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  • Accepted Manuscript: Yeast grown in continuous culture systems can detect mutagens with improved sensitivity relative to the Ames test
Title: Yeast grown in continuous culture systems can detect mutagens with improved sensitivity relative to the Ames test
Continuous culture systems allow for the controlled growth of microorganisms over a long period of time. Here, we develop a novel test for mutagenicity that involves growing yeast in continuous culture systems exposed to low levels of mutagen for a period of approximately 20 days. In contrast, most microorganism-based tests for mutagenicity expose the potential mutagen to the biological reporter at a high concentration of mutagen for a short period of time. Our test improves upon the sensitivity of the well-established Ames test by at least 20-fold for each of two mutagens that act by different mechanisms (the intercalator ethidium bromide and alkylating agent methyl methanesulfonate). To conduct the tests, cultures were grown in small, inexpensive continuous culture systems in media containing (potential) mutagen, and the resulting mutagenicity of the added compound was assessed via two methods: a canavanine-based plate assay and whole genome sequencing. In the canavanine-based plate assay, we were able to detect a clear relationship between the amount of mutagen and the number of canavanine-resistant mutant colonies over a period of one to three weeks of exposure. Whole genome sequencing of yeast grown in continuous culture systems exposed to methyl methanesulfonate demonstrated that quantification of mutations is more » possible by identifying the number of unique variants across each strain. However, this method had lower sensitivity than the plate-based assay and failed to distinguish the different concentrations of mutagen. In conclusion, we propose that yeast grown in continuous culture systems can provide an improved and more sensitive test for mutagenicity. « less
Authors:
; ; ; ;
Editors:
Rabilloud, Thierry
Award ID(s):
1556349
Publication Date:
NSF-PAR ID:
10280661
Journal Name:
PLOS ONE
Volume:
16
Issue:
3
Page Range or eLocation-ID:
e0235303
ISSN:
1932-6203
Sponsoring Org:
National Science Foundation
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