一种制备[β-32P]嘌呤核苷三磷酸的简单方法。
A simple method for the preparation of [beta-32P]purine nucleoside triphosphase.
作者信息
Furuichi Y, Shatkin A J
出版信息
Nucleic Acids Res. 1977 Oct;4(10):3341-55. doi: 10.1093/nar/4.10.3341.
A rapid, simple and inexpensive procedure is described for the preparation of purine ribo-and deoxyribonucleoside triphosphates specifically and highly radiolabeled with [32P]phosphate in the beta position. The method involves two successive enzymatic reactions: conversion of donor [gamma-32P]ATP in the presence of an excess of acceptor 5'-mononucleotide to the 5'-diphosphates by myokinase or guanosine 5'-monophosphate kinase followed by phosphorylation with pyruvate kinase to yield 5'-triphosphates.
描述了一种快速、简单且廉价的方法，用于制备在β位用[32P]磷酸盐特异性且高度放射性标记的嘌呤核糖核苷三磷酸和脱氧核糖核苷三磷酸。该方法涉及两个连续的酶促反应：在过量的受体5'-单核苷酸存在下，通过肌激酶或鸟苷5'-单磷酸激酶将供体[γ-32P]ATP转化为5'-二磷酸，然后用丙酮酸激酶进行磷酸化以产生5'-三磷酸。
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