Nanopore DNA sequencing with MspA

Proc Natl Acad Sci U S A. 2010 Sep 14;107(37):16060-5. doi: 10.1073/pnas.1001831107. Epub 2010 Aug 26.

Abstract

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.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Base Sequence
  • DNA / analysis*
  • DNA / chemistry
  • DNA / metabolism
  • Models, Molecular
  • Mycobacterium smegmatis / chemistry*
  • Mycobacterium smegmatis / metabolism
  • Nanostructures / chemistry*
  • Nucleic Acid Conformation
  • Porins / metabolism*
  • Porosity
  • Protein Structure, Tertiary
  • Sequence Analysis, DNA / methods*

Substances

  • Porins
  • mspA protein, Mycobacterium smegmatis
  • DNA