Table 1.
Key bacterial plasmids and strains used in this study
| Plasmids or strains | Description | Source or Reference |
|---|---|---|
| Plasmids | ||
| pGEMT-Easy | PCR cloning vector, Ampr | Promega |
| pCee | T-easy vector derivative containing Cj1376 and partial Cj1377c | (Zeng, et al., 2013) |
| pCee+Cm | pCee derivative with cat cassette inserted at PacI site between cee and cj1377c | This study |
| pRR | T-easy vector derivative containing C.jejuni NCTC 11168 ribosomal fragment | This study |
| pRR-CfrB | pRR derivative in which cfrB gene was inserted inside ribosomal region | This study |
| pRRE-CfrB | The erythromycin resistant cassette was attached at the end of cfrB in pRR-CfrB | This study |
| pRY107 | E. coli-Campylobacter shuttle vector, kanamycin resistant (Kmr) | (Yao, et al., 1993) |
| pTonB3 | pRY107 derivative containing the tonB3 and its promoter. | This study |
| pCfrA | pRY107 derivative containing cfrA plus its promoter region | (Zeng, et al., 2009) |
| pTonB3-CfrA | pCfrA derivative containing both intact tonB3 and cfrA genes | This study |
| pTonB3-CfrA-T | T-easy vector derivative containing the tonB3-cfrA locus | This study |
| pTonB3(tetO) | pTonB3-CfrA-T derivative with tetO cassette inserted in tonB3 | This study |
| Strains | ||
| C.jejuni | ||
| 81-176 | Human isolate | (Black, et al., 1988) |
| JL727 | 81-176 derivative for which 81-176 was chromosomally complemented with cee gene | This study |
| JL680 | 81-176 complemented with plasmid pTonB3-CfrA | This study |
| JL534 | 81-176 complemented with plasmid pCfrA | This study |
| 11168 | NCTC 11168, human isolate | (Parkhill, et al., 2000) |
| JL709 | 11168 derivative with cat cassette inserted between cee and cj1377c | This study |
| JL832 | 11168 derivative with tonB3 inactivated by insertion of tetO cassette | This study |
| JL324 | 11168 derivative, cfrA::cat | (Zeng, et al., 2009) |
| JL612 | JL324 derivative with chromosomal complementation of cfrB gene. Thus, this strain has genetically repaired CfrB-dependent pathway for FeEnt acquisition |
This study |
| JL845 | JL612 derivative, tonB2− cfrA− cfrB+ | This study |
| JL868 | JL845 derivative, tonB2− tonB3− cfrA− cfrB+ | This study |
| JL869 | JL845 derivative, tonB3− cfrA− cfrB+ | This study |
| E. coli | ||
| DH5α | F- Φ80lacZΔM15 Δ(lacZYA-argF)U169 recAl endAl hsdR17 (rk−, mk+) phoA supE44 thi-1 gyrA96 relAl λ- |
Invitrogen |
| JL48 | DH5α/pRK2013, help strain for conjugation. | (Zeng, et al., 2009, Xu, et al., 2010) |