Table 1.
Bacterial strains and plasmids used in this study.
| Bacterial strains/ Plasmids | Description | Source/ Reference |
|---|---|---|
| Strains | ||
| H. pylori G27 wild type | Clinical isolate, wild type | Xiang et al., 1995 |
| H. pylori G27 (hspR::Km) | G27 derivative; bp 66 to 334 of the hspR coding sequence replaced by a Kanamycin (Km) cassette. | Spohn and Scarlato, 1999 |
| H. pylori G27 (vacA::Pcbpwt-lux) | G27 derivative; containing the Pcbp wild type promoter region upstream of the luxC gene in the vacA locus; Cpr | This work |
| H. pylori G27 (vacA::Pcbpwt-lux, hspR::Km) | G27 derivative; containing the Pcbp wild type promoter region upstream of the luxC gene in the vacA locus and the hspR coding sequence (bp 66 to 334) replaced by a Kanamycin (Km) cassette; Cpr, Kmr | This work |
| H. pylori G27 (vacA::PcbpM1 + 2-lux, hspR::Km) | G27 derivative; containing the Pcbp HAIR-mutant promoter region upstream of the luxC gene in the vacA locus and the hspR coding sequence (bp 66 to 334) replaced by a Kanamycin (Km) cassette; Cpr, Kmr | This work |
| H. pylori G27 (vacA::PcbpM1 + 2-lux) | G27 derivative; containing the Pcbp HAIR-mutant promoter region upstream of the luxC gene in the vacA locus; Cpr | This work |
| E. coli DH5α | supE44 ΔlacU169 (φ80 lacZΔM15) hsdR17 recA1 endA1 gyrA96 thi-1 relA1 | Hanahan, 1983 |
| E. coli BL21(DE3) | hsdS gal (λcIts857 ind1 Sam7 nin5 lacUV5-T7 gene 1). | Studier et al., 1990 |
| Plasmids | ||
| pGEM-T-Easy | Cloning vector, Ampr | Promega |
| pGEM-T-Easy-RBS gro | pGEM-T-Easy derivative, containing a 147 bp DNA fragment corresponding to the region from 9,452–9,543 of H. pylori G27 genome amplified by PCR with oligonucleotides RBS GroF/RBS GroR. This region corresponds to a portion of the promoter region of HPG27_RS00075 (HP0011 according to 26695 annotation). | This work |
| pGEM-T-Easy-RBS hrc | pGEM-T-Easy derivative, containing a 252 bp DNA fragment corresponding to the region from 118,944–119,035 of H. pylori G27 genome amplified by PCR with oligonucleotides RBShrcF/PhrcF. This region corresponds to a portion of the promoter region of HPG27_RS00580 (HP0111 according to 26695 annotation). | This work |
| pGEM-T-Easy-HBS cbp | pGEM-T-Easy derivative, containing a 146 bp DNA fragment corresponding to the region from 433,049 to 433,140 of H. pylori G27 genome amplified by PCR with oligonucleotides HBSCbpF/HBSCbpF. This region corresponds to a portion of the promoter region and coding sequence of HPG27_RS02130 (HP1024 according to 26695 annotation). | This work |
| pGEM-T-Easy-HBSspeA | pGEM-T-Easy derivative, containing a 135 bp DNA fragment corresponding to the region from 1,035,104–1,035,195 of H. pylori G27 genome amplified by PCR with oligonucleotides HBSspeAF /HBSspeAR. This region corresponds to a portion of the coding sequence of HPG27_RS02130 (HP0422 according to 26695 annotation). | This work |
| pGEM-T-Easy-HBScbpM1 | pGEM-T-Easy derivative, containing a 91 bp DNA fragment corresponding to the region from 432,991–433,071 of H. pylori G27 genome generated annealing oligonucleotides Mut1F/Mut1R. This region corresponds to a portion of the promoter region and coding sequence of HPG27_RS02130 (HP1024 according to 26695 annotation). | This work |
