TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Descriptiona | Reference or source |
|---|---|---|
| Strains | ||
| Pseudomonas nitroreducens | ||
| TX1 | Wild-type strain TX1, Ampr | Laboratory collection |
| ΔaceA mutant | Strain TX1 with in-frame deletion of aceA gene, Ampr | This study |
| ΔaceB mutant | Strain TX1 with in-frame deletion of aceB gene, Ampr | This study |
| ΔfixC mutant | Strain TX1 with in-frame deletion of fixC gene, Ampr | This study |
| ΔglcE mutant | Strain TX1 with in-frame deletion of glcE gene, Ampr | This study |
| ΔaceA(pBaceA) mutant | ΔaceA mutant harboring plasmid pBaceA, Ampr, Gmr | This study |
| ΔaceB(pBaceB) mutant | ΔaceB mutant harboring plasmid pBaceB, Ampr, Gmr | This study |
| E. coli | ||
| DH5α | F− ϕ80dlacZΔM15 Δ(lacZYA-argF)U169 deoR recA1 endA1 hsdR17(rK− mK+) phoA supE44 λ− thi-1 gyrA96 | Laboratory collection |
| DH5α/λpir | λpir lysogen of DH5α | Laboratory collection |
| BW20767 | Containing pRL27; used as donor for pRL27 conjugations | Larsen et al. (21) |
| HB101 | Containing pRK2013; used as helper for pRL27 conjugations | Figurski and Helinski (22) |
| Plasmids | ||
| pGEM-T Easy | PCR cloning vector, Ampr | Promega |
| pGaceA | Ampr, pGEM-T Easy with 2.6-kb fragment, including aceA | This study |
| pGaceB | Ampr, pGEM-T Easy with 3.2-kb fragment, including aceB | This study |
| pRL27 | Kmr, mini-Tn5 transposon (oriR6K) delivery vector | Larsen et al. (21) |
| pRK2013 | Kmr, carrying RK2 transfer genes | Figurski and Helinski (22) |
| pK18mobsacB | Kmr, oriT(RP4) sacB lacZα Plac Pmbi, mobilization and counterselection | Schafer et al. (24) |
| pKaceA | pK18mobsacB containing a gene fragment with 98% deletion of internal region of aceA | This study |
| pKaceB | pK18mobsacB containing a gene fragment with 98% deletion of internal region of aceB | This study |
| pKfixC | pK18mobsacB containing a gene fragment with 98% deletion of internal region of fixC | This study |
| pKglcE | pK18mobsacB containing a gene fragment with 98% deletion of internal region of glcE (GenBank accession no. WP_017520229.1) | This study |
| pBBR1MCS-5 | Broad-host-range cloning vector, lacZ, Gmr | Kovach et al. (26) |
| pBaceA | Gmr, pBBR1-MCS5 with 2.6-kb fragment, including aceA | This study |
| pBaceB | Gmr, pBBR1-MCS5 with 3.2-kb fragment, including aceB | This study |
| pBglcE | Gmr, pBBR1-MCS5 with 1.5-kb fragment, including glcE | This study |
a
Kmr, Gmr, and Ampr indicate resistance to kanamycin, gentamicin, and ampicillin, respectively.