Table 3 Strains and plasmids used in this study
Strains or plasmids | Characteristics | Reference or source |
|---|---|---|
Strains | Â | Â |
S. cerevisiae | Â | Â |
BY4741 | MATa his3Δ0 leu2Δ0 met15Δ0 ura3Δ0 | [47] |
BAD | BY4741/δ::TEF1p-ago1-TPI1p-dcr1 | This study |
BAD-P | BAD/pRS416-TTrcx | This study |
SIZ1-kd | BAD/pRS416-TTrcx-siz1 | This study |
GCN4-kd | BAD/pRS416-TTrcx-gcn4 | This study |
siz1Δ | BAD/siz1Δ::leu2 | This study |
siz1Δ-P | siz1Δ/pRS416e | This study |
siz1Δ-SIZ1 | siz1Δ/pRS416e-siz1 | This study |
gcn4Δ | BAD/gcn4Δ::leu2 | This study |
siz2Δ | BAD/siz2Δ::leu2 | This study |
mms21Δ | BAD/siz1Δ::leu2 | This study |
cst9Δ | BAD/cst9Δ::leu2 | This study |
BAD-YKL071W | BAD/pRS416e-ykl071w | This study |
BAD-ZWF1 | BAD/pRS416e-zwf1 | This study |
BAD-MSN2 | BAD/pRS416e-msn2 | This study |
BAD-ALD6 | BAD/pRS416e-ald6 | This study |
BAD-ADH7 | BAD/pRS416e-adh7 | This study |
BAD-ARI1 | BAD/pRS416e-ari1 | This study |
siz1Δ-GCN4-kd | siz1Δ/pRS416-TTrcx-gcn4 | This study |
HZ848 | MATα, ade2-1, Δura3, his3-11, 15, trp1-1, leu2-3, 112, and can1-100 | [23] |
HZ848-siz1Δ | HZ848/siz1Δ::ura3 | This study |
W303a | MATa; ura3-1; trp1Δ 2; leu2-3,112; his3-11,15; ade2-1; can1-100 | [24] |
W303a-siz1Δ | W303a/siz1Δ::hygromycin B | This study |
E. coli | Â | Â |
DH5α | General cloning host | Takara |
WM1788 | Cloning host | Provided by Professor William Metcalf |
Plasmids | Â | Â |
pRS416 | Yeast centromere with URA3 marker | [48] |
pRS425-TEF1p-PmeI-PGK1t | Yeast gene expression vector | [19] |
pRS416e | Derived from pRS416, with TEF1 promoter and PGK1 terminator added | This study |
pRS-delta-KanMX-LoxP-TEF1p-AGO1-PGK1t-TPI1p-DCR1-GPD1t | Helper plasmid for integration of S. castellii RNAi pathway into delta-site | [19] |
pRS416-TTrc | Derived from pRS416, with convergent promoters to produce dsRNA | [19] |
pRS416-TTrcx | Derived from pRS416-TTrc, with XhoI restriction recognition sequence instead of BamHI | This study |
pRS416-TTrcx-siz1 | Derived from pRS416-TTrcx, with gene SIZ1 fragment added | This study |
pRS416-TTrcx-gcn4 | Derived from pRS416-TTrcx, with gene GCN4 fragment added | This study |
pRS415 | Yeast centromere with LEU2 marker | [48] |
pUG6 | The loxP-KanMX-loxP disruption module | Euroscarf |
pUG72 | The loxP-URA3-loxP disruption module | Euroscarf |
pLHCX | Template for amplification of hygromycin B resistance gene | Clontech |
pXZ5 | Derived from pUG72, with hygromycin B resistance gene expression cassette instead of ura3 | This study |
pRS416e-siz1 | Derived from pRS416e, with SIZ1 gene cassette added | This study |
pRS416e-ykl071w | Derived from pRS416e, with YKL071W gene cassette added | This study |
pRS416e-zwf1 | Derived from pRS416e, with ZWF1 gene cassette added | This study |
pRS416e-msn2 | Derived from pRS416e, with MSN2 gene cassette added | This study |
pRS416e-ald6 | Derived from pRS416e, with ALD6 gene cassette added | This study |
pRS416e-adh7 | Derived from pRS416e, with ADH7 gene cassette added | This study |
pRS416e-ari1 | Derived from pRS416e, with ARI1 gene cassette added | This study |