TABLE 1.
Strains and plasmids
| Strain or plasmid | Relevant characteristic(s) | Reference or source |
|---|---|---|
| Strains | ||
| E. coli TOP10 | F−mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 recA1 deoR araD139 Δ(ara-leu)7697 alU galK rpsL(Strr) endA1 nupG | Invitrogen |
| N. gonorrhoeae | ||
| FA19 | Wild type (TbpA+ TbpB+) | 43 |
| FA6905 | TbpA+ TbpB− (ΔtbpB) | 22 |
| FA6747 | TbpA− (tbpA::mTn3cat) TbpB+ | 20 |
| FA6815 | TbpA− TbpB− (tbpB::Ω) | 1 |
| MCV601 | FA19 derivative; LbpB− (lbpB::Ω) | 7 |
| MCV250 | TbpA E118A TbpB+ | This study |
| MCV251 | TbpA E118A TbpB− (ΔtbpB) | This study |
| MCV252 | TbpA E120A TbpB+ | This study |
| MCV253 | TbpA E120A TbpB− (ΔtbpB) | This study |
| MCV254 | TbpA Y121A TbpB+ | This study |
| MCV255 | TbpA Y121A TbpB− (ΔtbpB) | This study |
| MCV256 | TbpA E122A TbpB+ | This study |
| MCV257 | TbpA E122A TbpB− (ΔtbpB) | This study |
| MCV258 | TbpA E120A Y121A TbpB+ | This study |
| MCV259 | TbpA E120A Y121A TbpB− (ΔtbpB) | This study |
| MCV260 | TbpA E120A Y121A E122A TbpB+ | This study |
| MCV261 | TbpA E120A Y121A E122A TbpB− (ΔtbpB) | This study |
| Plasmids | ||
| pCR2.1 TOPO | Kanr Ampr | Invitrogen |
| pHSS6-GCU | Vector containing gonococcal uptake sequence (Kanr) | 24 |
| pUNCH411 | pBS-SK(+) containing entire tbpA gene | 19 |
| pVCU250 | pCR2.1 TOPO containing tbpA gene amplified with oVCU334 and oVCU335; E118A; PvuI restriction site | This study |
| pVCU251 | pHSS6-GCU containing EcoRI fragment from pVCU250 | This study |
| pVCU252 | pCR2.1 TOPO containing tbpA gene amplified with oVCU334 and oVCU335; E120A; NdeI restriction site | This study |
| pVCU253 | pHSS6-GCU containing EcoRI fragment from pVCU252 | This study |
| pVCU254 | pCR2.1 TOPO containing tbpA gene amplified with oVCU334 and oVCU335; Y121A; HaeIII restriction site | This study |
| pVCU255 | pHSS6-GCU containing EcoRI fragment from pVCU254 | This study |
| pVCU256 | pCR2.1 TOPO containing tbpA gene amplified with oVCU334 and oVCU335; E122A; HpyCH4V restriction site | This study |
| pVCU257 | pHSS6-GCU containing EcoRI fragment from pVCU256 | This study |
| pVCU258 | pCR2.1 TOPO containing tbpA gene amplified with oVCU334 and oVCU335; E120A Y121A; PstI restriction site | This study |
| pVCU259 | pHSS6-GCU containing EcoRI fragment from pVCU258 | This study |
| pVCU260 | pCR2.1 TOPO containing tbpA gene amplified with oVCU334 and oVCU335; E120A Y121A E122A; PstI and AluI restriction sites | This study |
| pVCU261 | pHSS6-GCU containing EcoRI fragment from pVCU260 | This study |