TABLE 1.
Yeast strains and plasmids
| Strain or plasmid | Genotype or relevant features | Source or reference |
|---|---|---|
| S. cerevisiae | ||
| XMP10-7B | MATa ura3 ade1 hom3 | This work |
| GRF167 | MATα ura3-167 his3-Δ200 | 2 |
| SG15 | MATα ura3-52 trp1 gal2 | S. Holmberg |
| SG183 | MATα ura3-52 trp1 cha1 gal2 | S. Holmberg |
| HT2 | MATα ura3-52 trp1 hom3::TRP1 gal2 | I. Velasco |
| Plasmids | ||
| pYH3 | URA3, 2μm, Aps Tcr, HOM3 | 7 |
| pMP1 | URA3, Apr, HOM3 | This work |
| pMR3-R21 | URA3, Apr, HOM3-R2 | 21 |
| pCGS42 | URA3, 2μm, Apr Tcr | F. del Rey |
| YCp50 | URA3, ARS, CEN, Apr Tcr | 25 |
| pYES2 | URA3, 2μm, Apr, PGAL1 | J. M. Pardo |
| pTK120 | URA3, ARS, CEN, Apr, PCHA1 | S. Holmberg |
| YEP/HSE2 | 2μm, Apr PCYC1-HSE2 | A. Rodríguez |
| YEp356, GPH-lacZ | URA3, 2μm, Apr, PGPH1 | J. François |
| pEGAL-H3 or -R2a | pYES2 containing a 1.9-kb HindIII fragment (HOM3 or HOM3-R2) from pMP1 or pMR3-R21, respectively | This work |
| pCGAL-H3 or -R2a | YCp50 containing a 2.4-kb SpeI fragment (PGAL1::HOM3 or HOM3-R2) from pEGAL-H3 or -R2, respectively | This work |
| pCHA-H3 or -R2a | YCp50 containing a 1.9-kb BamHI-EcoRI fragment (HOM3 or HOM3-R2) from pEHSE-H3 or -R2, respectively, and a 0.7-kb BamHI fragment (PCHA1) from pTK120 | This work |
| pEHSE-H3 or -R2a | pCGS42 containing a 0.8-kb SalI-HindIII fragment (PCYC1-HSE2) from YEP/HSE2+Hb and a 1.9-kb HindIII fragment (HOM3 or HOM3-R2) from pMP1 or pMR3-R21, respectively | This work |
| pCHSE-H3 or -R2ac | YCp50 containing a 1.9-kb BamHI-EcoRI fragment (HOM3 or HOM3-R2) from pEHSE-H3 or -R2, respectively, and a 0.8-kb SalI-BglII fragment (PCYC1-HSE2) from YEP/HSE2 | This work |
| pGPH-H3 or -R2a | YCp50 containing a 0.4-kb EcoRI-HindIII fragment (PGPH1) from YEp356, GPH-lacZ, and a 1.9-kb HindIII fragment (HOM3 or HOM3-R2) from pMP1-Hd or pMR3-R21-Hd, respectively | This work |
a
In these plasmids, the expression of the HOM3 alleles is under the control of the respective inducible promoter; the HindIII site used for cloning is 25 bp upstream from the ATG start codon of the HOM3 gene.
b
This plasmid was constructed by inserting a linker HindIII in the BamHI site of YEP/HSE2.
c
In these plasmids an additional ATG start codon from a lac gene present in the original promoter was deleted.
d
These plasmids are identical to pMP1 and pMR3-R21 except that the internal HindIII site in the HOM3 or HOM3-R2 open reading frame was removed by site-directed mutagenesis, without altering the amino acid sequence.