TABLE 1.
Bacterial strains, plasmids, and primers used in this study
| Strain, plasmid, or primer | Genotype or descriptiona | Source |
|---|---|---|
| Strains | ||
| BW25113 | lacIqrrnBT14 ΔlacZWJ16hsdR514 ΔaraBADAH33 ΔrhaBADLD78 | B. Wanner (3) |
| AW2A | BW25113 fixA | This work |
| AW1B | BW25113 fixB | This work |
| Plasmids | ||
| pKD4 | oriR[R6Kγ] Kanr Ampr | B. Wanner (3) |
| pKD46 | Temperature-sensitive derivative of pSC101, AmpraraC-ParaB λ recombination genes (gam-β-exo) | B. Wanner (3) |
| Primers | ||
| 1A | 5′-GCGTGATATCTGTAATTAACACCACCGATATGAACGACGTGTAGGCTGGAGCTGCTTC-3′ | This work |
| 2A | 5′-GAGAAAACGTGTTCATAGCATCCCCTGTAATTAAATGACCATATGAATATCCTCCTTAG-3′ | This work |
| 1B | 5′-CGCAAAGTCATTTAATTACAGGGGATGCTATGAACACGGTGTAGGCTGGAGCTGCTTC-3′ | This work |
| 2B | 5′-GCCAAAATGCAGCCTTGCCAGAGTGGATCAACGCGCTAACATATGAATATCCTCCTTAG-3′ | This work |
a
In primers 1A and 2A, underlined sequence is identical to the E. coli fixA DNA sequences. In primers 1B and 2B, underlined sequence is identical to the E. coli fixB DNA sequences. Boldface sequence is identical to the kanamycin cassette DNA sequences on pKD4.