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Analysing and meta-analysing time-series data of microbial growth and gene expression from plate readers

Responding to change is a fundamental property of life, making time-series data invaluable in biology. For microbes, plate readers are a popular, convenient means to measure growth and also gene expression using fluorescent reporters. Nevertheless, the difficulties of analysing the resulting data ca...

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Autores principales: Montaño-Gutierrez, Luis Fernando, Moreno, Nahuel Manzanaro, Farquhar, Iseabail L., Huo, Yu, Bandiera, Lucia, Swain, Peter S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9176753/
https://www.ncbi.nlm.nih.gov/pubmed/35617352
http://dx.doi.org/10.1371/journal.pcbi.1010138
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author Montaño-Gutierrez, Luis Fernando
Moreno, Nahuel Manzanaro
Farquhar, Iseabail L.
Huo, Yu
Bandiera, Lucia
Swain, Peter S.
author_facet Montaño-Gutierrez, Luis Fernando
Moreno, Nahuel Manzanaro
Farquhar, Iseabail L.
Huo, Yu
Bandiera, Lucia
Swain, Peter S.
author_sort Montaño-Gutierrez, Luis Fernando
collection PubMed
description Responding to change is a fundamental property of life, making time-series data invaluable in biology. For microbes, plate readers are a popular, convenient means to measure growth and also gene expression using fluorescent reporters. Nevertheless, the difficulties of analysing the resulting data can be a bottleneck, particularly when combining measurements from different wells and plates. Here we present omniplate, a Python module that corrects and normalises plate-reader data, estimates growth rates and fluorescence per cell as functions of time, calculates errors, exports in different formats, and enables meta-analysis of multiple plates. The software corrects for autofluorescence, the optical density’s non-linear dependence on the number of cells, and the effects of the media. We use omniplate to measure the Monod relationship for the growth of budding yeast in raffinose, showing that raffinose is a convenient carbon source for controlling growth rates. Using fluorescent tagging, we study yeast’s glucose transport. Our results are consistent with the regulation of the hexose transporter (HXT) genes being approximately bipartite: the medium and high affinity transporters are predominately regulated by both the high affinity glucose sensor Snf3 and the kinase complex SNF1 via the repressors Mth1, Mig1, and Mig2; the low affinity transporters are predominately regulated by the low affinity sensor Rgt2 via the co-repressor Std1. We thus demonstrate that omniplate is a powerful tool for exploiting the advantages offered by time-series data in revealing biological regulation.
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spelling pubmed-91767532022-06-09 Analysing and meta-analysing time-series data of microbial growth and gene expression from plate readers Montaño-Gutierrez, Luis Fernando Moreno, Nahuel Manzanaro Farquhar, Iseabail L. Huo, Yu Bandiera, Lucia Swain, Peter S. PLoS Comput Biol Research Article Responding to change is a fundamental property of life, making time-series data invaluable in biology. For microbes, plate readers are a popular, convenient means to measure growth and also gene expression using fluorescent reporters. Nevertheless, the difficulties of analysing the resulting data can be a bottleneck, particularly when combining measurements from different wells and plates. Here we present omniplate, a Python module that corrects and normalises plate-reader data, estimates growth rates and fluorescence per cell as functions of time, calculates errors, exports in different formats, and enables meta-analysis of multiple plates. The software corrects for autofluorescence, the optical density’s non-linear dependence on the number of cells, and the effects of the media. We use omniplate to measure the Monod relationship for the growth of budding yeast in raffinose, showing that raffinose is a convenient carbon source for controlling growth rates. Using fluorescent tagging, we study yeast’s glucose transport. Our results are consistent with the regulation of the hexose transporter (HXT) genes being approximately bipartite: the medium and high affinity transporters are predominately regulated by both the high affinity glucose sensor Snf3 and the kinase complex SNF1 via the repressors Mth1, Mig1, and Mig2; the low affinity transporters are predominately regulated by the low affinity sensor Rgt2 via the co-repressor Std1. We thus demonstrate that omniplate is a powerful tool for exploiting the advantages offered by time-series data in revealing biological regulation. Public Library of Science 2022-05-26 /pmc/articles/PMC9176753/ /pubmed/35617352 http://dx.doi.org/10.1371/journal.pcbi.1010138 Text en © 2022 Montaño-Gutierrez et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Montaño-Gutierrez, Luis Fernando
Moreno, Nahuel Manzanaro
Farquhar, Iseabail L.
Huo, Yu
Bandiera, Lucia
Swain, Peter S.
Analysing and meta-analysing time-series data of microbial growth and gene expression from plate readers
title Analysing and meta-analysing time-series data of microbial growth and gene expression from plate readers
title_full Analysing and meta-analysing time-series data of microbial growth and gene expression from plate readers
title_fullStr Analysing and meta-analysing time-series data of microbial growth and gene expression from plate readers
title_full_unstemmed Analysing and meta-analysing time-series data of microbial growth and gene expression from plate readers
title_short Analysing and meta-analysing time-series data of microbial growth and gene expression from plate readers
title_sort analysing and meta-analysing time-series data of microbial growth and gene expression from plate readers
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9176753/
https://www.ncbi.nlm.nih.gov/pubmed/35617352
http://dx.doi.org/10.1371/journal.pcbi.1010138
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