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1. 发明授权

US07262005B1 Methods of protein destabilization and uses thereof 有权
标题翻译：蛋白质不稳定的方法及其用途
公开(公告)号：US07262005B1
公开(公告)日：2007-08-28
申请号：US09498098
申请日：2000-02-04
申请人： Jeffrey Stack , Michael Whitney , Andrew B. Cubitt , Brian Pollok
发明人： Jeffrey Stack , Michael Whitney , Andrew B. Cubitt , Brian Pollok
IPC分类号： C12Q1/68
CPC分类号： C12Q1/485
摘要： This invention is directed towards methods of destabilizing proteins in living cells, and their use for the development of novel assays. In one embodiment, the invention comprises the use of non-cleavable multimerized ubiquitin fusion proteins to destabilize a target protein, such as a reporter moiety. In one aspect of this method the constructs also comprises a linker that operatively couples the reporter moiety to the multimerized ubiquitin fusion protein. In this embodiment, enzymatic modification of the linker results in a modulation of the coupling of the reporter protein to the multimerized ubiquitin domains resulting in a change in the stability of the reporter moiety. The level of the reporter moiety in the cell can then be used as a measure of the enzymatic activity in the cell. In another embodiment the invention provides for a generalized way of coordinately regulating the cellular concentration of a plurality of target proteins. In one aspect of this method, the target proteins are operatively coupled to a ubiquitin fusion protein via a linker containing a protease cleavage site. Cleavage of the linker by a protease results in uncoupling of the target protein from the multimerized ubiquitin construct, and results in an increase in the stability and concentration of the target protein.
摘要翻译：本发明涉及使活细胞中的蛋白质失稳的方法及其用于开发新的测定方法。在一个实施方案中，本发明包括使用不可切割的多聚化泛素融合蛋白来使目标蛋白如报道分子部分不稳定。在该方法的一个方面，构建体还包含使报道部分与多聚化泛素融合蛋白可操作地连接的接头。在该实施方案中，接头的酶修饰导致报道蛋白与多聚泛素结构域的偶联的调节，导致报道部分稳定性的变化。然后可以将细胞中的报告物部分的水平用作细胞中酶活性的量度。在另一个实施方案中，本发明提供了协调调节多种靶蛋白的细胞浓度的一般化方式。在该方法的一个方面，靶蛋白通过含有蛋白酶切割位点的接头与泛素融合蛋白可操作地偶联。通过蛋白酶切割接头导致靶蛋白与多聚化泛素构建体解偶联，并导致靶蛋白的稳定性和浓度增加。

2. 发明申请

US20050227345A1 Quorum sensing signaling in bacteria 审中-公开
公开(公告)号：US20050227345A1
公开(公告)日：2005-10-13
申请号：US11014621
申请日：2004-12-15
申请人： Marvin Whiteley , Kimberly Lee , E. Greenberg , Ute Muh
发明人： Marvin Whiteley , Kimberly Lee , E. Greenberg , Ute Muh
IPC分类号： C07K14/21 , C12N1/21 , C07H21/04 , C12N15/74
CPC分类号： C07K14/21
摘要： The invention provides methods for identifying a modulator of quorum sensing signaling in bacteria, and for identifying a quorum sensing controlled gene in bacteria. In addition, the invention provides quorum sensing controlled genetic loci in Pseudomas aeruginosa. Novel indicator strains and vectors for engineering the strains for use in the method of the invention are also provided.

3. 发明申请

US20020025940A1 Methods and compositions for sensitive and rapid, functional identification of genomic polynucleotides and use for cellular assays in drug discovery 审中-公开
标题翻译：用于灵敏和快速，功能鉴定基因组多核苷酸的方法和组合物，并用于药物发现中的细胞测定
公开(公告)号：US20020025940A1
公开(公告)日：2002-02-28
申请号：US09772114
申请日：2001-01-26
申请人： Aurora Biosciences Corporation
发明人： Michael A. Whitney , Kleanthis Xanthopoulos , David Nelson , Paul Negulescu , Frank Craig , J. Gordon Foulkes
IPC分类号： A61K031/70
CPC分类号： A61K31/70
摘要： The invention provides for methods and compositions for identifying proteins or chemicals that directly or indirectly modulate a genomic polynucleotide and methods for identifying active genomic polynucleotides. Generally, the method comprises inserting an adeno-associated virus derived expression construct having a reporter gene into an eukaryotic genome, usually non-yeast, contained in at least one living cell, contacting the cell with a predetermined concentration of a modulator, and detecting reporter gene expression in the cell.
摘要翻译：本发明提供用于鉴定直接或间接调节基因组多核苷酸的蛋白质或化学物质的方法和组合物以及用于鉴定活性基因组多核苷酸的方法。通常，该方法包括将具有报告基因的腺相关病毒衍生的表达构建体插入包含在至少一个活细胞中的真核基因组（通常为非酵母），使细胞与预定浓度的调节剂接触，并检测报告基因细胞中的基因表达。

