Table 5.
Ability of URE2 homologs to induce or cure [URE3] in S. cerevisiae
| URE2 gene | Plasmid | Induction, USA+/106 cells | Plasmid | Curing, USA+/USA− |
|---|---|---|---|---|
| Vector | pH317 | 22 | pH316 | 39/1 |
| S. cerevisiae | pH739 | 7,700 | pH740 | 40/0 |
| S. bayanus | pH661 | 52 | pH679 | 40/0 |
| S. paradoxus | pH660 | 6,300 | pH678 | 40/0 |
| C. glabrata | pH659 | <1 | pH677 | 0/40 |
| A. gossypii | pH656 | 1 | pH674 | 0/40 |
| C. kefyr-1 | pH713 | 2 | pH711 | 0/40 |
| C. kefyr-2 | pH714 | 2 | pH712 | 0/40 |
| C. albicans | pH563 | 6 | pH672 | 0/40 |
| C. maltosa | pH657 | <0.1 | pH675 | 0/40 |
| C. lipolytica | pH658 | 22 | pH676 | 40/0 |
Strain YHE711 (MATα ura2 leu2∷hisG) was transformed with 2μ plasmids carrying URE2 homologs under control of the GAL1 promoter. Individual transformants were grown to saturation in leucine dropout medium containing 2% galactose and 1% rafinose and plated in 10-fold dilutions onto USA plates to assay [URE3] induction. For curing, centromeric plasmids were transformed into YHE64. Transformants were confirmed to still carry [URE3], then grown to single colonies on YPAGal2%Raf2% to overexpress the Ure2p homolog. Leu+ colonies were grown as patches three times on dextrose, then tested for USA phenotype.