Single-nucleotide discrimination in immobilized DNA oligonucleotides with a biological nanopore.
The sequencing of individual DNA strands with nanopores is under investigation as a rapid, low-cost platform in which bases are identified in order as the DNA strand is transported through a pore under an electrical potential. Although the preparation of solid-state nanopores is improving, biological nanopores, such as alpha-hemolysin (alphaHL), are advantageous because they can be precisely manipulated by genetic modification. Here, we show that the transmembrane beta-barrel of an engineered alphaHL pore contains 3 recognition sites that can be used to identify all 4 DNA bases in an immobilized single-stranded DNA molecule, whether they are located in an otherwise homopolymeric DNA strand or in a heteropolymeric strand. The additional steps required to enable nanopore DNA sequencing are outlined.
Stoddart
David
D
Department of Chemistry, University of Oxford, Oxford OX1 3TA, United Kingdom.
Heron
Andrew J
AJ
Mikhailova
Ellina
E
Maglia
Giovanni
G
Bayley
Hagan
H
D000818:Animals; D001483:Base Sequence; D004247:DNA / Q000235:genetics*; D004277:DNA, Single-Stranded / Q000235:genetics; D023281:Genomics; D006460:Hemolysin Proteins / Q000235:genetics; D008969:Molecular Sequence Data; D053758:Nanoparticles / Q000737:chemistry; D049329:Nanostructures; D036103:Nanotechnology / Q000379:methods*; D009841:Oligonucleotides / Q000235:genetics*; D015202:Protein Engineering / Q000379:methods; D011817:Rabbits; D017422:Sequence Analysis, DNA / Q000295:instrumentation / Q000379:methods*; D012689:Sequence Homology, Nucleic Acid
D004277:DNA, Single-Stranded; D006460:Hemolysin Proteins; D009841:Oligonucleotides; D004247:DNA