TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Relevant characteristics | Source or reference |
|---|---|---|
| Strains | ||
| E. coli | ||
| DH5α | supE44 ΔlacU169 (ϕ80dlacZΔM15) recA1 endA1 hsdR17 thi-1 gyrA96 relA1 | Novagen |
| BL21(DE3) | F− ompT hsdS (rB− mB−) gal dcm lacY1(DE3) | Novagen |
| S17-1 | Donor strain for conjugation, Tpr Smr recA hsdR thi pro RP4-2-Tc::Mu-Km::Tn7 λpir | 25 |
| K-12 substrain W3110 | Source of d-malate dehydrogenase gene | 20 |
| P. alcaligenes | ||
| SponMu | Spontaneous mutant of P. alcaligenes NCIMB 9867 | NCIMB |
| SponMuΔhbzF | hbzF gene-disrupted mutant of SponMu (Kmr) | This study |
| SponMuΔhbzG | hbzG gene-disrupted mutant of SponMu (Kmr) | This study |
| SponMu-hbzG::dmlA | Derivative constructed from SponMuΔhbzG in which hbzG was replaced by dmlA | This study |
| Plasmids | ||
| pET28a | Expression vector, Kmr | Novagen |
| pEX18Tc | Tcr sacB+, gene replacement vector with a multiple-cloning site from pUC18 | 23 |
| pTnMod-OKm | Kmr, source of neomycin phosphotransferase II gene (nptII) | 24 |
| pET28a-hbzIJ | Kmr, pET28a derivative for expression of hbzIJ | This study |
| pET28a-hbzG | Kmr, pET28a derivative for expression of hbzG | This study |
| pET28a-dmlA | Kmr, pET28a derivative for expression of dmlA | This study |
| pHbzF::NptII | Tcr Kmr, hbzF gene-knockout vector containing DNA fragments homologous to the upstream and downstream regions of hbzF and nptII from pTnMod-Okm | This study |
| pHbzG::NptII | Tcr Kmr, hbzG gene-knockout vector containing DNA fragments homologous to the upstream and downstream regions of hbzG and nptII from pTnMod-OKm | This study |
| pHbzG::DmlA | Tcr, vector containing DNA fragments homologous to the upstream and downstream regions of hbzG and dmlA from E. coli K-12 for replacement of hbzG by dmlA | This study |