TABLE 1.
Bacterial strains and plasmids
| Strain or plasmid | Properties | Source or reference |
|---|---|---|
| Plasmids | ||
| pUP1 | Template for pIDN plasmids; contains 10-bp neisserial DUS in polylinker; Kmr | 10 |
| pMOB | Source of polylinker for pIDN plasmids; pUC origin; Apr; multicloning site (MCS); T3 and T7 promoters | 39 |
| pHSS23 | Source of original ermC gene | 44 |
| pJD1145 | Source of improved ermC gene for pIDN construction | This work |
| pNH5 | Forward orientation of ermC from pJD1145 in pUP1; Emr Kmr | This work |
| pNH5-1 | Reverse orientation of ermC from pJD1145 in pUP1; Emr Kmr | This work |
| pNH9 | BamHI-PvuII deletion of Kmr in pNH5 | This work |
| pNH9-1 | BamHI-PvuII deletion of Kmr in pNH5-1 | This work |
| pNH9-6 | pMOB polylinker in forward orientation in pNH9 (Emr) | This work |
| pNH9-7 | pMOB polylinker in reverse orientation in pNH9 (Emr) | This work |
| pNH9-8 | pMOB polylinker in forward orientation in pNH9-1 (Emr) | This work |
| pNH9-9 | pMOB polylinker in reverse orientation in pNH9-1 (Emr) | This work |
| pIDN1 | IDM vector: ermC in forward orientation, polylinker in forward orientation | This work |
| pIDN2 | IDM vector: ermC in forward orientation, polylinker in reverse orientation | This work |
| pIDN3 | IDM vector: ermC in reverse orientation, polylinker in forward orientation | This work |
| pIDN4 | IDM vector: ermC in reverse orientation, polylinker in reverse orientation | This work |
| pJD1175 | 642-bp EcoRV-Sau3AI fragment of traH from MS11A cloned into pIDN1 | This work |
| pJD1177 | 642-bp EcoRV-Sau3AI fragment of traH from MS11A cloned into pIDN3 | This work |
| pKS43 | 479-bp Sau3AI fragment of traG from MS11A cloned into pIDN1 | This work |
| pKS53 | 479-bp Sau3AI fragment of traG from MS11A cloned into pIDN3 | This work |
| pNH10-1M | AgeI-NsiI fragment of traG from PCR of MS11A with 23F and EndL6R in XmaI and PstI sites of pNH9-9 | This work |
| pHH1 | HindIII deletion of pNH10-1M | This work |
| pHH2 | ScaI-EcoRV deletion of pNH10-1M | This work |
| pHH3 | ScaI-Ecl136II deletion of pNH10-1M | This work |
| pNH11 | Blunted PCR product of MS11A with 27F and TraH2R in EcoRV site of pNH9-9; contains 5′ end of traH and upstream sequence | This work |
| pGCC6 | Plasmid for complementation in the gonococcal chromosome; lac promoter/operator; lacI Cmr Kmr | 23 |
| pHH15 | PCR product of MS11A with TraH-up and TraH-down, cut with NsiI and EcoRV and cloned into the NsiI and PmeIsites of pGCC6 | This work |
| N. gonorrhoeae | ||
| MS11A | Wild-type N. gonorrhoeae | 32 |
| KS1 | MS11A transformed by IDM with pJD1175, polar mutant of traH (Emr) | This work |
| KS58 | MS11A transformed by IDM with pKS43, polar mutant of traG (Emr) | This work |
| KS16 | MS11A transformed by IDM with pJD1177, nonpolar mutant of traH (Emr) | This work |
| KS59 | MS11A transformed by IDM with pKS53, nonpolar mutant of traG (Emr) | This work |
| HH500 | MS11A transformed by IDM with pNH10-1M (Emr) | This work |
| HH501 | MS11A transformed by IDM with pHH1 (Emr) | This work |
| HH502 | MS11A transformed by IDM with pHH2 (Emr) | This work |
| HH503 | MS11A transformed by IDM with pHH3 (Emr) | This work |
| JD1510 | MS11A atlA::ermC (insertion-deletion mutation) | 6 |
| JD1603 | MS11A transformed by IDM with pNH11 (Emr) | This work |
| JD1545 | MS11A cnp::cat recA6 (Cmr) | 7 |
| MS11-Spc | MS11A Spr | 33 |
| HH507 | JD1545 transformed by IDM with pNH10-1M (traG recA6 Cmr Emr) | This work |
| HH508 | JD1545 transformed by IDM with pNH11 (recA6 Cmr Emr) | This work |
| HH518 | KS16 transformed with pHH15; traH expression in trans (Cmr Emr) | This work |