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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. 发明授权

US07087388B1 Compositions and methods for identifying modulators of transducisomes, a new class of therapeutic targets 有权
标题翻译：用于鉴定转导体调节剂的组合物和方法，一类新的治疗靶点
公开(公告)号：US07087388B1
公开(公告)日：2006-08-08
申请号：US09462517
申请日：1998-07-15
申请人： Charles S. Zuker , John D. Mendlein , Yumei Sun , Susan Tsunoda , Jimena Sierralta
发明人： Charles S. Zuker , John D. Mendlein , Yumei Sun , Susan Tsunoda , Jimena Sierralta
IPC分类号： G01N33/53 , C07K14/00
CPC分类号： G01N33/5041 , G01N2333/726 , G01N2333/912 , G01N2333/916
摘要： The invention provides cells and methods for identifying modulators of signal transduction, based on transducisome proteins that coordinate and assemble many types of signal transduction proteins. A transducisome is a PDZ domain containing protein that binds at least one signal transduction protein or a PDZ domain containing protein with at least one signal transduction protein bound. Examples of transducisome proteins include INAD, GRIP and other recently identified multi-PDZ domain proteins. Examples of signal transduction proteins include GPCRs, tyrosine kinase receptors, tyrosine phosphatase receptors, ion channels, phospholipases, adenylate cyclases, kinases and G-proteins. Also provided are methods for identifying modulators of signal transduction, proteins (and polynucleotides encoding the same) corresponding to transducisomes, modified transducisomes or defective transducisomes to use in assays of signal transduction, and a screening assay system for detecting protein-protein interactions.
摘要翻译：本发明提供用于鉴定信号转导调节剂的细胞和方法，其基于协调和组合许多类型的信号转导蛋白的传导蛋白。传导蛋白是含有蛋白质的PDZ结构域，其结合至少一个信号转导蛋白或含有至少一个信号转导蛋白结合的含有PDZ结构域的蛋白质。传导蛋白的实例包括INAD，GRIP和其他最近鉴定的多PDZ结构域蛋白。信号转导蛋白的实例包括GPCR，酪氨酸激酶受体，酪氨酸磷酸酶受体，离子通道，磷脂酶，腺苷酸环化酶，激酶和G蛋白。还提供了用于鉴定信号转导调节剂的方法，对应于用于信号转导测定的传导蛋白，修饰的传导蛋白或有缺陷的传导蛋白的蛋白质（和编码它们的多核苷酸）的方法，以及用于检测蛋白质 - 蛋白质相互作用的筛选测定系统。

9. 发明申请

US20050112773A1 Multiwell scanner and scanning method 审中-公开
标题翻译： Multiwell扫描仪和扫描方法
公开(公告)号：US20050112773A1
公开(公告)日：2005-05-26
申请号：US10981821
申请日：2004-11-05
申请人： T. Vuong
发明人： T. Vuong
IPC分类号： B01J19/00 , C40B60/14 , G01N21/25 , G01N21/64 , G01N21/76 , G01N35/02 , G01N21/00
CPC分类号： G01N21/6452 , B01J19/0046 , B01J2219/00315 , B01J2219/00317 , B01J2219/00707 , B01J2219/0072 , C40B60/14 , G01N21/253 , G01N21/76 , G01N35/028
摘要： A multiwell plate scanner comprises a detector for measuring an attribute of a sample which is scanned continuously over wells of a multiwell plate. A signal obtained during the scan may be sampled and digitized based on detector position over the multiwell plate. The scanner is also disclosed for scanning microarrays, bio-chips and areas of samples not having physical separations. The scanner may be used in a high throughput screening system comprising a storage and retrieval module, a sample distribution module, a reagent distribution module, and a detector which incorporates the scanner. The screening system may further comprise a transport module and a data processing and integration module for transporting samples between the components of the system and for controlling system operation. In a further aspect, the present invention comprises methods and apparatus to compensate for variations, inconsistencies and other differences in the dimensions, configuration and/or materials of the platform in which an assay is performed. Another aspect of the invention is a system and method for performing an assay to detect the effect of a reagent on a target. The present invention is also directed to compositions and therapeutics identified by the disclosed methods. A further aspect of the present invention is a method of testing a therapeutic for therapeutic activity and toxicology by identifying a compound 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：多孔板扫描器包括用于测量在多孔板的孔上连续扫描的样品的属性的检测器。在扫描期间获得的信号可以基于多孔板上的检测器位置被采样和数字化。还公开了用于扫描微阵列，生物芯片和不具有物理分离的样品区域的扫描仪。扫描仪可以用于包括存储和检索模块，样品分配模块，试剂分配模块和结合扫描仪的检测器的高通量筛选系统中。筛选系统还可以包括传送模块和数据处理和集成模块，用于在系统的组件之间传送样本并用于控制系统操作。在另一方面，本发明包括用于补偿其中进行测定的平台的尺寸，构型和/或材料的变化，不一致性和其它差异的方法和装置。本发明的另一方面是用于进行测定以检测试剂对靶标的影响的系统和方法。本发明还涉及通过所公开的方法鉴定的组合物和治疗剂。本发明的另一方面是通过使用本发明的方法鉴定化合物并在体内模型中监测治疗剂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. 发明申请

US20030170767A1 Fluorescent protein sensors of post-translational modifications 审中-公开
标题翻译：荧光蛋白传感器的翻译后修饰
公开(公告)号：US20030170767A1
公开(公告)日：2003-09-11
申请号：US10293580
申请日：2002-11-12
申请人： Aurora Biosciences Corporation
发明人： Andrew B. Cubitt
IPC分类号： C12Q001/48 , C12Q001/37 , C07H021/04 , C12P021/02 , C12N005/06 , C07K014/435
CPC分类号： C07K14/43595
摘要： The present invention includes a fluorescent compound that can detect an activity. such as an enzymatic activity, and exhibits quenching. The fluorescent compound can include a fluorescent protein, such as an Aequorea-related green fluorescent protein. The fluorescent compound can include a substrate site for an enzymatic activity such as a kinase activity, a phosphatase activity, a protease activity, and a glycosylase activity The fluorescent compound of the present invention can be used to detect such enzymatic activities in samples, such as biological samples, including cells. The present invention also includes nucleic acids that encode the fluorescent compounds of the present inventions, and cells that include such nucleic acids or fluorescent compounds.
摘要翻译：本发明包括能够检测活性的荧光化合物。例如酶活性，并显示淬灭。荧光化合物可以包括荧光蛋白，例如与水母相关的绿色荧光蛋白。荧光化合物可以包括酶活性的底物位点，例如激酶活性，磷酸酶活性，蛋白酶活性和糖基化酶活性。本发明的荧光化合物可用于检测样品中的这种酶活性，例如生物样品，包括细胞。本发明还包括编码本发明的荧光化合物的核酸和包括这种核酸或荧光化合物的细胞。

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