Table 1.
Strains and plasmids used
| Strain name | Relevant characteristics | Reference |
|---|---|---|
| Yeast strains | ||
| BY4741 | MATa his3Δ1 leu2Δ0 met15Δ0 ura3Δ0 (S288C) | (59) |
| BY4743 | MATa/MATα his3-Δ1/his3-Δ1 leu2-Δ0/leu2-Δ0 lys2-Δ0/lys2-Δ0 ura3-0/ura3-0 | (59) |
| VDC5118 | YGN63 transformed with plasmid pYES-DEST52; Ura+, Leu+ | This study |
| VDC5119 | YGN63 transformed with plasmid pYES-DEST52; Ura+, Leu+ | This study |
| VDC5120 | YGN63 transformed with plasmid pBY135; Ura+, Leu+ | This study |
| VDC5121 | YGN63 transformed with plasmid pBY135; Ura+, Leu+ | This study |
| VDC5122 | YGN63 transformed with plasmid pBY136; Ura+, Leu+ | This study |
| VDC5123 | YGN63 transformed with plasmid pBY136; Ura+, Leu+ | This study |
| VDC5124 | YGN63 transformed with plasmid pBY137; Ura+, Leu+ | This study |
| VDC5125 | YGN63 transformed with plasmid pBY137; Ura+, Leu+ | This study |
| VDC5126 | YGN63 transformed with plasmid pBY138; Ura+, Leu+ | This study |
| VDC5127 | YGN63 transformed with plasmid pBY138; Ura+, Leu+ | This study |
| VDC5300 | Derivative of BY4743; SUA5/sua5-Δ1::LEU2 | This study |
| VDC5309 | Derivative of BY4741 transformed with pYES-DEST52::yrdCEc; Ura+ | This study |
| VDC5416 | Derivative of VDC5309; sua5-Δ1::LEU2; Ura+ Leu+ | This study |
| VDC5417 | Derivative of VDC5309; SUA5; Ura+ Leu+ | This study |
| VDC5440 | Derivative of VDC5416 after FOA treatment; sua5-Δ1::LEU2;Ura− Leu+ (cured of pYES-DEST52::yrdCEc), clone 14 | This study |
| VDC5441 | Derivative of VDC5416 after FOA treatment; sua5-Δ1::LEU2;Ura− Leu+ (cured of pYES-DEST52::yrdCEc), clone 15 | This study |
| YJN63 | MATα CYC I cyc7-67 ura3-52 leu2-3,112 cyh2 sua5::LEU2 | (4) |
| YMH13 | MATα cyc1–1019 cyc7-67 ura3–52 leu2–3,112 cyh2 | (4) |
| E. coli strains | ||
| BL21(DE3) | F− ompT hsdSB(rB−, mB−) gal dcm (DE3) | |
| E. coli TOP10 | F− mcrA Δ(mrr−hsdRMS−mcrBC) ϕ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(ara-leu) 7697 galU galK rpsL (StrR) endA1 nupG | Invitrogen |
| MG1655 | Coli Genetic Stock Center | |
| NM1100 | IN(rrnD−rrnE)1, rph-1, att PLCI857 Red αβγTetR | (60) |
| VDC5173 | Derivative of NM1100 ΔyrdCEc-1::KmR/pBAD24::yrdCEc; KmR/AmpR | This study |
| VDC5174 | Derivative of NM1100 ΔyrdCEc-1::KmR/pBAD24::yrdCEc; KmR/AmpR | This study |
| VDC5217 | MG1655 transformed with plasmid pBY125; AmpR | This study |
| VDC5218 | MG1655 transformed with plasmid pBY125; AmpR | This study |
| VDC5219 | MG1655 transformed with plasmid pBY126; AmpR | This study |
| VDC5220 | MG1655 transformed with plasmid pBY126; AmpR | This study |
| VDC5221 | MG1655 transformed with plasmid pBY128; AmpR | This study |
| VDC5222 | MG1655 transformed with plasmid pBY128; AmpR | This study |
| VDC5223 | MG1655 transformed with plasmid pBY130; AmpR | This study |
| VDC5224 | MG1655 transformed with plasmid pBY130; AmpR | This study |
| VDC5227 | MG1655 transformed with plasmid pBAD24; AmpR | This study |
| VDC5228 | MG1655 transformed with plasmid pBAD24; AmpR | This study |
| VDC5241 | MG1655 transformed with plasmid pBY166; AmpR | This study |
| VDC5333 | MG1655 transformed with plasmid pYC101.1; AmpR | This study |
| VDC5333 | MG1655 transformed with plasmid pYC101.4; AmpR | This study |
| VDC5335 | MG1655 transformed with plasmid pYC102.3; AmpR | This study |
| VDC5336 | MG1655 transformed with plasmid pYC102.3; AmpR | This study |
| VDC5337 | MG1655 transformed with plasmid pYC103.1; AmpR | This study |
| VDC5337 | MG1655 transformed with plasmid pYC103.2; AmpR | This study |
| VDC5339 | MG1655 transformed with plasmid pYC104.2; AmpR | This study |
