TABLE 1.
Strains and plasmids used in this study
| Strain | Genotype or description | Reference or source |
|---|---|---|
| N. meningitidis strains | ||
| NMB | B:2b:P1.2,5:L2 (CDC8201085) | 45 |
| IR3261 | NMB hpuB::erm hmbRoff | 38 |
| IR3287 | NMB hpuB::erm hmbRon | 38 |
| NMB310 | NMB misS::aphA-3 | 52 |
| YT391 | NMB310 complemented with Plac::misS at the lctP-aspC locus | 42 |
| SZT1001/YT0336 | NMB misR::aphA-3 | 51 |
| C1001 | NMB misR::aphA-3 complemented with Ptrc::misR in the iga locus | 51 |
| SZT1003 | NMB misRS::aphA-3 | 51 |
| SZT1004 | IR3261 misR::aphA-3 | This study |
| SZT1005 | IR3261 misS::aphA-3 | This study |
| SZT1006 | IR3261 misRS::aphA-3 | This study |
| SZT1009 | IR3287 misR::aphA-3 | This study |
| SZT1010 | IR3287 misS::aphA-3 | This study |
| GMT101 | PhmbR::lacZ | This study |
| GMT107 | PhmbR::lacZ ΔmisS | This study |
| O101 | lacZ-erm cassette inserted into BamHI site of hemO | This study |
| O101s | hemO::lacZ ΔmisS | This study |
| R103 | lacZ-erm cassette inserted into HincII site of hmbR | This study |
| R103s | hmbR::lacZ ΔmisS | This study |
| O1101 | PhemO1::lacZ | This study |
| O1101s | PhemO1::lacZ ΔmisS | This study |
| O2002 | PhemO2::lacZ | This study |
| O2002s | PhemO2::lacZ ΔmisS | This study |
| O305 | PhemO::lacZ with 918-bp promoter fragment | This study |
| O305s | PhemO::lacZ with 918-bp promoter fragment, ΔmisS | This study |
| O501 | PhemO::lacZ with 1,104-bp promoter fragment | This study |
| O501s | PhemO::lacZ with 1,104-bp promoter fragment, ΔmisS | This study |
| R1203 | PhemO::lacZ with 1,220-bp promoter fragment | This study |
| R1203s | PhmbR::lacZ with 1,220-bp promoter fragment, ΔmisS | This study |
| E. coli strains | ||
| DH5α | Cloning strain | New England Biolabs |
| TOP10 | Cloning strain | Invitrogen |
| Plasmids | ||
| pUC18k | Source of aphA-3 cassette | 32 |
| pCR2.1 | TOPO cloning vector | Invitrogen |
| pSmartLCKan | Blunt-end cloning vector | Lucigen |
| pYT328 | Cloning vector for chromosomal lacZ fusion | 52 |
| pYT338 | 5′ sequence (YT174-YT185) and 3′ sequence (YT186-YT141) of misR inserted into HindIII-KpnI site and EcoRI site of pCR2.1 | This study |
| pSZ001 | aphA-3 cassette inserted into KpnI-BamHI site of pYT338 | This study |
| pGM1 | PCR product obtained with hmbR-pF2 and hmbR-PR in pCR2.1 | This study |
| pGM2 | EcoRI fragment of pGM1 inserted into EcoRI site of pYT328 | This study |
| pPK-hemO | PCR fragment obtained with hemO-pF1-ERI and hmbR-pR-ERI cloned into pSmartLCKan | This study |
| phemO101 | lacZ-erm cassette inserted into BamHI site of pPK-hemO | This study |
| pPK-hmbR | PCR fragment obtained with hmbR-pF2 and hmbR-F3R cloned into pCR2.1 | This study |
| phmbR103 | lacZ-erm cassette inserted into HincII site of pPK-hmbR | This study |
| pPK-hemO1101 | PCR fragment obtained with hemO-pF1-ER and hemO-pR1-ER inserted into pYT328 | This study |
| pPK-hemO2002 | PCR fragment obtained with hemO-pF2-ER and hemO-pR1-ER inserted into pYT328 | This study |
| pPK-hemO305 | PCR fragment obtained with hemO-pF1-ER and hemO-pR3-ER inserted into pYT328 | This study |
| pPK-hemO501 | PCR fragment obtained with hemO-pF1-ER and hemO-pR5-ER inserted into pYT328 | This study |
| pPK-hmbR1203 | PCR fragment obtained with hemO-pF1-ER and hmbR-pR-ER inserted into pYT328 | This study |