Table 1 Strains and plasmids used in this work
Strain/Plasmid | Genotype or relevant features | Reference |
|---|---|---|
Strain: | ||
X. dendrorhous | ||
UCD 67–385 | Wild type, diploid strain. | ATCC 24230 |
385-cyp61(−/−) | Homozygote transformant derived from UCD 67–385 with both CYP61 alleles interrupted, one with a hygromycin B resistance cassette and the other with a zeocin resistance cassette. | [16] |
385-erg10(+/−) | Heterozygous mutant of UCD 67–385 with one allele of ERG10 replaced by a hygromycin B resistance cassette. | This work |
385-pot1(+/−) | Heterozygous mutant of UCD 67–385 with one allele of POT1 replaced by a hygromycin B resistance cassette. | This work |
385-pot1(−/−) | Homozygous mutant of UCD 67–385 with both POT1 alleles replaced by a hygromycin B resistance cassette. | This work |
385-ERG10 | Heterozygote transformant derived from UCD 67–385 containing an additional ERG10 allele and a hygromycin B resistance cassette integrated at locus int. | This work |
385-POT1 | Heterozygote transformant derived from UCD 67–385 containing an additional POT1 allele and a hygromycin B resistance cassette integrated at locus int. | This work |
385-Vexp2 | Heterozygote transformant derived from UCD 67–385 containing an empty over-expressing cassette (without an inserted ORF) and a hygromycin B resistance cassette integrated at locus int. | This work |
S. cerevisiae | ||
s288C | MATα, SUC2, gal2, mal, mel, flo1, flo8-1, hap1, ho, bio1, bio6 | [38] |
ATCC 4022800 (Sc-ERG10/erg10) | MATa/MATalpha his3delta1/his3delta1 leu2delta0/leu2delta0 lys2delta0/+ met15delta0/+ ura3delta0/ura3delta0 deltaERG10 | [39] |
Sc-ERG10sc | Strain Sc-ERG10/erg10 carrying plasmid YEpNP-10sc. | This work |
Sc-cERG10xd | Strain Sc-ERG10/erg10 carrying plasmid YEpNP-c10Xd. | This work |
Sc-cPOT1xd | Strain Sc-ERG10/erg10 carrying plasmid YEpNP-cPOT1Xd. | This work |
Plasmid: | ||
pBluescript SK- (pBS) | ColE1 replication origin, AmpR, LacZ for blue-white colony screening. | Stratagene |
pBS-PT-ERG10xd | pBS with 620 bp of DNA upstream of ERG10 gene and 412 bp downstream with a SmaI site between them. | This work |
pBS-PT-POT1xd | pBS with 559 bp of DNA upstream of POT1 gene and 560 bp downstream with a SmaI site between them. | This work |
pMN-hph | Plasmid containing an hygromycin B resistance cassette for X. dendrorhous. | [12] |
pPHT-Erg10xd | pBS -PT-ERG10Xd with a hygromycin B resistance cassette in the SmaI site. | This work |
pPHT-POT1xd | pBS-PT-POT1Xd with a hygromycin B resistance cassette in the SmaI site. | This work |
pPZT-Erg10xd | pBS-PT-ERG10Xd with a zeocin resistance cassette in the SmaI site. | This work |
YEp-ACT4 | pBR322 and 2 micron replication origins, AmpR, LEU2, promoter ACT4. | [32] |
YEp-NP | YEpACT4 with a TDH3 terminator next to pACT4. | [28] |
YEpNP-10sc | YEp-NP with ERG10 DNA from S. cerevisiae between pACT4 and tTDH3. | This work |
YEpNP-c10xd | YEp-NP with ERG10 from X. dendrorhous cDNA between pACT4 and tTDH3. | This work |
YEpNP-cPOT1xd | YEp-NP with POT1 from X. dendrorhous cDNA between pACT4 and tTDH3 | This work |
pXdVexp2 | X. dendrorhous expression vector: pBS bearing the X. dendrorhous ubiquitin promoter [GenBank: KJ140285] and GPD terminator [Genbank:Y08366] with a BamHI site between them to insert the gene to express and the hygromycin B cassette for selection, flanked by non-encoding genomic [GenBank: KJ140286] regions to target the construction integration in the genome. | [14] |
pXdVexp2-cERG10xd | pXdVexp2 containing the cDNA version of the ERG10 gene from X. dendrorhous | This work |
pXdVexp2-cPOT1xd | pXdVexp2 containing the cDNA version of the POT1 gene from X. dendrorhous | This work |