Table 2.
PCR primers used in this studya
| Primer | Sequence (5′–3′) | Comments |
|---|---|---|
| SigGapraF | CCG ACA TCT GTC GGA CGG CCG CAG TAA GCT CCA ACC ATG att ccg ggg atc cgt cga cc | Aprar cassette amplification for sigG replacement, forward |
| SigGapraR | CAT GTA CGG ACG CGT TTT CCG CGC CAT CGG TAG GAG TCA tgt agg ctg gag ctg ctt c | Aprar cassette amplification for sigG replacement, reverse |
| SigGtestF | CGGAATGAAGACTCTGCTC | External sigG primer, forward |
| SigGtestR | GTGTGGGACACGGTAGAAC | External sigG primer, reverse |
| SigGintF | GTGTGGGACACGGTAGAAC | Internal sigG primer, forward |
| SigGintR | ATCAGGGTGCCGATCGAAG | Internal sigG primer, reverse |
a
The 5′ end of SigGapraF contained 39 nucleotides (uppercase letters) corresponding to the sense strand upstream of sigG ending in the start codon. The 5′ end of SigGapraR contained 39 nucleotides (uppercase letters) corresponding to the antisense strand ending in the stop codon. The 3′ ends of the PCR primers (lowercase letters) matched the 20-nucleotide and 19-nucleotide extensions, respectively, of the aprA-containing disruption cassette sequence.