TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Relevant characteristicsa | Source or referenceb |
|---|---|---|
| Strains | ||
| E. coli XL1-Blue | recA1 endA gyrA96 thi1 hsdR17 (rk− mk+) supE44 relA1 λ lac[F′ proAB laclqZΔM15 Tn10(Tcr)] | 5 |
| S. cerevisiae G175 | MATα ADE2 MET his3 leu2 ura3 trp1 TAG+ SE+; parental strain of H1246 | ScanBi |
| S. cerevisiae H1246 | MATα are1-Δ::HIS3 are2-Δ::LEU2 dga1-Δ::KanMX4 Iro1-Δ::TRP1 ADE2 ura3 TAG− SE− | ScanBi, 27 |
| Plasmids | ||
| pBluescript KS | AprlacPOZ′ | Stratageneb |
| pKS::atfA | atfA (formerly designated as wax/dgat) from A. calcoaceticus ADP1 with artificial Shine-Dalgarno sequence for E. coli cloned as 1.4-kbp EcoRI-BamHI fragment into pBluescript KS− colinear to lacZ′ | 12 |
| pESC-URA | E. coli-S. cerevisiae shuttle vector; URA3 GAL1 GAL10 Apr | Stratageneb |
| pESC-URA::atfA | atfA (formerly designated as wax/dgat) from A. calcoaceticus ADP1 with artificial Kozak initiation sequence cloned as 1.4-kbp BamHI-SalI fragment into pESC-URA colinear to the GAL1 promoter | This study |
a
TAG, TAG accumulation; SE, sterol ester accumulation; Apr, ampicillin resistance.
b
ScanBi, Scandinavian Biotechnology Research (Alnarp, Sweden). Stratagene is located in La Jolla, Calif.