Reporter-Based Synthetic Genetic Array Analysis: A Functional Genomics Approach for Investigating Transcript or Protein Abundance Using Fluorescent Proteins in Saccharomyces cerevisiae
| Autor: | Hendrikje Göttert, Brenda J. Andrews, Adam P. Rosebrock, Mojca Mattiazzi Usaj |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Genetics Reporter gene medicine.diagnostic_test biology Saccharomyces cerevisiae Mutant Computational biology Synthetic genetic array biology.organism_classification Yeast Flow cytometry 03 medical and health sciences 030104 developmental biology 0302 clinical medicine medicine Functional genomics Gene 030217 neurology & neurosurgery |
| Zdroj: | Methods in Molecular Biology ISBN: 9781493973057 |
| DOI: | 10.1007/978-1-4939-7306-4_40 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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