TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Genotype | Reference or source |
|---|---|---|
| Bacterial strains | ||
| E. coli | ||
| DH5α | supE44 ΔlacU169 (φ80 lacZΔM15) hsdR17 recA1 endA1 gyrA96 thi-1 relA1β | 23 |
| BL21(DE3) | hsdS gal (λcIts857 ind1 Sam7 nin5 lacUV5-T7 gene 1) | 38 |
| H. pylori | ||
| G27 | Clinical isolate; wild type | 44 |
| ΔnikR::Km strain | G27 derivative with nikR partially deleted; Kmr | 29 |
| G27vac::PA3 lacZ strain | G27 derivative containing the wild-type PfecA3 promoter-lacZ fusion in the vacA locus; Kmr | This study |
| G27vac::PA3.1 lacZ strain | G27 derivative containing the OP-I mutant PfecA3.1-lacZ fusion in the vacA locus; Kmr | This study |
| G27vac::PA3.15 lacZ strain | G27 derivative containing the OP-I mutant PfecA3.15-lacZ fusion in the vacA locus; Kmr | This study |
| G27vac::PA3.16 lacZ strain | G27 derivative containing the OP-I mutant PfecA3.16-lacZ fusion in the vacA locus; Kmr | This study |
| G27vac::PA3.17 lacZ strain | G27 derivative containing the OP-II mutant PfecA3.17-lacZ fusion in the vacA locus; Kmr | This study |
| G27vac::PA3.19 lacZ strain | G27 derivative containing the OP-II mutant PfecA3.19-lacZ fusion in the vacA locus; Kmr | This study |
| G27vac::PA3.21 lacZ strain | G27 derivative containing the OP-II mutant PfecA3.21-lacZ fusion in the vacA locus; Kmr | This study |
| ΔnikR::Km vac::PA3.1 lacZ strain | ΔnikR strain derivative containing the OP-I mutant PfecA3.1-lacZ fusion in the vacA locus; Kmr Cpr | This study |
| ΔnikR::Km vac::PA3.15 lacZ strain | ΔnikR strain derivative containing the OP-I mutant PfecA3.15-lacZ fusion in the vacA locus; Kmr Cpr | This study |
| ΔnikR::Km vac::PA3.16 lacZ strain | ΔnikR strain derivative containing the OP-I mutant PfecA3.16-lacZ fusion in the vacA locus; Kmr Cpr | This study |
| Plasmids | ||
| pBluescript SK (pBS) | General cloning vector; Ampr | Stratagene |
| pA3 | pBluescript derivative containing the wild-type fragment of the fecA3 promoter region, amplified by PCR with primers A3Fnew and A3Rnew | 12 |
| pA3.1 | pA3 derivative containing 265 bp of fecA3 promoter region with a SmaI site in OP-I generated by PCR amplification of the wild-type construct with primers A3Fnew-A3.12 and A3Rnew-A3.11 and subsequent ligation at the SmaI site | This study |
| pA3.15 | pA3 derivative containing 265 bp of fecA3 promoter region with a SmaI site in OP-I generated by PCR amplification of the wild-type construct with primers A3Fnew-A3.14 and A3Rnew-A3.13 and subsequent ligation at the SmaI site | This study |
| pA3.16 | pA3 derivative containing 265 bp of fecA3 promoter region with a SmaI site in OP-I generated by PCR amplification of the wild-type construct with primers A3Fnew-A3.16 and A3Rnew-A3.15 and subsequent ligation at the SmaI site | This study |
| pA3.17 | pA3 derivative containing 265 bp of fecA3 promoter region with a SmaI site in OP-II generated by PCR amplification of the wild-type construct with primers A3Fnew-A3.18 and A3Rnew-A3.17 and subsequent ligation at the SmaI site | This study |
| pA3.19 | pA3 derivative containing 265 bp of fecA3 promoter region with a SmaI site in OP-II generated by PCR amplification of the wild-type construct with primers A3Fnew-A3.20 and A3Rnew-A3.19 and subsequent ligation at the SmaI site | This study |
| pA3.21 | pA3 derivative containing 265 bp of fecA3 promoter region with a SmaI site in the OP-II generated by PCR amplification of the wild-type construct with primers A3Fnew-A3.22 and A3Rnew-A3.21 and subsequent ligation at the SmaI site | This study |
| pBS-CAT | pBluescript derivative containing the chloramphenicol resistance cassette from Campylobacter coli; Ampr Cpr | 41 |
| pVac::Km | pGemZ derivative containing a kanamycin cassette | 16 |
| pVac::PA3lacZ | pVac::Km derivative containing the transcriptional fusion PfecA3::lacZ; Kmr | This study |
| pVac::PA3.1lacZ | pVac::Km derivative containing the transcriptional fusion PfecA3.1::lacZ; Kmr Cpr | This study |
| pVac::PA3.15lacZ | pVac::Km derivative containing the transcriptional fusion PfecA3.15::lacZ; Kmr Cpr | This study |
| pVac::PA3.16lacZ | pVac::Km derivative containing the transcriptional fusion PfecA3.16::lacZ; Kmr Cpr | This study |
| pVac::PA3.17lacZ | pVac::Km derivative containing the transcriptional fusion PfecA3.17::lacZ; Kmr | This study |
| pVac::PA3.19lacZ | pVac::Km derivative containing the transcriptional fusion PfecA3.19::lacZ; Kmr | This study |
| pVac::PA3.21lacZ | pVac::Km derivative containing the transcriptional fusion PfecA3.21::lacZ; Kmr | This study |
| pNKTB | pGemT derivative containing the nikR-exbB intergenic region; Ampr | 15 |
| pET15b | IPTGa-inducible, N-terminal His6-tagged recombinant protein overexpression vector; Ampr | Novagen |
| pET15b-nikR | pET15b derivative, containing the nikR coding sequence cloned in frame within the NdeI/BamHI restriction sites | 15 |
a
IPTG, isopropyl-β-d-thiogalactopyranoside.