Table 1. Strains and plasmids used in this study.
| Strain or plasmid | Descriptiona | Reference |
|---|---|---|
| pUC18-pilQ2-kan | Plasmid containing bp 1333-2196 of pilQ2, a kan resistance cassette, and an additional 300 bp of downstream sequence |
This study |
| pUC18-pilQFA19-kan | Same as above without the mutation at Glu-666 | This study |
| pUNCH290 | Plasmid containing pilQ disrupted by the Ω fragment | (Chen et al., 2004) |
| pMO/porIB-G120K-erm | Plasmid containing the mature coding sequence of PIB with a G120K mutation, an erm resistance cassette, and additional 300 bp of downstream sequence |
(Olesky et al., 2002) |
| pSY6 | Plasmid containing gonococcal DNA fragment encoding nalidixic acid resistance |
(Stein et al., 1991) |
| FA19 | Penicillin-susceptible laboratory strain | (Maness and Sparling, 1973) |
| FA6140 | Penicillin-resistant clinical isolate | (Danielsson et al., 1986) |
| FA19 penA4 | FA19 × FA6140 DNA (0.03 μg/ml penb) | (Ropp et al., 2002) |
| FA19 penA4 pilQ2 | FA19 penA4 X pUC18-pilQ2-kan (50 μg/ml kan) | This study |
| FA19 penA4 pilQ::Ω | FA19 penA4 X pUNCH290 (100 μg/ml spt) | This study |
| FA19 penA4 mtrR | FA19 penA4 X FA6140 DNA (1 mg/ml Triton X-100) | (Ropp et al., 2002) |
| FA19 penA mtrRpilQ2 | FA19 penA4 mtrR X pUC18-pilQ2-kan (50 μg/ml kan) | This study |
| FA19 penA mtrR pilQ::Ω | FA19 penA4 mtrR X pUNCH290 (100 μg/ml spt) | This study |
| FA19 penB | FA19 X pMO/porIB-G120K-erm (6 μg/ml erm) | This study |
| FA19 penB pilQ2 | FA19 penB X pUC18-pilQ2-kan (50 μg/ml kan) | This study |
| FA19 mtrR penB | FA19 mtrR X porIB-G120K-erm (6 μg/ml erm) | This study |
| FA19 mtrR penB pilQ2 | FA19 mtrR penB X pUC18-pilQ2-kan (50 μg/ml kan) | This study |
| PR100 | FA19 penA4 mtrR penB5 | (Ropp et al., 2002) |
| PR100 pilQ2 | PR100 X pUC18-penC-kan (50 μg/ml kan) | This study |
| PR100 pilQ::Ω | PR100 X pUNCH290 (100 μg/ml spt) | This study |
| FA6140 pilQ2 | FA6140 X pUC18-pilQ2-kan (50 μg/ml kan) | This study |
| FA6140 pilQ::Ω | FA6140 X pUNCH290 (100 μg/ml spt) | This study |
| RM11.2 recA6 | FA1090 with IPTG-regulatable recA allele | (Long et al., 1998) |
| RM11.2 recA6 pilQFA19 | RM11.2 recA6 X pUC18-pilQFA19-kan (50 μg/ml kan) | This study |
| RM11.2 recA6 pilQ2 | RM11.2 recA6 X pUC18-pilQ2-kan (50 μg/ml kan) | This study |
| SZ3 | RM11.2 recA6 penA4 mtrR penB5 | This study |
| SZ3 ΔpilE | pilE deletion strain of SZ3 | This study |
| SZ3 pilT::erm | SZ3 X RM11.2 recA6 pilT::erm (6 μg/ml erm) | This study |
| SZ3 ΔpilP | SZ3 X pUC18-ΔpilP (0.5 μg/ml pen) | This study |
| SZ3 pilQ2 | SZ3 X pUC18-pilQ2-kan (50 μg/ml kan) | This study |
| SZ3 pilQ::Ω | SZ3 X pUNCH290 (100 μg/ml spt) | This study |
a
Strains were constructed by homologous recombination as described in Materials and Methods and are shown as parental strain X donor DNA (selection agent). All relevant genotypes were confirmed by DNA sequencing.
b
The abbreviations are: pen, penicillin; kan, kanamycin; spt, spectinomycin; erm, erythromycin.