TABLE 1.
Strains and plasmids used in this study
| Name | Genotype/description | Reference/source |
|---|---|---|
| S. cerevisiae | ||
| SEY6210 | MATα leu2-3,112 ura3-52 his3-200 trp1-901 lys2-801 suc2-9 | Ref. 26 |
| BHY10 | SEY6210 CPY-Invertase::LEU2 (pBHY11) | Ref. 27 |
| MBY3 | SEY6210 vps4Δ::TRP1 | Ref. 28 |
| GOY223 | BHY10 vps9Δ::HIS3 | Ref. 3 |
| DNY516 | BHY10 muk1Δ::KAN | This study |
| DNY517 | BHY10 vps9Δ::HIS3 muk1Δ::KAN | This study |
| DNY206 | BHY10 vps21Δ::KAN | Ref. 3 |
| DNY471 | BHY10 vps21Δ0 ypt52Δ0 | Ref. 3 |
| E. coli | ||
| TOP10F′ | F′ lacIq, Tn10(TetR) mcrA Δ(mrr-hsdRMS-mcrBC) ϕ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(ara-leu)7697 galU galK rpsL (StrR) endA1 nupG | Invitrogen |
| BL21(DE3) | F − ompT gal dcm lon hsdSB (rB − mB −)λ (DE3 [lacI lacUV5-T7 ind1 sam7 nin5]) | Stratagene/Agilent |
| Plasmids | ||
| pRS416 | URA3 AmpR CEN | Ref. 29 |
| pRS426 | URA3 AmpR 2μ | Ref. 29 |
| pDN216 | MUK1 (pRS426) | This study |
| pGO36 | GFP (pRS416) | Ref. 30 |
| pGO45 | GFP-CPS1 (pRS426) | Ref. 30 |
| pDN219 | GFP-MUK1 (pRS416) | This study |
| pDN251 | URA3 LoxP::CEN::LoxP PGK1pr::RLuc 0::FLuc | Ref. 3 |
| pDN252 | PGK1pr::RLuc SNA3-FLuc (pDN251) | Ref. 3 |
| pDN312 | PGK1pr::RLuc STE3-FLuc (pDN251) | This study |
| pDN268 | PGK1pr::RLuc MUP1-FLuc (pDN251) | This study |
| pSna3-GFP | SNA3-GFP (pRS416) | Ref. 31 |
| pCHL642 | URA3 AmpR MUP1-GFP (pRS416) | S. Emr (Cornell) |
| pDN615 | LEU2 AmpR LoxP::CEN::LoxP (pRS415) | This study |
| pDN274.2 | mCherry-VPS9 (pDN615) | This study |
| AMP219 | AmpR GST-VPS21 (pParallel-GST) | Ref. 20 |
| AMP220 | AmpR GST-YPT52 (pParallel-GST) | Ref. 20 |
| AMP217 | AmpR GST-YPT53 (pParallel-GST) | Ref. 20 |
| AMP218 | AmpR GST-YPT7 (pParallel-GST) | Ref. 20 |
| AMP1434 | AmpR GST-MUK1 (pFB HTB) | This study |
| AMP84 | AmpR His6-Gyp1TBC (pET22) | Ref. 20 |