Tethered DNA hairpins facilitate electrochemical detection of DNA ligation.
A novel electrochemical assay for DNA ligase activity is described. The assay exploits the properties of DNA hairpins tethered at one terminus to a gold electrode and labelled at the other with a ferrocene group for rapid characterisation of DNA status by cyclic voltammetry. Successful ligation of 'nicked' DNA hairpins is indicated by retention of the ferrocene couple when exposure to DNA ligase is followed by conditions that denature the hairpin. The results demonstrate the simplicity of integrating electrochemical detection with hairpin based biosensors and illustrate a new approach to the assay of DNA ligases, of which the NAD(+)-dependent enzymes represent a potential broad spectrum antibacterial drug target.
Zauner
Gerhild
G
School of Biological Sciences, University of East Anglia, Norwich NR4 7TJ, UK.
Wang
Yating
Y
Lavesa-Curto
Manuel
M
MacDonald
Andrew
A
Mayes
Andrew G
AG
Bowater
Richard P
RP
Butt
Julea N
JN
D015374:Biosensing Techniques; D000072481:DNA Ligase ATP; D011088:DNA Ligases / Q000032:analysis*; D004563:Electrochemistry; D011489:Protein Denaturation
D011088:DNA Ligases; D000072481:DNA Ligase ATP