TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Genotype or relevant description | Source or reference |
|---|---|---|
| Strains | ||
| FA19 | Wild type | 45 |
| KH12 | FA19 mtrC::Kan | 33 |
| KH14 | FA19 mtrD::Kan | 32 |
| FA6807 | FA19 fetA::Ω (Strr Spcr) | 5 |
| MCV650 | FA19 tonB::Ω (Strr Spcr) | 30 |
| MCV841 | FA19 pilQ::Ω (Strr Spcr) | 21 |
| MCV904 | FA19 transformed by pVCU912 (Ermr) | This study |
| MCV906 | FA19 transformed by pVCU913 (Ermr) | This study |
| MCV917 | FA19 fetA::Ω tonB::Kan | This study |
| MCV920 | FA19 mtrC::Kan tonB::Ω | This study |
| MCV921 | FA19 mtrD::Kan tonB::Ω | This study |
| MCV925 | FA19 tdfF::Ω | This study |
| MCV926 | FA19 tdfG::Ω | This study |
| MCV927 | FA19 tdfH::Ω | This study |
| MCV928 | FA19 tdfJ::Ω | This study |
| Plasmids | ||
| pCR2.1 | Kanr Ampr | Invitrogen |
| pHSS6GCU | Plasmid containing the gonococcal uptake sequence Kanr | 24 |
| pHSX-ermC-lacIOP | Kanr Ermr | 61 |
| pUC18K | Vector containing the aphA3 cassette (Kanr) | 43 |
| pUNCH290 | PCR2.1 containing the Ω cassette in the HincII site of pilQ1 | 14 |
| pVCU690 | pCR2.1 containing tonBΔ160-194 allele | 39 |
| pVCU693 | pCR2.1 containing the Ω cassette in the AgeI site of tonB | 31 |
| pVCU704 | pCR2.1 containing the Ω cassette in the PflMI site of tdfF | 31 |
| pVCU705 | pCR2.1 containing the Ω cassette in the StuI site of tdfG | 31 |
| pVCU706 | pCR2.1 containing the Ω cassette in the HincII site of tdfH | 31 |
| pVCU707 | pCR2.1 containing the Ω cassette in the HincII site of tdfJ | 31 |
| pVCU907 | pHSS6GCU containing the fbpA allele from pVCU911 | This study |
| pVCU908 | pHSS6GCU fbpA upstream and coding | This study |
| pVCU909 | pCR2.1 fbpA upstream and coding region | This study |
| pVCU910 | pHSS6GCU containing fbpA upstream and coding region from pVCU909 | This study |
| pVCU911 | pET22b(+) containing FA19 fbpA allele | 53 |
| pVCU912 | pHSS6GCU fbpA ermC lacIP (−RBS) | This study |
| pVCU913 | pHSS6GCU fbpA ermC lacIP | This study |
| pVCU915 | MCV690 containing the aphA3 cassette in the AgeI site of tonB | This study |