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 |