High-Copy Yeast Library Construction and High-Copy Rescue Genetic Screen in Saccharomyces cerevisiae

Fanli Zeng*, David G. Quintana

*Autor correspondiente de este trabajo

Producción científica: Capítulo de libroCapítuloInvestigaciónrevisión exhaustiva

3 Citas (Scopus)

Resumen

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.

Idioma originalInglés
Título de la publicación alojadaMethods in Molecular Biology
Páginas77-83
Número de páginas7
Volumen2196
DOI
EstadoPublicada - 6 sept 2020

Serie de la publicación

NombreMethods in Molecular Biology
Volumen2196
ISSN (versión impresa)1064-3745
ISSN (versión digital)1940-6029

Huella

Profundice en los temas de investigación de 'High-Copy Yeast Library Construction and High-Copy Rescue Genetic Screen in Saccharomyces cerevisiae'. En conjunto forman una huella única.

Citar esto