尿素促进单链 DNA 和 RNA 通过α-溶血素纳米孔的易位。
Urea facilitates the translocation of single-stranded DNA and RNA through the alpha-hemolysin nanopore.
机构信息
Department of Chemistry, University of Oxford, Oxford, United Kingdom.
出版信息
Biophys J. 2010 May 19;98(9):1856-63. doi: 10.1016/j.bpj.2009.12.4333.
Abstract
The staphylococcal alpha-hemolysin (alphaHL) protein nanopore is under investigation as a fast, cheap detector for nucleic acid analysis and sequencing. Although discrimination of all four bases of DNA by the alphaHL pore has been demonstrated, analysis of single-stranded DNAs and RNAs containing secondary structure mediated by basepairing is prevented because these nucleic acids cannot be translocated through the pore. Here, we show that a structured 95-nucleotide single-stranded DNA and its RNA equivalent are translocated through the alphaHL pore in the presence of 4 M urea, a concentration that denatures the secondary structure of the polynucleotides. The alphaHL pore is functional even in 7 M urea, and therefore it is easily stable enough for analyses of challenging DNA and RNA species.
摘要
金黄色葡萄球菌α-溶血素（alphaHL）蛋白纳米孔被研究作为一种快速、廉价的核酸分析和测序探测器。尽管已经证明 alphaHL 孔可以区分 DNA 的所有四个碱基，但由于这些核酸不能通过孔转移，因此无法对含有碱基配对介导的二级结构的单链 DNA 和 RNA 进行分析。在这里，我们表明，在 4 M 脲存在的情况下，一种结构为 95 个核苷酸的单链 DNA 及其 RNA 等价物可以穿过 alphaHL 孔，4 M 脲的浓度会使多核苷酸的二级结构变性。alphaHL 孔在 7 M 脲中仍然具有功能，因此它很容易稳定到足以分析具有挑战性的 DNA 和 RNA 种类。
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