Table 4 Strains and plasmids used in this study.
Strain | Description | Source |
|---|---|---|
N. gonorrhoeae strains | ||
F62 | Parental strain | Laboratory stocks |
RUG7001 | F62 proAB paniA::lacZ | [43] |
RUG7022 | F62 proAB paniA::lacZ fnr | [6] |
JCGC201 | F62 proAB pccp::lacZ | [5] |
JCGC202 | F62 proAB pccp::lacZ fnr | [5] |
JCGC502 | F62 fnr- 3xFLAG KanR | This work |
E. coli strains | ||
RV | Parental strain | Laboratory stocks |
JCB386 | Δ (nirB-cycG) pnirB::lacZ | [44] |
JCB3861 | JCB386 narXL | This work |
JCB3862 | JCB386 narXL narQ | This work |
JCB3863 | JCB386 narXL narQ narP | This work |
JCB391 | RV narXL narQ pcnB | This work |
JCB12 | RV narX narQ frdA::lacZ | [10] |
Plasmids | ||
pGCFNR3 | Gonococcal fnr, under the regulation of the E. coli fnr promoter, cloned into pBR322. | [5] |
pGCFNRi | pGCFNR3 with Kpn I and Xho I restriction sites engineered downstream of the fnr gene | This work |
pGCFNR-FLAG | pGCFNR3 with a 3xFLAG tag and Kanamycin resistance cassette inserted downstream of the fnr gene | This work |
pSUB11 | Epitope tagging plasmid carrying 3xFLAG tag and kanamycin resistance cassette. | [36] |
pGCNarQP | Gonococcal narQP genes under the control of the E. coli fnr promoter cloned into pBR322. | This work |
pBADgcQ | Gonococcal narQ gene under the control of the araBAD promoter cloned into pBAD myc-His A. | This work |
pBADecQ | E. coli narQ gene under the control of the araBAD promoter cloned into pBAD myc-His A | This work |
pRNW18 | pBADEcQ containing R54K substitution in narQ | This work |
pRNW19 | pBADEcQ containing N48E I49E substitution in narQ | This work |
pRNW20 | pBADEcQ containing N48E I49E R54K substitution in narQ | This work |
pRNW29 | pBADEcQ containing D43A E45S A46V N48E I49E substitution in narQ | This work |
pRNW30 | pBADEcQ containing D43A E45S A46V substitution in narQ | This work |
pRNW31 | pBADEcQ containing D43A E45S A46V R54K substitution in narQ | This work |
pRNW32 | pBADEcQ containing D43A E45S A46V N48E I49E R54K substitution in narQ | This work |
pRNW33 | pBADEcQ containing D43A E45S A46V S52N S57A substitution in narQ | This work |
pRNW34 | pBADEcQ containing D43A E45S A46V N48E I49E S52N R54K S57A substitution in narQ | This work |
p7150 | E. coli nirB promoter cloned into the lacZ reporter vector pRW50. | [19] |
pRNW15 | E. coli napF promoter cloned into the lacZ reporter vector pRW50. | This work |