Urea Facilitates the Translocation of Single-Stranded DNA and RNA Through the α-Hemolysin Nanopore
Urea Facilitates the Translocation of Single-Stranded DNA and RNA Through the α-Hemolysin Nanopore
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DOI：
10.1016/j.bpj.2009.12.4333
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发表时间：
2010-05-05
影响因子：
3.4
通讯作者：
Bayley, Hagan
中科院分区：
文献类型：
--
作者：
Japrung, Deanpen;Henricus, Marsiyana;Bayley, Hagan
葡萄球菌α-溶血素（α HL）蛋白纳米孔作为用于核酸分析和测序的快速、廉价检测器正在研究中。尽管已经证明了通过α HL孔对DNA的所有四种碱基的区分，但是通过碱基配对介导的含有二级结构的单链DNA和RNA的分析被阻止，因为这些核酸不能通过孔移位。在这里，我们表明，在4 M尿素（该浓度使多核苷酸的二级结构变性）的存在下，结构化的95个核苷酸的单链DNA及其RNA等效物通过α HL孔移位。即使在7 M尿素中，α HL孔也是功能性的，因此它很容易足够稳定，用于分析具有挑战性的DNA和RNA物质。
The staphylococcal a-hemolysin (alpha HL) 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 alpha HL 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 alpha HL pore in the presence of 4 M urea, a concentration that denatures the secondary structure of the polynucleotides. The alpha HL pore is functional even in 7 M urea, and therefore it is easily stable enough for analyses of challenging DNA and RNA species.