4. 发明授权

US07824850B2 Methods of protein destabilization and uses thereof 有权
标题翻译：蛋白质不稳定的方法及其用途
公开(公告)号：US07824850B2
公开(公告)日：2010-11-02
申请号：US11821562
申请日：2007-06-22
申请人： Jeffrey Stack , Michael Whitney , Andrew B. Cubitt , Brian Pollok
发明人： Jeffrey Stack , Michael Whitney , Andrew B. Cubitt , Brian Pollok
IPC分类号： C12Q1/00 , C07K14/00
CPC分类号： C12Q1/485
摘要： This invention is directed towards methods of destabilizing proteins in living cells, and their use for the development of novel assays. In one embodiment, the invention comprises the use of non-cleavable multimerized ubiquitin fusion proteins to destabilize a target protein, such as a reporter moiety. In one aspect of this method the constructs also comprises a linker that operatively couples the reporter moiety to the multimerized ubiquitin fusion protein. In this embodiment, enzymatic modification of the linker results in a modulation of the coupling of the reporter protein to the multimerized ubiquitin domains resulting in a change in the stability of the reporter moiety. The level of the reporter moiety in the cell can then be used as a measure of the enzymatic activity in the cell. In another embodiment the invention provides for a generalized way of coordinately regulating the cellular concentration of a plurality of target proteins. In one aspect of this method, the target proteins are operatively coupled to a ubiquitin fusion protein via a linker containing a protease cleavage site. Cleavage of the linker by a protease results in uncoupling of the target protein from the multimerized ubiquitin construct, and results in an increase in the stability and concentration of the target protein.
摘要翻译：本发明涉及使活细胞中的蛋白质失稳的方法及其用于开发新的测定方法。在一个实施方案中，本发明包括使用不可切割的多聚化泛素融合蛋白来使目标蛋白如报道分子部分不稳定。在该方法的一个方面，构建体还包含使报道部分与多聚化泛素融合蛋白可操作地连接的连接体。在该实施方案中，接头的酶修饰导致报道蛋白与多聚泛素结构域的偶联的调节，导致报道部分稳定性的变化。然后可以将细胞中的报告物部分的水平用作细胞中酶活性的量度。在另一个实施方案中，本发明提供了协调调节多种靶蛋白的细胞浓度的一般化方式。在该方法的一个方面，靶蛋白通过含有蛋白酶切割位点的接头与泛素融合蛋白可操作地偶联。通过蛋白酶切割接头导致靶蛋白与多聚化泛素构建体解偶联，并导致靶蛋白的稳定性和浓度增加。

5. 外观设计

USD428157S Platform for a multi-well field 失效
公开(公告)号：USD428157S
公开(公告)日：2000-07-11
申请号：US81969
申请日：1998-01-12
申请人： Peter J. Coassin , Andrew A. Pham , Peter N. Pham
设计人： Peter J. Coassin , Andrew A. Pham , Peter N. Pham

