†Department of Materials Science and Engineering, Seoul National University, Seoul 151-742, Korea.
‡Graduate School of Biotechnology and Department of Food Science and Biotechnology, Kyung Hee University, Yongin 446-701, Korea.
ACS Nano. 2015 May 26;9(5):5289-98. doi: 10.1021/acsnano.5b00784. Epub 2015 May 6.
Solid-state nanopore has been widely studied as an effective tool to detect and analyze small biomolecules, such as DNA, RNA, and proteins, at a single molecule level. In this study, we demonstrate a rapid identification of the location of zinc finger protein (ZFP), which is bound to a specific locus along the length of a double-stranded DNA (dsDNA) to a single protein resolution using a low noise solid-state nanopore. When ZFP labeled DNAs were driven through a nanopore by an externally applied electric field, characteristic ionic current signals arising from the passage of the DNA/ZFP complex and bare DNA were detected, which enabled us to identify the locations of ZFP binding site. We examined two DNAs with ZFP binding sites at different positions and found that the location of the additional current drop derived from the DNA/ZFP complex is well-matched with a theoretical one along the length of the DNA molecule. These results suggest that the protein binding site on DNA can be mapped or that genetic information can be read at a single molecule level using solid-state nanopores.
固态纳米孔已被广泛研究为一种有效的工具，用于在单分子水平上检测和分析小分子生物分子，如 DNA、RNA 和蛋白质。在这项研究中，我们使用低噪声固态纳米孔，快速识别锌指蛋白 (ZFP) 的位置，该蛋白与双链 DNA (dsDNA) 上的特定位置结合。当 ZFP 标记的 DNA 在外加电场的驱动下穿过纳米孔时，检测到来自 DNA/ZFP 复合物和裸 DNA 通过的特征离子电流信号，这使我们能够识别 ZFP 结合位点的位置。我们检查了两条具有不同位置 ZFP 结合位点的 DNA，并发现来自 DNA/ZFP 复合物的额外电流下降的位置与 DNA 分子长度上的理论位置非常匹配。这些结果表明，使用固态纳米孔可以绘制 DNA 上的蛋白质结合位点，或者可以在单分子水平上读取遗传信息。
