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    Methods for identifying inhibitors of chlorophyll synthase
    9.
    发明申请
    Methods for identifying inhibitors of chlorophyll synthase 审中-公开
    鉴定叶绿素合酶抑制剂的方法

    公开(公告)号：US20050130118A1

    公开(公告)日：2005-06-16

    申请号：US10737014

    申请日：2003-12-16

    申请人： Abubakr Aslamkhan ,  Lining Guo ,  Rao Mulpuri ,  Neil Hoffman ,  Susanne Kjemtrup ,  Jocelyn Kearney ,  Cory Christensen ,  Keith Davis ,  Adel Zayed ,  Robert Ascenzi ,  Douglas Boyes

    发明人： Abubakr Aslamkhan ,  Lining Guo ,  Rao Mulpuri ,  Neil Hoffman ,  Susanne Kjemtrup ,  Jocelyn Kearney ,  Cory Christensen ,  Keith Davis ,  Adel Zayed ,  Robert Ascenzi ,  Douglas Boyes

    IPC分类号： C12Q1/00 ,  G01N33/573

    CPC分类号： G01N33/573

    摘要： The present inventors have discovered that antisense suppression of a chlorophyll synthase (CS) gene results in plants exhibiting one or more of chlorosis, reduced growth, and altered development. Thus, the present inventors have discovered that the protein encoded by the CS gene is essential for normal plant growth and development, and is useful as a target for the identification of compounds as antibiotics and herbicides, especially herbicides. The present invention is directed to methods for identifying inhibitors of a CS enzyme by incubating a CS polypeptide with a chlorophyllide and a phopholipid substrate in the presence and absence of a test compound under conditions suitable for the CS enzyme activity, adding a solution to the incubation reactions comprising a water immiscible organic solvent, a water-soluble alcohol, and a water-soluble dye that absorbs in the range of one or both the excitation and emission wavelength ranges of the chlorophyllide substrate, and measuring the fluorescence of the incubation reactions at from about 650 to 750 nm, using from about 425 to 445 nm as excitation wavelength, wherein a decrease in the fluorescence in the presence of the test compound indicates that the compound is a CS inhibitor.

    摘要翻译： 本发明人已经发现，叶绿素合酶（CS）基因的反义抑制导致植物表现出一种或多种褪绿，生长减少和改变发育。 因此，本发明人已经发现，由CS基因编码的蛋白质对于正常的植物生长和发育是必需的，并且可用作鉴定作为抗生素和除草剂，特别是除草剂的化合物的靶标。 本发明涉及通过在适合于CS酶活性的条件下，在存在和不存在测试化合物的情况下，将CS多肽与叶绿素和磷脂底物一起温育来鉴定CS酶抑制剂的方法，向培养基中加入溶液 包括与水不溶混的有机溶剂，水溶性醇和在所述叶绿素底物的激发和发射波长范围中的一个或两个的范围内吸收的水溶性染料的反应，以及测量从 约650至750nm，使用约425至445nm作为激发波长，其中在测试化合物存在下荧光的降低表明该化合物是CS抑制剂。