RNA适体的研制及其芯片电泳配体结合试验。

Current chemical genomics Pub Date : 2012-01-01 Epub Date: 2012-01-24 DOI:10.2174/1875397301206010001
Ken-Ichi Ohno, Chikara Nakata, Yoshihiro Sano, Fumiko Nishikawa, Satoshi Nishikawa, Hidetoshi Arakawa
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引用次数: 3

摘要

采用微芯片电泳(ME)结合荧光检测,通过靶分子的指数富集(SELEX)轮来估计配体在每个系统进化中的适配体结合活性。这种方法是一种无放射性同位素、快速和简单的平台,电泳分离似乎是寡核苷酸分子适配体的有效技术。我们试图通过上述方法获得促性腺激素特异性RNA适配体。结果,基于它们之间构象差异的适配体的峰被分离并在电泳图上检测。此外,未结合的核酸适体的峰强度随着SELEX轮的进展而降低,表明该方法获得了高亲和力的RNA适体。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

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Development of RNA aptamer and its ligand binding assay on microchip electrophoresis.

Microchip electrophoresis (ME) coupled with fluorescence detection was used to estimate the binding activity of aptamer in each systematic evolution of ligands by exponential enrichment (SELEX) round for a target molecule. This approach is a non-radioisotopic, rapid and simple platform, and electrophoretic separation appears to be an effective technique for aptamers of oligonucleotide molecules. We tried to obtain gonadotropin-specific RNA aptamer by the above approach. As a result, the peaks of aptamers based on the conformational differences between them were separated and detected on the electropherograms. Moreover, the intensity of peak of unbound aptamer was decreased with progression through the SELEX rounds, suggesting that RNA aptamer with high affinity was obtained by the proposed method.

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