molecule of interest. Further provided is a modified anthozoan luciferase protein and a decapod crustacean luciferase protein containing an insertion of one or more heterologous amino acid sequences, at least one of which directly or indirectly interacts with a molecule of interest.
摘要翻译：提供了一种修饰的荧光素酶蛋白，其是用于分子的传感器，包括cAMP，cGMP，钙，其螯合剂，激酶或磷酸酶。还提供了循环置换的anthozoan萤光素酶蛋白和十足配甲壳素萤光素酶蛋白，任选地含有一个或多个异源氨基酸序列，其中至少一个直接或间接与感兴趣的分子相互作用。进一步提供了一种修饰的抗虫虫萤光素酶蛋白质和含有一个或多个异源氨基酸序列插入物的十足甲壳蛋白萤光素酶蛋白质，其中至少一种与感兴趣的分子直接或间接相互作用。

5. 发明授权

EP3508570B1 MUTANT PROTEASE BIOSENSORS WITH ENHANCED DETECTION CHARACTERISTICS 有权
公开(公告)号：EP3508570B1
公开(公告)日：2020-07-22
申请号：EP19155964.0
申请日：2011-05-11
申请人： Promega Corporation
发明人： Binkowski, Brock , Butler, Braeden L. , Encell, Lance P. , Fan, Frank , Hook, Brad , Otto, Paul , Vidugiris, Gediminas , Wigdal, Susan , Zimmerman, Kristopher
IPC分类号： C12N9/02 , C12Q1/66 , A61K49/00 , C12Q1/37 , A01K67/027 , C12N5/00 , C12N15/00

6. 发明公开

EP3467119A1 MUTANT PROTEASE BIOSENSORS WITH ENHANCED DETECTION CHARACTERISTICS 有权
公开(公告)号：EP3467119A1
公开(公告)日：2019-04-10
申请号：EP18205893.3
申请日：2012-11-12
申请人： Promega Corporation
发明人： Binkowski, Brock , Butler, Braeden L. , Encell, Lance P. , Fan, Frank , Hook, Brad , Otto, Paul , Risma, Kimberly , Vidugiris, Gediminas , Wigdal, Susan , Zimmerman, Kristopher
IPC分类号： C12Q1/37 , C12Q1/66
摘要： A polynucleotide encoding a biosensor polypeptide comprising a modified circularly-permuted thermostable luciferase and a linker linking the C-terminal portion of the thermostable luciferase to the N-terminal portion of the thermostable luciferase. The modified circularly-permuted thermostable luciferase is modified relative to a parental circularly-permuted thermostable luciferase. The linker contains a sensor region capable of interacting with a target molecule in a cell. The modified circularly-permuted thermostable luciferase has an enhanced response after interaction of the biosensor with the target molecule relative to the parental circularly-permuted thermostable luciferase in the presence of the target molecule. Alternatively, the modified circularly-permuted thermostable luciferase has an enhanced response after interaction of the biosensor with the target molecule relative to the modified circularly-permuted thermostable luciferase in the absence of the target molecule.

7. 发明授权

EP2990479B1 MUTANT PROTEASE BIOSENSORS WITH ENHANCED DETECTION CHARACTERISTICS 有权
公开(公告)号：EP2990479B1
公开(公告)日：2019-03-27
申请号：EP15189725.3
申请日：2011-05-11
申请人： Promega Corporation
发明人： Binkowski, Brock , Butler, Braeden , Encell, Lance P. , Fan, Frank , Hook, Brad , Otto, Paul , Vidugiris, Gediminas , Wigdal, Susan , Zimmerman, Kristopher
IPC分类号： C12N9/02 , C12Q1/66 , A61K49/00 , C12Q1/37 , A01K67/027 , C12N5/00 , C12N15/00

8. 发明公开

EP2990479A1 MUTANT PROTEASE BIOSENSORS WITH ENHANCED DETECTION CHARACTERISTICS 有权
标题翻译： MUTANTE PROTEASE-BIOSENSOREN MIT VERBESSERTEN ERKENNUNGSEIGENSCHAFTEN
公开(公告)号：EP2990479A1
公开(公告)日：2016-03-02
申请号：EP15189725.3
申请日：2011-05-11
申请人： Promega Corporation
发明人： Binkowski, Brock , Butler, Braeden , Encell, Lance P. , Fan, Frank , Hook, Brad , Otto, Paul , Vidugiris, Gediminas , Wigdal, Susan , Zimmerman, Kristopher
IPC分类号： C12N9/02 , C12Q1/66 , A61K49/00 , C12Q1/37 , A01K67/027 , C12N5/00 , C12N15/00
CPC分类号： A61K49/0045 , A01K67/0275 , A61K49/0013 , C12N9/0069 , C12Q1/37 , C12Q1/66 , C12Y113/12007
摘要： The invention relates to methods of detecting the presence or activity of a target molecule in a sample comprising a) contacting the sample with i) a polynucleotide encoding a biosensor polypeptide comprising a modified circularly-permuted thermostable luciferase and a linker linking the C-terminal portion of the thermostable luciferase to the N-terminal portion of the thermostable luciferase, the modified circularly-permuted thermostable luciferase modified relative to a parental circularly-permuted thermostable luciferase, the linker comprising a sensor region capable of interacting with a target molecule in a cell, and wherein the modified circularly-permuted thermostable luciferase has an enhanced response after interaction of the biosensor with the target molecule relative to at least one of: A) the parental circularly-permuted thermostable luciferase in the presence of the target molecule, or B) the modified circularly-permuted thermostable luciferase in the absence of the target molecule; and ii) a substrate for the modified thermostable luciferase; and b) detecting luminescence in the sample.
摘要翻译：本发明涉及检测样品中靶分子的存在或活性的方法，其包括：a）使样品与i）编码包含修饰的环状置换的热稳定荧光素酶的生物传感器多肽的多核苷酸和连接C末端部分的接头所述热稳定荧光素酶相对于亲本循环置换的热稳定性荧光素酶修饰的经修饰的环状置换的热稳定荧光素酶，所述连接体包含能够与细胞中的靶分子相互作用的传感器区，并且其中所述经修饰的环状置换的热稳定性荧光素酶相对于所述生物传感器与所述靶分子相互作用之后相对于以下至少一种具有增强的响应：A）所述亲本循环置换的热稳定性荧光素酶在所述靶分子的存在下，或B）在没有靶标的情况下修饰的环状置换的热稳定性荧光素酶分子; 和ii）用于修饰的热稳定荧光素酶的底物; 和b）检测样品中的发光。

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