An activator/repressor dual system allows tight tetracycline-regulated gene expression in budding yeast
MetadataShow full item record
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.
Is part ofNucleic acids research, 1998, vol. 26, núm. 4, p. 942-947
European research projects
Showing items related by title, author, creator and subject.
Prokaryotic and eukaryotic monothiol glutaredoxins are able to perform the functions of Grx5 in the biogenesis of Fe/S clusters in yeast mitochondria Molina Navarro, Maria Micaela; Casas Herranz, Celia; Piedrafita Llorens, Lídia; Bellí i Martínez, Gemma; Herrero Perpiñán, Enrique (Elsevier, 2003)The Saccharomyces cerevisiae monothiol glutare- doxin Grx5 participates in the mitochondrial biogenesis of iron–sulfur clusters. Grx5 homologues exist in organisms from bacteria to humans. Chicken (cGRX5) and human ...
Osmotic stress causes a G1 cell cycle delay and downregulation of Cln3/Cdc28 activity in Saccharomyces cerevisiae Bellí i Martínez, Gemma; Garí Marsol, Eloi; Aldea, Martí; Herrero Perpiñán, Enrique (Blackwell Science, 2001)Moderate hyperosmotic stress on Saccharomyces cerevisiae cells produces a temporary delay at the G1 stage of the cell cycle. This is accompanied by transitory downregulation of CLN1, CLN2 and CLB5 transcript levels, ...
Herrero Perpiñán, Enrique; Bellí i Martínez, Gemma; Casas Herranz, Celia (Bentham Science Publishers, 2010)Glutaredoxins are defined as thiol disulfide oxidoreductases that reduce disulfide bonds employing reduced glutathione as electron donor. They constitute a complex family of proteins with a diversity of enzymatic and ...