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    • 10. 发明申请
    • FLUORESCENCE ASSAY FOR DNA MODIFYING ENZYMES
    • DNA修饰酶的荧光测定
    • WO2002072891A1
    • 2002-09-19
    • PCT/US2002/007413
    • 2002-03-11
    • UNIVERSITY OF CALIFORNIAREICH, Norbert, O.ALLAN, Barrett, W.LINDSTROM, William, Jr.PUNTZKE, Aaron, P.
    • REICH, Norbert, O.ALLAN, Barrett, W.LINDSTROM, William, Jr.PUNTZKE, Aaron, P.
    • C12Q1/68
    • C12Q1/6837C12Q2565/107
    • A method of assaying compounds for their ability to effect enzymes including enhancing or inhibiting the effect of those enzymes on double stranded DNA sequences is disclosed. The method comprises providing a modified nucleotide sequence comprised of a base analogue which analogue is characterized by increased fluorescence when moved out of its normal helical position, the sequence having a complimentary sequence hybridized thereto to provide a double stranded sequence. The modified sequence containing the base analogue is brought into contact with the enzyme which enzyme is characterized by effecting the 3-dimensional position of the analogue within the sequence. The enzyme is brought into contact with the sequences in the presence of a compound being assayed. By knowing the amount of increased fluorescence the enzyme would normally have on the sequence is possible to determine the inhibitory or enhancing effect of the compound on the enzyme.
    • 公开了一种测定化合物其影响酶的能力的方法,包括增强或抑制这些酶对双链DNA序列的影响。 所述方法包括提供由碱基类似物组成的修饰的核苷酸序列,所述碱基类似物的特征在于当其从其正常螺旋位置移出时增加荧光,所述序列具有与其杂交的互补序列以提供双链序列。 使含有碱基类似物的修饰的序列与该酶的酶接触,酶的特征在于在序列内实现类似物的3维位置。 在待测化合物存在下使酶与序列接触。 通过了解酶通常对该序列具有的荧光增加的量可以确定化合物对酶的抑制或增强的作用。