TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Genotype or descriptiona | Reference |
|---|---|---|
| Strains | ||
| X90 | F′ lacIq lac′ pro′/ara Δ(lac-pro) nal1 argE(amb) rifr thi-1 | 42 |
| CH12 | X90 (DE3) | 35 |
| CH113 | X90 (DE3) ssrA::cat, CmR | 35 |
| CH2016 | CH12 Δrna ΔslyD::kan, KanR | 43 |
| CH2017 | CH113 ΔslyD::kan, KanR | 21 |
| CH3574 | CH113 Δrnc-38::kan, KanR | 21 |
| CH4514 | CH12 ΔslyD::kan, KanR | 21 |
| CH4516 | X90 (DE3) ssrA (his6) ΔslyD::kan, Kan R | 21 |
| CH7189 | CH12 ΔarfA::kan, KanR | 21 |
| CH7330 | X90 (DE3) ΔarfA::kan ssrA::cat pCH450-arfA(Eco), KanR CmR TetR | This study |
| CH8230 | CH12 Δrnc-38::kan, KanR | 21 |
| CH8674 | CH113 ΔarfA::kan pCH450-arfA(Ngo), CmR KanR TetR | This study |
| CH8676 | CH113 ΔarfA::kan pCH450-arfA(Hin), CmR KanR TetR | This study |
| Plasmids | ||
| pCH201 | pACYC184 derivative that expresses tmRNA (His6), TetR | 33 |
| pET-his6-arfA | pET21 derivative that overexpresses E. coli his6-arfA, AmpR | 21 |
| pCH450-arfA | Arabinose-inducible expression of E. coli ArfA under control of PBAD promoter, TetR | 21 |
| pET-his6-arfA(Ngo) | pET21 derivative that overexpresses E. coli his6-arfA, AmpR | This study |
| pCH450-arfA(Ngo) | Arabinose-inducible expression of N. gonorrhoeae ArfA under control of PBAD promoter, TetR | This study |
| pCH450-arfA(Hin) | Arabinose-inducible expression of H. influenzae ArfA under control of PBAD promoter, TetR | This study |
| pFG21-trxA-arfA′(Eco) | pET21 derivative that overexpresses flag-trxA fused to codons Gly49--Ser69 of E. coli arfA, AmpR | This study |
| pFG21-trxA-arfA′(recode) | pET21 derivative that overexpresses flag-trxA fused to synonymously recoded Gly49--Ser69 of E. coli arfA, AmpR | This study |
| pFG21-trxA-arfA′(Vfi) | pET21 derivative that overexpresses flag-trxA fused to codons Lys58--Thr65 (and 38 nt from the 3′-UTR) of V. fischeri arfA, AmpR | This study |
| pFG21-trxA-arfA′(Pmu) | pET21 derivative over-expresses flag-trxA fused to codons Glu61--Leu79 of P. multocida arfA, AmpR | This study |
| pFG21-trxA-arfA(Eco) | pET21 derivative that overexpresses flag-trxA fused to codons Ser2--Cys72 of E. coli arfA, AmpR | This study |
| pFG21-trxA-arfA(Ngo) | pET21 derivative that overexpresses flag-trxA fused to codons Gly2--Leu73 of N. gonorrhoeae arfA, AmpR | This study |
| pFG21-trxA-arfA(Hin) | pET21 derivative that over-expresses flag-trxA fused to codons Arg2--Phe69 of H. influenzae arfA, AmpR | This study |
| pFG21-trxA-arfA(Pmi) | pET21 derivative that overexpresses flag-trxA fused to codons Ser2--Phe67 of P. mirabilis arfA, AmpR | This study |
| pCH450-flag-arfA | Arabinose-inducible expression of FLAG-ArfA(Eco) under control of PBAD promoter, TetR | 21 |
| pCH450-flag-arfA(Ngo) | Arabinose-inducible expression of FLAG-ArfA(Ngo) under control of PBAD promoter, TetR | This study |
| pCH450-flag-arfA(Ngo-fs) | Arabinose-inducible expression of FLAG-ArfA(Ngo) from gene with a frameshift in the hairpin coding sequence, TetR | This study |
| pFG501-trxA-arfA(Eco) | IPTG-inducible expression of FLAG-TrxA-ArfA(Eco) under control of Ptrc promoter, AmpR | This study |
| pFG501-trxA-arfA(Ngo) | IPTG-inducible expression of FLAG-TrxA-ArfA(Ngo) under control of Ptrc promoter, AmpR | This study |
| pFG501-trxA-arfA(Ngo-fs) | IPTG-inducible expression of FLAG-TrxA-ArfA(Ngo) from gene with a frameshift in the hairpin coding sequence, AmpR | This study |
| pTrc99A-flag-arfA(Eco) | IPTG-inducible expression of FLAG-ArfA(Eco) under control of Ptrc promoter, AmpR | This study |
| pTrc99A-flag-arfA(recode) | IPTG-inducible expression of FLAG-ArfA(Eco) from recoded construct under control of Ptrc promoter, AmpR | This study |
| pTrc99A-flag-arfA(Ngo) | IPTG-inducible expression of FLAG-ArfA(Ngo) under control of Ptrc promoter, AmpR | This study |
| pET21P-his6-rnc(Eco) | pET21 derivative for overproduction of E. coli RNase III carrying an N-terminal His6 tag, AmpR | This study |
| pET21P-his6-rnc(Ngo) | pET21 derivative for overproduction of N. gonorrhoeae RNase III carrying an N-terminal His6 tag, AmpR | This study |
| pCH450-his6-rnc(Ngo) | Arabinose-inducible expression of N. gonorrhoeae RNase III carrying an N-terminal His6 tag, TetR | This study |
a The abbreviations used are: AmpR, ampicillin resistant; CmR, chloramphenicol resistant; KanR, kanamycin resistant; TetR, tetracycline resistant; UTR, untranslated region.