TABLE 1.
Bacterial strains and plasmids
| Strain or plasmid | Relevant genotype or phenotype | Reference or source |
|---|---|---|
| Strains | ||
| E. coli | ||
| DH5α | λ− φ80dlacZΔM15 Δ(lacZYA-argF)U169 recA1 endA hsdR17(rK− mK−) supE44 thi-1 gyrA relA1 | Laboratory collection |
| XL10-Gold | Tetr Δ(mcrA)183 Δ(mcrCB-hsdSMR-mrr)173 endA1 supE44 thi-1 recA1 gyrA96 relA1 lac Hte [F[′] proAB lacIqZΔM15 Tn10(Tetr) Amy Camr] | Stratagene |
| MB596 | DH5α pletS | This work |
| MB597 | DH5α pletST311M | This work |
| MB598 | DH5α pletS::thyA | This work |
| MB610 | DH5α pletSH307Q | This work |
| MB540 | DH5α pBluescript KS+ with thyA | Laboratory collection |
| L. pneumophila | ||
| MB110 | Lp02 wild type; thyA hsdR rpsL | 5 |
| MB355 | Lp02 pflaG | 27 |
| MB416 | letS-36::Kan | 28 |
| MB417 | letS::kan pflaG | 28 |
| MB599 | letS::thyA | This work |
| MB600 | letS(T311M) | This work |
| MB605 | letS(T311M) pflaG | This work |
| MB611 | letS(H307Q) | This work |
| Plasmids | ||
| pGEM-T | Multiple cloning site within coding region of β-lactamase α fragment linearized with single-T overhangs; 3 kb; Ampr | Promega |
| pflaG | 150-bp flaA promoter fragment fused to GFP; encodes thymidylate synthetase; 10.5 kb; Ampr | 27 |
| pletS | pGEM-T containing 3.1-kb letS fragment PCR amplified from Lp02 chromosome and ligated into T overhangs; 6.1 kb; Ampr | This work |
| pletST311 M | pletS with a ACC to ATG change; 6.1 kb; Ampr | This work |
| pletSH307Q | pletS with a CAT to CAA change; 6.1 kb; Ampr | This work |
| pletS::thyA | pletS with 1.8 kb thyA fragment inserted into EcoRI site at base 436 of letS; 7.9 kb; Ampr | This work |
| pMB540 | pBluescript KS+ with thyA | Laboratory collection |