. 2019 Dec 5;61(2):229–243. doi: 10.1194/jlr.RA119000431

TABLE 1.

Strains and plasmids used in this work

Strain/Plasmid	Description	Reference or Source
Strain
E. coli
DH5α	Amp^S. Used for molecular cloning and plasmid maintenance.	(27)
S. cerevisiae
S288c	Haploid reference strain used for plasmid construction by DNA assembler.	ATCC 204508
X. dendrorhous
CBS 6938	Wild-type strain (Zeo^S, Hyg^S, and Nat^S).	ATCC 96594
CBS.cyp61⁻	Mutant (Zeo^R, Hyg^S, and Nat^S) derived from CBS 6938. The single CYP61 locus was interrupted by the zeocin resistance cassette.	This work
CBS.Δstp1	Mutant (Zeo^S, Hyg^R, and Nat^S) derived from CBS 6938. The single STP1 locus was replaced by the hygromycin B resistance cassette.	This work
CBS.Δstp1/STP1	Mutant (Zeo^R, Hyg^S, and Nat^S) derived from CBS.Δstp1, with the native STP1 allele reintegrated at the STP1 native locus and followed by the zeocin resistance cassette.	This work
CBS.cyp61⁻.Δstp1	Mutant (Zeo^R, Hyg^R, and Nat^S) derived from CBS.cyp61^-. The single STP1 locus was replaced by the hygromycin B resistance cassette.	This work
CBS.FLAG.SRE1	Mutant (Zeo^R, Hyg^S, and Nat^S) derived from CBS 6938. The native SRE1 gene was replaced by a gene variant that expresses the Sre1 protein fused to the 3xFLAG epitope at its N-terminus, followed by the zeocin resistance cassette.	This work
CBS.FLAG.SRE1.Δstp1	Mutant (Zeo^R, Hyg^R, and Nat^S) derived from CBS.FLAG.SRE1. The single STP1 locus was replaced by the hygromycin B resistance cassette.	This work
CBS.Δstp1.FLAG.SRE1N	Mutant (Zeo^R, Hyg^R, and Nat^S) derived from CBS.Δstp1. The native SRE1 gene was replaced by a gene variant that expresses the N-terminal domain of Sre1 (Sre1N) fused to the 3xFLAG epitope at its N-terminus, followed by the zeocin resistance cassette.	This work
CBS.FLAG.SRE1N	Mutant (Zeo^R, Hyg^S, and Nat^S) derived from CBS 6938. The native SRE1 gene was replaced by a gene version that expresses Sre1N fused to the 3xFLAG epitope at its N-terminal, followed by the zeocin resistance cassette.	(23)
CBS.cyp61⁻.FLAG.SRE1	Mutant (Zeo^R, Hyg^R, and Nat^S) derived from CBS.cyp61⁻. The native SRE1 gene was replaced by a gene variant that expresses the Sre1 protein fused to the 3xFLAG epitope at its N-terminus, followed by the hygromycin B resistance cassette.	This work
CBS.cyp61⁻.FLAG.SRE1.Δstp1	Mutant (Zeo^R, Hyg^R, and Nat^R) derived from CBS.cyp61⁻.FLAG.SRE1. The single STP1 locus was replaced by the nourseothricin resistance cassette.	This work
CBS.sre1⁻	Mutant (Zeo^R, Hyg^S, and Nat^S) derived from CBS 6938. Gene SRE1 was partially deleted (approximately 90% of the coding region was replaced by the zeocin resistance cassette).	(23)
CBS.ΔS1P-Cn	Mutant (Zeo^S, Hyg^R, and Nat^S) derived from CBS 6938. The single CDZ97435 locus (putative homolog of the potential S1P homolog identified in C. neoformans) was replaced by the hygromycin B resistance cassette.	This work
CBS.ΔS1P-Hs	Mutant (Zeo^S, Hyg^R, and Nat^S) derived from CBS 6938. The single CED82778 locus (putative homolog of H. sapiens S1P encoding gene) was replaced by the hygromycin B resistance cassette.	This work
CBS.cyp61⁻.ΔS1P-Cn	Mutant (Zeo^S, Hyg^R, and Nat^S) derived from CBS.cyp61⁻. The single CDZ97435 locus was replaced by the hygromycin B resistance cassette.	This work
CBS.cyp61⁻.ΔS1P-Hs	Mutant (Zeo^S, Hyg^R, and Nat^S) derived from CBS.cyp61⁻. The single CED82778 locus was replaced by the hygromycin B resistance cassette.	This work
Plasmid
pBluescript SK- (pBS)	Cloning vector (ColE1 ori, Amp^R, blue-white colony selection).	Agilent Technologies Inc.
pMN-hph	pBS containing the hygromycin B resistance cassette (1.8 kb) used for X. dendrorhous transformant selection at the EcoRV site.	(30)
