TABLE 2.
Strains and plasmids used in this study
| Bacterial strain or plasmid | Relevant characteristic(s) | Source or reference |
|---|---|---|
| E. coli | ||
| DH5α | Invitrogen | |
| DH5αAF | Cmr Apr; pSK+cagA and pACYC184 × cagF | This study |
| DH5αA | Apr; pSK+cagA | This study |
| H. pylori | ||
| G27 | Wild-type clinical isolate strain | Xiang et al., 1995 (57) |
| G27/A-FL | Kanr, cagAPR-cagA-FLAG-kanamycin resistance cassette inserted at recA | This study |
| G27/F-FL | Kanr, cagAPR-cagF-FLAG-kanamycin resistance cassette inserted at recA | This study |
| G27/8-FL | Kanr, cagAPR-orf8-FLAG-kanamycin resistance cassette inserted at recA | |
| G27ΔcagA | Kanr | This study |
| G27ΔcagF | This study | |
| G27ΔcagF/F-FL | Kanr, insertion at recA | This study |
| G27ΔvirB9 | This study | |
| Plasmids | ||
| pSK+recxorf8 | Kmr Apr; vector for H. pylori transformation of orf8-FLAG | This study |
| pSK+recxcagA | Kmr Apr; vector for H. pylori transformation of cagA-FLAG | This study |
| pSK+recxcagF | Kmr Apr; vector for H. pylori transformation of cagF-FLAG | This study |
| pACYC184 | Tcr Cmr | New England Biolabs |
| pACYC184 × cagF | Cmr vector for expression of CagF-FLAG in E. coli under control of cagAPR (insertion of gene product at XhoI and BamHI restriction sites) | This study |
| pSK+cagA | Apr; cagAPR and cagA gene (insertion of gene product at XhoI and XbaI restriction sites) | This study |
| pMA1 | pSK + vector with CagF up- and downstream sequences (upstream region cloned into KpnI and XhoI, downstream cloned into XbaI and NotI) | This study |
| pMA2 | pMA1; Kmr and sacB insertion (insertion between up- and downstream regions of cagF at XhoI and XbaI restriction sites) | This study |