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The alpha hemolysin nanopore was the ï¬rst application of a biological channel as a tool for nucleic acid analysis (1). The hemolysin heptamer spontaneously inserts into a lipid bilayer membrane, and an ionic current through the pore is produced when a voltage is applied across the membrane. The large electric field generated in the pore by the voltage captures linear ionized polymers such as nucleic acids that are present in the solution bathing the pore, followed by translocation of the molecule through the pore. The nucleic acid strand transiently blocks ionic current during translocation, and modulations of the current provide information about the structure and composition of the molecule (2). We recently reported that movement of the DNA molecule in the pore can be controlled by varying the applied voltage (3). Furthermore, if a DNA-processing enzyme such as an exonuclease or polymerase is present, the enzyme-DNA complex is drawn to the pore. Activity of the enzyme, including single turnover events, can be monitored by fluctuations of the ionic current that reflect single-molecule dynamics on millisecond timescales (4-8, see ref. 9 for review). Our expectation is that the combined enzymatic and voltage control of a DNA molecule in the nanopore will be able to sequence DNA hundreds of nucleotides in length, at rates of 50 - 100 bases per second.
1. Kasianowicz J. et al. 1996. Proc Natl Acad Sci USA 93:13770-13773.
2. Akeson M. et al. 1999. Biophys J 77:3227-3233.
3. Wilson N. 2009. ACS Nano. 3:995-1003.
4. Hornblower B. et al. 2007. Nat Methods 4: 315-317.
5. Benner S. et al. 2007. Nat Nanotechnol 2:718-24.
6. Hurt N. et al. 2009. J Am Chem Soc 131:3772-3778.
7. Olasagasti, F. et al. 2010. Nat Nanotechnol 5:798-806.
8. Lieberman et al. J Am Chem Soc ePub Dec 1, 2010.
9. Deamer D. 2010. Annu Rev Biophys 39:79-90.