Table 1.
Plasmids and strains used in this study
| Strain or plasmid | Relevant characteristicsa | Reference or source |
|---|---|---|
| Plasmids | ||
| pTnMax5 | lacIqtnpR tnpA res orifdcatGC, Cmr | 17 |
| pZERO-2 | Cloning vector, MCS in lacZ′ neo, Kmr | Invitrogen |
| p0058PCAT | pZERO-2, ΔHP0058::Pcat, CmrKmr | This study |
| p0058StrepPCAT | pZERO-2 carrying 500 bp of the C terminus of HP0058 fused with a Strep tag, a chloramphenicol resistance cassette, and 500 bp of the N terminus of ccrp59 | This study |
| p1142PCAT | pZERO-2, ΔHP1142::Pcat, CmrKmr | This study |
| pASK-IBA7 | Expression vector, tetR Ptetbla, Apr | IBA |
| pIBA7-1142 | pASK-IBA7 carrying the HP1142 coding sequence under the control of the tetpromoter cloned in the BsaI site | This study |
| pIBA7-0059 | pASK-IBA7 carrying the HP0059 coding sequence under the control of the tetpromoter cloned in the BsaI site | 30 |
| pETDuet-1 | bla | Novagen |
| pETDuet-1143 | pETDuet-1 carrying the HP1143 coding sequence under the control of the T7promoter cloned between the NcoI and BamHI sites | 30 |
| pETDuet-0058 | pETDuet-1 carrying the HP0058 coding sequence under the control of the T7promoter cloned between the EcoRI and PstI sites | This study |
| Strains | ||
| E. coli strains | ||
| BL21 | F−dcm ompT hsdS(rB−mB−) gal | Stratagene |
| BL21-Ccrp58 | BL21 carrying plasmid pETDuet-0058 | This study |
| BL21-Ccrp59 | BL21 carrying plasmid pIBA7-0059 | 30 |
| BL21-Ccrp1143 | BL21 carrying plasmid pETDuet-1143 | 30 |
| BL21-Ccrp1142 | BL21 carrying plasmid p pIBA7-1142 | This study |
| DH5α | F−φ80dlacZΔM15 Δ(lacZYA-argF)U169 deoR recA1 endA1 hsdR17(rK−mK+) phoA supE44λ−thi-1 gyrA96 relA1 | Bethesda Research Laboratories |
| H. pyloristrains | ||
| 26695 | wt, containing the entire cagPAI | 27 |
| KE88-3887 | Piglet-passaged strain 26695 | 15 |
| KE-Ccrp58Strep | KE88-3887, Ccrp58 fused to Strep tag, Cmr | This study |
| KE-59PCAT | KE88-3887, ΔHP0059::Pcat, Cmr | 30 |
| KE-1143PCAT | KE88-3887, ΔHP1143::Pcat, Cmr | 30 |
| KE-58PCAT | KE88-3887, ΔHP0058::Pcat, Cmr | This study |
| KE-1142PCAT | KE88-3887, ΔHP1142::Pcat, Cmr | This study |
| 26695-59PCAT | 26695, ΔHP0059::Pcat, Cmr | 30 |
| 26695-1143PCAT | 26695, ΔHP1143::Pcat, Cmr | 30 |
| 26695-58PCAT | 26695, ΔHP0058::Pcat, Cmr | This study |
| 26695-1142PCAT | 26695, ΔHP1142::Pcat, Cmr | This study |
a
MCS, multiple cloning site; wt, wild type; PAI, pathogenicity island.