TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Descriptiona [parental strain] | Source or reference |
|---|---|---|
| Strains | ||
| S. enterica serovar Typhimurium | ||
| χ3761 | Wild-type isolate UK-1, highly virulent for chicks and mice | 21, 98 |
| χ4700 | Δ(galE-uvrB)-1005 [χ3761] | 90 |
| χ9226 | ΔrelA198::araC PBADlacI TT [χ3761] | This study |
| χ9411 | Δlrp-13 (lrp-null mutation) [χ3761] | This study |
| χ9448 | lrp-1281 [ΔPlrp::Ptrclrp, constitutive expression of Lrp; lrp(Con)] [χ3761] | This study |
| χ9449 | ΔrelA198::araC PBADlacI TT ΔaraBAD23 lrp-1281 [χ9509] | This study |
| χ9509 | ΔrelA198::araC PBADlacI TT ΔaraBAD23 [χ9226] | This study |
| χ9858 | PhilA::pYA4607 (PhilA-lacZ); Ampr Gmr [χ3761] | This study |
| χ9859 | Δlrp-13 PhilA::pYA4607 (PhilA-lacZ), Ampr Gmr [χ9411] | This study |
| χ9860 | lrp-1281 PhilA::pYA4607 (PhilA-lacZ); Ampr Gmr [χ9448] | This study |
| χ9861 | PinvF::pYA4608 (PinvF-lacZ); Ampr Gmr [χ3761] | This study |
| χ9862 | Δlrp-13 PinvF::pYA4608 (PinvF-lacZ); Ampr Gmr [χ9411] | This study |
| χ9863 | lrp-1281 PinvF::pYA4608 (PinvF-lacZ); Ampr Gmr [χ9448] | This study |
| χ9864 | PssrA::pYA4609 (PssrA-lacZ); Ampr Gmr [χ3761] | This study |
| χ9865 | Δlrp-13 PssrA::pYA4609 (PssrA-lacZ); Ampr Gmr [χ9411] | This study |
| χ9866 | lrp-1281 PssrA::pYA4609 (PssrA-lacZ); Ampr Gmr [χ9448] | This study |
| χ9867 | PmurA::pYA4610 (PmurA-lacZ); Ampr Gmr [χ3761] | This study |
| χ9868 | Δlrp-13 PmurA::pYA4610 (PmurA-lacZ); Ampr Gmr [χ9411] | This study |
| χ9869 | lrp-1281 PmurA::pYA4610 (PmurA-lacZ); Ampr Gmr [χ9448] | This study |
| E. coli | ||
| MGN-617 (χ7213) | thr-1 leuB6 fhuA21 lacY1 glnV44 recA1 ΔasdA4 thi-1 RP4-2-Tc::Mu [λ-pir]; Kmr | 81 |
| BL21(DE3) | F−ompT hsdSB(rB− mB−) gal dcm (DE3) | Invitrogen |
| Plasmids | ||
| pBluescript SK(−) | Cloning vector (pUC ori), Ampr | Stratagene |
| pCHSUI-1 | Positive selection suicide vector (R6K ori) for gene replacement; Tcr | 5 |
| pET-14b | T7 promoter-based expression vector (pBR ori) with His tag; Ampr | Invitrogen |
| pET SUMO | T7 promoter-lac operator-based expression vector (pBR ori) with lacI; Kmr | Invitrogen |
| pYA3337 | asd-based cloning vector (pSC101 ori) with Ptrc promoter, Asd+ | 19 |
| pYA4073 | Derivative of pYA3337 with lrp expressed from Ptrc, Asd+ | This study |
| pYA4124 | Derivative of pET SUMO containing His tag from pET-14b; Kmr | This study |
| pYA4185 | Derivative of pYA4124 for His-tagged Lrp expression; Kmr | This study |
| pYA4212 | Derivative of pCHSUI-1 for replacement of Plrp region with Ptrc; Tcr | This study |
| pKD3 | Template plasmid for λ Red recombination system; Ampr Cmr | 24 |
| pKD46 | Expressing Red (γ, β, exo), temperature sensitive; Ampr | 24 |
| pCP20 | Expressing FLP, temperature sensitive; Ampr | 24 |
| pSG3 | A suicide vector (R6K ori) for construction of promoter-lacZ fusion into chromosome by single-crossover insertion; Ampr Gmr | 4 |
| pYA4607 | Derivative of pSG3 for insertion of the PhilA-lacZ fusion into the chromosome; Ampr Gmr | This study |
| pYA4608 | Derivative of pSG3 for insertion of the PinvF-lacZ fusion into the chromosome; Ampr Gmr | This study |
| pYA4609 | Derivative of pSG3 for insertion of the PssrA-lacZ fusion into the chromosome; Ampr Gmr | This study |
| pYA4610 | Derivative of pSG3 for insertion of the PmurA-lacZ fusion into the chromosome; Ampr Gmr | This study |
a
Tcr, tetracycline resistance; Gmr, gentamicin resistance; Ampr, ampicillin resistance; Kmr, kanamycin resistance.