Full metadata
Title
Adaptation of Bacterial Comet Assays to Detect Antimicrobial-mediated DNA Strand Breaks in Escherichia coli
Description
This study was conducted as part of an underlying initiative to elucidate the mechanism of action of natural antibacterial clay minerals for application as therapeutic agents for difficult-to-treat infections such as methicillin-resistant Staphylococcus aureus (MRSA)-derived skin lesions and Buruli ulcer. The goal of this investigation was to determine whether exposure to the leachate of an antibacterial clay mineral, designated as CB, produced DNA double-strand breaks (DSBs) in Escherichia coli. A neutral comet assay for bacterial cells was adapted to assess DSB levels upon exposure to soluble antimicrobial compounds. Challenges involved with the adaptation process included comet visualization and data collection. To appropriately account for antimicrobial-mediated strand fragmentation, suitable control reactions comprised of exposures to water, ethanol, kanamycin, and bleomycin were developed and optimized for the assay. Bacterial exposure to CB resulted in significantly longer comet lengths compared to negative control exposures, suggesting that CB killing activity involves the induction of DNA DSBs. The results of this investigation further characterize the antimicrobial mechanisms associated with a particular clay mineral mixture. The adapted comet assay protocol described herein functions as an effective tool to assess double-strand fragmentation resulting from exposure to soluble antimicrobial compounds and to visually compare results from experimental and control reactions.
Date Created
2012-12
Contributors
- Solanky, Dipesh (Author)
- Haydel, Shelley (Thesis director)
- Stout, Valerie (Committee member)
- Adusumilli, Sarojini (Committee member)
- Barrett, The Honors College (Contributor)
- College of Liberal Arts and Sciences (Contributor)
Topical Subject
Resource Type
Extent
39 pages
Language
Copyright Statement
In Copyright
Primary Member of
Series
Academic Year 2012-2013
Handle
https://hdl.handle.net/2286/R.I.15688
Level of coding
minimal
Cataloging Standards
System Created
- 2017-10-30 02:50:57
System Modified
- 2021-08-11 04:09:57
- 2 years 9 months ago
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