Hybrid pore formation by directed insertion of α-haemolysin into solid-state nanopores.
Hybrid pore formation by directed insertion of α-haemolysin into solid-state nanopores.
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DOI：
10.1038/nnano.2010.237
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发表时间：
2010-12
影响因子：
38.3
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中科院分区：
文献类型：
--
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纳米孔对于基因组筛选和测序技术具有巨大的潜力。迄今为止，大多数研究集中在金黄色葡萄球菌孔形成蛋白α溶血素（α HL）和固态（SS）膜中的人工孔。虽然生物孔提供原子级精确的结构和遗传工程潜力，但SS孔提供耐用性，尺寸和形状控制以及可集成性。然而，每种系统也有明显的局限性：α HL很难整合，因为它依赖于精密的脂质双层进行机械支撑，并且精确尺寸的SS孔的制造仍然具有挑战性。在这里，我们表明，这些限制可以通过将单个α HL孔插入SS-纳米孔来克服。附着在蛋白质孔上的双链DNA通过电泳易位被穿入SS纳米孔中。在我们的尝试中观察到30 - 40%的蛋白质插入，并且单链DNA的易位表明杂交纳米孔保持功能。所得到的混合结构提供了一个平台，以创建用于基因组分析（包括测序）的晶片级器件阵列。
Nanopores hold great potential for genomic screening and sequencing technologies. Thus far, most studies have concentrated on the Staphylococcus aureus pore-forming protein alpha hemolysin (αHL) and artificial pores in solid-state (SS) membranes. While biological pores offer an atomically precise structure and genetic engineering potential, SS-pores offer durability, size and shape control and integratability. Each system, however, also has significant limitations: αHL is difficult to integrate because it relies on delicate lipid bilayers for mechanical support, and the fabrication of SS-pores at precise dimensions remains challenging. Here we show that these limitations may be overcome by inserting a single αHL pore into a SS-nanopore. A double-stranded DNA attached to a protein pore is threaded into a SS-nanopore by electrophoretic translocation. Protein insertion is observed in 30-40% of our attempts and translocation of single-stranded DNA demonstrates that the hybrid nanopore remains functional. The resulting hybrid structure offers a platform to create wafer-scale device arrays for genomic analysis including sequencing.
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影响因子：
19.6
作者：
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通讯作者：
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影响因子：
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影响因子：
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作者：
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通讯作者：
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