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    Methods and compositions for interaction trap assays
    63.
    发明申请
    Methods and compositions for interaction trap assays 有权
    相互作用阱测定法的方法和组成

    公开(公告)号：US20050064477A1

    公开(公告)日：2005-03-24

    申请号：US10915233

    申请日：2004-08-10

    Applicant: J. Joung ,  Jeffrey Miller ,  Carl Pabo

    Inventor： J. Joung ,  Jeffrey Miller ,  Carl Pabo

    IPC: C12N15/10 ,  C12Q1/68 ,  G01N33/50

    CPC classification number: C12N15/1055 ,  C12Q1/6897 ,  G01N33/50 ,  G01N33/5008 ,  G01N33/502 ,  C12Q2565/201

    Abstract: The present invention provides methods and compositions for interaction trap assays for detecting protein-protein, protein-DNA, or protein-RNA interactions. The methods and compositions of the invention may also be used to identify agents which may agonize or antagonize a protein-protein, protein-DNA, or protein-RNA interactions. In certain embodiments, the interaction trap system of the invention is useful for screening libraries with greater than 107 members. In other embodiments, the interaction trap system of the invention is used in conjunction with flow cytometry. The invention further provides a means for simultaneously screening a target protein or nucleic acid sequence for the ability to interact with two or more test proteins or nucleic acids.

    Abstract translation: 本发明提供了用于检测蛋白质 - 蛋白质，蛋白质-DNA或蛋白质 - RNA相互作用的相互作用阱测定法的方法和组合物。 本发明的方法和组合物还可用于鉴定可能激动或拮抗蛋白质 - 蛋白质，蛋白质-DNA或蛋白质 - RNA相互作用的试剂。 在某些实施方案中，本发明的相互作用捕获系统可用于筛选具有大于10 7个成员的文库。 在其它实施方案中，本发明的相互作用阱系统结合流式细胞术使用。 本发明还提供了用于同时筛选靶蛋白或核酸序列以获得与两种或多种测试蛋白或核酸相互作用的能力的方法。

    Mammalian two-hybrid system for screening for modulators of the accumulation of metabolic products
    65.
    发明授权
    Mammalian two-hybrid system for screening for modulators of the accumulation of metabolic products 有权
    哺乳动物双杂交系统用于筛选代谢产物积累的调节剂

    公开(公告)号：US06787321B1

    公开(公告)日：2004-09-07

    申请号：US09687593

    申请日：2000-10-13

    Applicant: Osamu Tetsu ,  Kenichi Wakita ,  Frank McCormick

    Inventor： Osamu Tetsu ,  Kenichi Wakita ,  Frank McCormick

    IPC: G01N3353

    CPC classification number: C12N15/1055 ,  A61K48/00 ,  A61K48/0058 ,  C12Q1/6897 ,  C12Q2565/201

    Abstract: This invention utilizes a two-hybrid system to screen for agents that modulate the ability of a cell to degrade or to accumulate a metabolic product or to selective kill a cell or to selectively express a gene or cDNA in a cell that has a defect in its ability to degrade or to accumulate a metabolic product. One embodiment provides a mammalian cell comprising a nucleic acid encoding a peptide binding domain and an effector gene; a first chimeric protein comprising a nucleic acid binding domain that binds the peptide binding domain attached to the metabolic product or to a ligand that binds to the metabolic product; and a second chimeric protein comprising an expression control protein attached to the metabolic product or to the ligand that binds to the metabolic product such that when the first chimeric protein comprises the metabolic product, the second chimeric protein comprises the ligand and when the first chimeric protein comprises the ligand, the second chimeric protein comprises the metabolic product. The cell is contacted with a test agent and in alteration of expression of the effector gene is detected (if present) where a change in expression of the effector gene in response to the test agent indicates that the test agent modulates the ability of the cell to accumulate or degrade the metabolic product.

    Abstract translation: 本发明利用双杂交系统筛选调节细胞降解能力或积累代谢产物或选择性杀死细胞或选择性地在其中具有缺陷的细胞中表达基因或cDNA的试剂 降解或积累代谢产物的能力。 一个实施方案提供了哺乳动物细胞，其包含编码肽结合结构域和效应基因的核酸; 第一嵌合蛋白，其包含结合与所述代谢产物连接的肽结合结构域的核酸结合结构域或与所述代谢产物结合的配体; 以及第二嵌合蛋白，其包含与代谢产物连接的表达控制蛋白或与所述代谢产物结合的配体，使得当所述第一嵌合蛋白包含所述代谢产物时，所述第二嵌合蛋白包含所述配体，并且当所述第一嵌合蛋白 包含配体，第二嵌合蛋白包含代谢产物。 细胞与测试试剂接触，检测效应基因表达的改变（如果存在），其中响应于测试试剂的效应基因的表达变化表明测试试剂调节细胞的能力 积累或降解代谢产物。

    Methods and compositions for interaction trap assays
    66.
    发明申请
    Methods and compositions for interaction trap assays 审中-公开

    公开(公告)号：US20030044787A1

    公开(公告)日：2003-03-06

    申请号：US09858852

    申请日：2001-05-16

    Inventor： J. Keith Joung ,  Jeffrey Miller ,  Carl O. Pabo

    IPC: C12Q001/68 ,  C07H021/04

    CPC classification number: G01N33/502 ,  C12N15/1055 ,  C12Q1/6897 ,  G01N33/50 ,  G01N33/5008 ,  C12Q2565/201

    Abstract: The present invention provides methods and compositions for interaction trap assays for detecting protein-protein, protein-DNA, or protein-RNA interactions. The methods and compositions of the invention may also be used to identify agents which may agonize or antagonize a protein-protein, protein-DNA, or protein-RNA interaction. In certain embodiments, the interaction trap system of the invention is useful for screening libraries with greater than 107 members. In other embodiments, the interaction trap system of the invention is used in conjunction with flow cytometry. The invention further provides a means for simultaneously screening a target protein or nucleic acid sequence for the ability to interact with two or more test proteins or nucleic acids.

