TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Description or genotypea | Reference or source |
|---|---|---|
| Strains | ||
| V. cholerae | ||
| JL2 | N16961 ΔtcpA mtlA-FLAG Smr | Laboratory strain |
| JL55 | N16961 ΔtcpA mtlA-FLAG ΔmtlR Smr | 23 |
| JL142 | N16961 ΔtcpA mtlA-FLAG/pJML01 Smr Apr | 30 |
| JL463 | N16961 ΔtcpA mtlA-FLAG PmtlA_ΔCRPbs Smr | This study |
| JL467 | N16961 ΔtcpA mtlA-FLAG PmtlA_−10mut Smr | This study |
| JL494 | N16961 ΔtcpA mtlA-FLAG ΔVC2338 (−235) Smr | This study |
| JL495 | N16961 ΔtcpA mtlA-FLAG ΔVC2338 (−235) PmtlA500-lacZ(Ec) Smr | This study |
| JL499 | N16961 ΔtcpA mtlA-FLAG ΔVC2338 (−235) PmtlS500-lacZ(Ec); Smr | This study |
| JL546 | N16961 ΔtcpA mtlA-FLAG/pmtlA5UTR Smr Apr | This study |
| E. coli | ||
| DH5α | F− Δ(lacZYA-argF)U169 recA1 endA1 hsdR17 supE44 thi-1 gyrA96 relA1 | Laboratory strain |
| DH5αλpir | F− Δ(lacZYA-argF)U169 recA1 endA1 hsdR17 supE44 thi-1 gyrA96 relA1 λ::pir | Laboratory strain |
| SM10λpir | thi recA thr leu tonA lacY supE RP4-2-Tc::Mu λ::pir | Laboratory strain |
| TOP10 | F– mcrA Δ(mrr-hsdRMS-mcrBC) ϕ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(ara leu)7697 galU galK rpsL endA1 nupG Smr | Invitrogen |
| Plasmids | ||
| pCVD442 | oriR6K mobRP4 sacB Apr | 58 |
| pJML01 | pBAD24 derivative with +1 start site of transcription after NheI site; Apr | 22 |
| pmtlA5UTR | pBAD24 derivative that expresses the entire 5′ UTR of mtlA; Apr | This study |
| pJL1 | pCVD442 derivative with 2.2-kb HpaI-digested VC2338 (V. cholerae lacZ) cloned into SmaI site of pCVD442; Apr | 59 |
| pJL1::lacZ(Ec) | pJL1 derivative with RBS and coding region of E. coli lacZ inserted into the VC2338 fragment of pJL1 in an antisense orientation; Apr | This study |
a
Smr, streptomycin resistance; Apr, ampicillin resistance.