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Títol:
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An activator/repressor dual system allows tight tetracycline-regulated gene expression in budding yeast
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Autor/a:
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Bellí i Martínez, Gemma; Garí Marsol, Eloi; Piedrafita Llorens, Lídia; Aldea, Martí; Herrero Perpiñán, Enrique
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Notes:
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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. |
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Matèries:
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-Llevat de cervesa
-Tetraciclina -- Aspectes genètics
-Saccharomyces cerevisiae
-Fongs -- Genètica |
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Drets:
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(c) Oxford University Press
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Tipus de document:
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article
publishedVersion |
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Publicat per:
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Oxford University Press
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Compartir:
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