Time course and steady-state analysis of oleate induction

For the time course of oleate induction, cells were grown in YPBG (YPB, 3% [wt/vol] glycerol) overnight to a density of 3 × 10⁷ cells/ml, pelleted by centrifugation, transferred to YPBO (YPB, 0.12% [wt/vol] oleate, 0.2% [wt/vol] Tween 40), and grown for 9 h to a density of 3 × 10⁷ cells/ml. Cells were harvested at 0, 0.5, 1, 3, 6, and 9 h after induction. For steady-state profiles of gene expression, cells were grown in YPBO, YPBG, or YPBD for 26 h to a final cell density of ∼3 × 10⁷ cells/ml. After harvesting, cells were frozen in liquid nitrogen and stored at –80°C.