6. 发明授权

US5928888A Methods and compositions for sensitive and rapid, functional identification of genomic polynucleotides and secondary screening capabilities 失效
标题翻译：用于敏感和快速，基因组多核苷酸功能鉴定和二级筛选能力的方法和组合物
公开(公告)号：US5928888A
公开(公告)日：1999-07-27
申请号：US719697
申请日：1996-09-26
申请人： Michael A. Whitney
发明人： Michael A. Whitney
IPC分类号： C12N15/10 , C12Q1/34 , C12Q1/68 , G01N33/50 , C12Q1/02 , C12Q1/00 , G01N33/566
CPC分类号： C12N15/1082 , C12N15/1051 , C12Q1/34 , C12Q1/6897 , G01N33/5008 , G01N33/502 , G01N33/5041 , G01N33/505 , G01N33/5073 , G01N2333/986 , G01N2500/00 , G01N2500/10
摘要： The invention provides for a methods and compositions for identifying proteins or compounds that directly or indirectly modulate a genomic polynucleotide and methods for identifying active genomic polynucleotides. Generally, the method comprises inserting a BL (beta-lactamase) expression construct into an eukaryotic genome, usually non-yeast, contained in at least one living cell, contacting the cell with a predetermined concentration of a modulator, and detecting BL activity in the cell.
摘要翻译：本发明提供用于鉴定直接或间接调节基因组多核苷酸的蛋白质或化合物的方法和组合物以及用于鉴定活性基因组多核苷酸的方法。通常，该方法包括将BL（β-内酰胺酶）表达构建体插入包含在至少一个活细胞中的真核基因组（通常为非酵母），使细胞与预定浓度的调节剂接触，并检测BL 细胞。

7. 发明授权

US5910287A Low background multi-well plates with greater than 864 wells for fluorescence measurements of biological and biochemical samples 失效
标题翻译：具有大于864个孔的低背景多孔板用于生物和生化样品的荧光测量
公开(公告)号：US5910287A
公开(公告)日：1999-06-08
申请号：US868049
申请日：1997-06-03
申请人： Peter J. Cassin , Alec Tate Harootunian , Andrew A. Pham , Roger Y. Tsien
发明人： Peter J. Cassin , Alec Tate Harootunian , Andrew A. Pham , Roger Y. Tsien
IPC分类号： B01L3/00 , B01L9/00 , C40B60/14
CPC分类号： B01L3/5085 , B01J2219/00317 , B01J2219/00659 , B01J2219/00707 , B01J2219/0072 , B01L2200/12 , B01L2300/0829 , B01L2300/0893 , B01L2300/12 , B01L9/523 , C40B60/14 , Y10S436/80 , Y10S436/809
摘要： The invention provides for multi-well plates with greater than 864 wells that comprise a layer of cycloolefin having low fluorescence and high transmittance. These multi-well plates are particularly well suited for fluorescence measurements.
摘要翻译：本发明提供了具有大于864个孔的多孔板，其包含具有低荧光和高透射率的环烯烃层。这些多孔板特别适用于荧光测量。

8. 发明申请

US20110191873A1 Methods of Protein Destabilization and Uses Thereof 审中-公开
标题翻译：蛋白质不稳定的方法及其用途
公开(公告)号：US20110191873A1
公开(公告)日：2011-08-04
申请号：US12938179
申请日：2010-11-02
申请人： Jeffrey Stack , Michael Whitney , Andrew B. Cubitt , Brian Pollok
发明人： Jeffrey Stack , Michael Whitney , Andrew B. Cubitt , Brian Pollok
IPC分类号： A01K67/00 , C12N5/10 , C07H21/04 , C12N1/00 , C07K19/00 , A01H5/00 , C12Q1/02 , C12Q1/37 , C12Q1/42 , C12Q1/48 , C12Q1/66 , C12N5/07 , C12N5/071 , C12N5/04
CPC分类号： C12Q1/485
摘要： This invention is directed towards methods of destabilizing proteins in living cells, and their use for the development of novel assays. In one embodiment, the invention comprises the use of non-cleavable multimerized ubiquitin fusion proteins to destabilize a target protein, such as a reporter moiety. In one aspect of this method the constructs also comprises a linker that operatively couples the reporter moiety to the multimerized ubiquitin fusion protein. In this embodiment, enzymatic modification of the linker results in a modulation of the coupling of the reporter protein to the multimerized ubiquitin domains resulting in a change in the stability of the reporter moiety. The level of the reporter moiety in the cell can then be used as a measure of the enzymatic activity in the cell. In another embodiment the invention provides for a generalized way of coordinately regulating the cellular concentration of a plurality of target proteins. In one aspect of this method, the target proteins are operatively coupled to a ubiquitin fusion protein via a linker containing a protease cleavage site. Cleavage of the linker by a protease results in uncoupling of the target protein from the multimerized ubiquitin construct, and results in an increase in the stability and concentration of the target protein.
摘要翻译：本发明涉及使活细胞中的蛋白质失稳的方法及其用于开发新的测定方法。在一个实施方案中，本发明包括使用不可切割的多聚化泛素融合蛋白来使目标蛋白如报道分子部分不稳定。在该方法的一个方面，构建体还包含使报道部分与多聚化泛素融合蛋白可操作地连接的连接体。在该实施方案中，接头的酶修饰导致报道蛋白与多聚泛素结构域的偶联的调节，导致报道部分稳定性的变化。然后可以将细胞中的报告物部分的水平用作细胞中酶活性的量度。在另一个实施方案中，本发明提供了协调调节多种靶蛋白的细胞浓度的一般化方式。在该方法的一个方面，靶蛋白通过含有蛋白酶切割位点的接头与泛素融合蛋白可操作地偶联。通过蛋白酶切割接头导致靶蛋白与多聚化泛素构建体解偶联，并导致靶蛋白的稳定性和浓度增加。

