针对甜味剂阿斯巴甜的修饰DNA适配体。

Modified DNA aptamers against sweet agent aspartame.

作者信息

Saitoh Hiroshi, Nakamura Akiko, Kuwahara Masayasu, Ozaki Hiroaki, Sawai Hiroaki

机构信息

Department of Applied Chemistry, Faculty of Engineering, Gunma University, Kiryu, Gunma 376-8515, Japan.

出版信息

Nucleic Acids Res Suppl. 2002(2):215-6. doi: 10.1093/nass/2.1.215.

DOI:10.1093/nass/2.1.215

PMID:12903182

Abstract

We obtained a modified DNA aptamer against sweetener, aspartame, by in vitro selection method. The modified DNA was prepared from dATP, dGTP, dCTP and a modified dTTP bearing a terminal amino group at C-5 position in place of thymidine by PCR using a hyper thermophilic DNA polymerase, KOD Dash DNA polymerase. The synthetic 102-mer DNA with a 60-mer random region was used as an initial template for the PCR. The PCR-amplified modified DNA library was applied to an aspartame-agarose column, and then the bound modified DNA was eluted from the column for the affinity chromatography selection. Repeating the procedure, we selected the modified DNA aptamer against aspartame.

摘要

我们通过体外筛选方法获得了一种针对甜味剂阿斯巴甜的修饰DNA适配体。修饰DNA由dATP、dGTP、dCTP和一种在C-5位带有末端氨基取代胸腺嘧啶的修饰dTTP，使用超嗜热DNA聚合酶KOD Dash DNA聚合酶通过PCR制备。具有60个碱基随机区域的合成102聚体DNA用作PCR的初始模板。将PCR扩增的修饰DNA文库应用于阿斯巴甜琼脂糖柱，然后从柱上洗脱结合的修饰DNA用于亲和色谱筛选。重复该过程，我们筛选出了针对阿斯巴甜的修饰DNA适配体。