TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Genotype or relevant properties | Reference(s) or source |
|---|---|---|
| N. meningitidis strains | ||
| Wild-type parent strains (ST, serogroup, immunotype) | ||
| NIID280 (2032, untypeable, L5) | lpt3 lpt6 lgtG+lot3+ | 36 |
| H44/76 (32, B, L3) | lpt3+lpt6 lgtG lot3a | 18, 36, 37 |
| NIID311 (5, A, L2) | lpt3 lpt6+lgtG+lot3+ | 36 |
| Mutants | ||
| NIID280 derivatives | ||
| HT1125 | NIID280 ΔsiaB-D::kan | 36 |
| HT1219 | NIID280 ΔlptA::spc | This study |
| HT1222 | NIID280 ΔsiaB-D::kan ΔlptA::spc | This study |
| HT1277 | NIID280 ΔsiaB-D::kan ΔlptA::spc ggt::lptA+-cat | This study |
| HT1278 | NIID280 ΔsiaB-D::kan ΔlptA::spc ggt::cat | This study |
| HT1238 | NIID280 ΔsiaB-D::kan pilE::ermC | This study |
| HT1249 | NIID280 ΔsiaB-D::kan pilE::ermC ΔlptA::spc | This study |
| HT1178 | NIID280 pilC1::spc | 36 |
| HT1182 | NIID280 ΔpilC2::ermC | 36 |
| HT1186 | NIID280 pilC1::spc ΔpilC2::ermC | 36 |
| H44/76 derivatives | ||
| HT1034 | H44/76 ΔsiaB-D::kan | 36 |
| HT1378 | H44/76 ΔlptA::spc | This study |
| HT1243 | H44/76 siaB-D::kan ΔlptA::spc | This study |
| HT1235 | H44/76 siaB-D::kan pilE::ermC | This study |
| HT1245 | H44/76 siaB-D::kan ΔlptA::spc pilE::ermC | This study |
| HT1234 | H44/76 siaB-D::kan Δopc::cat | This study |
| HT1244 | H44/76 siaB-D::kan ΔlptA::spc Δopc::cat | This study |
| HT1236 | H44/76 siaB-D::kan pilE::ermC Δopc::cat | This study |
| HT1246 | H44/76 siaB-D::kan ΔlptA::spc Δopc::cat pilE::ermC | This study |
| HT1295 | H44/76 lptA+-FLAG-cat | This study |
| HT1015 | H44/76 pilE::ermC | This study |
| HT1002 | H44/76 Δopc::cat | This study |
| NIID311 derivatives | ||
| HT1336 | NIID311 ΔsacAB::kan | 36 |
| HT1380 | NIID311 ΔlptA::spc | This study |
| HT1359 | NIID311 sacAB::kan ΔlptA::spc | This study |
| E. coli strains | ||
| JM109 | endA1 gyrA96 hsdR17(rK− mK+) mcrB+recA1 relA1 supE44 thi-1 Δ(lac-proAB) F′ [traD36 proAB lacIqZΔM15] | Takara Bio |
| BL21(DE3) | F−ompT hsdSB (rB− mB−) gal dcm (DE3) | Novagen |
| Plasmids | ||
| pET24a | Expression vector; Kmr | Novagen |
| pHT463 | Derivative of pET24a expressing lptA gene; Kmr | This study |
| pMW118 | Cloning vector; Apr | Nippon Gene |
| pHT432 | Derivative of pMW118 carrying an lptA gene; Apr | This study |
| pHT433 | Derivative of pHT432 carrying a ΔlptA::Spcr gene; Apr Spcr | This study |
| pHT473 | Derivative of pHT432 carrying a cat gene at the SphI site; Apr Cmr | This study |
| pTWV228 | Cloning vector; Apr | Takara Bio |
| pHT195 | Derivative of pTWV228 carrying a ggt gene; Apr | 38 |
| pHT507 | Derivative of pHT195 carrying lptA+- cat genes at the BstXI site; Apr Cmr | This study |
| pHT508 | Derivative of pHT195 carrying a cat gene at the BstXI site; Apr Cmr | This study |
| pGEM-T | PCR product cloning vector; Apr | Promega |
| pHT515 | Derivative of pGEM-T carrying a 1-kb downstream region of the lptA gene; Apr | This study |
| pHT516 | Derivative of pGEM-T carrying a 3′ part of the lptA gene with a FLAG epitope; Apr | This study |
| pHT518 | Derivative of pHT516 carrying a 1-kb downstream region of the lptA gene; Apr | This study |
| pHT519 | Derivative of pHT518 carrying a cat gene at the HpaI site; Apr Cmr | This study |
| pUC118 | Cloning vector; Ampr | Takara Bio |
| pHT148 | Derivative of pUC118 carrying a Δopc::ermC gene; Apr Ermr | 37 |
| pHT10 | Derivative of pHT148 carrying a Δopc::cat gene; Apr Cmr | This study |
| pHT434 | Derivative of pHT432 deleted of 0.7 kb of the NruI-SphI DNA fragment; Apr Spcr | This study |
| pGSS33 | Broad-host-range vector; Apr Cmr Strr Tetr | 37 |
| pSTV28 | Cloning vector; Cmr | Takara Bio |
| pHT261 | Derivative of pGSS33 carrying a cat gene and multicloning site of pSTV28 at the SacI and SalI sites; Cmr | This study |
| pHT435 | Derivative of pHT261 carrying an lptA gene; Cmr | This study |