Process 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 ethylenediamene in ethanol, which avoids hydrolysis of the methylphosphonate linkages. Two types of oligomers are synthesized: those containing only methylphosphonate linkages, d-Np(Np).sub.n N, and those which terminate with a 5' nucleotide residue, dNp(Np).sub.n N. The latter oligomers can be phosphorylated by polynucleotide kinase, and are separated by polyacrylamide gel electrophoresis according to their chain length. Piperidine 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.
在聚
苯乙烯聚合物支持上合成去氧核苷
甲基膦酸酯的过程包括将5'-二甲氧基三苯甲基去氧核苷3'-
甲基膦酸酯缩合。寡聚物从支持物中移除,并通过
乙二胺在
乙醇中处理以
水解碱基保护基团,避免
水解
甲基膦酸酯键。合成两种类型的寡聚物:仅含
膦酸甲酯键的寡聚物,d-Np(Np)。sub.n N,以及以5'核苷酸残基为终止的寡聚物,dNp(Np)。sub.n N。后者的寡聚物可以被多核苷酸激酶
磷酸化,并根据其链长通过聚
丙烯酰胺凝胶电泳进行分离。
吡啶随机裂解寡聚物的
甲基膦酸酯键,并生成一系列长度与原始寡聚物长度相对应的较短寡聚物。酸处理引入的缺失突变位点会自发
水解,形成以自由3'和5'-OH基团为终止的寡聚物。这些反应可用于表征寡聚物。