TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Genotypea | Reference or source |
|---|---|---|
| E. coli | ||
| DH5α | Δ(argF-lac)U169 ϕ80dlacZ58(M15) ΔphoA8 glnV44 deoR481 gyrA96 recA1 endA1 hsdR17 | Laboratory stock |
| BL21 (Tuner) | F− ompT hsdSB(rB− mB−) gal dcm lacY1 | Novagen |
| Rosetta(DE3) | F− ompT hsdSB(rB− mB−) gal dcm (DE3) pRARE | Novagen |
| H. pylori | ||
| 26695 | Wild-type | 38 |
| HpS42 | Kmr; 26695 ΔcfaS | This work |
| HpS43 | Kmr Cmr; 26695 ΔcfaS::cfaS | This work |
| Hp159 | Drug-resistant clinical isolate | Laboratory stock |
| HpS44 | Kmr; Hp159 ΔcfaS | This work |
| HpS45 | Kmr Cmr; Hp159 ΔcfaS::cfaS | This work |
| NSH57 | Mouse-adapted derivative of G27 strain | 39 |
| HpS46 | Kmr; NSH57 ΔcfaS | This work |
| HpS47 | Kmr Cmr; NSH57 ΔcfaS::cfaS | This work |
| HpS48 | Kmr; 26695 transformed with plasmid pHel3 | This work |
| HpS49 | Kmr; 26695 transformed with plasmid pBHKP380 | This work |
| Plasmids | ||
| pBluescript SK(+) | Ampr; cloning vector | Stratagene |
| pQE-2 | Ampr; T5 promoter-based expression vector | Qiagen |
| pHel3 | Kmr; E. coli/H. pylori shuttle vector | 43 |
| pBHKP109 | Ampr Kmr; PCR-amplified aphA3 inserted between PstI and BamHI sites of pBluescript SK(+) | This work |
| pBHKP202 | Ampr Cmr; PCR-amplified catGC inserted between PstI and BamHI sites of pBluescript SK(+) | This work |
| pBHKP203 | Ampr Kmr; PCR-amplified HpcfaSup inserted between PstI and EcoRI sites plus PCR-amplified HpcfaSdn inserted between BamHI and XbaI sites of BHKP109 | This work |
| pBHKP252 | Cmr; PCR-amplified Hp0203up inserted between KpnI and XhoI sites plus PCR-amplified Hp0204dn inserted between BamHI and XbaI sites of BHKP202 | This work |
| pBHKP388 | Cmr; PCR-amplified H. pylori cfaS (containing its promoter) inserted between XhoI and PstI sites of BHKP252 | This work |
| pBHKP380 | Kmr; PCR-amplified H. pylori cfaS (containing its promoter) inserted between KpnI and XhoI sites of pHel3 | This work |
| pBHKP292 | Ampr; PCR-amplified E. coli mtn inserted between NdeI and SalI sites of pQE-2 | This work |
| pBHKP338 | Ampr; PCR-amplified B. subtilis luxS inserted between KpnI and SalI sites of pQE-2 | This work |
| pBHKP389 | Ampr; PCR-amplified H. pylori cfaS inserted between NdeI and SalI sites of pQE-2 | This work |
a
Kmr, kanamycin resistance; Cmr, chloramphenicol resistance; Ampr, ampicillin resistance.