生物素标记试剂的合成方法改进及其在DNA合成中的应用

摘要

A biotin-labelled reagent, [1-N-(4,4'-dimethoxytrityl)-biotinyl-6-aminohexyl]-2-cyano-ethyl-N,N-diisopropylaminophosphoramidite(3) , was synthesized in total yield of 51.0% from bi-otion, 4,4'-dimethoxytrityl chloride, 6-aminohexanol and 2-cyanoethyl-N,N-diisopropylamin-ophos-phoramidite by improvement method. The structure was confirmed by 1H NMR, 13C NMR, 31P NMR and MS. 3 was used to produce 5'-biotinylated oligonucleotides by automated solid-phase synthesis. The structure was confirmed by MS.%以生物素,4,4′-二甲氧基三苯基氯甲烷,6-氨基-1-己醇,2-氰基乙氧基-双(N,N-二异丙基)亚磷酰胺等为原料,采用改进方法合成了生物素标记试剂——[1-N-(4,4′-二甲氧基三苯甲基)-生物素-6-氨基己基]-2-氰乙氧基-N,N-二异丙基亚磷酰胺(3),总收率51.0％,其结构经1H NMR,13C NMR,31p NMR和MS确证.通过固相亚磷酰胺三酯法,应用3合成了一段含有十个碱基的5′-端生物素标记的寡核苷酸链,其结构经MS确证.