Nanopore DNA sequencing with MspA

被引：446

作者：

Derrington, Ian M. ^{[1
]}

Butler, Tom Z. ^{[1
]}

Collins, Marcus D. ^{[1
]}

Manrao, Elizabeth ^{[1
]}

Pavlenok, Mikhail ^{[2
]}

Niederweis, Michael ^{[2
]}

Gundlach, Jens H. ^{[1
]}

机构：

[1] Univ Washington, Dept Phys, Seattle, WA 98195 USA

[2] Univ Alabama Birmingham, Dept Microbiol, Birmingham, AL 35294 USA

来源：

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA | 2010年 / 107卷 / 37期

基金：

美国国家卫生研究院;

关键词：

bionanotechnology; next generation sequencing; single-molecule; stochastic sensing; protein pore; DISCRIMINATION; HOMOPOLYMERS; MOLECULES; CHALLENGES;

D O I：

10.1073/pnas.1001831107

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

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.

引用

页码：16060 / 16065

页数：6

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