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    • 10. 发明授权
    • Sulfonamide-derivative compounds for tagging nucleic acid probes
    • 用于标记核酸探针的磺酰胺衍生化合物
    • US5210203A
    • 1993-05-11
    • US879593
    • 1992-05-07
    • Gary F. MussoSoumitra GhoshEmil T. Kaiser
    • Gary F. MussoSoumitra GhoshEmil T. Kaiser
    • C07F7/18C07H21/00C12Q1/68
    • C07H21/00C07F7/1852C07F7/1856C12Q1/6816
    • Nucleic acid hybridization probes are provided which comprise an N.sup.4 -(substituted amino)cytosine moiety, wherein the substituted amino group comprises a tag moiety, whereby the probe is detected. Methods of preparing probes of the invention, intermediates used in such methods, and methods of using the probes of the invention in hybridization assays are also provided. Typical tag moieties employed with the probes of the invention are biotinyl, aminothiadiazole and fluorescein derivatives, connected to N.sup.4 -amino groups of modified cytosines of the probe through linker moieties. Probes tagged with biotin are typically detected by binding to the biotinyl moieties, through a streptavidin or avidin molecule, a reporter group which includes streptavidin or avidin and then detecting a signal due to the reporter group. Probes tagged with aminothiadiazole derivatives are typically detected by essentially the same method as those tagged with biotinyl but employing as reporter group one which binds to the derivative through a carbonic anhydrase molecule. Probes tagged with fluorescein derivatives are detected by a fluorescence spectroscopic method without binding of a reporter group to the tag.
    • 提供了包含N4-(取代氨基)胞嘧啶部分的核酸杂交探针,其中取代的氨基包括标签部分,由此检测探针。 还提供了制备本发明的探针的方法,在这些方法中使用的中间体以及在杂交测定中使用本发明的探针的方法。 与本发明的探针一起使用的典型标签部分是通过接头部分连接到探针的修饰胞嘧啶的N4-氨基的生物素,氨基噻二唑和荧光素衍生物。 用生物素标记的探针通常通过结合生物素部分,通过链霉抗生物素蛋白或抗生物素蛋白分子,包括链霉抗生物素蛋白或抗生物素蛋白的报道基团检测,然后检测由于报告基团引起的信号。 用氨基噻二唑衍生物标记的探针通常用与生物素标记的那些基本上相同的方法来检测,但采用作为通过碳酸酐酶分子与衍生物结合的报告基团。 用荧光素衍生物标记的探针通过荧光光谱法检测,而不将报道基团与标签结合。