利用蛋白纳米孔对固定化寡核苷酸进行单个 RNA 碱基识别。
Individual RNA base recognition in immobilized oligonucleotides using a protein nanopore.
机构信息
Department of Chemistry, University of Oxford, Oxford, OX1 3TA, United Kingdom.
出版信息
Nano Lett. 2012 Nov 14;12(11):5637-43. doi: 10.1021/nl3027873. Epub 2012 Oct 19.
Abstract
Protein nanopores are under investigation as key components of rapid, low-cost platforms to sequence DNA molecules. Previously, it has been shown that the α-hemolysin (αHL) nanopore contains three recognition sites, capable of discriminating between individual DNA bases when oligonucleotides are immobilized within the nanopore. However, the direct sequencing of RNA is also of critical importance. Here, we achieve sharply defined current distributions that enable clear discrimination of the four nucleobases, guanine, cytosine, adenine, and uracil, in RNA. Further, the modified bases, inosine, N(6)-methyladenosine, and N(5)-methylcytosine, can be distinguished.
摘要
蛋白质纳米孔作为快速、低成本的 DNA 分子测序平台的关键组成部分正在受到研究。以前已经表明，α-溶血素（αHL）纳米孔包含三个识别位点，当寡核苷酸固定在纳米孔内时，能够区分单个 DNA 碱基。然而，直接对 RNA 进行测序也至关重要。在这里，我们实现了清晰的电流分布，能够清晰地区分 RNA 中的四个核碱基，鸟嘌呤、胞嘧啶、腺嘌呤和尿嘧啶。此外，还可以区分修饰碱基，肌苷、N(6)-甲基腺嘌呤和 N(5)-甲基胞嘧啶。
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