Incorporation of terminal phosphorothioates into oligonucleotides
Incorporation of terminal phosphorothioates into oligonucleotides
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DOI：
10.1093/nar/26.21.4983
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发表时间：
1998-11-01
影响因子：
14.9
通讯作者：
Eckstein, F
中科院分区：
文献类型：
--
作者：
Alefelder, S;Patel, BK;Eckstein, F
人们付出了大量的努力来研究寡核苷酸的结构和功能，并且一些方法依赖于报告基团与寡核苷酸的连接。我们在这里报告了将 3'-和 5'-末端硫代磷酸酯引入七聚寡核苷酸中，并用几个报告基团对其进行合成后修饰。末端硫代磷酸酯的合成基于核糖核苷亚磷酰胺分别在第一个或最后一个核苷酸处的偶联，在硫化后，通过邻位羟基的连续氧化和p-消除来去除该核糖核苷亚磷酰胺，产物形成率为95％左右，通过在Hg2+凝胶上电泳分析，硫代磷酸酯与磷酸酯封端的寡脱氧核苷酸的比率通常为85/15。描述了末端硫代磷酸酯与胆固醇、bimane 缀合和巯基交换的反应性实例。
Considerable effort has been directed towards studying the structure and function of oligonucleotides and several approaches rely on the attachment of reporter groups to oligonucleotides. We report here the introduction of 3'- and 5'-terminal phosphorothioates into heptameric oligonucleotides and their postsynthetic modification with several reporter groups. The synthesis of terminal phosphorothioates is based on the coupling of a ribonucleoside phosphoramidite at the first or last nucleotide, respectively, which, after sulphurization, is removed by sequential oxidation of the vicinal hydroxyl groups and then p-elimination, Product formation is of the order of 95%, The ratio of phosphorothioate- versus phosphate-terminated oligodeoxynucleotides as analysed by electrophoresis on a Hg2+ gel is in general 85/15. Examples for the reactivity of the terminal phosphorothioates for conjugation with cholesterol, bimane and for sulphydryl exchange are described.