Nanopore analysis of nucleic acids bound to exonucleases and polymerases.
When a voltage is imposed across a thin membrane containing a nanoscopic pore, the electric field generated within the pore captures linear ionized polymers, such as nucleic acids, that are present in the solution bathing the pore. The nucleic acid molecule transiently blocks ionic current as it is translocated through the pore, and modulations of the current provide information about the structure and dynamic motion of the molecule. Altering the imposed voltage allows movement of the DNA molecule in the pore to be controlled. If a DNA-processing enzyme such as an exonuclease or polymerase is present, the enzyme-DNA complex is also drawn to the pore, and further modulations of the ionic current reflect enzyme function at the single-molecule level on millisecond timescales. The combined enzymatic and voltage control of a DNA molecule in the nanopore can be used to sequence the DNA.
Deamer
David
D
Department of Biomolecular Engineering, University of California, Santa Cruz, California 95064, USA. deamer@soe.ucsc.edu
D004926:Escherichia coli / Q000201:enzymology; D005092:Exonucleases / Q000378:metabolism*; D006460:Hemolysin Proteins / Q000737:chemistry*; D049329:Nanostructures / Q000737:chemistry*; D009713:Nucleotidyltransferases / Q000378:metabolism; D017422:Sequence Analysis, DNA
D006460:Hemolysin Proteins; D009713:Nucleotidyltransferases; D005092:Exonucleases