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
Descripción
Sumario: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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