TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Descriptiona | Source or reference |
|---|---|---|
| Strains | ||
| S. putrefaciens | ||
| CN32 | Wild type | ATCC |
| 3591Z | CN32 derivative with lacZ transcriptionally fused into the bpfA operon | This study |
| CN32FLAG-FlrA | CN32 derivative with FLAG translationally fused into the 5′ end of flrA | This study |
| 3591Z ΔflrA | 3591Z derivative with deletion of Sputcn32_2580 | This study |
| 3591Z ΔflhG | 3591Z derivative with deletion of Sputcn32_2560 and flhG | This study |
| 3591Z ΔflrA ΔflhG | 3591Z derivative with double deletion of flrA and flhG | This study |
| CN32 ΔflrA | CN32 derivative with deletion of flrA | This study |
| CN32 ΔflhG | CN32 derivative with deletion of flhG | This study |
| CN32 ΔflrA ΔflhG | CN32 derivative with double deletion of flrA and flhG | This study |
| CN32 ΔbpfA | CN32 derivative with deletion of Sputcn32_3591 | 24 |
| CN32 ΔflrA ΔbpfA | CN32 derivative with double deletion of flrA and bpfA | This study |
| CN32 ΔflhG ΔbpfA | CN32 derivative with double deletion of flhG and bpfA | This study |
| E. coli | ||
| JM109 | General cloning host for plasmid manipulation | New England Biolabs |
| MG1655 | Derivative of E. coli K-12 | ATCC |
| WM3064 | Auxotrophic to DAP; used for conjugation | 48 |
| BL21(DE3) | Expression host carrying the T7 RNA polymerase | Novagen |
| Plasmids | ||
| pRL27 | Tn5 delivery vector, Kmr | 41 |
| pRE112 | Suicide vector, Cmr | 49 |
| pRE-flrA | pRE112 derivative to delete flrA | This study |
| pRE-flhG | pRE112 derivative to delete flhG | This study |
| pRE-PbpfA-lacZ | pRE112 derivative to construct 3591Z | This study |
| pRE-flag-flrA | pRE112 derivative to construct CN32FLAG-FlrA | This study |
| pET-28a(+) | Bacterial expression vector | Novagen |
| pET-FlrA | pET-28a(+) derivative to express His-tagged FlrA | This study |
| pJN105 | Broad-host-range vector carrying the araBAD promoter | 50 |
| pflrA | pJN105 derivative to express FlrA | This study |
| pflhG | pJN105 derivative to express FlhG | This study |
| pBBR1MCS-5 | Broad-host-range plasmid | 51 |
| PbpfA-lacZ | pBBR1MCS5 derivative bearing the wild-type bpfA promoter fused with lacZ | This study |
| PbpfA-lacZ-M1 | PbpfA-lacZ derivative with a mutation in FlrA box 1 | This study |
| PbpfA-lacZ-M2 | PbpfA-lacZ derivative with a mutation in FlrA box 2 | This study |
| PbpfA-lacZ-M1M2 | PbpfA-lacZ derivative with a mutation in FlrA box 1 and box 2 | This study |
a
DAP, diaminopimelic acid.