Revealing Antibiotic Tolerance of the <i>Mycobacterium smegmatis</i> Xanthine/Uracil Permease Mutant Using Microfluidics and Single-Cell Analysis
To reveal rare phenotypes in bacterial populations, conventional microbiology tools should be advanced to generate rapid, quantitative, accurate, and high-throughput data. The main drawbacks of widely used traditional methods for antibiotic studies include low sampling rate and averaging data for po...
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-06-01
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| Series: | Antibiotics |
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| Online Access: | https://www.mdpi.com/2079-6382/10/7/794 |
| _version_ | 1797528308154368000 |
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| author | Meltem Elitas Neeraj Dhar John D. McKinney |
| author_facet | Meltem Elitas Neeraj Dhar John D. McKinney |
| author_sort | Meltem Elitas |
| collection | DOAJ |
| description | To reveal rare phenotypes in bacterial populations, conventional microbiology tools should be advanced to generate rapid, quantitative, accurate, and high-throughput data. The main drawbacks of widely used traditional methods for antibiotic studies include low sampling rate and averaging data for population measurements. To overcome these limitations, microfluidic-microscopy systems have great promise to produce quantitative single-cell data with high sampling rates. Using <i>Mycobacterium smegmatis</i> cells, we applied both conventional assays and a microfluidic-microscopy method to reveal the antibiotic tolerance mechanisms of wild-type and <i>msm2570::Tn</i> mutant cells. Our results revealed that the enhanced antibiotic tolerance mechanism of the <i>msm2570::Tn</i> mutant was due to the low number of lysed cells during the antibiotic exposure compared to wild-type cells. This is the first study to characterize the antibiotic tolerance phenotype of the <i>msm2570::Tn</i> mutant, which has a transposon insertion in the <i>msm2570</i> gene—encoding a putative xanthine/uracil permease, which functions in the uptake of nitrogen compounds during nitrogen limitation. The experimental results indicate that the <i>msm2570::Tn</i> mutant can be further interrogated to reveal antibiotic killing mechanisms, in particular, antibiotics that target cell wall integrity. |
| first_indexed | 2024-03-10T09:56:24Z |
| format | Article |
| id | doaj.art-10e7b22e8bae41ffbafb38865a721746 |
| institution | Directory Open Access Journal |
| issn | 2079-6382 |
| language | English |
| last_indexed | 2024-03-10T09:56:24Z |
| publishDate | 2021-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Antibiotics |
| spelling | doaj.art-10e7b22e8bae41ffbafb38865a7217462023-11-22T02:15:57ZengMDPI AGAntibiotics2079-63822021-06-0110779410.3390/antibiotics10070794Revealing Antibiotic Tolerance of the <i>Mycobacterium smegmatis</i> Xanthine/Uracil Permease Mutant Using Microfluidics and Single-Cell AnalysisMeltem Elitas0Neeraj Dhar1John D. McKinney2Faculty of Engineering and Natural Sciences, Sabanci University, 34956 Istanbul, TurkeySchool of Life Sciences, École Polytechnique Fédérale de Lausanne (EPFL), 1015 Lausanne, SwitzerlandSchool of Life Sciences, École Polytechnique Fédérale de Lausanne (EPFL), 1015 Lausanne, SwitzerlandTo reveal rare phenotypes in bacterial populations, conventional microbiology tools should be advanced to generate rapid, quantitative, accurate, and high-throughput data. The main drawbacks of widely used traditional methods for antibiotic studies include low sampling rate and averaging data for population measurements. To overcome these limitations, microfluidic-microscopy systems have great promise to produce quantitative single-cell data with high sampling rates. Using <i>Mycobacterium smegmatis</i> cells, we applied both conventional assays and a microfluidic-microscopy method to reveal the antibiotic tolerance mechanisms of wild-type and <i>msm2570::Tn</i> mutant cells. Our results revealed that the enhanced antibiotic tolerance mechanism of the <i>msm2570::Tn</i> mutant was due to the low number of lysed cells during the antibiotic exposure compared to wild-type cells. This is the first study to characterize the antibiotic tolerance phenotype of the <i>msm2570::Tn</i> mutant, which has a transposon insertion in the <i>msm2570</i> gene—encoding a putative xanthine/uracil permease, which functions in the uptake of nitrogen compounds during nitrogen limitation. The experimental results indicate that the <i>msm2570::Tn</i> mutant can be further interrogated to reveal antibiotic killing mechanisms, in particular, antibiotics that target cell wall integrity.https://www.mdpi.com/2079-6382/10/7/794antibioticsconventionalmicrobiologymicrofluidicsmicroscopy<i>Mycobacterium smegmatis</i> |
| spellingShingle | Meltem Elitas Neeraj Dhar John D. McKinney Revealing Antibiotic Tolerance of the <i>Mycobacterium smegmatis</i> Xanthine/Uracil Permease Mutant Using Microfluidics and Single-Cell Analysis Antibiotics antibiotics conventional microbiology microfluidics microscopy <i>Mycobacterium smegmatis</i> |
| title | Revealing Antibiotic Tolerance of the <i>Mycobacterium smegmatis</i> Xanthine/Uracil Permease Mutant Using Microfluidics and Single-Cell Analysis |
| title_full | Revealing Antibiotic Tolerance of the <i>Mycobacterium smegmatis</i> Xanthine/Uracil Permease Mutant Using Microfluidics and Single-Cell Analysis |
| title_fullStr | Revealing Antibiotic Tolerance of the <i>Mycobacterium smegmatis</i> Xanthine/Uracil Permease Mutant Using Microfluidics and Single-Cell Analysis |
| title_full_unstemmed | Revealing Antibiotic Tolerance of the <i>Mycobacterium smegmatis</i> Xanthine/Uracil Permease Mutant Using Microfluidics and Single-Cell Analysis |
| title_short | Revealing Antibiotic Tolerance of the <i>Mycobacterium smegmatis</i> Xanthine/Uracil Permease Mutant Using Microfluidics and Single-Cell Analysis |
| title_sort | revealing antibiotic tolerance of the i mycobacterium smegmatis i xanthine uracil permease mutant using microfluidics and single cell analysis |
| topic | antibiotics conventional microbiology microfluidics microscopy <i>Mycobacterium smegmatis</i> |
| url | https://www.mdpi.com/2079-6382/10/7/794 |
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