TABLE 1.
Bacterial strains and plasmids
| Strain or plasmid | Relevant genetic markers or characteristic(s) | Source or referencea |
|---|---|---|
| Strains | ||
| DH5α | supE44 ΔlacU169(φ80lacZΔM15) hsdR17 recA1 endA1 gyrA96 thi-1 relA | Bethesda Research Laboratories |
| W3110 | F− λ− IN(rrnD-rrnE) rph-1 | CGSC |
| AB1133 | thr-1 leuB6 (gpt-proA)66 hisG4 argE3 thi-1 rfbD1 lacY1 ara-14 galK2 xyl-5 mtl-1 mgl-1 rpsL31 kdgK51 supE44 | CGSC |
| EC100 | F−mcrA Δ(mrr-hsdRMS-mcrBC) φ80dlacZΔM15 ΔlacX74 recA1 endA1 araD139 Δ(ara leu)7697 galU galK λ−rpsL nupG | Epicenter |
| EC100D | As EC100 but pir+ (dihydrofolate reductase) | Epicenter |
| KM32 | argE3 his-4 leuB6 proA2 thr-1 ara-14 galK2 lacY1 mtl-1 xyl-1 thi-1 rpsL-31 tsx-33 supE44 Δ(recC ptr recB recD)::Ptac-gam-bet-exo cat | 38 |
| PR4246 | As W3110 but yiaH::KAN-2 [PR4254 (P1) × W3110] | This study |
| PR4254 | As KM32 but yiaH::KAN-2 | This study |
| PR4275 | EC100/pJun3 | This study |
| PR4276 | As PR4275 but yiaH::KAN-2 | This study |
| PR4279 | EC100D/pRL170 | This study |
| PR4300 | PR4246/pRL180 | This study |
| Plasmids | ||
| pGEM-T Easy | PCR cloning vector | Promega |
| pQE30 | Expression vector | QIAGEN |
| pBR322 | Cloning vector | Promega |
| pJun1 | 3.73-kb BamHI-HindIII fragment of pRL105b cloned into pBR322 | This study |
| pJun3 | Wild-type wec gene cluster on a 12.78-kb EcoRI-BamHI fragment cloned into pBR322 | This study |
| pRL170 | Rescue plasmid containing yiaH::KAN-2 mutant allele | This study |
| pRL171 | Wild-type yiaH gene on a 1.14-kb PCR fragment cloned into pGEM-T Easy | This study |
| pRL172 | yiaH::KAN-2 on a 3.14-kb PCR fragment cloned into pGEM-T Easy | This study |
| pRL179 | Wild-type yiaH on a 1.10-kb fragment cloned in pGEM-T Easy | This study |
| pRL180 | Wild-type yiaH on a 1.10-kb BamHI-HindIII fragment cloned into pQE30 | This study |
a
CGSC, E. coli Genetic Stock Center and Mary Berlyn, Yale University, New Haven, Conn.
b
See reference 34.