| VDC5340 | MG1655 transformed with plasmid pYC104.4; AmpR | This study |
| VDC5342 | MG1655 transformed with plasmid pBY167; AmpR | |
| VDC5350 | Derivative of VDC5217 obtain from P1 transduction of the ΔyrdCEc-1::KmR allele of VDC5173; KmR, AmpR | This study |
| VDC5351 | Derivative of VDC5219 obtain from P1 transduction of the ΔyrdCEc-1::KmR allele of VDC5173; KmR, AmpR | This study |
| VDC5354 | Derivative of VDC5217 obtain from P1 transduction of the ΔyrdCEc-1::KmR allele of VDC5173; KmR, AmpR | This study |
| VDC5355 | Derivative of VDC5219 obtain from P1 transduction of the ΔyrdCEc-1::KmR allele of VDC5173; KmR, AmpR | This study |
| VDC5358 | Derivative of VDC5223 obtain from P1 transduction of the ΔyrdCEc-1::KmR allele of VDC5173; KmR, AmpR | This study |
| VDC5359 | Derivative of VDC5223 obtain from P1 transduction of the ΔyrdCEc-1::KmR allele of VDC517; KmR, AmpR | This study |
| VDC5362 | Derivative of VDC5242 obtain from P1 transduction of the ΔyrdCEc-1::KmR allele of VDC5173; KmR, AmpR | This study |
| VDC5363 | Derivative of VDC5242 obtain from P1 transduction of the ΔyrdCEc-1::KmR allele of VDC5173; KmR, AmpR | This study |
| VDC5366 | Derivative of VDC5333 obtain from P1 transduction of the ΔyrdCEc-1::KmR allele of VDC5173; KmR, AmpR | This study |
| VDC5367 | Derivative of VDC5333 obtain from P1 transduction of the ΔyrdCEc-1::KmR allele of VDC5173; KmR, AmpR | This study |
| VDC5370 | Derivative of VDC5337 obtain from P1 transduction of the ΔyrdCEc-1::KmR allele of VDC5173; KmR, AmpR | This study |
| VDC5371 | Derivative of VDC5337 obtain from P1 transduction of the ΔyrdCEc-1::KmR allele of VDC5173; KmR, AmpR | This study |
| Plasmids | ||
| pBN200 | pET21a:: yrdCEc | This Study |
| pBN204 | pYES::SUA5; URA3 | This study |
| pBY125 | pBAD24:: ywlCBs cloned as NcoI/XhoI fragment; AmpR | This study |
| pBY126 | pBAD24:: yrdCEc cloned as NcoI/XhoI fragment; AmpR | This study |
| pBY128 | pBAD24:: yciOEc cloned as NcoI/XhoI fragment; AmpR | This study |
| pBY130 | pBAD24:: SUA5 cloned as NcoI/PstI fragment; AmpR | This study |
| pBY135.1 | pYesDEST52:: yrdCEc; AmpR URA3 | This study |
| pBY136.1 | pYesDEST52:: yciOEc; AmpR URA3 | This study |
| pBY137.1 | pYesDEST52:: ywlCBs; AmpR URA3 | This study |
| pBY138.1 | pYesDEST52:: yrdCMm; AmpR URA3 | This study |
| pBY140 | pCR2.1::UpyrdCEc; AmpR (999 base pairs of upstream region of yrdCEc cloned in pCR2.1) | This study |
| pBY141.6 | pCR2.1::UpyrdCEc::DnyrdCEc; AmpR (999 base pairs of upstream and downstream region of yrdCEc cloned in pCR2.1) | This study |
| pBY146.1 | pGEM-T easy:: UpyrdCEc::DnyrdCEc; AmpR | This study |
| pBY152.1 | pGEM-T easy::UpyrdCEc::KmR::DnyrdCEc (ΔyrdCEC-1::KmR cassette) | |
| pBY166 | pBAD24:: ycdCMm; AmpR | This study |
| pBY167 | pBAD24:: ycdCMm; AmpR | This study |
| pYC101.1 | Derived from site directed mutagenesis of pBY126; yrdCEcArg110→Ala; AmpR | This study |
| pYC101.4 | Derived from site directed mutagenesis of pBY126; yrdCEcArg110→Ala; AmpR | This study |
| pYC102.2 | Derived from site directed mutagenesis of pBY126; yrdCEcArg52→Ala; AmpR | This study |
| pYC102.3 | Derived from site directed mutagenesis of pBY126; yrdCEcArg52→Ala; AmpR | This study |
| pYC103.1 | Derived from site directed mutagenesis of pBY126; yrdCEcLys50→Ala; AmpR | This study |
| pYC103.2 | Derived from site directed mutagenesis of pBY126; yrdCEcLys53→Ala; AmpR | This study |
| pYC104.2 | Derived from site directed mutagenesis of pBY126; yrdCEcLys50→Ala; AmpR | This study |
| pYC104.4 | Derived from site directed mutagenesis of pBY126; yrdCEcLys50→Ala; AmpR | This study |