期刊
ANALYTICAL CHEMISTRY
卷 85, 期 10, 页码 4961-4967出版社
AMER CHEMICAL SOC
DOI: 10.1021/ac400058z
关键词
-
资金
- New Energy and Industrial Technology Development Organization (NEDO) of Japan
- Japan Society for the Promotion of Science (JSPS) [25350962]
- JSPS for Young Scientists
- Grants-in-Aid for Scientific Research [11J10396, 25350962] Funding Source: KAKEN
Chemically modified DNA aptamers specific to human alpha-thrombin were obtained from oligodeoxyribonucleotide (ODN) libraries by using a capillary electrophoresis-systematic evolution of ligands by exponential enrichment (CE-SELEX) method. These libraries contained 2'-O,4'-C-methylene-bridged/linked bicyclic ribonucleotides (B/L nucleotides) in the primer region and/or C5-modified thymidine Migration time bearing N-6-ethyladenine (t) in the nonprimer region. Modified DNA aptamers showed high binding affinities to the target, with dissociation constants (K-d) values in the range of subnanomolar to several ten nanomolar levels. The introduction of base modification significantly suppressed the frequency of G-quadruplex motifs, which are often seen in thrombin-binding DNA aptamers. The resulting alternatives contained the 10-mer consensus sequence t(5)Gt(2)G(2), which is frequently found in modified DNA aptamers with subnanomolar protein binding affinities. Furthermore, some base- and sugar-modified DNA aptamers with the 12-mer consensus sequence t(2)G(2)tC(A/G)A(2)G(2)t displayed binding activities that were dependent on the presence of B/L nucleotides in the primer region. Such aptamers were interestingly not recovered from a natural DNA library or from DNA libraries modified with either B/L nucleotides or t's. This emerging characteristic binding property will enable the creation of a direct selection methodology for DNA-based molecular switches that are triggered by chemical conversion of B/L nucleotides introduced to constant sequence regions in ODN libraries.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据