An activator/repressor dual system allows tight tetracycline-regulated gene expression in budding yeast

We have developed an activator/repressor expression system for budding yeast in which tetracyclines control in opposite ways the ability of tetR-based activator and repressor molecules to bind tetO promoters. This combination allows tight expression of tetO-driven genes, both in a direct (tetracycli...

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Detalles Bibliográficos
Autores: Bellí i Martínez, Gemma, Garí Marsol, Eloi, Piedrafita Llorens, Lídia, Aldea, Martí, Herrero Perpiñán, Enrique
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:1998
País:España
Institución:Varias* (Consorci de Biblioteques Universitáries de Catalunya, Centre de Serveis Científics i Acadèmics de Catalunya)
Repositorio:Recercat. Dipósit de la Recerca de Catalunya
OAI Identifier:oai:recercat.cat:10459.1/47199
Acceso en línea:https://doi.org/10.1093/nar/26.4.942
http://hdl.handle.net/10459.1/47199
Access Level:acceso abierto
Palabra clave:Llevat de cervesa
Tetraciclina -- Aspectes genètics
Saccharomyces cerevisiae
Fongs -- Genètica
Descripción
Sumario:We have developed an activator/repressor expression system for budding yeast in which tetracyclines control in opposite ways the ability of tetR-based activator and repressor molecules to bind tetO promoters. This combination allows tight expression of tetO-driven genes, both in a direct (tetracycline-repressible) and reverse (tetracycline-inducible) dual system. Ssn6 and Tup1, that are components of a general repressor complex in yeast, have been tested for their repressing properties in the dual system, using lacZ and CLN2 as reporter genes. Ssn6 gives better results and allows complete switching-off of the regulated genes, although increasing the levels of the Tup1-based repressor by expressing it from a stronger promoter improves repressing efficiency of the latter. Effector-mediated shifts between expression and non-expression conditions are rapid. The dual system here described may be useful for the functional analysis of essential genes whose conditional expression can be tightly controlled by tetracyclines.