Detection of Local Protein Structures along DNA Using Solid-State Nanopores
Detection of Local Protein Structures along DNA Using Solid-State Nanopores
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DOI：
10.1021/nl903631m
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发表时间：
2010-01-01
期刊：
影响因子：
10.8
通讯作者：
Dekker, Cees
中科院分区：
文献类型：
--
作者：
Kowalczyk, Stefan W.;Hall, Adam R.;Dekker, Cees
纳米孔已经成功地用作快速检测单个生物聚合物，特别是DNA的新工具。当分子被外部施加的电场驱动通过纳米孔时，其引起跨孔电流的特征性暂时变化。在这里，我们研究的易位的DNA与离散补丁的DNA修复蛋白RecA沿着其长度连接。利用RecA包被的DNA和裸DNA产生非常不同的电流阻断特征的事实，我们证明了可以使用固态纳米孔沿着单个分子的长度绘制蛋白质的位置沿着，这是在高速下实现的，并且没有任何染色。我们目前获得的空间分辨率约为8 nm，或5 RecA蛋白结合到15个碱基对的DNA，我们讨论了可能的扩展到单个蛋白质的分辨率。这些结果是基因组筛选的关键第一步，因为它们证明了用固态纳米孔以高分辨率沿着DNA阅读信息的可行性。
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.