Department of Chemistry, Graduate School of Science, Kyoto University, Sakyo-ku, Kyoto, Japan.
Institute for Integrated Cell-Material Sciences, Kyoto University, Sakyo-ku, Kyoto, Japan.
Methods Mol Biol. 2023;2651:241-250. doi: 10.1007/978-1-0716-3084-6_17.
To study the physical properties of molecules and their reaction processes, direct visualization of target molecules is one of the straightforward methods. Atomic force microscopy (AFM) enables the direct imaging of biomolecules under physiological conditions at nanometer-scale spatial resolution. In addition, using the DNA origami technology, the precise placement of target molecules in a designed nanostructure has been achieved, and the detection of the molecules at the single-molecule level has been realized. DNA origami is applied for visualizing the detailed movement of molecules combining with high-speed AFM (HS-AFM), which enables the analysis of the dynamic movement of biomolecules in a subsecond time resolution.Here, we describe the combination of the DNA origami system with HS-AFM for the imaging of rotation of dsDNA originated from B-Z transition. The rotation of dsDNA during B-Z transition is directly visualized in a DNA origami using the HS-AFM. These target-oriented observation systems serve to the detailed analysis of DNA structural changes in real time at molecular resolution.
为了研究分子的物理性质及其反应过程,直接观察目标分子是一种直接的方法。原子力显微镜(AFM)能够在纳米级空间分辨率下对生物分子在生理条件下进行直接成像。此外,利用 DNA 折纸技术,可以将目标分子精确地放置在设计好的纳米结构中,并实现了对单个分子的检测。DNA 折纸技术被应用于结合高速原子力显微镜(HS-AFM)对分子的详细运动进行可视化,这使得我们能够在亚秒级时间分辨率下分析生物分子的动态运动。在这里,我们描述了 DNA 折纸系统与 HS-AFM 的结合,用于成像源于 B-Z 转变的双链 DNA 的旋转。通过 HS-AFM 在 DNA 折纸中直接观察到双链 DNA 在 B-Z 转变过程中的旋转。这些面向目标的观察系统可用于在分子分辨率下实时、详细地分析 DNA 结构变化。