@inbook{c840b6a72ae6466789d9b91ce6d0a2c4,
title = "High-Copy Yeast Library Construction and High-Copy Rescue Genetic Screen in Saccharomyces cerevisiae",
abstract = "High-copy rescue genetic screening is a powerful strategy for the identification of suppression genetic interactions in the model eukaryotic organism Saccharomyces cerevisiae (budding yeast). The strain carrying the mutant allele of interest is transformed with a genomic library cloned in a high-copy plasmid. Each clone carries a genomic fragment insertion of around 10 kb, typically containing one to three complete genes under their own promoters. The high-copy vector favors the accumulation of high levels of the corresponding protein, aimed at suppressing the mutant phenotype. Typically, high-copy genetic screens select for viable clones under conditions restrictive or lethal for the query mutant strain. Here, we describe in detail the procedure to generate a high-copy genomic library and a protocol for rescue genetic screening and identification of the suppressor clones.",
keywords = "2 μm, Budding yeast, Dosage suppression, Genetic screen, High-copy genomic library, High-copy suppressor, Saccharomyces cerevisiae, Transformation, Genetic, Plasmids/genetics, Genomic Library, Gene Dosage, Saccharomyces cerevisiae/genetics, Phenotype, Genetic Testing/methods, Genes, Fungal",
author = "Fanli Zeng and Quintana, {David G.}",
note = "Funding Information: This work was supported by the National Natural Science Foundation of China to FZ (grant No. 31801039) and by the Ministry of Economy and Competitiveness of Spain (MINECO) and the European Regional Development Fund (FEDER) to DGQ (grant BFU2015-68493-P). Publisher Copyright: {\textcopyright} Springer Science+Business Media, LLC, part of Springer Nature 2021. Copyright: Copyright 2020 Elsevier B.V., All rights reserved.",
year = "2020",
month = sep,
day = "6",
doi = "10.1007/978-1-0716-0868-5_7",
language = "English",
isbn = "9781071608678",
volume = "2196",
series = "Methods in Molecular Biology",
pages = "77--83",
booktitle = "Methods in Molecular Biology",
}