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    Inducible fluorescence assay
    1.
    发明授权
    Inducible fluorescence assay 有权
    诱导荧光测定

    公开(公告)号：US07635598B2

    公开(公告)日：2009-12-22

    申请号：US11177998

    申请日：2005-07-08

    申请人： Ronald M. Cook ,  Eliana Saxon Armstrong ,  Hans E. Johansson

    发明人： Ronald M. Cook ,  Eliana Saxon Armstrong ,  Hans E. Johansson

    IPC分类号： G01N33/533 ,  G01N33/53 ,  C12Q1/26 ,  C12N1/21 ,  C12P21/00

    CPC分类号： C12Q1/26 ,  C09B11/08 ,  C12Q1/6818 ,  C12Q1/6897 ,  G01N33/542 ,  G01N2333/90209 ,  Y10S436/903 ,  Y10T436/13

    摘要： In one aspect, the invention provides fluorescent probes and assays. The probes include a fluorophore-quencher pair that undergoes a switch from dark to fluorescent in response to a reaction of the quencher. The switch of the probe from dark to fluorescent is typically mediated by an enzyme that acts directly or indirectly on the quencher, interfering with its ability to quench fluorescence emission from the fluorophore.In another aspect, the invention provides a reporter gene assay system and methods of using this system. The assay system includes a fluorophore-quencher probe and an enzyme that acts directly or indirectly on the quencher, increasing the fluorescent emission of the fluorophore. In still other aspects, the invention provides nucleic acid constructs and cells expressing the peptide products of these constructs.In assays of the invention, the presence of a target substance is detected by the switching of fluorescence mediated by the change in oxidation state of the quencher. The assay systems are of use in numerous assay formats, e.g., confirming the expression of the enzyme that acts on the quencher, and/or detecting a species conjugated to the enzyme.

    摘要翻译： 一方面，本发明提供了荧光探针和测定法。 探针包括响应于猝灭剂的反应经历从暗变为荧光的荧光团 - 猝灭剂对。 探针从黑暗到荧光的切换通常由直接或间接作用于猝灭剂的酶介导，干扰其从荧光团中淬灭荧光发射的能力。 另一方面，本发明提供报道基因测定系统和使用该系统的方法。 测定系统包括荧光团猝灭剂探针和直接或间接作用于猝灭剂的酶，增加荧光团的荧光发射。 在其它方面，本发明提供了表达这些构建体的肽产物的核酸构建体和细胞。 在本发明的测定中，通过由猝灭剂的氧化态的变化介导的荧光的转换来检测目标物质的存在。 测定系统可用于许多测定形式，例如确认作用于猝灭剂的酶的表达和/或检测与酶缀合的物质。

    STABILIZED NUCLEIC ACID DARK QUENCHER-FLUOROPHORE PROBES
    2.
    发明申请
    STABILIZED NUCLEIC ACID DARK QUENCHER-FLUOROPHORE PROBES 有权
    稳定的核酸深色探针 - 荧光探针

    公开(公告)号：US20110282041A1

    公开(公告)日：2011-11-17

    申请号：US13193517

    申请日：2011-07-28

    申请人： Ronald M. Cook ,  Matt Lyttle

    发明人： Ronald M. Cook ,  Matt Lyttle

    IPC分类号： C07H19/10 ,  C07H21/00 ,  C07H19/06

    CPC分类号： C12Q1/6876 ,  C07D219/04 ,  C07H19/06 ,  C07H19/073 ,  C09B31/02

    摘要： The present invention provides a new class of solids supports for synthesis of modified oligomers of nucleic acids, and nucleic acid probes that have a format expediently synthesized on the new supports. Exemplary solid supports include at least one quencher bound through a linker to the solid support. Various exemplary embodiments include a moiety that stabilizes a duplex, triplex or higher order aggregation (e.g., hybridization) of nucleic acids of which the oligomer of the invention is a component. Other components of the solid support include moieties that stabilize aggregations of nucleic acids, e.g., intercalators, minor groove binding moieties, bases modified with a stabilizing moiety (e.g., alkynyl moieties, and fluoroalkyl moieties), and conformational stabilizing moieties, such as those described in commonly owned U.S. Patent Application Publication No. 2007/0059752.

