Kavli Institute of Nanoscience, Delft University of Technology, Lorentzweg 1, Delft, The Netherlands.
Nano Lett. 2010 Jan;10(1):324-8. doi: 10.1021/nl903631m.
Nanopores have been successfully employed as a new tool to rapidly detect single biopolymers, in particular DNA. When a molecule is driven through a nanopore by an externally applied electric field, it causes a characteristic temporary change in the trans-pore current. Here, we examine the translocation of DNA with discrete patches of the DNA-repair protein RecA attached along its length. Using the fact that RecA-coated DNA and bare DNA yield very different current-blockade signatures, we demonstrate that it is possible to map the locations of the proteins along the length of a single molecule using a solid-state nanopore. This is achieved at high speed and without any staining. We currently obtain a spatial resolution of about 8 nm, or 5 RecA proteins binding to 15 base pairs of DNA, and we discuss possible extensions to single protein resolution. The results are a crucial first step toward genomic screening, as they demonstrate the feasibility of reading off information along DNA at high resolution with a solid-state nanopore.
纳米孔已被成功用作快速检测单生物聚合物（特别是 DNA）的新工具。当分子在外加电场的驱动下穿过纳米孔时，它会导致跨孔电流的特征性暂时变化。在这里，我们研究了带有离散 DNA 修复蛋白 RecA 补丁的 DNA 的易位。利用 RecA 涂层 DNA 和裸 DNA 产生非常不同的电流阻断特征这一事实，我们证明了使用固态纳米孔可以沿单个分子的长度定位蛋白质的位置。这是在高速和无任何染色的情况下实现的。我们目前获得的空间分辨率约为 8nm，即 5 个 RecA 蛋白结合到 15 个碱基对的 DNA 上，我们还讨论了提高到单蛋白分辨率的可能途径。这些结果是朝着基因组筛选迈出的关键的第一步，因为它们证明了使用固态纳米孔以高分辨率读取 DNA 上信息的可行性。
