Chapter title |
Reporter-Based Synthetic Genetic Array Analysis: A Functional Genomics Approach for Investigating Transcript or Protein Abundance Using Fluorescent Proteins in Saccharomyces cerevisiae
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Chapter number | 40 |
Book title |
Genome Instability
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Published in |
Methods in molecular biology, January 2018
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DOI | 10.1007/978-1-4939-7306-4_40 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7305-7, 978-1-4939-7306-4
|
Authors |
Hendrikje Göttert, Mojca Mattiazzi Usaj, Adam P. Rosebrock, Brenda J. Andrews |
Abstract |
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. |
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