    摘要翻译： 本发明提供了用于合成核酸修饰低聚物的新类固体支持物，以及具有在新载体上方便合成的形式的核酸探针。 示例性的固体支持物包括通过连接体与固体支持物结合的至少一种猝灭剂。 各种示例性实施方案包括稳定其本发明的低聚物是组分的核酸的双链体，三链体或更高级聚集（例如杂交）的部分。 固体支持物的其它组分包括稳定核酸聚集的部分，例如嵌入剂，小槽结合部分，用稳定部分修饰的碱基（例如炔基部分和氟烷基部分）和构象稳定部分，例如描述的那些 在共同拥有的美国专利申请公开No.2007 / 0059752中。

    Oxidized polyethylene or polypropylene particulate supports
    3.
    发明授权
    Oxidized polyethylene or polypropylene particulate supports 失效
    氧化聚乙烯或聚丙烯颗粒载体

    公开(公告)号：US5679539A

    公开(公告)日：1997-10-21

    申请号：US377808

    申请日：1995-01-24

    申请人： Derek Hudson ,  Ronald M. Cook

    发明人： Derek Hudson ,  Ronald M. Cook

    IPC分类号： B01D15/32 ,  B01D15/36 ,  B01J20/32 ,  C07K1/04 ,  C07K17/08 ,  C12N11/08 ,  C12P21/06 ,  C07K17/06 ,  C12N11/06 ,  G01N33/549

    CPC分类号： B01D15/325 ,  B01D15/361 ,  B01J20/0229 ,  B01J20/261 ,  B01J20/262 ,  B01J20/265 ,  B01J20/28004 ,  B01J20/28009 ,  B01J20/28026 ,  B01J20/287 ,  B01J20/289 ,  B01J20/3021 ,  B01J20/3092 ,  B01J20/3246 ,  B01J20/3248 ,  B01J20/3251 ,  B01J20/3272 ,  B01J20/328 ,  C07K1/042 ,  C07K17/08 ,  C12N11/08 ,  B01J2220/46 ,  B01J2220/54 ,  B01J2220/58 ,  Y10S530/815 ,  Y10S530/816

    摘要： Polyolefin particles are chemically modified by oxidation to provide a large surface area and high loading. The particles result in low back pressure in column systems, and are economical to manufacture. The particles are useful as supports in a wide range of applications including general organic as well as biopolymer synthesis, library methods, purification processes and enzyme mediated processes. In a preferred embodiment, polyethylene or polypropylene particles are oxidized in a solution containing trifluoroacetic acid or trifluoromethane sulfonic acid, chromium trioxide and sulfuric acid to provide the particles with a chemically reactive irregular surface and open channels that extend below the surface and up to essentially the length of the radius of the particles resulting in increased surface area and decreased density. The particles have pendant functional groups produced by the oxidation and/or by subsequent chemical reaction. The pendant functional groups may be attached to spacer arms that are attached to the surface of the particles. Biological material such as proteins, enzymes and whole cells may be immobilized on the particles.

    摘要翻译： 聚烯烃颗粒通过氧化进行化学改性以提供大的表面积和高负载量。 颗粒在柱系统中导致低背压，并且制造经济。 该颗粒可用作广泛应用的载体，包括一般有机物以及生物聚合物合成，文库方法，纯化方法和酶介导的方法。 在优选的实施方案中，聚乙烯或聚丙烯颗粒在含有三氟乙酸或三氟甲磺酸，三氧化铬和硫酸的溶液中被氧化，以使颗粒具有化学反应性不规则表面和在表面下方延伸至基本上为 颗粒半径的长度导致表面积增加和密度降低。 颗粒具有通过氧化和/或随后的化学反应产生的侧链官能团。 侧链官能团可以连接到附着到颗粒表面的间隔臂上。 生物材料如蛋白质，酶和全细胞可以固定在颗粒上。

