Table 1.
Bacterial strains, plasmids and oligonucleotides used in this study.
| Strain, plasmid or primer | Phenotype, genotype or sequence | Reference or source |
|---|---|---|
| E.coli strains | ||
| DH5α | F- endA1 hsdRI7 (rk - mk +) supE44 thi-1λ- recA1 gyrA96 (Nalr) relA1 Δ (lacZYA-argF)U169 deoR φ 80dlacZ Δ M15 | Lab collection |
| DH5αλpir | DH5α transduced with λpir | [69] |
| L. pneumophila strains | ||
| JR32 | Virulent L. pneumophila serogroup 1, strain Philadelphia, salt-sensitive isolate of AM511 | [43] |
| LpΔclpP | JR32 with clpP deletion | This study |
| LpΔclpP-pclpP | LpΔclpP containing pclpP | This study |
| JR32-pBC | JR32 containing pBC(gfp)Pmip | This study |
| LpΔclpP-pBC | LpΔclpP containing pBC(gfp)Pmip | This study |
| LpΔdotA | JR32 with dotA deletion | Lab collection |
| Plasmids | ||
| pRE112 | Mobilizable suicide vector for construction of gene knockouts in G- bacteria, oriT oriV sacB Cm | [69] |
| pMD18-T | cloning vector, Ap | TaKaRa |
| pBC(gfp)Pmip | ColE1 ori Cm Pmip gfpmut2 | [70] |
| pREΔclpP | pRE112::clpP for clpP deletion | This study |
| pclpP | pBC(gfp)Pmip containing clpP under the control of mip promoter | This study |
| Primers | ||
| PXC-F1 | AGAGAGCTCCTGCCAGTAGGTCCTATAAG | This study |
| PXC-R1 | TATGACATACAAGTTGCTGGACATTCTAC | This study |
| PXC-F2 | CAACTTGTATGTCATAGGAACGCTCACC | This study |
| PXC-R2 | GATGGTACCTGGGAAAATTGACAAACCGT | This study |
| PXH-clpPF | TGGTGGAAGCTTTAGGAGTATCTAGCAAAGTTATAAGTC | This study |
| PXH-clpPR | TGGTGGTCTAGATGAGAAAAAAGGAGAGTAAGC | This study |
*Abbreviations: Ap, ampicillin resistant; Cm, chloramphenicol resistant; sacB, sucrose sensitive.