Table 3.
Strains and plasmids used in this study
| Strains or plasmids | Characteristics | Reference or source |
|---|---|---|
| Strains |
|
|
|
S. cerevisiae |
|
|
| 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 |
|
|
| DH5α |
General cloning host |
Takara |
| WM1788 |
Cloning host |
Provided by Professor William Metcalf |
| Plasmids |
|
|
| 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 |