| pGEM-T-Easy-HBScbpM2 | pGEM-T-Easy derivative, containing a 146 bp DNA fragment corresponding to the region from 433,049–433,140 of H. pylori G27 genome amplified by all around PCR with oligonucleotides Mut2F/Mut2R and using as DNA template the plasmid pGEM-T-Easy-HBScbp. This region corresponds to a portion of the promoter region and coding sequence of HPG27_RS02130 (HP1024 according to 26695 annotation). | This work |
| pGEM-T-Easy-HBScbpM1 + 2 | pGEM-T-Easy derivative, containing a 146 bp DNA fragment corresponding to the region from 433,049–433,140 of H. pylori G27 genome amplified by all around PCR with oligonucleotides Mut2F/Mut2DR and using as DNA template the plasmid pGEM-T-Easy-HBScbp. This region corresponds to a portion of the promoter region and coding sequence of HPG27_RS02130 (HP1024 according to 26695 annotation). | This work |
| pGEM-T-Easy-HBScbpM3 | pGEM-T-Easy derivative, containing a 91 bp DNA fragment corresponding to the region from 432,991–433,071 of H. pylori G27 genome generated annealing oligonucleotides Mut3F/Mut3R. This region corresponds to a portion of the promoter region and coding sequence of HPG27_RS02130 (HP1024 according to 26695 annotation). | This work |
| pGEM-T-Easy-HBScbpM4 | pGEM-T-Easy derivative, containing a 91 bp DNA fragment corresponding to the region from 432,991–433,071 of H. pylori G27 genome generated annealing oligonucleotides Mut4F/Mut4R. This region corresponds to a portion of the promoter region and coding sequence of HPG27_RS02130 (HP1024 according to 26695 annotation). | This work |
| pGEM-T-Easy-HBScbpM5 | pGEM-T-Easy derivative, containing a 91 bp DNA fragment corresponding to the region from 432,991–433,071 of H. pylori G27 genome generated annealing oligonucleotides Mut5F/Mut5R. This region corresponds to a portion of the promoter region and coding sequence of HPG27_RS02130 (HP1024 according to 26695 annotation). | This work |
| pVCC | Vector carrying the luxCDABE cassette | Vannini et al., 2014 |
| pVAC::Km | Cloning Vector, Kmr | Delany et al., 2002b |
| pVAC::CAT | pVAC::Km derivative, carrying a BglII/BamHI cat cassette from pBS::cat (Vannini et al., 2012). | This work |
| pVAC-Pcbpwt-lux | pVAC-CAT derivative, containing a 146 bp DNA fragment amplified by PCR with oligonucleotides HBSCbpFEco/HBSCbpRBamHI, encompassing the Pcbp wt promoter region and a 1,000 bp DNA fragment amplified by PCR with oligonucleotides LuxF/LuxR of the luxC gene. | This work |
| pVAC-PcbpM1+2-lux | pVAC-CAT derivative, containing a 146 bp DNA fragment amplified by PCR with oligonucleotides HBSCbpFEco/HBSCbpRBamHI (using as DNA template the plasmid pGEM-T-Easy-HBScbpM1 + 2), encompassing the Pcbp M1 + 2 promoter region and a 1,000 bp DNA fragment amplified by PCR with oligonucleotides LuxF/LuxR of the luxC gene. | This work |
| pGEM3-hspR::Km | pGEM3 vector carrying the Campylobacter coli Kanamycin cassette flanked by a 1067 bp fragment comprising the cbpA gene (HPG27_RS02130) and a 716 bp fragment comprising the 5′ region of the rarA gene (HPG27_RS02120). | Spohn and Scarlato, 1999 |
| pET22b | Expression vector, allow C-terminal histidine-tag gene fusion; Ampr | Novagen |
| pET22b-HspR | pET22b derivative, containing the HspR coding sequence amplified by PCR | Spohn and Scarlato, 1999 |