Single-nucleotide discrimination in immobilized DNA oligonucleotides with a biological nanopore.

Journal article

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.

Authors: Stoddart, D; Heron, A; Mikhailova, E; Maglia, G; Bayley, H

• Publication status: Published
• Journal: Proceedings of the National Academy of Sciences of the United States of America
• Volume: 106
• Issue: 19
• Pages: 7702-7707
• Publication date: 2009-05-01
• DOI: 10.1073/pnas.0901054106
• EISSN: 1091-6490
• ISSN: 0027-8424
• Language: English
• Keywords: DNA; Nanotechnology; Sequence Analysis, DNA; Base Sequence; Hemolysin Proteins; Molecular Sequence Data; Protein Engineering; Nanostructures; DNA, Single-Stranded; Rabbits; Nanoparticles; Genomics; Animals; Oligonucleotides; Sequence Homology, Nucleic Acid