    Stabilized nucleic acid dark quencher-fluorophore probes
    4.
    发明授权
    Stabilized nucleic acid dark quencher-fluorophore probes 有权
    稳定的核酸暗猝灭剂 - 荧光团探针

    公开(公告)号：US08674094B2

    公开(公告)日：2014-03-18

    申请号：US13193517

    申请日：2011-07-28

    申请人： Ronald M. Cook ,  Matt Lyttle

    发明人： Ronald M. Cook ,  Matt Lyttle

    IPC分类号： C07F9/6512 ,  C07F9/655

    CPC分类号： C12Q1/6876 ,  C07D219/04 ,  C07H19/06 ,  C07H19/073 ,  C09B31/02

    摘要： The present invention provides a new class of solids supports for synthesis of modified oligomers of nucleic acids, and nucleic acid probes that have a format expediently synthesized on the new supports. Exemplary solid supports include at least one quencher bound through a linker to the solid support. Various exemplary embodiments include a moiety that stabilizes a duplex, triplex or higher order aggregation (e.g., hybridization) of nucleic acids of which the oligomer of the invention is a component. Other components of the solid support include moieties that stabilize aggregations of nucleic acids, e.g., intercalators, minor groove binding moieties, bases modified with a stabilizing moiety (e.g., alkynyl moieties, and fluoroalkyl moieties), and conformational stabilizing moieties, such as those described in commonly owned U.S. Patent Application Publication No. 2007/0059752.

    摘要翻译： 本发明提供了用于合成核酸修饰低聚物的新类固体支持物，以及具有在新载体上方便合成的形式的核酸探针。 示例性的固体支持物包括通过连接体与固体支持物结合的至少一种猝灭剂。 各种示例性实施方案包括稳定其本发明的低聚物是组分的核酸的双链体，三链体或更高级聚集（例如杂交）的部分。 固体支持物的其它组分包括稳定核酸聚集的部分，例如嵌入剂，小槽结合部分，用稳定部分修饰的碱基（例如炔基部分和氟烷基部分）和构象稳定部分，例如描述的那些 在共同拥有的美国专利申请公开No.2007 / 0059752中。

    Chemically programmable immunity
    5.
    发明授权
    Chemically programmable immunity 有权
    化学可编程免疫

    公开(公告)号：US08604184B2

    公开(公告)日：2013-12-10

    申请号：US12774194

    申请日：2010-05-05

    申请人： Kary B. Mullis ,  Jeevalatha Vivekananda ,  Johnathan Lloyd Kiel ,  Ronald M. Cook

    发明人： Kary B. Mullis ,  Jeevalatha Vivekananda ,  Johnathan Lloyd Kiel ,  Ronald M. Cook

    IPC分类号： A61K48/00 ,  C07H21/04

    CPC分类号： A61K31/7088 ,  Y02A50/471 ,  Y02A50/473

    摘要： Methods and compositions for immediately immunizing an individual against any molecule or compound are provided. The present invention is directed to an immunity linker with at least two sites; (1) at least one first binding site that binds to an immune response component in an individual, and (2) at least one second binding site that binds specifically to a desired compound or molecule, the target. The second binding sites are preferably thiolated aptamers that have the benefit of increased stability, resistance to degradation and longer circulating half life. Methods of making and using pharmaceutical compositions including immunity linker molecules having a thiolated aptamer are also provided.

    摘要翻译： 提供了立即将个体免疫任何分子或化合物的方法和组合物。 本发明涉及具有至少两个位点的免疫接头; （1）结合个体中的免疫应答组分的至少一个第一结合位点，和（2）至少一个第二结合位点，其特异性结合所需化合物或分子，靶标。 第二结合位点优选为具有增加的稳定性，耐降解性和更长的循环半衰期益处的硫醇化适体。 还提供了制备和使用药物组合物的方法，包括具有巯基适体的免疫接头分子。

    Stabilized nucleic acid dark quencher-fluorophore probes
    8.
    发明授权
    Stabilized nucleic acid dark quencher-fluorophore probes 有权
    稳定的核酸暗猝灭剂 - 荧光团探针

