Rapid discrimination of DNA sequences using an engineered nanopore
2015 International Conference on Manipulation, Manufacturing and Measurement on the Nanoscale (3M-NANO)(2015)
摘要
Nanopores have been used for the development of biosensors for various substances and are currently under intense investigation as a potential tool for the entire genome sequencing. It has been demonstrated that an α-hemolysin protein pore covalently attached with a β-cyclodextrin adapter can differentiate single bases[1]; furthermore, single base discrimination could be achieved when a single-stranded DNA (ssDNA) is immobilized in the α-hemolysin pore[2]. Here, we show that when an unlabelled/unmodified double-stranded DNA (dsDNA) translocates through a properly engineered α-hemolysinnanopore, a sequence of sub-state blockage events is observed. Since different sequences of sub-state modulation events are produced by different DNA strands, they offer the potential as characteristic fingerprints to identify and discriminate the precise sequences of dsDNA samples.
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关键词
a-hemolysin,nanopores,DNA sequencing,base identification,protein engineering
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