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电化学 microRNA 检测的新见解:分子卡尺,p19 蛋白。

A new insight into electrochemical microRNA detection: a molecular caliper, p19 protein.

机构信息

Faculty of Engineering and Architecture, Department of Biomedical Engineering, Izmir Katip Celebi University, Turkey.

出版信息

Biosens Bioelectron. 2013 Oct 15;48:165-71. doi: 10.1016/j.bios.2013.04.011. Epub 2013 Apr 20.

Abstract

microRNA (miRNA) has drawn a great attention in biomedical research due to its functions on biological processes. Detection of miRNAs is a big challenge since the amount present in real samples is very low and the length of them is short. In this study, for the first time an electrochemical biosensor for detection of mir21 using the oxidation signal of protein 19 (p19) as a molecular caliper was designed. The proposed method enables detection of mir21 in direct, rapid, sensitive, inexpensive and label-free way. Binding specificity of the p19 to 20-23 base pair length double stranded RNA (dsRNA) and direct/water-mediated intermolecular contacts between the fusion protein and miRNA allows detection of miRNA-antimiRNA hybrid structure. The detection of mir21 was achieved in picomole sensitivity through the changes of intrinsic p19 oxidation signals observed at +0.80 V with Differential Pulse Voltammetry (DPV) and the specifity of the designed sensor was proved by control studies.

摘要

微小 RNA(miRNA)因其在生物过程中的功能而在生物医学研究中受到极大关注。由于真实样本中存在的 miRNA 数量非常低,而且它们的长度很短,因此 miRNA 的检测是一个巨大的挑战。在这项研究中,首次设计了一种基于电化学生物传感器的方法,该方法利用蛋白 19(p19)的氧化信号作为分子卡尺来检测 mir21。该方法可直接、快速、灵敏、廉价、无需标记地检测 mir21。p19 与 20-23 碱基对长度双链 RNA(dsRNA)的结合特异性以及融合蛋白与 miRNA 之间的直接/水介导的分子间接触允许检测 miRNA-反 miRNA 杂交结构。通过差分脉冲伏安法(DPV)观察到的固有 p19 氧化信号的变化,以皮摩尔灵敏度检测到 mir21,并且通过控制研究证明了设计传感器的特异性。

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