The pheromone response pathway of the yeast Saccharomyces cerevisiae is a well-established model for the study of G proteins and mitogen-activated protein kinase (MAPK) cascades. Our longstanding ability to combine sophisticated genetic approaches with established functional assays has provided a thorough understanding of signalling mechanisms and regulation. In this report, we compare new and established methods used to quantify pheromone-dependent MAPK phosphorylation, transcriptional induction, mating morphogenesis, and gradient tracking. These include both single-cell and population-based assays of activity. We describe several technical advances, provide example data for benchmark mutants, highlight important differences between newer and established methodologies, and compare the advantages and disadvantages of each as applied to the yeast model. Quantitative measurements of pathway activity have been used to develop mathematical models and reveal new regulatory mechanisms in yeast. It is our expectation that experimental and computational approaches developed in yeast may eventually be adapted to human systems biology and pharmacology.

中文翻译：

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酵母信息素途径的定量分析。

酵母酿酒酵母的信息素反应途径是研究G蛋白和丝裂原活化蛋白激酶（MAPK）级联反应的公认模型。我们将复杂的遗传学方法与已建立的功能分析相结合的长期能力，对信号传导机制和调控提供了透彻的了解。在本报告中，我们比较了用于量化信息素依赖性MAPK磷酸化，转录诱导，交配形态发生和梯度跟踪的新方法和已建立的方法。这些包括单细胞和基于群体的活性测定。我们描述了数项技术进步，提供了基准突变体的示例数据，重点介绍了新方法与已建立方法之间的重要差异，并比较应用于酵母模型的每种方法的优缺点。途径活性的定量测量已用于开发数学模型并揭示酵母中的新调节机制。我们期望酵母中开发的实验和计算方法最终可以适应人类系统生物学和药理学。