TABLE 2.
Bacterial strains and plasmids
| Strain or plasmid | Descriptiona | Reference or source |
|---|---|---|
| E. coli strains | ||
| MG1655 | Wild-type K-12 | 58 |
| MG1655ΔydeA | K-12 MG1655ΔydeA::FRT | This work |
| BL21 | F− ompT hsdSB (rB− mB−) gal dcm | 59 |
| C41 (DE3) | Derived from BL21(DE3): F− ompT hsdSB (rB− mB−) gal dcm (DE3) | 60 |
| EG235 | C41 (DE3) ΔhipBA::kan, pMG25::gltX (optimized SD), pBAD33::His6::hipA (SD8 and start codon ATG) | Laboratory collection |
| JW2395 | AG1 [recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1] carrying pCA24N::gltX, GltX purification plasmid encoding N-terminally His6-tagged gltX, from ASKA collection | 55 |
| Plasmids | ||
| pBAD33 | p15 araC PBAD, Cmr | 61 |
| pNDM220 | Mini-R1 lacIq PA1/04/03, Ampr | 56 |
| pCP20 | pSC101 rep(Ts), Ampr Cmr | 47 |
| pBR322 | pMB1 rop, Ampr Tetr | 62 |
| pMG25 | pUC lacIq PA1/O4/O3, Ampr | Laboratory collection |
| pEG25 | pMG25 derivative that has reduced leakiness of the IPTG-inducible PA1/O4/O3 promoter | Laboratory collection |
| pEG::gltX | pMG25::gltX, optimized SD | Laboratory collection |
| pEG::His6hipA | pBAD33::His6hipA, HipA purification plasmid harboring N-terminally His6-tagged hipA | Laboratory collection |
| pET-15b | pBR322 lacI PT7, Ampr | Novagen |
| pKG127 | pUC57::hipBSTO127 | Genscript |
| pSVN1 | pBAD33::hipTO127, start codon GTG | This work |
| pSVN42 | pEG25::hipTO127::His6, optimized SD, HipTO127 purification plasmid C-terminally His6-tagged hipTO127 | This work |
| pSVN46 | pEG25::trpSHis6, optimized SD, TrpS purification plasmid harboring C-terminally His6-tagged trpS | This work |
| pSVN49 | pEG25::trpSS197DHis6, optimized SD, TrpSS197D purification plasmid harboring C-terminally His6-tagged trpSS197D |
This work |
| pSVN52 | pEG25::trpSS197AHis6, optimized SD, TrpSS197A purification plasmid harboring C-terminally His6-tagged trpSS197A |
This work |
| pSVN60 | pUC57::His6-TEVhipB::hipS::hipTO127, optimized SDs for all genes | Genscript |
| pSVN61 | pUC57::hipB::hipS::hipTO127::His6, optimized SDs for all genes used for TA complex purification via C-terminally His6-tagged hipTO127 |
Genscript |
| pSVN37 | pEG25::trpS, optimized SD | This work |
| pSVN44 | pBAD33::hipBSO127, optimized SD for hipBO127 | This work |
| pSVN94 | pET-15b::His6-TEVhipB::hipS::hipTD233QO127, optimized SDs for all genes | This work |
| pSVN103 | pNDM220::trpS, optimized SD | This work |
| pSVN109 | pNDM220::hipSO127, optimized SD | This work |
| pSVN110 | pNDM220::hipBSO127, optimized SDs | This work |
| pSVN111 | pNDM220::hipBO127, optimized SD | This work |
| pSVN113 | pUC57::hipBSTHi | Genscript |
| pSVN114 | pUC57::hipBSTTa | Genscript |
| pSVN116 | pNDM220::hipTO127, start codon GTG | This work |
| pSVN122 | pNDM220::hipBHi, optimized SD | This work |
| pSVN123 | pNDM220::hipSHi, optimized SD | This work |
| pSVN124 | pNDM220::hipBSHi, optimized SD for hipBHi, overlapping hipBHi and hipSHi | This work |
| pSVN126 | pNDM220::hipBTa, optimized SD | This work |
| pSVN127 | pNDM220::hipSTa, optimized SD | This work |
| pSVN128 | pNDM220::hipBSTa, optimized SD for hipBTa, overlapping hipBTa and hipSTa | This work |
| pSVN129 | pBAD33::hipTTa, optimized SD, start codon GTG | This work |
| pSVN135 | pBAD33::hipTHi, optimized SD | This work |
| pSVN138 | pNDM220::hipBSTa, optimized SDs | This work |
| pSVN139 | pNDM220::hipBSHi, optimized SDs | This work |
| pAH1 | pNDM220::hipA | A. Harms |
| pAH2 | pNDM220::relE | A. Harms |
a
SD, Shine-Dalgarno sequence.