TABLE 3.
Strains and plasmids used in this study
| Strains | Relevant characteristicsa | Source or reference |
|---|---|---|
| Strains | ||
| E. coli | ||
| TOP10 | F– mcrA Δ(mrr-hsdRMS-mcrBC)ϕ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(ara-leu)7697 galU galK rpsL (Strr) endA1 nupG | Invitrogen |
| M15(pREP4) | thi lac ara gal mtl F′ recA+ uvr+ lon+; pREP4 plasmid; Kanr | Qiagen |
| BL21(DE3)pLysS | F– ompT hsdSB(rB– mB–) gal dcm (DE3)/pLysS Cmr | Novagen |
| S17-1 | recA endA thi hsdR RP4-2-Tc::Mu-Kan::Tn7 Tpr Smr | 66 |
| R. sphaeroides | ||
| WS8N | Wild type; spontaneous Nalr | 64 |
| SltF1 | WS8N derivative ΔsltF(1-336)::aadA Fla– Spcr Nalr | 48 |
| SP13 | WS8N derivative fleQΔ::Kan | 42 |
| Plasmids | ||
| pTZ19R | Cloning vector; Apr ; pUC derivative | Pharmacia |
| pRK415 | pRK404 derivative, for expression in R. sphaeroides, lacZ mob+ Tcr | 65 |
| pBBMCS53 | Transcriptional uidA fusion vector; Gmr | 67 |
| pET19-b | Expression vector 10×His N terminal; Apr | Novagen |
| pQE30 | Expression vector 6×His N terminal; Apr | Qiagen |
| pQE60 | Expression vector 6×His C terminal; Apr | Qiagen |
| pGT001 | 1.4-kb PstI fragment containing sltF wild type cloned into pTZ19R | 40 |
| pRKSltF | sltF wild type cloned into the EcoRI/HindIII sites of pRK415 | 48 |
| pRKrevSltF | sltF wild type cloned into the HindIII/EcoRI sites of pRK415 | This study |
| pBG313 | 3.8 kb BamHI fragment from WS8-N cloned in pTZ19R | 40 |
| Up120sltFprev | pBBMCS53 derivative, sltFp3′-5′–uidA gene fusion | This study |
| Up120sltFpfw | pBBMCS53 derivative, sltFp5′-3′–uidA gene fusion | This study |
| pRS1sltF | sltF cloned into the SacI/HindIII sites of pQE30 | 49 |
| pQE30sltFΔ47 | ΔsltF(510-651) cloned into the SacI/HindIII sites of pQE30 | 41 |
| pQE30sltFΔ48/Δ6 | ΔsltF(651-795) cloned into the SacI/HindIII sites of pQE30 | 49 |
| pQE30sltFΔ95 | ΔsltF(510-795) cloned into the SacI/HindIII sites of pQE30 | 41 |
| pRSJ | flgJ wild type cloned into the NcoI/BglII sites of pQE60 | 48 |
| pRSFliE | fliE wild type cloned into the SacI/HindIII sites of pQE30 | 71 |
| pRSFlgB | flgB wild type cloned into the BamHI/HindIII sites of pQE30 | 71 |
| pRSFlgC | flgC wild type cloned into the NdeI/BamHI sites of pET19-b | 71 |
| pRSFlgF | flgF wild type cloned into the KpnI/HindIII sites of pQE30 | 71 |
| pRSFlgG | flgG wild type cloned into the NdeI/BamHI sites of pET19-b | 71 |
| pRSY5 | cheY5 wild type cloned into the BamHI/PstI sites of pQE30 | 72 |
a
Str, streptomycin; Ap, ampicillin; Nal, nalidixic acid; Kan, kanamycin; Gm, gentamicin; Spc, spectinomycin; Tc, tetracycline; Cm, chloramphenicol.