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Fluorescent protein expression as a proxy of bacterial fitness in a high throughput assay

View ORCID ProfileRudolf O Schlechter, Evan J Kear, Daniela M Remus, View ORCID ProfileMitja NP Remus-Emsermann
doi: https://doi.org/10.1101/2020.12.01.399113
Rudolf O Schlechter
1School of Biological Sciences, University of Canterbury, Christchurch, New Zealand
2Biomolecular Interaction Centre, University of Canterbury, Christchurch, New Zealand
3Bioprotection Research Core, University of Canterbury, Christchurch, New Zealand
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  • ORCID record for Rudolf O Schlechter
Evan J Kear
1School of Biological Sciences, University of Canterbury, Christchurch, New Zealand
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Daniela M Remus
4Protein Science & Engineering, Callaghan Innovation, School of Biological Sciences, University of Canterbury, Christchurch, New Zealand
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Mitja NP Remus-Emsermann
1School of Biological Sciences, University of Canterbury, Christchurch, New Zealand
2Biomolecular Interaction Centre, University of Canterbury, Christchurch, New Zealand
3Bioprotection Research Core, University of Canterbury, Christchurch, New Zealand
5Institute of Biology, Freie Universität Berlin, Berlin, Germany
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  • ORCID record for Mitja NP Remus-Emsermann
  • For correspondence: mitja-remus.emsermann@fu-berlin.de
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Abstract

Bacterial growth is classically assessed by measuring the increase in optical density of pure cultures in shaken liquid media. Measuring growth using optical density has severe limitations when studying multistrain interactions as it is not possible to measure the growth of individual strains within mixed cultures. Here we demonstrated that constitutively expressed fluorescent proteins can be used to track the growth of individual strains in different liquid media. Fluorescence measurements were highly correlated with optical density measurements and cell counts. This allowed us to assess bacterial growth not only in pure cultures, but also in mixed bacterial cultures and determine the impact of competitors on a focal strain, thereby assessing relative fitness. Furthermore, we were able to track the growth of two different strains simultaneously by using fluorescent proteins with differential excitation and emission wavelengths. Bacterial densities measured by fluorescence yielded more consistent data between technical replicates than optical density measurements. Our setup employs fluorescent microplate readers that allow for high throughput and replication.

Importance We expand on an important limitation of the concept of measuring bacterial growth which is classically limited to one strain at a time. By adopting this approach, it is possible to measure growth of several bacterial strains simultaneously in high temporal resolution and in a high throughput manner. This is important to investigate bacterial interactions such as competition and facilitation.

Competing Interest Statement

The authors have declared no competing interest.

Footnotes

  • Author Evan Kear was added to the author list. Author affiliations were updated. Additional experiments were performed and analysed leading to changed and additional figures in the main text and supplemental material.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted May 13, 2021.
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Fluorescent protein expression as a proxy of bacterial fitness in a high throughput assay
Rudolf O Schlechter, Evan J Kear, Daniela M Remus, Mitja NP Remus-Emsermann
bioRxiv 2020.12.01.399113; doi: https://doi.org/10.1101/2020.12.01.399113
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Fluorescent protein expression as a proxy of bacterial fitness in a high throughput assay
Rudolf O Schlechter, Evan J Kear, Daniela M Remus, Mitja NP Remus-Emsermann
bioRxiv 2020.12.01.399113; doi: https://doi.org/10.1101/2020.12.01.399113

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