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An electrochemical one-step system for assaying methyltransferase activity based on transport of a quantum dot signaling tracer

Authors
Journal
Biosensors and Bioelectronics
0956-5663
Publisher
Elsevier
Publication Date
Volume
49
Identifiers
DOI: 10.1016/j.bios.2013.04.036
Keywords
  • Dna Methylation
  • Methyltransferase
  • Epigenetic Analysis
  • Electrochemical Biosensor
  • Quantum Dot
Disciplines
  • Biology
  • Chemistry
  • Physics

Abstract

Abstract A one-step, electrochemical method for assaying methyltransferase (MTase) activity, based on the convective transport of a quantum dot (QD) signaling tracer, has been developed. The assay chip used in this system was prepared by modifying a gold matrix with CdSe/ZnS QD-tagged dsDNA, which contains a specific methylation site (5′–GATC-3′) recognized by MTase. Treatment of the chip with DNA adenine methylation (Dam) MTase, generates a methylated sequence (5′-GAmTC-3′) within the dsDNA. The methylated dsDNA is then subjected to a cleavage reaction, induced by DpnI, which leads to release from the gold matrix of a DNA fragment tethered to a QD. Detection of the released QD, using square wave anodic stripping voltammetry (SWASV) on a glassy carbon (GC) electrode, enables the reliable quantitation of the methylated DNA. Because it is accomplished in a simple and convenient one step and does not require any complicated secondary or tedious washing steps, the new assay method holds great promise for epigenetic analysis in facility-limited environments or point-of-care testing (POCT) applications.

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