    公开(公告)号：US08466266B2

    公开(公告)日：2013-06-18

    申请号：US12416901

    申请日：2009-04-01

    申请人： Ronald M. Cook ,  Matt Lyttle

    发明人： Ronald M. Cook ,  Matt Lyttle

    IPC分类号： C07C245/02

    CPC分类号： C12Q1/6876 ,  C07D219/04 ,  C07H19/06 ,  C07H19/073 ,  C09B31/02

    摘要： The present invention provides a new class of solids supports for synthesis of modified oligomers of nucleic acids, and nucleic acid probes that have a format expediently synthesized on the new supports. Exemplary solid supports include at least one quencher bound through a linker to the solid support. Various exemplary embodiments include a moiety that stabilizes a duplex, triplex or higher order aggregation (e.g., hybridization) of nucleic acids of which the oligomer of the invention is a component. Other components of the solid support include moieties that stabilize aggregations of nucleic acids, e.g., intercalators, minor groove binding moieties, bases modified with a stabilizing moiety (e.g., alkynyl moieties, and fluoroalkyl moieties), and conformational stabilizing moieties, such as those described in commonly owned U.S. Patent Application Publication No. 2007/0059752.

    摘要翻译： 本发明提供了用于合成核酸修饰低聚物的新类固体支持物，以及具有在新载体上方便合成的形式的核酸探针。 示例性的固体支持物包括通过连接体与固体支持物结合的至少一种猝灭剂。 各种示例性实施方案包括稳定其本发明的低聚物是组分的核酸的双链体，三链体或更高级聚集（例如杂交）的部分。 固体支持物的其它组分包括稳定核酸聚集的部分，例如嵌入剂，小槽结合部分，用稳定部分修饰的碱基（例如炔基部分和氟烷基部分）和构象稳定部分，例如描述的那些 在共同拥有的美国专利申请公开No.2007 / 0059752中。

    STABILIZED NUCLEIC ACID DARK QUENCHER-FLUOROPHORE PROBES
    10.
    发明申请
    STABILIZED NUCLEIC ACID DARK QUENCHER-FLUOROPHORE PROBES 有权
    稳定的核酸深色探针 - 荧光探针

    公开(公告)号：US20090259030A1

    公开(公告)日：2009-10-15

    申请号：US12416901

    申请日：2009-04-01

    申请人： Ronald M. Cook ,  Matt Lyttle

    发明人： Ronald M. Cook ,  Matt Lyttle

    IPC分类号： C07C245/02 ,  C07H21/00 ,  C07D241/46

    CPC分类号： C12Q1/6876 ,  C07D219/04 ,  C07H19/06 ,  C07H19/073 ,  C09B31/02

    摘要： The present invention provides a new class of solids supports for synthesis of modified oligomers of nucleic acids, and nucleic acid probes that have a format expediently synthesized on the new supports. Exemplary solid supports include at least one quencher bound through a linker to the solid support. Various exemplary embodiments include a moiety that stabilizes a duplex, triplex or higher order aggregation (e.g., hybridization) of nucleic acids of which the oligomer of the invention is a component. Other components of the solid support include moieties that stabilize aggregations of nucleic acids, e.g., intercalators, minor groove binding moieties, bases modified with a stabilizing moiety (e.g., alkynyl moieties, and fluoroalkyl moieties), and conformational stabilizing moieties, such as those described in commonly owned U.S. Patent Application Publication No. 2007/0059752.

    摘要翻译： 本发明提供了用于合成核酸修饰低聚物的新类固体支持物，以及具有在新载体上方便合成的形式的核酸探针。 示例性的固体支持物包括通过连接体与固体支持物结合的至少一种猝灭剂。 各种示例性实施方案包括稳定其本发明的低聚物是组分的核酸的双链体，三链体或更高级聚集（例如杂交）的部分。 固体支持物的其它组分包括稳定核酸聚集的部分，例如嵌入剂，小槽结合部分，用稳定部分修饰的碱基（例如炔基部分和氟烷基部分）和构象稳定部分，例如描述的那些 在共同拥有的美国专利申请公开No.2007 / 0059752中。