将末端硫代磷酸酯掺入寡核苷酸中。
Incorporation of terminal phosphorothioates into oligonucleotides.
作者信息
Alefelder S, Patel B K, Eckstein F
机构信息
Max-Planck-Institut für experimentelle Medizin, Hermann-Rein-Strasse 3, D-37075 Göttingen, Germany.
出版信息
Nucleic Acids Res. 1998 Nov 1;26(21):4983-8. doi: 10.1093/nar/26.21.4983.
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 post-synthetic 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 beta-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.
人们在研究寡核苷酸的结构和功能方面投入了大量精力，有几种方法依赖于将报告基团连接到寡核苷酸上。我们在此报告将3'-和5'-末端硫代磷酸酯引入七聚体寡核苷酸及其用几种报告基团进行的合成后修饰。末端硫代磷酸酯的合成基于分别在第一个或最后一个核苷酸处偶联核糖核苷亚磷酰胺，硫化后，通过邻位羟基的顺序氧化然后β-消除将其除去。产物形成率约为95%。通过在Hg2+凝胶上进行电泳分析，硫代磷酸酯末端与磷酸酯末端的寡脱氧核苷酸的比例通常为85/15。描述了末端硫代磷酸酯与胆固醇、双硫腙缀合以及巯基交换反应性的实例。
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