Table 1 Strains and plasmids used in this work
Strain/plasmid | Description | Reference or source |
|---|---|---|
Strains | ||
E. coli | ||
DH5α | AmpS | [37] |
X. dendrorhous | ||
UCD 67–385 | Diploid wild-type strain (HygS and ZeoS) | [3]; ATCC 24230 |
385cyc8 − | HygR and ZeoR homozygous mutant (cyc8 −/cyc8 − ) that derives from UCD 67–385. A CYC8 allele was interrupted by a module that confers resistance to hygromycin B and the other allele by a module that confers resistance to zeocin | This work |
385tup1 − | HygR and ZeoR homozygous mutant (tup1 −/tup1 − ) that derives from UCD 67–385. A TUP1 allele was interrupted by a module that confers resistance to hygromycin B and the other allele by a module that confers resistance to zeocin | This work |
S. cerevisiae | ||
S288c | Haploid reference strain | ATCC 204508 |
YBR112C BY4741 (Sccyc8 −) | Haploid cyc8 − mutant strain derived from S288c. Genotype: MATa his3delta1 leu2delta0 met15delta0 ura3delta0 deltaCYC8 | [38]; ATCC 4007161 |
Sccyc8 −+YEpNP-CYC8 | Sccyc8 − strain carrying YEpNP-CYC8 (vector to express the X. dendrorhous CYC8 gene) | This work |
Sccyc8 −+YEpNP | Sccyc8 − strain carrying YEpNP (empty vector) | This work |
YCR084C BY4741 (Sctup1 −) | Haploid tup1 − mutant strain derived from S288c. Genotype: MATa his3delta1 leu2delta0 met15delta0 ura3delta0 deltaTUP1 | [38]; ATCC 4007198 |
Sctup1 −+YEpNP-TUP1 | Sctup1 − strain carrying YEpNP-TUP1 (vector to express the X. dendrorhous TUP1 gene) | This work |
Sctup1 − + YEpNP | Sctup1 − strain carrying YEpNP (empty vector) | This work |
Plasmids | ||
pBluescript II SK (−) XR | Cloning vector. (AmpR, pUC ori, white/blue colony selection) | Stratagene, La Jolla, CA, USA |
pMN-hph | pBluescript II SK (−) XR containing at the EcoRV site a 1.8 kb module that confers resistance to hygromycin B | [39] |
pIR-zeo | pBluescript II SK (−) XR containing at the EcoRV site a 1.2 kb module that confers resistance to zeocin | [7] |
pgCYC8 | pBluescript II SK (−) XR containing at the EcoRV site a 2.4 kb X. dendrorhous genomic fragment of the CYC8 gene | This work |
pgCYC8-Hyg | pgCYC8 containing at the BmgBI site the hygromycin B resistance module disrupting the CYC8 gene fragment | This work |
pgCYC8-Zeo | pgCYC8 containing between PmlI and BmgBI sites the zeocin resistance module replacing 527 bp of the CYC8 gene fragment | This work |
pgTUP1 | pBluescript II SK(−)XR containing at EcoRV site a 2.7 kb X. dendrorhous genomic fragment of the TUP1 gene | This work |
pgTUP1-Hyg | pgTUP1 containing the hygromycin B resistance module replacing a 2.0 kb EcoRV fragment inside the TUP1 gene fragment | This work |
pgTUP1-Zeo | pgTUP1 containing the zeocin resistance module replacing a 2.0 kb EcoRV fragment inside the TUP1 gene fragment | This work |
YEpNP | S. cerevisiae expression vector modified from YEpAct4 [40]. LEU2 gene auxotrophy marker. It has a XbaI restriction site between the Act4 constitutive promoter and the TDH3t terminator | [41] |
YEpNP-CYC8 | S. cerevisiae expression vector derived from YEp-NP containing at the XbaI site between the Act4 promoter and the TDH3t terminator the X. dendrorhous CYC8 cDNA | This work |
YEpNP-TUP1 | S. cerevisiae expression vector derived from YEp-NP containing at the XbaI site between the Act4 promoter and the TDH3t terminator the X. dendrorhous TUP1 cDNA | This work |