In vitro selection of DNA aptamers that bind L-Tyrosinamide

被引:85
作者
Vianini, E [1 ]
Palumbo, M [1 ]
Barbara, G [1 ]
机构
[1] Univ Padua, Dept Pharmaceut Sci, I-35131 Padua, Italy
关键词
D O I
10.1016/S0968-0896(01)00054-2
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We have applied SELEX (Systematic Evolution of Ligands by EXponential enrichment), a combinatorial method that employs biopolymers for drug discovery, to identify single stranded DNA sequences able to bind L-Tyrosinamide, a simple mimic of Tyrosine, an amino acid essential to the catalytic activity of several enzymes of pharmaceutical interest. After 15 SELEX cycles using L-Tyrosinamide immobilized on an affinity chromatography column, the percentage of aptamers specifically eluted from the affinity column with free L-Tyrosinamide was 55% of the total. Aptamers were subcloned and sequenced, allowing the identification of a highly conserved consensus sequence, and showed a K-d value for L-Tyrosinamide of 45 muM. The identified aptamer sequence will constitute the basis for further in vitro evolution protocols and structure-based drug design. (C). 2001 Elsevier Science Ltd. All rights reserved.
引用
收藏
页码:2543 / 2548
页数:6
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