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

STAR Protocols

Jorrit M. Enserink¹  Ignacio Garcia²  Lucía Ramos-Alonso³  Pierre Chymkowitch³ 

[1] Centre for Cancer Cell Reprogramming, Institute of Clinical Medicine, Faculty of Medicine, University of Oslo, 0318 Oslo, Norway;Department of Molecular Cell Biology, Institute for Cancer Research, Oslo University Hospital, 0379 Oslo, Norway;Section for Biochemistry and Molecular Biology, Faculty of Mathematics and Natural Sciences, University of Oslo, 0316 Oslo, Norway;

关键词: Cell Biology;    Model Organisms;    Molecular Biology;    Gene Expression;    Signal Transduction;   

DOI  :  

来源: DOAJ

【 摘 要 】

Summary: 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).

【 授权许可】

Unknown