Table 2.
Bacterial plasmids and strains
| Plasmid/Strain | Description | Reference |
|---|---|---|
| Plasmids | ||
| pIDN1 | Cloning vector (EmR), 2 kb | Hamilton et al. (2001) |
| pIDN2 | Cloning vector (EmR), 2 kb | Hamilton et al. (2001) |
| pIDN3 | Cloning vector (EmR), 2 kb | Hamilton et al. (2001) |
| pKH37 | Complementation vector (CmR), 6.5 kb | Kohler et al. (2007) |
| pKS94 | pIDN2 containing ∼7 kb of the GGI from traI’ to traB’, 9 kb | Hamilton et al. (2001) |
| pWSP3 | ‘yaf-traI’; ∼0.8 kb BamHI-MfeI fragment of pKS94 in pIDN2, 2.8 kb | This study |
| pWSP4 | traIY200F/Y201F (TAT to TTT); mutagenesis PCR of pWSP3, 2.8 kb | This study |
| pWSP5 | traIY93F (TAT to TTT); mutagenesis PCR of pWSP3, 2.8 kb | This study |
| pWSP14 | Full-length traI; PCR amplified from MS11, ClaI and SacI digested in pIDN1, 4.7 kb | This study |
| pWSP15 | traI 3′ end Flag tag; PCR of pWSP14 with primers containing the Flag tag; GAC TAC AAG GAC GAC GAC AAG, 4.7 kb | This study |
| pWSP30 | ‘traI-ltgX’; ∼5.2 kb PvuII-SpeI fragment of pKS94 | This study |
| pWSP31 | IR::cat; PmeI-FspI cat-containing fragment from pKH37 in StuI-digested pWSP30, ∼8.1 kb | This study |
| pWSP33 | traI 3′ end Flag & His10 tags; PCR of pWSP15 with primers containing the His10 tag, 4.7 kb | This study |
| pWSP34 | traI 3′ end Flag and His10 tags; ∼2.7 kb ClaI-SacI fragment of pWSP33 in pKH37, 9 kb | This study |
| pWSP36 | Full-length traI; PCR amplified from MS11 ClaI- and SacI-digested in pKH37, 9 kb | This study |
| pWSP37 | yaf-ltgX non-coding region; PCR amplified, ∼750 bp in pIDN1, 2.4 kb | This study |
| pWSP38 | 0.9 kb EcoRV-NheI ermC-containing fragment from pIDN3 in PsiI-digested pWSP37, 4.5 kb | This study |
| pWSP42 | traIY200F (TAT to TTC); mutagenesis PCR of pWSP3, 2.8 kb | This study |
| pWSP43 | traIY201F (TAT to TTC); mutagenesis PCR of pWSP3, 2.8 kb | This study |
| pWSP44 | traIR99S (AGG to AGC); mutagenesis PCR of pWSP3, 2.8 kb | This study |
| pWSP45 | traIH106S (CAT to TCG); mutagenesis PCR of pWSP3, 2.8 kb | This study |
| pWSP46 | traIH108S (CAT to AGC); mutagenesis PCR of pWSP3, 2.8 kb | This study |
| pWSP47 | traI’; ∼2.9 kb BseRI-XhoI fragment of pWSP14 | This study |
| pWSP48 | traIH106A/H108A (CAT to AGC); mutagenesis PCR of pWSP3, 2.8 kb | This study |
| pWSP49 | traIH161S (CAC to AGT); mutagenesis PCR of pWSP47, 2.9 kb | This study |
| pWSP50 | traID162N (GAC to AAC); mutagenesis PCR of pWSP47, 2.9 kb | This study |
| pWSP51 | traIY212F (TAT to TTT); mutagenesis PCR of pWSP47, 2.9 kb | This study |
| pWSP52 | traID120N (GAC to AGT); mutagenesis PCR of pWSP47, 2.9 kb | This study |
| pWSP54 | traIL6K/L12K (CTC and CTA to AAG); mutagenesis PCR of pWSP3, 2.8 kb | This study |
| pWSP58 | yaf-ltgX non-coding region in pKH37, 8.2 kb | This study |
| pSJ001 | traIΔ261-850; PCR amplified from MS11, BspHI and XhoI digested in pET15b | This study |
| pBSKftsQ | Plasmid encoding FtsQ | van der Laan et al. (2004) |
| pET147 | Plasmid encoding proOmpA | van der Wolk et al. (1997) |
| Strains | ||
| MS11 | Wild-type N. gonorrhoeae | Swanson et al. (1971) |
| ND500 | Gonococcal genetic island deletion in MS11 | Hamilton et al. (2005) |
| WSP4 | MS11 transformed with pWSP4; traIY200F/Y201F | This study |
| WSP5 | MS11 transformed with pWSP5; traIY93F | This study |
| WSP31 | MS11 transformed with pWSP31; IR::cat | This study |
| WSP34 | E. coli overexpressing TraIFLAG/His10 (C-terminus tags) | This study |
| WSP36 | MS11 transformed with pWSP36; traI+OE from an inducible construct on the chromosome | This study |
| WSP38 | MS11 transformed with pWSP38; traI+OE from native site | This study |
| WSP42 | MS11 transformed with pWSP42; traIY200F | This study |
| WSP43 | MS11 transformed with pWSP43; traIY201F | This study |
| WSP44 | MS11 transformed with pWSP44; traIR99S | This study |
| WSP45 | MS11 transformed with pWSP45; traIH106S | This study |
| WSP46 | MS11 transformed with pWSP46; traIH108S | This study |
| WSP48 | MS11 transformed with pWSP48; traIH106A/H108A | This study |
| WSP49 | MS11 transformed with pWSP49; traIH161S | This study |
| WSP50 | MS11 transformed with pWSP50; traID162N | This study |
| WSP51 | MS11 transformed with pWSP51; traIY212F | This study |
| WSP52 | MS11 transformed with pWSP52; traID120N | This study |
| WSP54 | MS11 transformed with pWSP54; traIL6K/L12K | This study |
| HH532 | TraI C-terminus truncation in MS11; traIΔ349-850 | Hamilton et al. (2005) |
| WSP532 | HH532 transformed with pWSP36; traI C-terminus truncation complement | This study |
| WSP3604 | WSP4 transformed with pWSP36; traIY200F/Y201F complement | This study |
| WSP3605 | WSP5 transformed with pWSP36; traIY93F complement | This study |
| WSP3643 | WSP43 transformed with pWSP36; traIY201F complement | This study |
| WSP3854 | WSP54 transformed with pWSP38; traIL6K/L12KOE from native site | This study |
| WSP5831 | WSP31 transformed with pWSP58; IR::cat complement | This study |
| MS11Sp | Spectinomycin resistant, recipient strain (MS11 background) | Dillard and Seifert (2001) |
| JD1545 | MS11 recA6 cnp::cat, donor strain (MS11 background) | Dillard and Seifert (2001) |