TABLE 1.
Strains, plasmids, and primers used in this study
| Strain/plasmid/primer | Description | Source or reference |
|---|---|---|
| Strainsa | ||
| MG1655 | F− λ−ilvG-negative rfb50 rph1 | 16 |
| LA01 | MG1655 ΔpflB::FRT ΔfrdA::FRT-Kan-FRT; sequential deletion of pflB and frdA in MG1655 | This study |
| LA02 | MG1655 Δpta::FRT ΔadhE::FRT ΔfrdA::FRT-Kan-FRT; sequential deletion of pta, adhE, and frdA in MG1655 | This study |
| LA01Δdld | MG1655 ΔpflB::FRT ΔfrdA::FRT Δdld::FRT-Kan-FRT; dld deletion in LA01 | This study |
| LA02Δdld | MG1655 Δpta::FRT ΔadhE::FRT ΔfrdA::FRT Δdld::FRT-Kan-FRT; dld deletion in LA02 | This study |
| Plasmids | ||
| pCP20 | reppSC101ts Apr CmrcI857 l PRflp+ | 5 |
| pZSblank | Blank plasmid created by removing Citrobacter freundii dhaKL from pZSKLcf and self ligating the plasmid (tetR, oriR SC101*, cat) | 47a |
| pZSKLMgldA | E. coli dhaKLM and gldA under the control of PLtetO-1 (tetR, oriR SC101*, cat) | 47a |
| pZSKLcf | C. freundii dhaKL under the control of PLtetO-1 (tetR, oriR SC101*, cat) | 2 |
| pZSglpK | E. coli glpK gene control of PLtetO-1 (tetR, oriR SC101*, cat) | This study |
| pZSglpKglpD | E. coli glpK and glpD under the control of PLtetO-1 (tetR, oriR SC101*, cat) | This study |
| pZSldhA | E. coli ldhA under the control of PLtetO-1 (tetR, oriR SC101*, cat) | This study |
| Primersb | ||
| v-pflB | AAATCCACTTAAGAAGGTAGGTG | This study |
| TCGTGGAGCCTTTATTGTAC | ||
| v-frdA | ACCCTGAAGTACGTGGCTG | This study |
| GCACCACCTCAATTTTCAGG | ||
| v-pta | GCTGTTTTGTAACCCGCC | This study |
| GCAGCGCAAAGCTGCGG | ||
| v-adhE | CAAATCATCACCGCACTGAC | This study |
| CCTTAACTGATCGGCATTG | ||
| v-dld | TGATATTTTTTCGCCACCACAAG | This study |
| AAACAAAAAGCCGCCCAAATG | ||
| c-glpK | ACTGCAGGTACCATGACTGAAAAAAAATATATCGTTG | This study |
| TGCTACCTGCAGTTATTCGTCGTGTTCTTCCCAC | ||
| c-glpD | TCGATCCTGCAGGAAAGTGAATGAGGGCAGCATG | This study |
| ACTTGTGGATCCTTACGACGCCAGCGATAACC | ||
| c-ldhA | CATTAAAGAGGAGAAAGGTACCATGAAACTCGCCG | This study |
| GATGCCTCTAGCACGCGTTTAAACCAGTTCGTT |
a
Deletions were moved into each strain in the order they appear in the description column.
b
v and c indicate the primer sequences (5′ to 3′) that were used for verification purposes during the creation of disruption mutants and cloning purposes, respectively. The forward sequence follows the reverse sequence in each case. Genes or operons deleted or cloned are apparent from primer names.