TABLE 1.

Strains and plasmids used in this study

Strain or plasmidRelevant characteristicsaSource or reference
Strains
    P. putida strains
        DOT-T1ERifr17
        DOT-T1EVTRifr Kmr Telr ttgV::Km ttgT::kilA/telAB27
    E. coli strains
        BL21(DE3)Carries T7 RNA polymerase under the control of the lacUV5 promoterNovagen
        DH5αsupE44 Δ(lacZYA-argF) U169 deoR (φ80lacZΔM15) hsdR17(rK mK) recA1 endA1 gyrA (Nalr) thi-1 relA1 Δ(lacIZYΔ-argF)28
        JM109F′ traD36 lacIq Δ(lacZ)M15 proA+B+/e14 (McrA) Δ(lac-proAB) thi gyrA96 (Nalr) endA1 hsdR17(rK mK) relA1 supE44 recA129
Plasmids
    pET28b(+)Kmr; protein expression vectorNovagen
    pANA126Kmr; pET28b(+) derivative vector used to produce His6-TtgV22
    pMP220Tcr; promoterless lacZ expression vector26
    pMPD1Tcr; ttgD::′lacZ fusion cloned in pMP22027
    pANA96Tcr; ttgG::′lacZ fusion cloned in pMP22022
    pGEM-TApr; cloning vectorPromega
    pMBL-TApr; cloning vectorPromega
    pGG1Apr; pUC18 bearing an 8-kb BamHI fragment with ttgGHI and ttgVW22
    pT1-B6Apr; pUC18 bearing an 6.8-kb BamHI fragment with ttgDEF and ttgT22
    pBBRNpBBR-MCS5 derivative with an additional NdeI restriction siteThis work
    pBBRN::ttgVGmr; pBBRN derivative vector expressing TtgVThis work
    pBBRN::R47AGmr; pBBRN derivative vector expressing TtgV(R47A)This work
    pBBRN::V50AGmr; pBBRN derivative vector expressing TtgV(V50A)This work
    pBBRN::Q51AGmr; pBBRN derivative vector expressing TtgV(Q51A)This work
    pBBRN::E60AGmr; pBBRN derivative vector expressing TtgV(E60A)This work
    pET28b(+)::R47AKmr; pET28b(+) derivative vector used to produce His6-TtgV(R47A)This work
    pET28b(+)::S48AKmr; pET28b(+) derivative vector used to produce His6-TtgV(S48A)This work
    pET28b(+)::T49AKmr; pET28b(+) derivative vector used to produce His6-TtgV(T49A)This work
    pET28b(+)::V50AKmr; pET28b(+) derivative vector used to produce His6-TtgV(V50A)This work
    pET28b(+)::Q51AKmr; pET28b(+) derivative vector used to produce His6-TtgV(Q51A)This work
    pET28b(+)::R52AKmr; pET28b(+) derivative vector used to produce His6-TtgV(R52A)This work
    pET28b(+)::I53AKmr; pET28b(+) derivative vector used to produce His6-TtgV(I53A)This work
    pET28b(+)::I54AKmr; pET28b(+) derivative vector used to produce His6-TtgV(I54A)This work
    pET28b(+)::N55AKmr; pET28b(+) derivative vector used to produce His6-TtgV(N55A)This work
    pET28b(+)::L57AKmr; pET28b(+) derivative vector used to produce His6-TtgV(L57A)This work
    pET28b(+)::E58AKmr; pET28b(+) derivative vector used to produce His6-TtgV(E58A)This work
    pET28b(+)::E59AKmr; pET28b(+) derivative vector used to produce His6-TtgV(E59A)This work
    pET28b(+)::E60AKmr; pET28b(+) derivative vector used to produce His6-TtgV(E60A)This work
    pET28b(+)::F61AKmr; pET28b(+) derivative vector used to produce His6-TtgV(F61A)This work
    pET28b(+)::L62AKmr; pET28b(+) derivative vector used to produce His6-TtgV(L62A)This work
    pET28b(+)::V63AKmr; pET28b(+) derivative vector used to produce His6-TtgV(V63A)This work
    pET28b(+)::E64AKmr; pET28b(+) derivative vector used to produce His6-TtgV(E64A)This work
    pMBLT::pDEFApr; pMBLT derivative plasmid bearing the ttgD promoterThis work
    pMBLT::pDEF box 1Apr; pMBLT derivative plasmid bearing a mutant ttgD promoter in box 1This work
    pGEMT::pDEF box 3Apr; pGEMT derivative plasmid bearing a mutant ttgD promoter in box 3This work
    pMBLT::pDEF box 4Apr; pMBLT derivative plasmid bearing a mutant ttgD promoter in box 4This work
    pGEMT::pDEF box 5Apr; pGEMT derivative plasmid bearing a mutant ttgD promoter in box 5This work
    pMBLT::pGHIApr; pMBLT derivative plasmid bearing the ttgG promoterThis work
    pGEMT::pGHI box 1Apr; pGEMT derivative plasmid bearing a mutant ttgG promoter in box 1This work
    pMBLT::pGHI box 3Apr; pMBLT derivative plasmid bearing a mutant ttgG promoter in box 3This work
    pGEMT::pGHI box 4Apr; pGEMT derivative plasmid bearing a mutant ttgG promoter in box 4This work
    pGEMT::pGHI box 5Apr; pGEMT derivative plasmid bearing a mutant ttgG promoter in box 5This work
  • a Apr, Gmr, Kmr, Rifr, Tcr, and Telr stand for resistance to ampicillin, gentamicin, kanamycin, rifampin, tetracycline, and tellurite, respectively.