Fig. 5.
Functional analysis of human CoA Synthase gene (COASY) in S. cerevisiae. a For plasmid shuffling, strains LSY20 (cab4Δ + rescue plasmid pGE7 [ARS CEN URA3 CAB4]; upper part) and LSY21 (cab5Δ + rescue plasmid pGE9 [ARS CEN URA3 CAB5]; lower part) were transformed with the ARS CEN LEU2 plasmid pLS20 containing the human COASY gene activated by the MET25 promoter. Plasmids pGE8 (CAB4) and pGE10 (CAB5) served as positive controls, empty vector YCp111 as a negative control. FOA: 5-Fluoroorotic acid. b Stable biosynthesis of full-length hCoasy and truncated variant in transformants of S. cerevisiae. Expression plasmids pLS14 and pLS15 encoding HA-tagged length variants hCoasy1-564 and hCoasy30-564, respectively, were transformed into strain JS91.15–23. For comparison of expression efficiencies, S. cerevisiae Cab3 of similar size has been also analyzed (expression plasmid pJO3). Protein extracts were analyzed by immuno-blotting using anti-HA antibodies