Figure 2

Maximum yields of Fps1 are achieved by tuning BMS1 transcript number in a doxycycline-repressible system (yTHC). Fps1 yield in shake flasks is presented for the yTHCBMS1 overexpression strain cultured in 2 × CBS medium with varying amounts of doxycycline (the concentration in μg/mL is shown in parentheses next to each point), thereby varying the copies of BMS1 RNA/cell. The y-axis shows the factor improvement over the control, as for Figure 1. For comparison, data for the wild-type and the three deletion strains presented in Figure 1 (srb5Δ, spt3Δ and gcn5Δ) are also shown. Inset is a typical anti-HA-tag immunoblot showing Fps1 yields for 75 μg total wild-type membrane (lane 1), and 15 μg total yTHCBMS1 membranes extracted from shake-flask cultures grown in the presence of 0 (lane 2), 0.25 (lane 3), 0.5 (lane 4) and 10 μg/mL (lane 5) doxycycline. The markers (M) are 62, 98, 188 kDa ascending from the bottom of the gel upwards.