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哑铃状金-氧化铁磁性纳米粒子对 DNA 的原子力显微镜成像研究

Atomic Force Microscopy Imaging Study of Aligning DNA by Dumbbell-like Au-FeO Magnetic Nanoparticles.

机构信息

State Key Laboratory of Supramolecular Structure and Materials, College of Chemistry , Jilin University , Changchun 130012 , People's Republic of China.

出版信息

Langmuir. 2018 Dec 11;34(49):14875-14881. doi: 10.1021/acs.langmuir.8b01784. Epub 2018 Jul 30.

Abstract

Studies on nucleic acid structure and interactions between nucleic acid and its binding molecules are of great importance for understanding and controlling many important biological processes. Atomic force microscopy (AFM) imaging is one of the most efficient methods to disclose the DNA structure and binding modes between DNA and DNA-binding molecules. Long-chain DNA tends to form a random coiled structure, which prevents direct AFM imaging observation of the subtle structure formed by DNA itself or protein binding. Aligning DNA from the random coiled state into the extended state is not only important for applications in DNA nanotechnology but also for elucidating the interaction mechanism between DNA and other molecules. Here, we developed an efficient method based on the magnetic field to align long-chain DNA on a silicon surface. We used AFM imaging to study the alignment of DNA at the single-molecule level, showing that DNA can be stretched and highly aligned by the manipulation of magnetic nanoparticles tethered to one end of DNA and that the aligned DNA can be imaged clearly by AFM. In the absence of the magnetic field, the aligned DNA can relax back to a random coiled state upon rinsing. Such alignment and relaxation can be repeated many times, which provides an efficient method for the manipulation of individual DNA molecules and the investigation of DNA and DNA-binding molecule interactions.

摘要

核酸结构和核酸与其结合分子之间相互作用的研究对于理解和控制许多重要的生物过程具有重要意义。原子力显微镜(AFM)成像技术是揭示 DNA 结构和 DNA 与 DNA 结合分子之间结合模式的最有效方法之一。长链 DNA 往往形成随机卷曲结构,这使得直接 AFM 成像观察 DNA 本身或蛋白质结合形成的细微结构变得困难。将 DNA 从随机卷曲状态排列成伸展状态不仅对于 DNA 纳米技术的应用很重要,而且对于阐明 DNA 与其他分子之间的相互作用机制也很重要。在这里,我们开发了一种基于磁场的有效方法,用于在硅表面上排列长链 DNA。我们使用 AFM 成像技术在单分子水平上研究 DNA 的排列,结果表明,通过操纵连接在 DNA 一端的磁性纳米颗粒,可以拉伸和高度排列 DNA,并且可以通过 AFM 清晰地成像排列的 DNA。在没有磁场的情况下,排列的 DNA 在冲洗后可以松弛回随机卷曲状态。这种排列和松弛可以重复多次,为单个 DNA 分子的操纵以及 DNA 和 DNA 结合分子相互作用的研究提供了一种有效的方法。

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