TABLE 1.
Bacterial strains and plasmids
| Strain or plasmid | Relevant properties | Reference or source |
|---|---|---|
| L. pneumophila | ||
| Philadelphia-1 | Clinical isolate, serogroup 1 | CDCa |
| Lp02 | Philadelphia-1 SmR, r−thyA | 2 |
| JV918 | Lp02 ΔdotB | This study |
| JV3079 | Lp02 ΔdotB::Flag coding sequence fused to dotB | This study |
| E. coli | ||
| C600 | e14−(McrA−) supE44 thi-1 thr-1 leuB6 lacY1 tonA21 | Stratagene |
| XL1-Blue | recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac [F′ proAB laclqZΔM15 Tn10(Tetr)] | Stratagene |
| Plasmids | ||
| pQE30 | Expression vector for His-tagged proteins (Ampr) | Qiagen |
| pJB1540 | pQE30 with thrombin site | This study |
| pJB1572 | pJB1540 with dotB | This study |
| pJB2442 | pJB1540 with dotBK162Q | This study |
| pJB908 | RSF1010 cloning vector (AmprtdΔi) | This study |
| pJB1153 | pJB908 with dotB | This study |
| pJB1568 | pJB908 with dotBK162Q | This study |
| pJB1192 | pJB908 with His coding sequence fused to dotB | This study |
| pJB2444 | pJB908 with His coding sequence fused to dotBK162Q | This study |
| pSR47S | R6K suicide vector (KanrsacB) | 17 |
| pJB920 | pSR47S with dotB flanking regions | This study |
| pJB2911 | pSR47S with Flag coding sequence fused to dotB | This study |
a
CDC, Centers for Disease Control and Prevention (Atlanta, Ga.).