Figure 8.

Allele specific accumulation of GFP tagged proteins in yeast membranes expressed at 15 °C, 20 °C, 25 °C, 30 °C and 35 °C. Crude membranes from yeast strain PAP4920 expressing Pmr1-GFP, Pmc1-GFP, wt hSPCA1a-GFP or HHD hSPCA1a-GFP from the replicative vector pPAP4997 were analyzed for fluorescence as described in Materials and Methods. Yeast cultures were inoculated in standard minimal medium supplemented with 20 mM CaCl₂ in shake flasks at 15 °C, 20 °C, 25 °C, 30 °C or 35 °C at OD₄₅₀ = 0.05. Cultures were harvested at OD₄₅₀ = 1.0–1.5. GFP fluorescence was measured in equal amounts of purified yeast crude membranes containing 25 µg total membrane protein. Fluorescence was converted to pmol per mg crude membranes as described in Materials and Methods. Each meassurment was made in triplicate.