pIR-zeo	pBS containing the zeocin resistance cassette (1.2 kb) used for X. dendrorhous transformant selection at the EcoRV site.	(26)
pBS-nat	pBS containing the nourseothricin resistance cassette (1.4 kb) used for X. dendrorhous transformant selection at the EcoRV site. The constructed cassette contains the Streptomyces noursei nat1 gene regulated by the X. dendrorhous EF-1α gene promoter and GPD gene transcription terminator.	This work
pBS-ΔgSTP1^hph	pBS containing the 715 bp upstream and 620 bp downstream of the STP1 gene and the hygromycin B resistance cassette between them at the EcoRV site. Used to delete the X. dendrorhous STP1 gene by homologous recombination.	This work
pBS-ΔgSTP1^nat	pBS containing the 715 bp upstream and 620 bp downstream of the STP1 gene and the nourseothricin resistance cassette between them at the EcoRV site. Used to delete the X. dendrorhous STP1 gene by homologous recombination.	This work
pBS-gSTP1^up-down	pBS containing at the SmaI site, the X. dendrorhous STP1 gene and the zeocin resistance cassette at the AgeI site of the downstream region of the STP1 gene. Used to integrate the X. dendrorhous STP1 gene by homologous recombination in Δstp1 mutants.	This work
pXd-gSRE1N-zeo	Plasmid constructed by DNA assembler used to replace the X. dendrorhous SRE1 gene with the gene variant that expresses the Sre1N (Sre1 N-terminal domain) 3xFLAG-tagged at its N-terminal end, followed by the zeocin resistance cassette for transformant selection.	(23)
pXd-gSRE1-zeo	Plasmid constructed by DNA assembler used to replace the X. dendrorhous SRE1 gene with the gene variant that expresses Sre1 (full-length Sre1 protein) 3xFLAG-tagged at its N-terminal end. It contains 2,816 bp (from the start to the stop codon) of the genomic SRE1 fused to the 3xFLAG-encoding sequence at the 5′ end. This sequence is flanked by the upstream and downstream regions of the SRE1 gene to direct its integration at the SRE1 locus including a zeocin resistance cassette for transformant selection.	This work
pXd-gSRE1-hph	Plasmid constructed by DNA assembler used to replace the X. dendrorhous SRE1 gene with the gene variant that expresses Sre1 (full-length Sre1 protein) 3xFLAG-tagged at its N-terminal end, followed by the hygromycin B resistance cassette for transformant selection. It was constructed in the same way as pXd-gSRE1-zeo, but instead of zeocin, it contains the hygromycin B resistance cassette for transformant selection.	This work
pYES2	S. cerevisiae expression vector containing 2 μ origin. Used to amplify the 2 μ DNA by PCR, which was then used for plasmid construction by DNA assembler.	Thermo Fisher Scientific Inc.
pFA6	Yeast plasmid with kanamycin/geneticin (G418) resistance marker. Used to amplify the G418 marker by PCR, which was then used for plasmid construction by DNA assembler.	(65)
pFlagTEM1	Expression plasmid containing the FLAG-tagged lactamase-encoding sequence. Used to amplify the 3xFLAG epitope (3xDYKDDDDK)-encoding sequence, which was then used for plasmid construction by DNA assembler.	(66)
pBS.ΔS1P-Cn ^hph	pBS containing the 720 bp upstream and 659 bp downstream of the CDZ97435 locus and the hygromycin B resistance cassette between them at the EcoRV site. Used to delete the X. dendrorhous CDZ97435 locus by homologous recombination.	This work
pBS.ΔS1P-Hs ^hph	pBS containing the 725 bp upstream and 640 bp downstream of the CED82778 locus and the hygromycin B resistance cassette between them at the EcoRV site. Used to delete the X. dendrorhous CED82778 locus by homologous recombination.	This work
Amp^S, sensitive to ampicillin; Zeo^S/Zeo^R, sensitive/resistant to zeocin; Hyg^S/Hyg^R, sensitive/resistant to hygromycin B; Nat^S/Nat^R, sensitive/resistant to nourseothricin; ColE1 ori, replication origin of E. coli ColE1 plasmid, Amp^R, ampicillin resistance.