Fluorescent reporter genes have long been used to quantify various cell features such as transcript and protein abundance. Here, we describe a method, reporter synthetic genetic array (R-SGA) analysis, which allows for the simultaneous quantification of any fluorescent protein readout in thousands of yeast strains using an automated pipeline. R-SGA combines a fluorescent reporter system with standard SGA analysis and can be used to examine any array-based strain collection available to the yeast community. This protocol describes the R-SGA methodology for screening different arrays of yeast mutants including the deletion collection, a collection of temperature-sensitive strains for the assessment of essential yeast genes and a collection of inducible overexpression strains. We also present an alternative pipeline for the analysis of R-SGA output strains using flow cytometry of cells in liquid culture. Data normalization for both pipelines is discussed.
CITATION STYLE
Göttert, H., Mattiazzi Usaj, M., Rosebrock, A. P., & Andrews, B. J. (2018). Reporter-based synthetic genetic array analysis: A functional genomics approach for investigating transcript or protein abundance using fluorescent proteins in Saccharomyces cerevisiae. In Methods in Molecular Biology (Vol. 1672, pp. 613–629). Humana Press Inc. https://doi.org/10.1007/978-1-4939-7306-4_40
Mendeley helps you to discover research relevant for your work.