Nanopore DNA sequencing with MspA.
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.
Derrington
Ian M
IM
Department of Physics, University of Washington, Seattle, WA 98195-1560, USA.
Butler
Tom Z
TZ
Collins
Marcus D
MD
Manrao
Elizabeth
E
Pavlenok
Mikhail
M
Niederweis
Michael
M
Gundlach
Jens H
JH
D001483:Base Sequence; D004247:DNA / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D008958:Models, Molecular; D020102:Mycobacterium smegmatis / Q000737:chemistry* / Q000378:metabolism; D049329:Nanostructures / Q000737:chemistry*; D009690:Nucleic Acid Conformation; D018272:Porins / Q000378:metabolism*; D016062:Porosity; D017434:Protein Structure, Tertiary; D017422:Sequence Analysis, DNA / Q000379:methods*
D018272:Porins; C121838:mspA protein, Mycobacterium smegmatis; D004247:DNA
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