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Table 3 Plasmids and strains used and constructed in this study

From: Identification and functional characterization of the CYP51 gene from the yeast Xanthophyllomyces dendrorhous that is involved in ergosterol biosynthesis

  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