FIG. 1.
Effect of overexpression and repression of the ERG11 gene. (A) Construction of the controllable strains 97ERG11, 98ERG11, and 99ERG11. We generated 97ERG11, 98ERG11, and 99ERG11 by replacing the endogenous ERG11 promoter with a tetracycline-regulatable promoter as shown on the right. Its replacement was confirmed by PCR (on the left). (B) Northern blot analysis of 97ERG11, 98ERG11, and 99ERG11. The actin gene (ACT1) was used as an internal control to quantify the amount of RNAs. Minus signs show total RNA extracted from cells not treated with DOX; plus signs show total RNA extracted from DOX-treated cells. (C) Growth defects of 97ERG11, 98ERG11, and 99ERG11 produced by DOX. 97ERG11, 98ERG11, 99ERG11, and ATCC 2001 cells (105 cells each) were inoculated into YEPD medium and cultured at 37°C with or without DOX (10 μg/ml). The results for three independent experiments were averaged. OD660, optical density at 660 nm.