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    • 4. 发明授权
    • &ggr;-phosphoester nucleoside triphosphates
    • γ-磷酸酯核苷三磷酸
    • US06399335B1
    • 2002-06-04
    • US09441108
    • 1999-11-16
    • C. Cheng KaoTheodore WidlanskiWilliam VassiliouJeffrey Epp
    • C. Cheng KaoTheodore WidlanskiWilliam VassiliouJeffrey Epp
    • C12Q168
    • C12P19/34
    • The invention provides methods and compositions for polymerizing a particular nucleotide with a polymerase. In general, the method involves (a) forming a mixture of a polymerase and a nucleoside triphosphate (NTP) comprising &agr;, &bgr; and &ggr; phosphates and a &ggr;-phosphate phosphoester-linked functional group; and incubating the mixture under conditions wherein the polymerase catalyzes cleavage of the NTP between the &agr; and &bgr; phosphates, liberating a pyrophosphate comprising the functional group and polymerizing the resultant nucleoside monophosphate, i.e. incorporates the nucleoside monophosphate in a nascent polynucleotide. A variety of functional groups compatible with the polymerization reaction are provided.
    • 本发明提供了用聚合酶聚合特定核苷酸的方法和组合物。 通常,该方法包括(a)形成聚合酶和包含α,β和γ磷酸盐和γ-磷酸磷酸酯连接的官能团的核苷三磷酸(NTP)的混合物; 并且在所述聚合酶催化所述α和β磷酸盐之间的NTP裂解的条件下孵育所述混合物,释放包含所述官能团的焦磷酸盐并聚合所得核苷单磷酸酯,即在新生多核苷酸中掺入核苷单磷酸酯。 提供了与聚合反应相容的各种官能团。