Nanopore DNA sequencing with MspA
Nanopore DNA sequencing with MspA
复制标题
DOI：
10.1073/pnas.1001831107
复制
发表时间：
2010-09-14
影响因子：
11.1
通讯作者：
Gundlach, Jens H.
中科院分区：
文献类型：
--
作者：
Derrington, Ian M.;Butler, Tom Z.;Gundlach, Jens H.
纳米孔测序有可能成为一种直接、快速且廉价的DNA测序技术。纳米孔DNA测序最简单的形式利用了这样一种假设：单链DNA的单个核苷酸通过纳米孔时会独特地调节流经该孔的离子电流，从而使电流记录能够得出DNA序列。我们证明，当双链DNA暂时将核苷酸固定在孔的狭窄处时，通过改造后的耻垢分枝杆菌孔蛋白A（MspA）的离子电流能够区分所有四种DNA核苷酸，并分辨单链DNA中的单个核苷酸。让带有一系列双链区段的DNA通过MspA，为使用纳米孔的一种简单DNA测序方法提供了原理验证。这些发现凸显了MspA在纳米孔测序未来发展中的重要性。
Nanopore sequencing has the potential to become a direct, fast, and inexpensive DNA sequencing technology. The simplest form of nanopore DNA sequencing utilizes the hypothesis that individual nucleotides of single-stranded DNA passing through a nanopore will uniquely modulate an ionic current flowing through the pore, allowing the record of the current to yield the DNA sequence. We demonstrate that the ionic current through the engineered Mycobacterium smegmatis porin A, MspA, has the ability to distinguish all four DNA nucleotides and resolve single-nucleotides in single-stranded DNA when double-stranded DNA temporarily holds the nucleotides in the pore constriction. Passing DNA with a series of double-stranded sections through MspA provides proof of principle of a simple DNA sequencing method using a nanopore. These findings highlight the importance of MspA in the future of nanopore sequencing.