TABLE 1.
Plasmids and bacterial strains used in this studya
| Strain or plasmid | Relevant characteristic(s) | Antibiotic resistance | Reference or source |
|---|---|---|---|
| E. coli | |||
| DH10B | Cloning strain | Gibco BRL | |
| H. pylori | |||
| G27 | Wild type, type I | 12 and Nina Salama | |
| SS1 | Wild type, type I | 25 and Janie O'Rourke | |
| G27 cheW mutant | G27 ΔcheW::aphA3 | Kn | This study |
| SS1 cheW mutant | SS1 ΔcheW::aphA3 | Kn | This study |
| SS1 cheA mutant | SS1 ΔcheA::cat forward | Cm | This study |
| SS1 cheA mutant | SS1 ΔcheA::cat reverse | Cm | This study |
| LC143 | SS1 rdxA::cat | Cm, Mt | This study |
| LC144 | SS1 rdxA::aphA3 | Kn, Mt | This study |
| SS1 cheY mutant | SS1 ΔcheY::cat102 | Cm | This study |
| SS1 cheY complemented | SS1 ΔcheY::cat102 rdxA::cheY | Cm, Mt | This study |
| Plasmids | |||
| pBluescript KS+ (pBS) | Cloning vector | Ap | Stratagene |
| pBS-Kan | pBS with aphA3 from Campylobacter coli | Ap, Kn | Nina Salama |
| pBS-cat | pBS with cat gene from C. coli | Ap, Cm | 33 and Nina Salama |
| pKT10 | pBS with 2,653-bp H. pylori cheW and flanking sequence inserted into EcoRV site | Ap | This study |
| pKT11 | pKT10 with 334-bp deletion in cheW replaced by aphA3: ΔcheW::aphA3 | Ap, Kn | This study |
| pKT20 | pBS with 6,879-bp H. pylori cheA and flanking sequence inserted into EcoRV site | Ap | This study |
| pKT22 | pKT20 with 1,638-bp deletion in cheA and cat inserted in the opposite orientation of cheA | Ap, Cm | This study |
| pKT23 | pKT20 with 1,638-bp deletion in cheA and cat inserted in the same orientation as cheA | Ap, Cm | This study |
| pRdxA | pBC-SK− with 5′ and 3′ regions of H. pylori rdxA locus flanking a polylinker | Cm | 35 and Johannes Kusters |
| pLC292 | 0.75-kb KpnI-SacI fragment from | Ap | This study |
| pRdxA cloned into the KpnI-SacI sites of pBS-SK+ | |||
| pLC307 | 0.8-kb HincII fragment of pBS-cat cloned into the SmaI site of pLC292 | Ap, Cm | This study |
| pLC308 | 1.4-kb fragment from pBS-Kan using KANupstrm and KANdowns primers cloned into the SmaI site of pLC292 | Ap, Kn | This study |
| pKO126 | pBS::cheY | Ap | 3 |
| pKO127 | pKO126::cat-mut (cheY102) | Ap, Cm | This study |
| pKO140 | pLC292::cheY | Ap | This study |
| pCat-mut | pBScat mutagenized to place restriction sites (BamHI, SmaI, AscI) between the end of cat and its transcriptional terminators to create cat-mut | Ap, Cm | This study |
a
The following plasmids were constructed with both H. pylori SS1 and G27 genes cloned into them: pKT10, pKT11, pKT20, pKT22, pKT23. Antibiotic abbreviations are as follows: Ap, ampicillin; Cm, chloramphenicol; Kn, kanamycin; Mt, metronidazole.