Table 1 Primers, plasmids and strains used in this work
Name | Description | Reference |
|---|---|---|
E. coli strain | ||
Top 10 | F-mcr A Δ(mrr-hsd RMS-mcr BC) Φ80lac ZΔM15 Δlac X74 rec A1 ara D139 Δ(ara leu) 7697 gal U gal Krps L (StrR) end A1 nup G | Life Technology |
Primers for gene disruption | ||
Δpxa2-F | 5′- ATAATAATAC AATTAAAAGT TACCGAAGAA AGATTTTATA CAGCTGAAGC TTCGTACGC-3′ | This work |
Δpxa2-R | 5′- CAATTTATAC ATGATTTGGA TCCTCCTTTG GCTATGTATG GCATAGGCCA CTAGTGGATC TG-3′ | This work |
Primers for restriction endonuclease | ||
F-BamHI | 5′-CGGGATCC AT GTTGCATAAC TTGTTCGACA AGA-3′ | This work |
R-SalI | 5′-GCGTCGAC AG TAGAATCCAA ACCCAACAAT GGA-3′ | This work |
Plasmids | ||
PUG27 | loxP-kanMX-loxP disruption module plasmid | EUROSCARF |
pESC-leu | With GAL1 and GAL10 yeast promoters in opposite orientation, CYC1 and ADH1 terminator respectively | Agilent |
S. cerevisiae strains | ||
Wild type | MATa; his3Δ 1; leu2Δ 0; met15Δ 0; ura3Δ 0 | EUROSCARF |
Δpxa1 | BY4741; Mat a; his3D1; leu2D0; met15D0; ura3D0; YPL147w::kanMX4 | EUROSCARF |
Δpxa2 | BY4741; Mat a; his3D1; leu2D0; met15D0; ura3D0; YKL188c::kanMX4 | EUROSCARF |
Δpxa1&2 | BY4741; Mat a; his3D1; leu2D0; met15D0; ura3D0; YPL147w::kanMX4; YKL188c::his | This work |
WT-pESC | wild type carring pESC | This work |
Δpxa1-pESC | Δpxa1 carrying pESC | This work |
Δpxa2-pESC | Δpxa2 carrying pESC | This work |
Δpxa1&2-pESC | Δpxa1&2 carrying pESC | This work |
WT-9LHP | wild type carring 9LHP | This work |
Δpxa1-9LHP | Δpxa1 carrying 9LHP | This work |
Δpxa2-9LHP | Δpxa2 carrying 9LHP | This work |
Δpxa1&2-9LHP | Δpxa1&2 carrying 9LHP | This work |