2',4'-BNA/LNA 适体：使用全长度 2'-O、4'-C-亚甲基桥接/连接双环核糖核苷酸的基于 DNA 的文库进行 CE-SELEX。

2',4'-BNA/LNA aptamers: CE-SELEX using a DNA-based library of full-length 2'-O,4'-C-methylene-bridged/linked bicyclic ribonucleotides.

机构信息

Graduate School of Engineering, Gunma University, 1-5-1 Tenjin-cho, Kiryu, Gunma 376-8515, Japan.

出版信息

Bioorg Med Chem Lett. 2013 Mar 1;23(5):1288-92. doi: 10.1016/j.bmcl.2012.12.093. Epub 2013 Jan 16.

DOI:10.1016/j.bmcl.2012.12.093

Abstract

DNA-based aptamers that contain 2'-O,4'-C-methylene-bridged/linked bicyclic ribonucleotides (B/L nucleotides) over the entire length were successfully obtained using a capillary electrophoresis systematic evolution of ligands by exponential enrichment (CE-SELEX) method. A modified DNA library was prepared with an enzyme mix of KOD Dash and KOD mutant DNA polymerases. Forty 2'-O,4'-C-methylene bridged/locked nucleic acid (2',4'-BNA/LNA) aptamers were isolated from an enriched pool and classified into six groups according to their sequence. 2',4'-BNA/LNA aptamers of groups V and VI bound human thrombin with K(d) values in the range of several 10 nanomolar levels.

摘要

通过毛细管电泳指数富集配体系统进化（CE-SELEX）方法，成功获得了全长含有 2'-O，4'-C-亚甲基桥/连接双环核糖核苷酸（B/L 核苷酸）的基于 DNA 的适体。使用 KOD Dash 和 KOD 突变 DNA 聚合酶的酶混合物制备了修饰的 DNA 文库。从富集池中分离出 40 个 2'-O，4'-C-亚甲基桥/锁核酸（2'，4'-BNA/LNA）适体，并根据其序列分为六组。第 V 和第 VI 组的 2'，4'-BNA/LNA 适体与人凝血酶结合，K（d）值在几个 10 纳摩尔水平范围内。