    Methods for modulating transcriptional activation using mint proteins
    67.
    发明申请
    Methods for modulating transcriptional activation using mint proteins 失效
    使用薄荷蛋白调节转录激活的方法

    公开(公告)号：US20030036169A1

    公开(公告)日：2003-02-20

    申请号：US10227490

    申请日：2002-08-23

    Inventor： Thomas C. Sudhof ,  Thomas Biederer ,  Angela Ho ,  Xinran Liu

    IPC: C12P021/02 ,  C12N005/06 ,  C07H021/04 ,  C12N009/64

    CPC classification number: G01N33/5008 ,  A01K2217/05 ,  A01K2217/075 ,  C07K14/4702 ,  C07K14/4711 ,  C07K14/70567 ,  C07K2319/00 ,  C12Q1/6897 ,  G01N33/5023 ,  G01N33/5035 ,  G01N33/6896 ,  G01N2333/4709 ,  G01N2500/20 ,  G01N2800/2821 ,  C12Q2565/201

    Abstract: The present invention is directed to isolated nucleic acids encoding Mint protein variants having enhanced abilities to modulate the transcriptional activation mediated by the cytoplasmic tail of the amyloid precursor protein (APP) relative to wild-type Mint proteins. The present invention is further directed toward purified Mint protein variants having enhanced abilities to modulate the transcriptional activation mediated by the cytoplasmic tail of APP relative to wild-type Mint proteins. The present invention also encompasses methods of modulating transcriptional activation and methods of identifying compounds that modulate transcriptional activation, and vectors, as well as transfected cells and kits useful for modulating transcriptional activation or for the identification of compounds that can modulate transcriptional activation. The present invention further encompasses transgenic knockout mice with little or no expression of Mint 1, Mint 2 or Mint 3 proteins. Such reagents may be useful as candidate therapeutics for Alzheimer's disease (AD), or as models for the rational design of drugs useful for the treatment of AD.

    Abstract translation: 本发明涉及编码具有增强的相对于野生型薄荷蛋白调节由淀粉样蛋白前体蛋白（APP）的细胞质尾部介导的转录激活的能力的Mint蛋白变体的分离的核酸。 本发明还涉及具有相对于野生型薄荷蛋白调节由APP的细胞质尾巴介导的转录激活的能力的纯化的薄荷蛋白变体。 本发明还包括调节转录激活的方法和鉴定调节转录激活的化合物的方法，以及载体，以及用于调节转录激活或用于鉴定可调节转录激活的化合物的转染细胞和试剂盒。 本发明还包括具有很少或没有表达Mint1，Mint2或Mint3蛋白的转基因敲除小鼠。 这些试剂可用作阿尔茨海默病（AD）的候选治疗剂，或作为用于治疗AD的药物的合理设计的模型。

    Two hybrid assay that detects hiv-1 reverse transcriptase dimerization
    68.
    发明申请
    Two hybrid assay that detects hiv-1 reverse transcriptase dimerization 失效
    检测hiv-1逆转录酶二聚化的两种杂交测定法

    公开(公告)号：US20020197598A1

    公开(公告)日：2002-12-26

    申请号：US10009433

    申请日：2002-06-12

    Inventor： Stephen P. Goff ,  Gilda Tachedjian

    IPC: C12Q001/70 ,  C12N001/18 ,  C12N015/74 ,  C12N001/14 ,  C12N001/16

    CPC classification number: C12Y207/07049 ,  C07K2319/00 ,  C12N9/1276 ,  C12Q1/6897 ,  C12Q2565/201

    Abstract: This invention provides methods of determining whether a compound inhibits HIV-1 reverse transcriptase. This invention provides methods of determining whether a compound inhibits formation of a complex between a p66 and p51 subunit polypeptides of HIV-1 reverse transcriptase. This invention provides a method of determining whether a compound enhances formation of a complex between a p66 and p51 subunit polypeptides of HIV-1 reverse transcriptase. This invention provides methods of determining whether a compound inhibits formation of a complex between two p66 subunit polypeptides of HIV-1 reverse transcriptase. This invention provides methods of determining whether a compound enhances formation of a complex between two p66 subunit polypeptides of HIV-1 reverse transcriptase.

    Abstract translation: 本发明提供了确定化合物是否抑制HIV-1逆转录酶的方法。 本发明提供了确定化合物是否抑制HIV-1逆转录酶的p66和p51亚基多肽之间复合物形成的方法。 本发明提供了确定化合物是否增强HIV-1逆转录酶的p66和p51亚基多肽之间的复合物形成的方法。 本发明提供了确定化合物是否抑制HIV-1逆转录酶的两个p66亚基多肽之间复合物形成的方法。 本发明提供了确定化合物是否增强HIV-1逆转录酶的两个p66亚基多肽之间复合物形成的方法。

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