9. 发明申请

US20020155617A1 Low fluorescence assay platforms and related methods for drug discovery 有权
标题翻译：低荧光测定平台及药物发现的相关方法
公开(公告)号：US20020155617A1
公开(公告)日：2002-10-24
申请号：US10120644
申请日：2002-04-09
申请人： Aurora Biosciences Corporation
发明人： Andrew A. Pham , Peter J. Coassin , Alec Tate Harootunian , Harry Stylli , Roger Y. Tsien
IPC分类号： B01L003/00
CPC分类号： B01L3/5085 , B01J2219/00315 , B01J2219/00317 , B01J2219/00659 , B01J2219/00707 , B01J2219/0072 , B01L9/523 , B01L2300/021 , B01L2300/0829 , B01L2300/12 , C12M23/12 , C12Q1/00 , C40B60/14 , G01N1/30 , G01N21/03 , G01N21/6428 , G01N21/6452 , G01N33/5308 , G01N33/542 , G01N2021/0346
摘要： One aspect of the present invention is a multi-well platform for fluorescence measurements, comprising a plurality of wells within a frame, wherein the multi-well platform has low fluorescence background. Another aspect of the present invention is a system for spectroscopic measurements, comprising reagents for an assay and a multi-well platform for fluorescence measurements. A further aspect of the present invention is a method for detecting the presence of an analyte in a sample contained in a multi-well platform by detecting light emitted from the sample. Another aspect of the present invention is a method from identifying a modulator of a biological process or target in a sample contained in a multi-well platform by detecting light emitted from the sample Another aspect of the present invention is a composition identified by this method. A further aspect of the present invention is a method to identify a therapeutic. A further aspect of the present invention is a method of testing a therapeutic for therapeutic activity and toxicology by identifying a therapeutic using a method of the present invention and monitoring the toxicology and efficacy of the therapeutic in an in vivo model.
摘要翻译：本发明的一个方面是用于荧光测量的多孔平台，其包括框架内的多个孔，其中所述多孔平台具有低荧光背景。本发明的另一方面是用于光谱测量的系统，其包括用于测定的试剂和用于荧光测量的多孔平台。本发明的另一方面是通过检测从样品发射的光来检测多孔平台中包含的样品中分析物的存在的方法。本发明的另一方面是通过检测从样品发出的光来鉴定多孔平台中包含的样品中的生物过程或靶的调节剂的方法。本发明的另一方面是通过该方法鉴定的组合物。本发明的另一方面是鉴定治疗剂的方法。本发明的另一方面是通过使用本发明的方法鉴定治疗剂并监测治疗剂在体内模型中的毒理学和功效来测试治疗活性和毒理学治疗剂的方法。

10. 发明授权

US6004808A Promiscuous G-protein compositions and their use 失效
公开(公告)号：US6004808A
公开(公告)日：1999-12-21
申请号：US878801
申请日：1997-06-19
申请人： Paul Negulescu , Stefan Offermanns , Melvin Simon , Charles Zuker
发明人： Paul Negulescu , Stefan Offermanns , Melvin Simon , Charles Zuker
IPC分类号： C07K14/705 , C12N15/85 , G01N33/50 , C12N5/10
CPC分类号： G01N33/5008 , C07K14/705 , C12N15/85 , G01N33/5041 , C12N2830/002 , C12N2830/003
摘要： Disclosed are compositions and methods for their use, such as in identifying G-protein coupled receptors and ligands and compounds that modulate signal transduction. The compositions and methods employ promicuous G-proteins. Activation of the promiscous G-protein can be detected in a variety of assays, including assays in which activation is indicated by a change in fluorescence emission of a sample that contains the composition.

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