Single Molecule Bioelectronics and Their Application to Amplification-Free Measurement of DNA Lengths.
Single Molecule Bioelectronics and Their Application to Amplification-Free Measurement of DNA Lengths.
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DOI：
10.3390/bios6030029
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发表时间：
2016-06-24
期刊：
影响因子：
--
通讯作者：
Collins PG
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文献类型：
--
作者：
Gül OT;Pugliese KM;Choi Y;Sims PC;Pan D;Rajapakse AJ;Weiss GA;Collins PG
As biosensing devices shrink smaller and smaller, they approach a scale in which single molecule electronic sensing becomes possible. Here, we review the operation of single-enzyme transistors made using single-walled carbon nanotubes. These novel hybrid devices transduce the motions and catalytic activity of a single protein into an electronic signal for real-time monitoring of the protein’s activity. Analysis of these electronic signals reveals new insights into enzyme function and proves the electronic technique to be complementary to other single-molecule methods based on fluorescence. As one example of the nanocircuit technique, we have studied the Klenow Fragment (KF) of DNA polymerase I as it catalytically processes single-stranded DNA templates. The fidelity of DNA polymerases makes them a key component in many DNA sequencing techniques, and here we demonstrate that KF nanocircuits readily resolve DNA polymerization with single-base sensitivity. Consequently, template lengths can be directly counted from electronic recordings of KF’s base-by-base activity. After measuring as few as 20 copies, the template length can be determined with <1 base pair resolution, and different template lengths can be identified and enumerated in solutions containing template mixtures.
