Chasse Scott A, Dohlman Henrik G
Department of Biochemistry and Biophysics, The University of North Carolina at Chapel Hill, 27599-7260, USA.
Methods Enzymol. 2004;389:399-409. doi: 10.1016/S0076-6879(04)89024-4.
Gene deletion analysis is a powerful tool for resolving the contributions of individual open reading frames to the physiology of cells. Analysis of deletion phenotypes in conjunction with a specific pathway reporter can identify constituents of a physiological pathway and reveal potential effectors that regulate the pathway by quantifying the phenotypic responses of the mutant cells. This article describes a high-throughput method of analyzing a yeast gene deletion library for novel G-protein signaling modulators using a yeast pheromone pathway-specific reporter.
基因缺失分析是一种强大的工具,可用于解析单个开放阅读框对细胞生理学的贡献。结合特定的信号通路报告基因对缺失表型进行分析,能够通过量化突变细胞的表型反应来鉴定生理信号通路的组成成分,并揭示调节该信号通路的潜在效应分子。本文描述了一种高通量方法,该方法利用酵母信息素信号通路特异性报告基因,对酵母基因缺失文库进行分析,以寻找新型G蛋白信号调节剂。