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 | |
Vibrio cholerae | ||
S2CHK17 | V.cholerae O1 El Tor, isolated from Pakistan 2010 | |
AcfC mutant | S2CHK17 ΔAcfC, in frame deletion mutant | This study |
Plasmids | ||
pEXT20 | Expression vector, Ampr, IPTG-inducible | |
pDS132 | Suicide plasmid, Cnr | |
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 |
