title: Tethered DNA hairpins facilitate electrochemical detection of DNA ligation.
abstract: 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.
authors.affiliation: School of Biological Sciences, University of East Anglia, Norwich NR4 7TJ, UK.
authors.forename: Gerhild
authors.initials: G
authors.lastname: Zauner
authors.forename: Yating
authors.initials: Y
authors.lastname: Wang
authors.forename: Manuel
authors.initials: M
authors.lastname: Lavesa-Curto
authors.forename: Andrew
authors.initials: A
authors.lastname: MacDonald
authors.forename: Andrew G
authors.initials: AG
authors.lastname: Mayes
authors.forename: Richard P
authors.initials: RP
authors.lastname: Bowater
authors.forename: Julea N
authors.initials: JN
authors.lastname: Butt
mesh_terms: D015374:Biosensing Techniques; D000072481:DNA Ligase ATP; D011088:DNA Ligases / Q000032:analysis*; D004563:Electrochemistry; D011489:Protein Denaturation
chemical_list: D011088:DNA Ligases; D000072481:DNA Ligase ATP