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    POLYSOME-MEDIATED CELL TYPE-, TISSUE TYPE- OR CONDITION-ENHANCED TRANSCRIPT PROFILING
    47.
    发明申请
    POLYSOME-MEDIATED CELL TYPE-, TISSUE TYPE- OR CONDITION-ENHANCED TRANSCRIPT PROFILING 审中-公开
    多聚体介质细胞类型，组织类型或状态增强转录物

    公开(公告)号：US20100071086A1

    公开(公告)日：2010-03-18

    申请号：US12557449

    申请日：2009-09-10

    申请人： Peter P. Repetti ,  Oliver J. Ratcliffe ,  Luc J. Adam ,  T. Lynne Reuber ,  Hans E. Holtan

    发明人： Peter P. Repetti ,  Oliver J. Ratcliffe ,  Luc J. Adam ,  T. Lynne Reuber ,  Hans E. Holtan

    IPC分类号： C12N15/82 ,  A01H5/00 ,  C12N5/04 ,  C12N15/74

    CPC分类号： C12N15/8216

    摘要： In this invention, a method is described that allows for the efficient creation and identification of validated biological materials that greatly enhance the ability to perform polysome-mediated RNA profiling, such as constitutive, cell type-, tissue type-, or condition-enhanced RNA profiling. The method relies on the use of a tri-partite plant binary expression vector comprised of the following components: a) a DNA promoter element that drives expression of a sequence specific transcription activator protein such as a LexA:Gal4 fusion protein in a unique desired pattern, b) a DNA promoter element comprising a target site for the transcriptional activator protein, such as opLexA, fused to a nucleotide encoding an epitope tagged ribosomal component protein and c) a DNA promoter element comprising a target site for the transcriptional activator protein, such as opLexA, fused to a nucleotide encoding an in vivo reporter protein. By visualization of the co-regulated reporter, this method allows for in planta confirmation that the promoter element is driving expression, such as constitutive, cell type-, tissue type-, or condition-enhanced expression, of the tagged ribosomal protein in the desired cell or tissue types.

    摘要翻译： 在本发明中，描述了允许有效创建和鉴定验证的生物材料的方法，其极大地增强了进行多聚体介导的RNA分析的能力，例如组成型，细胞类型，组织型或条件增强型RNA 剖析。 该方法依赖于使用由以下组分组成的三分子植物二元表达载体：a）以独特的期望模式驱动序列特异性转录激活蛋白如LexA：Gal4融合蛋白的表达的DNA启动子元件 b）包含与编码表位标记的核糖体组分蛋白的核苷酸融合的转录激活蛋白的靶位点如opLexA的DNA启动子元件，和c）包含转录激活蛋白的靶位点的DNA启动子元件，例如 作为opLexA，与编码体内报道蛋白的核苷酸融合。 通过共同调节的报告者的可视化，这种方法允许在植物中确认启动子元件是驱动所需标记的核糖体蛋白质的组成型，细胞类型，组织型或条件增强表达的表达 细胞或组织类型。