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Designing a single-molecule biophysics tool for characterising DNA damage for techniques that kill infectious pathogens through DNA damage effects

Lookup NU author(s): Dr Adam WollmanORCiD


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© Springer International Publishing Switzerland 2016. Antibiotics such as the quinolones and fluoroquinolones kill bacterial pathogens ultimately through DNA damage. They target the essential type IIA topoisomerases in bacteria by stabilising the normally transient double-strand break state which is created to modify the supercoiling state of the DNA. Here we discuss the development of these antibiotics and their method of action. Existing methods for DNA damage visualisation, such as the comet assay and immunofluorescence imaging can often only be analysed qualitatively and this analysis is subjective. We describe a putative single-molecule fluorescence technique for quantifying DNA damage via the total fluorescence intensity of a DNA origami tile fully saturated with an intercalating dye, along with the optical requirements for how to implement these into a light microscopy imaging system capable of single-molecule millisecond timescale imaging. This system promises significant improvements in reproducibility of the quantification of DNA damage over traditional techniques.

Publication metadata

Author(s): Miller H, Wollman AJM, Leake MC

Editor(s): Mark C. Leake

Publication type: Book Chapter

Publication status: Published

Book Title: Biophysics of Infection

Year: 2016

Volume: 915

Pages: 115-127

Online publication date: 19/05/2016

Acceptance date: 02/04/2016

Series Title: Advances in Experimental Medicine and Biology

Publisher: Springer

Place Published: New York


DOI: 10.1007/978-3-319-32189-9_9

PubMed id: 27193541

Library holdings: Search Newcastle University Library for this item

ISBN: 9783319321899