TABLE 1.
Plasmids, strains, and primers used in this study
| Relevant or descriptive genotype or description | Source or sequence | |
|---|---|---|
| Plasmids | ||
| pLKS2 | Csd4 in pBluescript II SK+ | Ref. 8 |
| pLC292 | pRdxA with ampicillin resistance marker | Ref. 8 |
| pKB20 | Csd4 WT:3x-Flag (in pBluescript II SK+) | This work |
| pKB27 | Csd4 1–251:3x-Flag (in pBluescript II SK+) | This work |
| pKB29 | Csd4 1–343:3x-Flag (in pBluescript II SK+) | This work |
| pKB37 | Csd4 Q46A:3x-Flag (in pBluescript II SK+) | This work |
| pKB38 | Csd4 Q46H:3x-Flag (in pBluescript II SK+) | This work |
| pKB44 | rdxA::Csd4 WT:3x-Flag (in pLC292) | This work |
| pKB46 | rdxA::Csd4 Q46H:3x-Flag (in pLC292) | This work |
| pAC-C4H | Csd4 Q46H (in pET15) | This work |
| pAC-C4E | Csd4 Q46E (in pET15) | This work |
| pAC-C4A | Csd4 Q46A (in pET15) | This work |
| Strains | ||
| LSH100 | Wild-type H. pylori (G27 background) | Ref. 8 |
| LSH18 | csd4::catsacB | Ref. 8 |
| LSH108 | rdxA::kansacB | Ref. 2 |
| LSH122 | csd4::cat | Ref. 8 |
| TRH1 | Csd4 4x-Flag (at native locus) | This work |
| KBH19 | Csd4 WT:3x-Flag (at native locus) | This work |
| KBH33 | Csd4 Q46H:3x-Flag (at native locus) | This work |
| KBH35 | Csd4 1–343:3x-Flag (at native locus) | This work |
| KBH37 | Csd4 1–251:3x-Flag (at native locus) | This work |
| KBH42 | Csd4 Q46A:3x-Flag (at native locus) | This work |
| KBH47 | Csd4 Q46H:3x-Flag, rdxA::kansacB | This work |
| KBH54 | csd4::cat, rdxA::kansacB | This work |
| KBH60 | Csd4 Q46H:3x-Flag, rdxA::Csd4 Q46H:3x-Flag | This work |
| KBH64 | Csd4 WT:3x-Flag, rdxA::kansacB | This work |
| KBH65 | Csd4 WT:3x-Flag, rdxA::Csd4 WT:3x-Flag | This work |
| KBH66 | Csd4 WT:3x-Flag, rdxA::Csd4 Q46H:3x-Flag | This work |
| Primers | ||
| 53 | Csd4FLAG_UP_F | cgggccccccctcgaatgatagaagcttgcaaagcgtctt |
| 54 | Csd4FLAG_UP_R | acttttcaatctatatcttacttgtcg |
| 55 | Csd4FLAG_DN_F | tatagattgaaaagtgaatgggtgaatcttgccaaaagat |
| 56 | Csd4FLAG_DN_R | cgggctgcaggaattaacatgcatagctccatcaggtttca |
| 94 | Csd4-Q46A-SDM | tgttgcttttagcagggattgcaggcgatgagcctggggggt |
| 95 | Csd4-Q46H-SDM | ttgcttttagcagggattcatggcgatgagcctggggggtt |
| 85 | Csd4–3xF_UP_KpnI_F | gaattgggtaccgggccccc |
| 86 | Csd4–1–251_3xF_DNR | ataatccttatcgtcatcgtctttataatcaaaagggatattgagttgattgagtaa |
| 88 | Csd4–1–343_3xF_DNR | ataatccttatcgtcatcgtctttataatcaggatcaaactctatgtaaaaaggc |
| 89 | Csd4–3xF_DN_F | gattataaagacgatgacgataaggattat |
| 90 | Csd4–3xFDNSacI_R | aagctggagctccaccgcggt |
| 120 | Csd4–3xFl-BamHI | aggagggatccattggttttaaaatcgcttgtttcaac |
| 121 | Csd4–3xFl-EcoRI | ctcctgaattcttctaatgtatggcatgactatcctt |
| A1 | Csd4-Q46H-For | gttgcttttagcagggattcatggcgatgagcc |
| A2 | Csd4-Q46H-Rev | ggctcatcgccatgaatccctgctaaaagcaac |
| A3 | Csd4-Q46E-For | ttgttgcttttagcagggattgaaggcgatgagc |
| A4 | Csd4-Q46E-Rev | gctcatcgccttcaatccctgctaaaagcaacaa |
| A5 | Csd4-Q46A-For | aggctcatcgcctgcaatccctgctaaaagcaacaaatgg |
| A6 | Csd4-Q46A-Rev | ccatttgttgcttttagcagggattgcaggcgatgagcct |