TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Relevant characteristicsa | Source or reference |
|---|---|---|
| L. pneumophila | ||
| JR32 | Homogenous salt-sensitive isolate of AM511 (Philadelphia-1 Smr r− m+) | H. Shuman (57) |
| PB126 | JR32 katA::Gmr (katA null strain) | This work |
| PB140 | PB126 katB::ΩCm (katA katB double null) | This work |
| E. coli UM383 | HB101 katG17::Tn10(Tet) katE::Tn10(Kan) | P. Loewen |
| Plasmids | ||
| pAM6 | Bluescript KS with a 2.9-kb gentamicin resistance cassette cloned into the HindIII site | A. Marra and H. Shuman |
| pAM6::katA int | 875-bp EcoRI-EcoRI katA internal fragment (codon E67 to codon S358) cloned into the EcoRI site of pAM6 | This work |
| pJBZ281 | lacZ with polylinker upstream for constructing translational fusions, ColE1 ori(Kanr) | M. R. K. Alley (2) |
| pJBZ281::PkatA | 1,761-bp fragment of katA (1,733 nt upstream of ATG start through first nt of codon for Ala-10) cloned using BamHI and PstI into pJBZ281 (katA translational fusion plasmid) | This work |
| pMMB207αB | RSF1010 derivative, IncQ lacIq Cmr Ptac oriT MCS α complementation | L. Wiater and H. Shuman (57) |
| pMMB207αB::katB | katB ORF (288 nt upstream of translational ATG start through 220 nt downstream of TAA stop) cloned using PstI and HindIII into pMMB207αB | 5 |
| pMMB207αB-Km14 | RSF1010 derivative, IncQ lacIq Cmr Ptac oriT MCS α complementations, with kanamycin resistance cassette interrupting mobA at nt 3325 | G. Segal and H. Shuman (45) |
| pMMB207αB-Km14::katA | katA ORF (191 nt upstream of translational ATG start through 267 nt downstream of TAA stop) cloned into BamHI site of pMMB207αB-Km14 in opposite direction from Ptac | This work |
| pNPTS138 | Derivative of pLITMUS 38 cloning vector with nptI, RK2 oriT, and Bacillus subtilus sacB, Kanr | E. Quardokus and Y. Brun (42) |
| pNPTS138::katB::ΩCm | katB null allele (katB::ΩCm) cloned into BamHI site of pNPTS138 | 5 |
a
MCS, multiple cloning site.