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    • 1. 发明授权
    • Method and construct for screening for inhibitors of transcriptional
activation
    • 用于筛选转录激活抑制剂的方法和构建
    • US5747338A
    • 1998-05-05
    • US719577
    • 1996-09-25
    • Klaus GieseJaime Escobedo
    • Klaus GieseJaime Escobedo
    • C12Q1/68C12N5/10
    • C12Q1/6897
    • A collision construct is provided that contains a first regulatory sequence containing a first promoter linked to a reporter gene which is also linked to a second regulatory sequence containing a second promoter such that the reporter gene is under regulatory control of the first promoter. The direction of transcription under the first promoter is opposite the direction of transcription under the second promoter and activation of the second promoter interferes with reporter gene activity. The collision construct is used to screen inhibitors of the second regulatory sequence and the second promoter whereby a functional inhibitor causes an enhanced signal of the reporter gene. Vectors, host cells and kits containing the collision construct are provided, as well as methods for producing the collision construct, the vectors and host cells, and methods for screening candidate inhibitors for their ability to inhibit target promoter activities.
    • 提供了一种碰撞构建体,其包含含有与报告基因连接的第一启动子的第一调控序列,其也与含有第二启动子的第二调控序列连接,使得报道基因处于第一启动子的调控下。 第一启动子下的转录方向与第二启动子下的转录方向相反,第二启动子的活化会干扰报告基因的活性。 碰撞构建体用于筛选第二调节序列和第二启动子的抑制剂,由此功能性抑制剂引起报告基因的增强的信号。 提供载体,宿主细胞和含有碰撞构建体的试剂盒,以及用于产生碰撞构建体,载体和宿主细胞的方法以及用于筛选候选抑制剂以抑制靶启动子活性的能力的方法。
    • 5. 发明申请
    • Human platelet-derived growth factor receptors
    • 人血小板衍生生长因子受体
    • US20070178527A1
    • 2007-08-02
    • US11495096
    • 2006-07-28
    • Lewis WilliamsJaime Escobedo
    • Lewis WilliamsJaime Escobedo
    • G01N33/53C07H21/04C12P21/06C07K14/71
    • C07K14/71G01N33/74G01N2333/49
    • DNA sequences encoding human platelet-derived growth factor receptors (hPDGF-R), and expression constructs comprising sequences which encode a receptor that can be secreted or incorporated into the membrane of a mammalian cell. Peptide fragments with functions equivalent to the wild-type receptor, conferring a PDGF-sensitive mitogenic response on cells lacking the receptor are provided. The constructs can be used for enhancing PDGF response of cells, determining the regions involved in transducing the signal in response to PDGF binding, providing mutated analogs and evaluating drugs for their physiologic activity. Soluble fragments comprising PDGF receptor sequences are also provided, including important intracellular kinase insert sequences which interact with intracellular proteins.
    • 编码人血小板衍生生长因子受体(hPDGF-R)的DNA序列和包含编码可以分泌或并入哺乳动物细胞膜的受体的序列的表达构建体。 提供了功能相当于野生型受体的肽片段,赋予缺乏受体的细胞对PDGF敏感的促有丝分裂反应。 该构建体可用于增强细胞的PDGF应答,确定与PDGF结合相关的转导信号所涉及的区域,提供突变的类似物并评估药物的生理活性。 还提供了包含PDGF受体序列的可溶性片段,包括与细胞内蛋白质相互作用的重要细胞内激酶插入序列。