TABLE 1.
Bacterial strains and plasmids used in this study
| Bacterial strain/plasmid | Relevant characteristics | Source |
|---|---|---|
| L. pneumophila | ||
| 130b | O1; clinical isolate | Ref. 81 |
| 130b (pMIP) | 130b carrying pMIP | Ref. 12 |
| lpg1905::km | lpg1905 insertion mutant of 130b (KmR) | Ref. 12 |
| lpg1905::km (pMIP) | lpg1905::km carrying pMIP | Ref. 12 |
| lpg1905::km (pMIP:lpg1905) | lpg1905::km carrying pMIP:lpg1905 | Ref. 12 |
| lpg1905::km (pMIP:lpg1905E159A) | lpg1905::km carrying pMIP:lpg1905E159A | Ref. 12 |
| lpg1905::km (pMIP:lpg1905R122A) | lpg1905::km carrying pMIP:lpg1905R122A | This study |
| lpg1905::km (pMIP:lpg1905N168A) | lpg1905::km carrying pMIP:lpg1905N168A | This study |
| lpg1905::km (pMIP:lpg1905Q193A) | lpg1905::km carrying pMIP:lpg1905Q193A | This study |
| lpg1905::km (pMIP:lpg1905W384A) | lpg1905::km carrying pMIP:lpg1905W384A | This study |
| E. coli | ||
| XL-1 Blue | recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac (F′ proAB+ lacIqlacZΔM15 Tn10) (TetR) | Stratagene |
| BL21 (DE3) | F- ompT hsdSB(rB—mB-)gal dcm (DE3) | Novagen |
| Plasmids | ||
| pGEM-T Easy | 3015 bp, AmpR, high copy cloning | Promega |
| pGEM:lpg1905 | Full-length lpg1905 cloned into BamHI/HindIII | This study |
| pGEM:lpg1905R122A | Arg122 mutated to Ala in pGEM:lpg1905 | This study |
| pGEM:lpg1905N168A | Asn168 mutated to Ala in pGEM:lpg1905 | This study |
| pGEM:lpg1905Q193A | Gln193 mutated to Ala in pGEM:lpg1905 | This study |
| pGEM:lpg1905W384A | Trp384 mutated to Ala in pGEM:lpg1905 | This study |
| pRSETb | 2897bp, AmpR, E. coli expression vector with a bacteriophage T7 promoter and N-terminal polyhistidine (His6) tag | Invitrogen |
| pRSET:lpg1905 | Truncated enzymatically active form of lpg1905 cloned into EcoRI/HindIII (missing the first 34 amino acids corresponding to a putative signal peptide sequence) | Ref. 12 |
| pRSET:lpg1905E159A | Glu159 mutated to Ala in pRSET:lpg1905 | Ref. 12 |
| pRSET:lpg1905R122A | Arg122 mutated to Ala in pRSET:lpg1905 | This study |
| pRSET:lpg1905N168A | Asn168 mutated to Ala in pRSET:lpg1905 | This study |
| pRSET:lpg1905Q193A | Gln193 mutated to Ala in pRSET:lpg1905 | This study |
| pRSET:lpg1905W384A | Trp384 mutated to Ala in pRSET:lpg1905 | This study |
| pMIP | 9523 bp, pMMB207 with the promoter region of mip cloned into SacI/XbaI (CmR) | Ref. 37 |
| pMIP:lpg1905 | pMIP with full-length lpg1905 cloned into BamHI and HindIII | Ref. 12 |
| pMIP:lpg1905E159A | pMIP with full-length lpg1905E159A cloned into BamHI and HindIII | Ref. 12 |
| pMIP:lpg1905R122A | pMIP with full-length lpg1905R122A cloned into BamHI and HindIII | This study |
| pMIP:lpg1905N168A | pMIP with full-length lpg1905N168A cloned into BamHI and HindIII | This study |
| pMIP:lpg1905Q193A | pMIP with full-length lpg1905Q193A cloned into BamHI and HindIII | This study |
| pMIP:lpg1905W384A | pMIP with full-length lpg1905W384A cloned into BamHI and HindIII | This study |