Miller P S, Agris C H, Murakami A, Reddy P M, Spitz S A, Ts'o P O
Nucleic Acids Res. 1983 Sep 24;11(18):6225-42. doi: 10.1093/nar/11.18.6225.
An efficient procedure is described for synthesizing deoxyribonucleoside methylphosphonates on polystyrene polymer supports which involves condensing 5'-dimethoxytrityldeoxynucleoside 3'-methylphosphonates. The oligomers are removed from the support and the base protecting groups hydrolyzed by treatment with ethylenediamine in ethanol, which avoids hydrolysis of the methylphosphonate linkages. Two types of oligomers were synthesized: those containing only methylphosphonate linkages, d-Np(Np)nN, and those which terminate with a 5' nucleotide residue, dNp (Np)nN. The latter oligomers can be phosphorylated by polynucleotide kinase, and are separated by polyacrylamide gel electrophoresis according to their chain length. Piperdine randomly cleaves the oligomer methylphosphonate linkages and generates a series of shorter oligomers whose number corresponds to the length of the original oligomer. Apurinic sites introduced by acid treatment spontaneously hydrolyze to give oligomers which terminate with free 3' and 5' OH groups. These reactions may be used to characterize the oligomers.
本文描述了一种在聚苯乙烯聚合物载体上合成脱氧核糖核苷甲基膦酸酯的有效方法，该方法涉及缩合5'-二甲氧基三苯甲基脱氧核苷3'-甲基膦酸酯。通过在乙醇中用乙二胺处理，将低聚物从载体上除去并水解碱基保护基团，从而避免甲基膦酸酯键的水解。合成了两种类型的低聚物：仅含甲基膦酸酯键的低聚物，即d-Np(Np)nN，以及以5'核苷酸残基结尾的低聚物，即dNp (Np)nN。后一种低聚物可被多核苷酸激酶磷酸化，并根据其链长通过聚丙烯酰胺凝胶电泳进行分离。哌啶可随机切割低聚物的甲基膦酸酯键，并生成一系列较短的低聚物，其数量与原始低聚物的长度相对应。经酸处理引入的脱嘌呤位点会自发水解，产生以游离3'和5'羟基结尾的低聚物。这些反应可用于表征低聚物。
