TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Featuresa | Source or reference |
|---|---|---|
| Strains | ||
| E. coli | ||
| S17-1 (λpir) | Cloning host; λpir recA thi pro hsdR M+ RP4:2-Tc:Mu:Km Tn7TprStrr | 55 |
| JM109 | Cloning host; endA1 recA1 gyrA96 thiA hsdR17 relA1 supE44 Δ(lac-proAB) [F′ traD36 proAB+laqIqlacZ ΔM15] | 71 |
| M15 | Protein expression host; E. coli K-12 Strr F− ΔlacZ (pREP4) | Qiagen |
| E. cloacae | ||
| UW5 | Wild-type strain | 45 |
| J3 | UW5 ipdC::Kmr | 45 |
| J35 | UW5 tyrR::Tcr | This study |
| J51 | J35 ipdC::uidA; Tcr Gmr | This study |
| J55 | UW5 ipdC::uidA; Gmr | This study |
| Plasmids | ||
| pJP2 | Source of Tcr cassette | 50 |
| pGEM-T Easy | Cloning vector; Ampr | Promega |
| pG16 | 124-bp 16S rRNA fragment (amplified with 16S 1369F, and 1492R) in pGEM-T Easy; Ampr | This study |
| pGIC | 120-bp ipdC fragment (amplified with IPRT1F and -1R) in pGEM-T Easy; Ampr | This study |
| pJQ200SK | Low-copy suicide vector; GmrsacB | 52 |
| pJQ200TM | 1,177-bp tyrR fragment with 2.1-kb Tcr cassette insertion cloned into pJQ200SK; Gmr TcrsacB | This study |
| pJQS | pJQ200SK with sacB deletion; Gmr | This study |
| pJQSIPG | ipdC::uidA in pJQS; Gmr | This study |
| pREP4 | Expresses lac repressor in trans; Kmr | Qiagen |
| pQE30 | Protein expression plasmid; N-terminal His6 tag for protein purification; Ampr | Qiagen |
| pQEtyrR | 1,542-bp tyrR coding region in pQE30; Ampr | This study |
a
Tcr, tetracycline resistance; Ampr, ampicillin resistance; Strr, streptomycin resistance; Kmr, kanamycin resistance; Tpr, trimethoprim resistance; Gmr, gentamicin resistance.