一种用于合成在β位特异性标记有无机[32P]磷酸盐的核苷三磷酸的新方法。
A novel method for the synthesis of nucleoside triphosphates labelled with inorganic [32P]phosphate specifically in the beta-position.
作者信息
Abraham K A
出版信息
Biochem J. 1977 Mar 1;161(3):615-7. doi: 10.1042/bj1610615.
A method was developed for the introduction of [32p]Pi specifically into the beta-position of ATP and GTP. The method is based on two separate reactions involving (a) phosphorolysis of poly(A) or poly(G) [Soreq, Nudel, Salomon, Revel & Littauer (1974) J. Mol Biol. 88, 233-245] in the presence of [32P]Pi and (b) conversion of the labelled diphosphate into the corresponding triphosphate by transferring the active phosphate group from 1,3-diphosphoglycerate in a coupled reaction as decribed by Glynn & Chappell [(1964) Biochem. J. 90, 147-149]. Radioactivity in the beta- and gamma-phosphate groups of the labelled triphosphate was measured by using polynucleotide kinase. No detectable radioactivity was found in the gamma-phosphate group. The suitability of this method for the synthesis of other nucleoside triphosphates specifically labelled in the beta-position is discussed.
已开发出一种方法，可将[32P]Pi特异性引入ATP和GTP的β位。该方法基于两个独立的反应，包括（a）在[32P]Pi存在下对聚（A）或聚（G）进行磷酸解[索雷克、努德尔、萨洛蒙、雷维尔和利陶尔（1974年）《分子生物学杂志》88卷，233 - 245页]，以及（b）如格林和查佩尔所述[（1964年）《生物化学杂志》90卷，147 - 149页]，通过在偶联反应中从1,3 - 二磷酸甘油酸转移活性磷酸基团，将标记的二磷酸转化为相应的三磷酸。使用多核苷酸激酶测量标记三磷酸的β和γ磷酸基团中的放射性。在γ磷酸基团中未发现可检测到的放射性。讨论了该方法对于合成在β位特异性标记的其他核苷三磷酸的适用性。
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