TABLE 1.
Bacterial strains and plasmids
| Strain or plasmid | Genotype or propertya | Source or referenceb |
|---|---|---|
| Serovar Typhimurium | ||
| CS015 | phoP102::Tn10d-Cam | 32 |
| CS401 | phoN2 zxx-6251::Tn10d-Cam rpsL (Strr) | 21 |
| CS093 | 14028s; wild type | ATCC |
| CS022 | pho-24 (PhoP constitutive); a mutation in phoQ that results in increased net phosphorylation of PhoP and unregulated expression of PhoP-activated genes (18) | 33 |
| TG59 | CS401 ΔpgtE::Kan | This work |
| TG61 | CS022 ΔpgtE::Kan | This work |
| TG71 | TG61 with pBluescript KS+ | This work |
| TG73 | TG61 with pTG73 | This work |
| CS435 | pho-24 pagP::Tn10d-Tet phoN2 zxx-6251::Tn10d-Cam (also LG069) | 21 |
| TG66 | CS435 ΔpgtE::Kan | This work |
| TG172 | CS401 with pTG171 | This work |
| TG173 | CS015 with pTG171 | This work |
| TG174 | CS022 with pTG171 | This work |
| TG200 | CS022 pagA::MudJ | This work |
| CS404 | CS022 pagA::MudJ pagP::Tn10d-Tet | This work |
| E. coli | ||
| SM10 λpir | thi-1 thr-1 leuB6 supE44 tonA21 lacY1 recA::RP4-2-Tc::Mu | |
| DH5α | F− φ80dlacZΔM15 Δ(lacZYA-argF)U169 endA1 recA1 hsdR17 deoR thi-1 supE441 gyrA96 relA1 | Gibco-BRL |
| CC118 | araD139 Δ(ara,leu)7697 ΔlacX74 phoAΔ20 galE galK thi rpsE rpoB argE(Am) recA1 | 28 |
| Plasmids and transposons | ||
| pKAS32 | Ampr Strs (rpsL); a low-copy allelic exchange vector | 41 |
| pTG51 | Ampr Kanr; pgtE with an inserted Kanr cassette (39) replacing a portion of pgtE is ligated to KpnI-SacI fragment of pKAS32 | This work |
| pBluescript KS+ | Ampr; a high-copy cloning vector | Stratagene |
| pTG73 | Ampr; contains pgtE and 904 bp of upstream region (nt 671 to 2671; GenBank accession no. M21279) cloned into XbaI site of pBluescript KS+ | This work |
| pGPLFR03 | Ampr; firefly (f-luc) and vanilla (v-luc) double luciferase suicide vector | 30 |
| pTG171 | Ampr; contains a transcriptional fusion of pgtE (nt 5266 to 5641; GenBank no. M21279) to f-luc; ligated with an EcoRI-KpnI fragment of pGPLFR03 | This work |
| pWSK129 | Kanr; low-copy cloning vector | 51 |
| pTG82 | Kanr; contains pgtE and 904 bp of upstream region (nt 671 to 2671; GenBank no. M21279) cloned into XbaI site of pWSK129 | This work |
| pTG85 | Kanr; contains TnphoA/in insertion at codon 112 of pgtE in pTG82 | This work |
| pTG86 | Kanr; contains TnphoA/in insertion at codon 152 of pgtE in pTG82 | This work |
| pTG87 | Kanr; contains TnphoA/in insertion at codon 190 of pgtE in pTG82 | This work |
| TnphoA/in | Tn5 derivative; generates translational fusions to alkaline phosphatase (phoA) | 27 |
a
nt, nucleotide.
b
ATCC, American Type Culture Collection.