Single Molecule Bioelectronics and Their Application to Amplification-Free Measurement of DNA Lengths

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Gul O. T., Pugliese K. M., Choi Y., Sims P. C., Pan D., Rajapakse A. J., ...More

BIOSENSORS-BASEL, vol.6, no.3, 2016 (ESCI) identifier identifier identifier

  • Publication Type: Article / Article
  • Volume: 6 Issue: 3
  • Publication Date: 2016
  • Doi Number: 10.3390/bios6030029
  • Journal Name: BIOSENSORS-BASEL
  • Journal Indexes: Emerging Sources Citation Index (ESCI), Scopus
  • Keywords: DNA polymerase, carbon nanotube sensors, single molecule enzymology, DNA sequencing, WALLED CARBON NANOTUBES, I KLENOW FRAGMENT, POLYMERASE-I, ESCHERICHIA-COLI, KINETIC MECHANISM, ELECTRONIC MEASUREMENTS, CONFORMATIONAL-CHANGES, NUCLEOTIDE SELECTION, SUBSTRATE-BINDING, DNTP BINDING
  • Ankara Haci Bayram Veli University Affiliated: No

Abstract

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.