TABLE 1.
Bacterial strains, plasmids, and primers used in this study
| Nontypeable H. influenzae strain or plasmid | Genotype, relevant characteristic(s), and/or sequence (5′-3′)a | Source or reference |
|---|---|---|
| NTHI strains | ||
| 2019 | Wild type | Our laboratory |
| 2019Δnuc | Δnuc; Kan | This study |
| 2019Δnuc::nuc | Δnuc::nuc; Kan Sp | This study |
| Plasmids | ||
| pGEM-T | Promega | |
| pET151/D-TOPO | Amp | Life Technologies |
| pUC18K3 | Kan | This laboratory |
| p601.1-Sp2 | Sp | 10 |
| pCEC#27 | pUC18K3::Δnuc::kan; Kan | This study |
| pCEC#29 | pET151::nuc; Amp | This study |
| pCEC#31 | p601.1Sp2::nuc; Sp | This study |
| Primers | ||
| DNase 1 FRET assay primer | Cy3-CCGCGAGAAACCAAGCACAGAGCACCGAAGA-BHQ_2 | |
| qRT-PCR primers | ||
| nuc primers | ||
| Forward | AGGGTATGGCATGGGCTTAT | |
| Reverse | GTGCGAGCCTGTTCATATTG | |
| OMP p6 primers | ||
| Forward | GGTATTCFTGGGTGGTTGCGTATT | |
| Reverse | AGATCAGCAACAGAGTAACCGCCA |
a
Kan, kanamycin; Sp, spectinomycin; Amp, ampicillin; BHQ, black hole quencher.