Analysis of the pheromone signaling pathway by RT-qPCR in the budding yeast Saccharomyces cerevisiae

FUS3 and STE2 expression levels can be used as reporters for signaling through the pheromone pathway in the budding yeast Saccharomyces cerevisiae. Here, we describe an optimized protocol to measure the expression levels of FUS3 and STE2 using quantitative reverse transcription PCR (RT-qPCR). We des...

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Detalles Bibliográficos
Autores principales: Ramos-Alonso, Lucía, Garcia, Ignacio, Enserink, Jorrit M., Chymkowitch, Pierre
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8899044/
https://www.ncbi.nlm.nih.gov/pubmed/35265859
http://dx.doi.org/10.1016/j.xpro.2022.101210
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author Ramos-Alonso, Lucía
Garcia, Ignacio
Enserink, Jorrit M.
Chymkowitch, Pierre
author_facet Ramos-Alonso, Lucía
Garcia, Ignacio
Enserink, Jorrit M.
Chymkowitch, Pierre
author_sort Ramos-Alonso, Lucía
collection PubMed
description FUS3 and STE2 expression levels can be used as reporters for signaling through the pheromone pathway in the budding yeast Saccharomyces cerevisiae. Here, we describe an optimized protocol to measure the expression levels of FUS3 and STE2 using quantitative reverse transcription PCR (RT-qPCR). We describe the steps for comparing untreated and pheromone-treated yeast cells and how to quantify the changes in various deletion strains. The protocol can be applied to determine potential regulators of the pheromone pathway. For complete details on the use and execution of this protocol, please refer to Garcia et al. (2021).
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spelling pubmed-88990442022-03-08 Analysis of the pheromone signaling pathway by RT-qPCR in the budding yeast Saccharomyces cerevisiae Ramos-Alonso, Lucía Garcia, Ignacio Enserink, Jorrit M. Chymkowitch, Pierre STAR Protoc Protocol FUS3 and STE2 expression levels can be used as reporters for signaling through the pheromone pathway in the budding yeast Saccharomyces cerevisiae. Here, we describe an optimized protocol to measure the expression levels of FUS3 and STE2 using quantitative reverse transcription PCR (RT-qPCR). We describe the steps for comparing untreated and pheromone-treated yeast cells and how to quantify the changes in various deletion strains. The protocol can be applied to determine potential regulators of the pheromone pathway. For complete details on the use and execution of this protocol, please refer to Garcia et al. (2021). Elsevier 2022-03-03 /pmc/articles/PMC8899044/ /pubmed/35265859 http://dx.doi.org/10.1016/j.xpro.2022.101210 Text en © 2022 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Ramos-Alonso, Lucía
Garcia, Ignacio
Enserink, Jorrit M.
Chymkowitch, Pierre
Analysis of the pheromone signaling pathway by RT-qPCR in the budding yeast Saccharomyces cerevisiae
title Analysis of the pheromone signaling pathway by RT-qPCR in the budding yeast Saccharomyces cerevisiae
title_full Analysis of the pheromone signaling pathway by RT-qPCR in the budding yeast Saccharomyces cerevisiae
title_fullStr Analysis of the pheromone signaling pathway by RT-qPCR in the budding yeast Saccharomyces cerevisiae
title_full_unstemmed Analysis of the pheromone signaling pathway by RT-qPCR in the budding yeast Saccharomyces cerevisiae
title_short Analysis of the pheromone signaling pathway by RT-qPCR in the budding yeast Saccharomyces cerevisiae
title_sort analysis of the pheromone signaling pathway by rt-qpcr in the budding yeast saccharomyces cerevisiae
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8899044/
https://www.ncbi.nlm.nih.gov/pubmed/35265859
http://dx.doi.org/10.1016/j.xpro.2022.101210
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AT enserinkjorritm analysisofthepheromonesignalingpathwaybyrtqpcrinthebuddingyeastsaccharomycescerevisiae
AT chymkowitchpierre analysisofthepheromonesignalingpathwaybyrtqpcrinthebuddingyeastsaccharomycescerevisiae

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