Table 1.
Yeast strains
| No. strain | Construct/gene | Parent vector | MAT | ACC1 | URA3‡ | LEU2§ | HIS3‡ | TRP1‡ | Galactose dependence | ACCase content* |
|---|---|---|---|---|---|---|---|---|---|---|
| 1 MJ 1.12 | gyccwy† | pRS424 | a | − | − | + | − | + | + | 0.80 |
| 2 MJ 1.13 | gyccwy† | pRS424 | α | − | − | + | − | + | + | 0.88 |
| 3 MJ 6.8 | gyccwy† | pRS423 | a | − | − | + | + | − | + | 0.69 |
| 4 MJ 6.9 | gyccwy† | pRS423 | α | − | − | + | + | − | + | 0.78 |
| 5 MJ 9.11 | Yeast ACC1‡ | pRS426 | a | + | + | + | − | − | − | 1.12 |
| 6 MJ 9.12 | Yeast ACC1§ | None | α | + | − | (+)¶ | − | − | − | 3.76 |
| 7 MJ 9.13 | Yeast ACC1§ | None | α | + | − | − | − | − | − | 1.27 |
| 8 MJ 9.14 | Yeast ACC1‡ | pRS426 | a | + | + | + | − | − | − | 1.23 |
| 9 W303D | Yeast ACC1§ | None | a/α | + | − | + | − | − | − | 1.00 |
| 10 W303D-ACC1ΔLeu2 | Yeast ACC1§ | None | a/α | + | − | + | − | − | − | 0.59 |
All MJ strains were obtained by sporulation of W303D-ACC1ΔLeu2 transformed with an appropriate plasmid. The related haploid strains (1.12 and 1.13, 6.8 and 6.9, 9.11–9.14) described here were obtained from single tetrads. W303D relevant genotype (3): MATa/MATα, leu2-3,112/leu2-3,112 his3-11,15/his311,15 ade2-1/ade2-1 ura3-1/ura3-1 trp1-1/trp1-1 can1-100/can1-100 ACC1/ACC1; W303D-ACC1ΔLeu2 relevant genotype; MATa/MATα, leu2-3,112/leu2-3,112 his3-11,15/his311,15 ade2-1/ade2-1 ura3-1/ura3-1 trp1-1/trp1-1 can1-100/can1-100 ACC1/acc1::LEU2.
Relative amount of the 250-kDa biotinylated peptide estimated from band intensities on Western blots probed with [35S]streptavidin (e.g., Fig. 2) and normalized with respect to pyruvate carboxylase (PCase) as an internal standard. Averages of three measurements are shown.
GAL10 promoter/yeast ACC1 leader/cytosolic carboxylase wheat cDNA/yeast ACC1 tail (see Fig. 1).
Plasmid born.
Genomic.
Unexplained.