FIG. 2.

Functionality of GFP-tagged constructs. (A) GFP-tagged SEC4 and YPT1 constructs were transformed into SEC4Δ and YPT1Δ tester strains and streaked onto medium with (+) or without (−) 5-FOA at 25°C to assess functionality. This assay was performed in comparison to yeast transformed with empty vector as a negative control and wild-type SEC4 and YPT1 as positive controls. (B) ypt6Δ cells were assayed for survival at 37°C when transformed with empty vector-, YPT6-, or GFP-YPT6-containing plasmids. (C) ypt31Δ ypt31^ts cells were assayed for the ability of GFP-tagged YPT31 to rescue growth at 37°C compared to that of vector alone. (D) vps21Δ cells were assayed for the uptake of lucifer yellow CH into the vacuole in the presence (+) or absence (−) of GFP-Vps21p. (E) ypt7Δ cells with GFP-YPT7 (i), vector only (ii), and wild-type YPT7 (iii) constructs were assayed for vacuolar morphologies with FM4-64. DIC, differential interference contrast. (F) ypt10Δ cells with GFP-YPT10 (i), vector only (ii), and wild-type YPT10 (iii) expressed behind the copper-inducible promoter P_CUP1 were assayed for growth on media ± 0.7 mM CuSO₄.