Table 2.
Strains, plasmids and oligonucleotides used in this study.
| Strain, plasmid and oligonucleotides | Relevant characteristic (s) or sequence (5′ to 3′) | Source or reference |
|---|---|---|
| Strains | ||
| Escherichia coli | ||
| DH5α | F– Φ80lacZΔM15 Δ(lacZYA-argF) U169 recA1 endA1 hsdR17 (rK–, mK+) phoA supE44 λ– thi-1 gyrA96 relA1 | Invitrogen |
| LMG194 | F− ΔlacX74 galE thi rpsL ΔphoA (PvuII) Δara714 leu::Tn10 | Invitrogen |
| MFD λ-pir | MG1655 RP4-2-Tc::[ΔMu1::aac(3)IV-ΔaphA-Δnic35-ΔMu2::zeo] ΔdapA::(erm-pir) ΔrecA | 43 |
| Vibrio cholerae | ||
| S2CHK17 | V.cholerae O1 El Tor, isolated from Pakistan 2010 | 19 |
| AcfC mutant | S2CHK17 ΔAcfC, in frame deletion mutant | This study |
| Plasmids | ||
| pEXT20 | Expression vector, Ampr, IPTG-inducible | 38 |
| pDS132 | Suicide plasmid, Cnr | 35 |
| pEXT20-AcfC | pEXT20 with optimized RBS and AcfC open reading frame (xxxbp) | This study |
| pDS132-acfC | pDS132 with SOE PCR fragment (xxxbp) | This study |
| Oligonucleotides | ||
| VC0841A-SacI | AAGAGCTCTAATGCTGAAGCCATTGCATC | This study |
| VC0841B | TGTACTCTCCCTAAAATCACTTAAATGATAAACTTACTGATTAAATCATC | This study |
| VC0841C | GTGATTTTAGGGAGAGTACATGTACTC | This study |
| VC0841D-SacI | TTGAGCTCATCTGCTTAACTTTGGTAACTGG | This study |
| Internal primer F | TACAAATTGTTGATATCACAAATCAAGTAG | This study |
| Internal primer R | CAAAATAAAATAGTAATGCAAAGTGATAAAG | This study |
| VC0841RBS_EcoRIF | AAGAATTCAGGAGGTAAAACATATGTATTTCATGAAAAGTAAGAATCG | This study |
| VC0841RBS_XbaIR | TTTCTAGATCAATGATGATGATGATGATGCTTAAACCAGCCATAGTGCT | This study |