Table 1.
Strains and plasmids used in this study
| Strain or plasmid | Genotype or relevant characteristics | Source or reference |
|---|---|---|
| Strain | ||
| S. cerevisiae | ||
| RS453 | MATaade2-1 his3-11,15 leu2-3112 trp1-1 ura3-52 | (41) |
| SS1026 | pah1Δ::TRP1 derivative of RS453 | (31) |
| SS1132 | pah1Δ::TRP1 nem1Δ::HIS3 derivative of RS453 | (48) |
| E. coli | ||
| DH5⍺ | F- Φ80 lacZΔM15Δ (lacZYA-argF)U169 deoR recA1 endA1 hsdR17(rk- mk+) phoA supE44 λ−thi-1 gyrA96 relA1 | (95) |
| NiCo21(DE3)pLysSRARE2 | can::CBD fhuA2 [Ion] ompT gal (λ DE3) [dcm] amA::CBD sly::CBD glmS6Ala ΔhsdS λ DE3 = λ sBamHIo ΔEcoRI-B int::(lacI::PlacUV5::T7 gene1) i21 Δnin5 pLysSRARE2 | New England Biolabs |
| Plasmid | ||
| pET-15b | E. coli expression vector with N-terminal His6-tag fusion | Novagen |
| pGH313 | PAH1 coding sequence inserted into pET-15b | (5) |
| pGS108 | PAH1(Δ180–272) derivative of pGH313 | This study |
| pRS415 | Single-copy number E. coli/yeast shuttle vector with LEU2 | (105) |
| pGH315 | PAH1 inserted into pRS415 | (48) |
| pGH315-ΔN-NCRb | PAH1(Δ186–266) derivative of pGH315 | (63) |
| pYES2 | High-copy number E.coli/yeast shuttle vector with URA3 and GAL1 promoter | Thermo Fisher Scientific |
| pGH452 | PAH1-TAP in pYES2 with calmodulin binding peptide DNA sequence removed from the TAP tag | (64) |
| pGS104 | PAH1(Δ180–272) derivative of pGH452 | This study |
| YCplac33-SEC63-GFP | SEC63-GFP fusion inserted into the CEN/URA3 vector | (41) |