Silent inducible virus replicons and uses thereof
    8.
    发明授权
    Silent inducible virus replicons and uses thereof 有权
    沉默诱导型病毒复制子及其用途

    公开(公告)号:US6136538A

    公开(公告)日:2000-10-24

    申请号:US227556

    申请日:1999-01-08

    摘要: A recombinant cell stably transformed with a cDNA of a silent, inducible replicon encoding a recombinant protein is disclosed. Transcription of the replicon cDNA is under the control of a silent promoter inducible by a DNA virus and expression of the recombinant protein is dependent upon the presence of the DNA virus in the cell. The cell can be engineered to package the replicon upon infection by the DNA virus, leading to intercellular amplification of expression of the recombinant protein. Where the recombinant protein is a reporter gene product, the recombinant cell may be used in an assay for detecting DNA viruses. A kit for performing such an assay is also provided.

    摘要翻译: 公开了用编码重组蛋白的沉默的诱导型复制子的cDNA稳定转化的重组细胞。 复制子cDNA的转录受DNA病毒诱导的沉默启动子的控制,重组蛋白的表达取决于DNA病毒在细胞中的存在。 该细胞可被设计成在DNA病毒感染时包装复制子,导致重组蛋白质的表达的细胞间扩增。 当重组蛋白是报告基因产物时,重组细胞可用于检测DNA病毒的测定中。 还提供了用于进行这种测定的试剂盒。

    Detection of negative-strand RNA viruses
    9.
    发明授权
    Detection of negative-strand RNA viruses 有权
    检测负链RNA病毒

    公开(公告)号:US06270958B1

    公开(公告)日:2001-08-07

    申请号:US09253445

    申请日:1999-02-19

    IPC分类号: C12Q170

    摘要: A diagnostic assay for detecting a negative-strand RNA virus in a sample and a genetically engineered cell for use in the assay are disclosed. The cell expresses a heterologous DNA-dependent RNA polymerase that synthesizes a minigenome or miniantigenome of the RNA virus from a cDNA template present in the cell. The cell also expresses the nucleocapsid proteins of the negative-strand virus that are necessary for replication of the minigenome or miniantigenome. Infection of the cell by the negative-strand virus results in expression of a reporter gene product encoded by the miniantigenome.

    摘要翻译: 公开了用于检测样品中的负链RNA病毒的诊断测定法和用于测定中的遗传工程细胞。 细胞表达异源DNA依赖性RNA聚合酶,其从存在于细胞中的cDNA模板合成RNA病毒的小基因组或微型基因组。 细胞还表达了负链病毒的核衣壳蛋白,其是复制小基因组或微型基因组所必需的。 通过负链病毒感染细胞导致由本发明组编码的报告基因产物的表达。

    Indicator cell line for detecting RNA viruses and method therefor
    10.
    发明授权
    Indicator cell line for detecting RNA viruses and method therefor 失效
    用于检测RNA病毒的指示细胞系及其方法

    公开(公告)号:US5591579A

    公开(公告)日:1997-01-07

    申请号:US171214

    申请日:1993-12-21

    摘要: Cell lines and methods are disclosed for detecting the presence of RNA viruses in a specimen. The cell lines are stably transformed with a DNA molecule that includes a promoter capable of being recognized by the DNA dependent RNA polymerase of the cell capable of directing the transcription of a cDNA of a structurally defective RNA virus genome operably coupled to the promoter. The cDNA contains a structural coding sequence encoding a selected reporter gene product. The RNA molecules transcribed by the DNA dependent RNA polymerase are not capable of causing the translation of the reporter gene in the cell except when an active related virus that provides the necessary trans-acting enzymes to cause the increased replication of the RNA containing the reporter gene which is then translated into the reporter gene product is provided. Methods utilizing the cell lines of this invention to detect RNA viruses in a specimen by incubating the specimen with the cell line and assaying for expression of the reporter gene and a kit containing a supply of the cells and a supply of the reagents necessary for the detection of the reporter gene product are also provided.

    摘要翻译: 公开了用于检测样品中RNA病毒的存在的细胞系和方法。 细胞系被包括能够被能够引导结构有缺陷的RNA病毒基因的cDNA的转录的细胞的DNA依赖性RNA聚合酶识别的启动子的DNA分子稳定转化,所述基因可操作地连接到启动子上。 cDNA含有编码所选报道基因产物的结构编码序列。 由DNA依赖性RNA聚合酶转录的RNA分子不能引起报告基因在细胞中的翻译,除了当提供必需的反式作用酶以引起含有报告基因的RNA的复制增加的活性相关病毒时 然后将其翻译成报告基因产物。 利用本发明的细胞系的方法通过将样品与细胞系一起孵育并测定报告基因的表达和含有细胞供应的试剂盒和检测所必需的试剂的供应来检测标本中的RNA病毒 的报告基因产物也被提供。