TABLE 5.
Bacterial strains or plasmids used in this study
| Strain or plasmid | Genotype or description | Source |
|---|---|---|
| C. jejuni strains | ||
| 81-176 | Wild-type isolated from diarrheic patient | 87 |
| Δape1 | 81-176 ape1::aphA3;KmR | This study |
| ΔpatB | 81-176 patB::aphA3;KmR | This study |
| ΔpatA | 81-176 patA::aphA3;KmR | This study |
| Δoap | 81-176 oap::aphA3;KmR | This study |
| Δape1C | 81-176 Δape1 rrn::ape1 (from pRRC-0638) | This study |
| ΔpatBC | 81-176 ΔpatB rrn::patB (from pRRC-0639) | This study |
| ΔpatAC | 81-176 ΔpatA rrn::patA (from pRRC-0640) | This study |
| E. coli strains | ||
| DH5-α | F−, ϕ80d deoR lacZΔM15 endA1 recA1 hsdR17(rK-mK+) supE44 thi-1 gyrA96 relA1 Δ(lacZYA-argF) U169 | Invitrogen |
| BL21(λDE3) | F− ompT hsdSB(rB−, mB−) gal dcm [λ DE3] | Novagen |
| Plasmids | ||
| pGEM-T | High copy, linearized, T-tailed, Blue/White, ApR | Promega |
| pUC18-K2 | Source of non-polar aphA3 cassette; ApRKmR | 47 |
| pGEM-T-0638 | pGEM-T ligated to 0638 amplified with 0637-2 and 0639-5 (2113 bp); ApR | This study |
| pGEM-0638::aphA-3 | pGEM-T-0638 inverse PCR amplified with 0638-3 and 0638-2 (4098 bp) and ligated to aphA-3 (KpnI, HincII); ApR, KmR | This study |
| pGEM-T-0639 | pGEM-T ligated to 0639 amplified with 0639-1 and 0639-2 (2146 bp); ApR | This study |
| pGEM-0639::aphA-3 | pGEM-T-0639 inverse PCR amplified with 0639-3 and 0639-4 (4267 bp) and ligated to aphA-3 (KpnI, HincII); ApR, KmR | This study |
| pGEM-T-0640 | pGEM-T ligated to 0640 amplified with 0639-6 and 0641-4 (2396 bp); ApR | This study |
| pGEM-0640::aphA-3 | pGEM-T-0640 inverse PCR amplified with 0640-1 and 0640-2 (4168 bp) and ligated to aphA-3 (KpnI, HincII); ApR, KmR | This study |
| pGEM-0638–40::aphA-3 | pGEM-T ligated to 0637 fragment amplified with 0637-1 and 0638-1 (1483 bp), 0641 fragment amplified with 0641-1 and 0641-2 (822 bp), and aphA-3 (KpnI, HincII); ApR, KmR | This study |
| pRRC | C. jejuni rRNA spacer integration vector; CmR | 48 |
| pRRC-0638 | pRRC ligated to 0638 amplified with 0638-C1(NheI) and 0638-C2(MfeI) (1347 bp); CmR | This study |
| pRRC-0639 | pRRC ligated to 0639 amplified with 0639-C1(NheI) and 0639-C2(MfeI) (1276 bp); CmR | This study |
| pRRC-0640 | pRRC ligated to 0640 amplified with 0640-C1(NheI) and 0640-C2(MfeI) (1616 bp); CmR | This study |
| pET28a(+) | Commercial vector for expression of recombinant His6-tagged protein | Novagen |
| p0638-His6 | pET28a(+) ligated to ape1 amplified with 0638-eCF (NcoI) and 0638-eCR (EcoRI) (1121 bp) for expression of C-terminal His6-tagged 0638; KmR | This study |
| pHis6-0638 | pET28a(+) ligated to ape1 amplified with 0638-eNF (NheI) and 0638-eNR (EcoRI) (1116 bp); for expression of N-terminal His6-tagged 0638 KmR | This study |