Effects on secreted msGFP constructs of including the α-factor pro region after the α-factor signal sequence. (A)
S. cerevisiae cells, either expressing wild-type Erv29 (“WT”) or carrying an erv29Δ allele, were engineered to express msGFP fused to either the α-factor signal sequence alone (pre-αf-msGFP) or the complete α-factor secretion signal (pre-pro-αf-msGFP). Samples of the culture medium and cells were analyzed by SDS-PAGE, immunoblotting, and chemiluminescence to detect msGFP. (B) The strains expressing wild-type Erv29 plus the indicated msGFP constructs were imaged by fluorescence microscopy to detect GFP, and by differential interference contrast (DIC) microscopy to detect the cells. Representative cells are shown. Exposure times for the fluorescence images were 100 msec. Scale bar, 2 μm.