TABLE 1.
Relevant strains and plasmids used in this study
| Strain or plasmid | Relevant phenotype and/or genotype | Source or reference |
|---|---|---|
| Escherichia coli strains | ||
| TOP10F | F′ [laqIq Tn10(Tcr)] mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 recA1 deoR araD139 Δ(ara-leu) galU galK rpsL (Smr) endA1 nupG | Invitrogen |
| BL21 (DE3)/pLysS | F−ompT hsdSB (rB− mB−) gal dcm (DE3)/pLysS (Cmr) | Invitrogen |
| ER2566 | fhuA2 (lon) ompT lacZ::T7 gene1 gal sulA11 Δ(mcrC-mrr)114::IS10 R(mcr-73::mini- Tn10-Tets)2 R(zgb-210::Tn10)(Tets) endA1(dcm) | New England Biolabs |
| Haemophilus influenzae strains | ||
| Rd KW20 | Genome determined | 9 |
| AK01 | Strain Rd (sxy-1 Strepr) constitutive competent | 18 |
| SE01 | nadV, Nam+ | 42 |
| SE01ΔRNK | nadRΔRNK::cat, Nam+ Cmr | This study |
| SE01ΔRNK58 | nadRΔ58RNK::cat, Nam+ Cmr | This study |
| SE02ΔRNK | SE01ΔRNK nadR::pBADnadR-His, Kanr | This study |
| SE03ΔRNK | SE01ΔRNK nadR::pBADnadR(G238N), Kanr | This study |
| SE04ΔRNK | SE01ΔRNK nadR::pBADnadR(G238S), Kanr | This study |
| SE05ΔRNK | SE01ΔRNK nadR::pBADnadR(D238C), Kanr | This study |
| SE06ΔRNK | SE01ΔRNK nadR::pBADnadR(D238N), Kanr | This study |
| SE07ΔRNK | SE01ΔRNK nadR::pBADnadR(D238S), Kanr | This study |
| SE08ΔRNK | SE01ΔRNK nadR::pBADnadR(R238C), Kanr | This study |
| SE09ΔRNK | SE01ΔRNK nadR::pBADnadR(R238M), Kanr | This study |
| SE10ΔRNK | SE01ΔRNK nadR::pBADnadR(R238N), Kanr | This study |
| SE11ΔRNK | SE01ΔRNK nadR::pBADnadR(W256F), Kanr | This study |
| SE12ΔRNK | SE01ΔRNK nadR::pBADnadR(Y292I), Kanr | This study |
| SE13ΔRNK | SE01ΔRNK nadR::pBADnadR(K126A), Kanr | This study |
| SE14ΔRNK | SE01ΔRNK nadR::pBADnadR(K126T), Kanr | This study |
| Plasmids | ||
| pACYC184 | Cmr Tetr | New England Biolabs |
| pACYC184nadR(His6) | Tetr | This study |
| pMMnadR-His6 | Ampr; nadR-C′-terminal His tagged | 42 |
| pUCΔRNK | Cmr Ampr, nadR::cat::nadR′ | This study |
| pSSkan | Kanr Cmr | S. Schild |
| pSSkanΔcm | pSSkan, Cms Kanr | This study |
| pNadRkan | pSSkanΔcm, nadR+ | This study |
| pBAD18-Kan | Kanr | 16 |
| pBADnadR-His | Kanr, nadR-C′-terminal His tagged | This study |
| nadR point mutants | ||
| pBADnadR(G238N)-His6 | Kanr | This study |
| pBADnadR(G238S)-His6 | Kanr | This study |
| pBADnadR(D304C)-His6 | Kanr | This study |
| pBADnadR(D304N)-His6 | Kanr | This study |
| pBADnadR(D304S)-His6 | Kanr | This study |
| pBADnadR(R352C)-His6 | Kanr | This study |
| pBADnadR(R352M)-His6 | Kanr | This study |
| pBADnadR(R238N)-His6 | Kanr | This study |
| pBADnadR(W256F)-His6 | Ampr | This study |
| pBADnadR(Y292I)-His6 | Ampr | This study |
| pBADnadR(K126A)-His6 | Ampr | This study |
| pBADnadR(K126T)-His6 | Ampr | This study |
| pMMnadR(G238N)-His6 | Ampr | This study |
| pMMnadR(G238S)-His6 | Ampr | This study |
| pMMnadR(D304C)-His6 | Ampr | This study |
| pMMnadR(D304N)-His6 | Ampr | This study |
| pMMnadR(D304S)-His6 | Ampr | This study |
| pMMnadR(R352C)-His6 | Ampr | This study |
| pMMnadR(R352M)-His6 | Ampr | This study |
| pMMnadR(R238N)-His6 | Ampr | This study |
| pMMnadR(W256F)-His6 | Ampr | This study |
| pMMnadR(Y292I)-His6 | Ampr | This study |
| pMMnadR(K126A)-His6 | Ampr | This study |
| pMMnadR(K126T)-His6 | Ampr | This study |