Discrimination of oligonucleotides of different lengths with a wild-type aerolysin nanopore
Discrimination of oligonucleotides of different lengths with a wild-type aerolysin nanopore
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DOI：
10.1038/nnano.2016.66
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发表时间：
2016-08-01
影响因子：
38.3
通讯作者：
Long, Yi-Tao
中科院分区：
文献类型：
--
作者：
Cao, Chan;Ying, Yi-Lun;Long, Yi-Tao
蛋白质纳米孔提供了一种廉价的、无标记的分析单个寡核苷酸的方法。该方法的灵敏度在很大程度上取决于所采用的成孔蛋白的特性，并且通常依赖于已经化学修饰或并入分子马达的纳米孔。使用野生型生物纳米孔对寡核苷酸进行有效的高分辨率区分仍然难以实现。在这里，我们表明野生型气溶素纳米孔可以分辨2至10个碱基长的单个短寡核苷酸。气溶素的传感能力归因于气溶素的几何形状和纳米孔与寡核苷酸之间的静电相互作用。我们还表明，野生型气溶素纳米孔可以区分单个寡核苷酸与混合物，并可以监测核酸外切酶I对寡核苷酸的逐步切割。
Protein nanopores offer an inexpensive, label-free method of analysing single oligonucleotides. The sensitivity of the approach is largely determined by the characteristics of the pore-forming protein employed, and typically relies on nanopores that have been chemically modified or incorporate molecular motors. Effective, high-resolution discrimination of oligonucleotides using wild-type biological nanopores remains difficult to achieve. Here, we show that a wild-type aerolysin nanopore can resolve individual short oligonucleotides that are 2 to 10 bases long. The sensing capabilities are attributed to the geometry of aerolysin and the electrostatic interactions between the nanopore and the oligonucleotides. We also show that the wild-type aerolysin nanopores can distinguish individual oligonucleotides from mixtures and can monitor the stepwise cleavage of oligonucleotides by exonuclease I.