FIGURE 1:

Comparison of different depletion systems. (A) Schematic representation of the depletion cassettes used. The promoter is indicated by a broken arrow; DAA, destabilizing amino acid; tc, tetracycline aptamer. (B) Growth of strains with the standard TetO7 promoter and the new TetO7-Ubi-Leu cassette during depletion. Doxycycline or ethanol (solvent) was added to exponentially growing cultures to a final concentration of 2 μg/ml, and optical density was measured at the indicated time points. (C) Reversibility of the depletion in the TetO7-Ubi-Leu-Has1 strain. Doxycycline or ethanol (solvent) was added to exponentially growing yeast to a final concentration of 2 μg/ml at time 0. After 8 h, the doxycycline/ethanol was washed away, and the cultures were monitored until the exponential growth was restored. (D) Viability of cells after 8 h of depletion. Two hundred cells from the TetO7-Ubi-Leu-Has1 strain were grown for 8 h with or without doxycycline and then plated on YPD agar; growing colonies were counted after 2 d. For all experiments, a mean of three independent biological replicas is shown; error bars represent SDs. The y-axes in B and C are in logarithmic scale.