Relevant features or strain genotype | Source or Reference | |
---|---|---|
Plasmids: | ||
pBluescript SK- (pBS) | ColE1 ori; AmpR; cloning vector with blue-white selection. | Invitrogen |
pMN-hph | pBS containing at the EcoRV site a cassette of 1.8 kb bearing the E. coli-Hygromycin B resistance (hph) gene under EF-1 α promoter and GPD transcription terminator of X. dendrorhous. | [56] |
pBS-gCYP51 | pBS bearing the genomic version of the CYP51 gene from X. dendrorhous (4,318 bp). | This work |
pBS-cCYP51 | pBS bearing the ORF of the CYP51 gene from X. dendrorhous (1,653 bp). | This work |
pBS-cyp51/hph | pBS bearing the hygromycin B resistance cassette flanked by approximately 700 bp upstream and 600 pb downstream of the X. dendrorhous CYP51 locus. | This work |
YEp-Act4 | pBR322 and 2 micron ori, AmpR, LEU2 and containing the ACT promoter to regulate gene expression. | [48] |
YEpNP | YEp-Act4 bearing the S. cerevisiae TDH3 terminator. | This work |
YEpNP-cCYP51 | YEpNP bearing the X. dendrorhous CYP51 cDNA (1,653 bp) under the regulation of the ACT promoter and the TDH3 terminator. | This work |
YEpNP-gERG11 | YEpNP bearing the S. cerevisiae ERG11 gene (1,593 bp) under the regulation of the ACT promoter and the TDH3 terminator. | This work |
Strains: | ||
E. coli: | ||
DH-5α | F- φ80d lacZΔM15Δ (lacZY-argF) U169 deoR recA1 endA1 hsdR17(rk- mk+) phoA supE44 l- thi-1 gyrA96 relA1 | [47] |
S. cerevisiae: | ||
S288c | MATα, SUC2, gal2, mal, mel, flo1, flo8-1, hap1, ho, bio1, bio6. | [64] |
Meyen ex E.C Hansen YHR007C BY4743 (Sc-erg11+/−) | MATa/MATα, his3Δ1/ his3Δ1, leu2Δ0/leu2Δ0, lys2Δ0/+, met15Δ0/+, ura3Δ0/ura3Δ0, ΔERG11:KanMx. ATCC 26604. | ATCC 26604 |
Sc-cCYP51Xd | Diploid transformant derived from Sc-erg11 +/− containing the expression vector YEpNP-cCYP51. | This work |
Sc-gERG11Sc | Diploid transformant derived from Sc-erg11 +/− containing the expression vector YEpNP-gERG11. | This work |
Sc-hCYP51 | Sporulation product derived from Sc-erg11 +/− containing the expression vector YEpNP- cCYP51. | This work |
Sc-hERG11 | Sporulation product derived from Sc-erg11 +/− containing the expression vector YEpNP-gERG11. | This work |
X. dendrorhous: | ||
UCD 67-385 | ATCC 24230, wild-type. | ATCC 24230 |
CBS 6938 | ATCC 96594, wild-type. | ATCC 96594 |
CBS-CYP51 +/− | (CBS-CYP51/cyp51 hph). Transformant derived from CBS 6938 containing an allele of the CYP51 locus replaced by a hygromycin B resistance module. | This work |