TABLE 3.
Primers used in this studya
| Purpose of constructs | Oligonucleotide sequence (5′–3′) | Reference |
|---|---|---|
| Construction of Δhly strains | Forward: GGG AAT TCA ATT GTT GAT ACA ATG ACA TC | 88 |
| Reverse: GGC TGC AGG GTC TTT TTG GCT TGT GTA T | 88 | |
| Primers to amplify the hly ORF | Forward: CCG TCG GAT CCA TGA AAA AAA TAA TGC TAG TTT TTATTACAC | 88 |
| Reverse: ATC CGC GCT GCA GTT CGA TTG GAT TAT CTA CTT TAT TAC | 88 | |
| pET29b-inB6His (bp 106 to 1890) | Forward: AAC GTG CAT ATG GAG ACT ATC ACC GTG CCA ACG | This study |
| Reverse: ATT CTC GAG TTT CTG TGC CCT TAA ATT AGC TGC | This study | |
| Sequencing prfA mutants | Forward: CTA TCT GTT GCA GCT CTT CTT GG | This study |
| Reverse: CAG CTA ACA ATT GTT GTT ACT GCC | ||
| Confirm gus-neo insertion (prfA* mutants) | Forward: GCA GTC AAT TAA TAT GCC GAG CC | This study |
| Reverse: CGG ACC AAC TAA GTT TAT GTG G | This study | |
| Hydrolysis primers and probes for qPCR for gene target | ||
| inlA | Forward: GGC AAA GAA ACA ACC AAA GAA G | This study |
| Reverse: GGG CAT CAA ACC AAC CAA | This study | |
| Probe: AT TGA CTG AAC CAG CTA AGC CCG T | This study | |
| inlB | Forward: CCG AGC ACT TAA CAC ATT CTA C | This study |
| Reverse: TTA TCT GCT ACC GGG ACT TTA T | This study | |
| Probe: ATG TCA GCG CCA ATA AAG CTG GC | This study | |
| hly | Forward: CTG GTT TAG CTT GGG AAT GG | This study |
| Reverse: ATT TCG GAT AAA GCG TGG TG | This study | |
| Probe: TGA TGA CCG GAA CTT ACC ACT TGT GA | This study | |
| gap | Forward: TCA CAG CGC AAG ACA AAG | This study |
| Reverse: ACT GTT TCA GTT CCG TCT AAT G | This study | |
| Probe: TG TTA TCT CCG CTC CAG CAA CTG G | This study | |
| rpoB | Forward: TGT AAA ATA TGG ACG GCA TCG T | 90 |
| Reverse: GCT GTT TGA ATC TCA ATT AAG TTT GG | 90 | |
| Probe: CT GAT TCG CGC AAA ACT TCT ACG CG | 90 |
a
All probes have a 5' 6-FAM reporter dye and a 3' Iowa Black FQ quencher.