Department of Chemistry, University of Oxford, Oxford, United Kingdom.
Nano Lett. 2011 Feb 9;11(2):746-50. doi: 10.1021/nl1038874. Epub 2011 Jan 11.
Protein nanopores may provide a cheap and fast technology to sequence individual DNA molecules. However, the electrophoretic translocation of ssDNA molecules through protein nanopores has been too rapid for base identification. Here, we show that the translocation of DNA molecules through the α-hemolysin protein nanopore can be slowed controllably by introducing positive charges into the lumen of the pore by site directed mutagenesis. Although the residual ionic current during DNA translocation is insufficient for direct base identification, we propose that the engineered pores might be used to slow down DNA in hybrid systems, for example, in combination with solid-state nanopores.
蛋白质纳米孔可能为测序单个 DNA 分子提供一种廉价且快速的技术。然而，由于 ssDNA 分子在蛋白质纳米孔中的电泳迁移速度过快，因此无法进行碱基识别。在这里，我们通过定点突变在孔腔内引入正电荷，从而证明了 α-溶血素蛋白纳米孔中 DNA 分子的迁移可以被可控地减慢。尽管 DNA 迁移过程中的剩余离子电流不足以进行直接碱基识别，但我们提出，通过工程化的纳米孔可以减慢杂交体系中的 DNA，例如与固态纳米孔结合使用。
