TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Description | Source or reference |
|---|---|---|
| Strains | ||
| N. meningitidis | ||
| NMB | B:2b:P1.2,5:L2 (CDC8201085) | 27 |
| NMBmisR | NMB with misR::erm mutation | 31 |
| YT0310 | NMB with ΔmisS::aphA-3 mutation | This study |
| YT0263 | NMB with promoterless lacZ fusion in the chromosomal 988 locus | This study |
| YT0321 | NMB with misR(−504/+104)::lacZ fusion in the chromosomal 988 locus | This study |
| YT0322 | NMB with misR(−348/+104)::lacZ fusion in the chromosomal 988 locus | This study |
| YT0323 | NMB with misR(−205/+104)::lacZ fusion in the chromosomal 988 locus | This study |
| YT0324 | NMB with misR(−89/+104 with internal deletion between −8 and +25)::lacZ fusion in the chromosomal 988 locus | This study |
| YT0326 | NMB with misR(−205/+3)::lacZ fusion in the chromosomal 988 locus | This study |
| YT0329 | NMB with misR(−46/+104)::lacZ fusion in the chromosomal 988 locus | This study |
| YT0330 | NMB with misR(−103/+104)::lacZ fusion in the chromosomal 988 locus | This study |
| YT0332 | NMB with misR(−89/+104)::lacZ fusion in the chromosomal 988 locus | This study |
| E. coli | ||
| DH5α | Cloning strain | New England Biolabs |
| BL21(DE3) | Host strain for PT7-controlled expression | Novagen |
| TB1 | Host strain for MBP fusion protein expression | New England Biolabs |
| Plasmids | ||
| pCR2.1 | TA cloning vector, Apr, Kmr | Invitrogen |
| pET28a(+) | PT7-controlled expression vector, Kmr | Novagen |
| pMal-c2 | N-terminal MBP fusion vector | New England Biolabs |
| pUC18k | Source of aphA-3 cassette | 17 |
| pYT298 | MisR in pET28a | This study |
| PYT311 | MisS (residue 202 to 469) fused to MalE at N terminus in pMal-c2 | This study |
| PYT310 | aphA-3 cassette (EcoRI-HincII blunted) replacing EcoRV-HincII within misS fragment of YT140-YT67 in pCR2.1 | This study |
| pXZ015 | MisR-(His)6 fusion with MisR52DA mutation in pET28a | This study |
| pXZ019 | MBP-MisS fusion with MisS246HA mutation in pMal-c2 | This study |
| pYT328 | Cloning vector for lacZ fusion in a chromosomal intergenic region | This study |
| pYT321 | misR::lacZ carrying 608-bp (−504/+104) misR promoter region | This study |
| pYT322 | misR::lacZ carrying 452-bp (−348/+104) misR promoter region | This study |
| pYT323 | misR::lacZ carrying 309-bp (−205/+104) misR promoter region | This study |
| pYT324 | misR::lacZ carrying 193-bp (−89/+104 with internal deletion between −8 and +25) misR promoter region | This study |
| pYT326 | misR::lacZ carrying 208-bp (−205/+3) misR promoter region | This study |
| pYT329 | misR::lacZ carrying 150-bp (−46/+104) misR promoter region | This study |
| pYT330 | misR::lacZ carrying 207-bp (−103/+104) misR promoter region | This study |
| pYT332 | misR::lacZ carrying 193-bp (−89/+104) misR promoter region | This study |