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    Novel method for detecting and analyzing protein interactions in vivo
    2.
    发明申请
    Novel method for detecting and analyzing protein interactions in vivo 失效
    用于检测和分析体内蛋白质相互作用的新方法

    公开(公告)号：US20050084864A1

    公开(公告)日：2005-04-21

    申请号：US10507506

    申请日：2003-03-13

    申请人： Moritz Rossner ,  Rico Laage ,  Klaus-Armin Nave ,  Sylvia Gruenewald

    发明人： Moritz Rossner ,  Rico Laage ,  Klaus-Armin Nave ,  Sylvia Gruenewald

    IPC分类号： C12N15/10 ,  C12Q1/25 ,  C12Q1/37 ,  C40B30/04 ,  G01N33/542 ,  G01N33/68 ,  C12Q1/68 ,  G01N33/53 ,  G01N33/567

    CPC分类号： C40B30/04 ,  C12N15/1055 ,  C12Q1/25 ,  C12Q1/37 ,  G01N33/542 ,  G01N2500/00

    摘要： The invention relates to a method of detecting and analyzing protein interactions in a cell, which comprises the steps: d) provision of the activity of at least one enzyme from the group consisting of recombinases and proteases in the cell as a result of a protein interaction, e) continual generation of an active reporter protein in the cell in question as a result of the enzymic activity of step a) for a period of time which exceeds the duration of the protein interaction of step a), f) generation of a detection signal by the reporter proteins generated in b). The invention furthermore relates to reverse embodiments of the method above of detecting and analyzing protein interactions in a cell, with, as a result of the induced dissociation of a defined interaction between proteins, the activity of at least one enzyme from the group consisting of recombinases and proteases being provided in the cell and converted to a permanent detection signal of said cell. The invention moreover relates to cells expressing the protein components of the invention and to kits providing the protein components of the invention at the DNA level in the form of suitable expression vectors and, where appropriate, suitable transfectable or injectable cells. The cells provided may, where appropriate, express stably or transiently individual protein components of the invention.

    摘要翻译： 本发明涉及一种检测和分析细胞中蛋白质相互作用的方法，其包括以下步骤：d）作为蛋白质相互作用的结果，提供来自细胞中的重组酶和蛋白酶的至少一种酶的活性 e）作为步骤a）的酶活性的结果，在所述细胞中连续产生活性报道蛋白超过步骤a）的蛋白质相互作用的持续时间，f）产生检测 信号由b）中报告蛋白产生。 本发明还涉及上述方法的检测和分析细胞中的蛋白质相互作用的反向实施方案，由于蛋白质之间限定的相互作用的诱导解离的结果，来自重组酶的至少一种酶的活性 并且蛋白酶被提供在细胞中并被转换成所述细胞的永久检测信号。 此外，本发明涉及表达本发明的蛋白质组分的细胞和以适当的表达载体形式的DNA水平提供本发明的蛋白质组分并在合适的情况下提供合适的可转染或可注射细胞的试剂盒。 在适当的情况下，所提供的细胞可稳定地或瞬时地表达本发明的单个蛋白质组分。

    COMBINATORIAL SYNTHESIS AND USE OF LIBRARIES OF SHORT EXPRESSED NUCLEIC ACID SEQUENCES FOR THE ANALYSIS OF CELLULAR EVENTS
    5.
    发明申请
    COMBINATORIAL SYNTHESIS AND USE OF LIBRARIES OF SHORT EXPRESSED NUCLEIC ACID SEQUENCES FOR THE ANALYSIS OF CELLULAR EVENTS 审中-公开
    组合合成和使用短细胞核酸序列图案进行细胞活性分析

    公开(公告)号：US20110177973A1

    公开(公告)日：2011-07-21

    申请号：US13120616

    申请日：2009-09-24

    申请人： Moritz Rossner

    发明人： Moritz Rossner

    IPC分类号： C40B30/06 ,  C40B40/02

    CPC分类号： G01N33/5023 ,  C12N15/1065 ,  C12N15/1086 ,  C12Q1/6897

    摘要： The present invention is concerned with the provision of screening assays. More specifically it relates to a method to a method for determining whether an exogenous stimulus is capable of eliciting at least one biological response in a host cell comprising the steps of (i) applying the said stimulus to a host cell which comprises a plurality of different polynucleotide constructs each construct comprising a unique expressed tag (EXT) operatively linked to an expression control sequence whose activity can be exclusively modulated by one specific signalling pathway or cellular sensor implemented in said host cell, said signalling pathway or cellular sensor being involved in eliciting a biological response and being sensitive for the exogenous stimulus, (i) determining the amount of at least one expressed tag; and (iii) comparing the amount of the at least one unique expressed tag to a suitable reference amount, whereby it is determined whether the exogenous stimulus is capable of eliciting at least one biological response in the host cell.

    摘要翻译： 本发明涉及提供筛选测定。 更具体地说，本发明涉及一种用于确定外源刺激是否能够在宿主细胞中引发至少一种生物反应的方法的方法，包括以下步骤：（i）将所述刺激应用于宿主细胞，所述宿主细胞包含多种不同的 多核苷酸构建包含可操作地连接到表达控制序列的唯一表达标签（EXT）的每个构建体，其表达控制序列的活性可以由在所述宿主细胞中实施的一种特定信号传导途径或细胞传感器专一调节，所述信号传导途径或细胞传感器涉及引发 生物反应并对外源刺激敏感，（i）确定至少一个表达标签的量; 和（iii）将至少一个独特表达标签的量与合适的参考量进行比较，由此确定外源刺激是否能够在宿主细胞中引发至少一种生物反应。