TABLE 2.
Primers used for the construction of the ampC knockout mutants
| Primer | Sequence (5′-3′)a | Locationb | PCR product size (bp) |
|---|---|---|---|
| AmpC-F-ERI | TCGAATTCGCGCGCAGGGCGTTCAG | −186 to −169 | 415 |
| AmpC-I-R-HDIII | TCAAGCTTCGTCCTCTTACGAGGCCAGc | 210 to 229 | |
| AmpC-I-F-HDIII | TCAAGCTTCAGGGCAGCCGCTTCGAC | 366 to 384 | 432 |
| AmpC-R-BHI | TCGGATCCCAGGTTGGCATCGACGAAG | 779 to 798 |
a
Restriction sites for endonucleases are underlined.
b
Relative to the ampC start codon.
c
A T→A substitution (boldfaced) was introduced to generate an early stop codon in the ampC sequence and therefore avoid the potential effect on fitness of the truncated AmpC enzyme.