TABLE 1.
Plasmids and strains used
| Plasmid or strain | Relevant properties | Reference or source |
|---|---|---|
| Plasmids | ||
| pGP704-sacB28 | Used for sucrose-based selection, Apr | Gary Schoolnik |
| pGPVC0928 | pGP704-sacB28 containing the SOEing PCR product of rbmA | This study |
| pACYC177 | Low-copy-number cloning vector, Apr | New England Biolabs |
| prbmA | pACYC177 harboring wild-type copy of rbmA | This study |
| pBAD/Myc-His B | Arabinose-inducible expression vector with C-terminal Myc epitope and six-His tags | Invitrogen |
| prbmA-myc | pBAD/Myc-His B containing the coding region of rbmA | This study |
| pRS415 | lacZ-based cloning vector for transcriptional fusion studies, Apr | 50 |
| prbmA::lacZ | pRS415 containing rbmA upstream regulatory fragment fused to lacZ | This study |
| pCC11 | pRS415 containing vpsA upstream regulatory fragment fused to lacZ | 8 |
| pCC12 | pRS415 containing vpsL upstream regulatory fragment fused to lacZ | 8 |
| pGP704::Tn7-GFP | Mini-Tn7 vector harboring a constitutively expressed gfp cassette | Gary Schoolnik |
| pUX-BF13 | Helper plasmid containing transposase gene for mini-Tn7 | Gary Schoolnik |
| Strains | ||
| V. cholerae 92A1552 | ||
| FY_Vc_0001 | O1 E1 Tor, Inaba, smooth, Rifr | 60 |
| FY_Vc_0002 | O1 E1 Tor, Inaba, rugose, Rifr | 60 |
| FY_Vc_0004 | Rugose ΔlacZ | 8 |
| FY_Vc_0005 | RΔvpsT ΔlacZ | 8 |
| FY_Vc_0006 | RΔvpsR ΔlacZ | 8 |
| FY_Vc_0007 | RΔvpsR ΔvpsT ΔlacZ | 8 |
| FY_Vc_102 | SΔrbmA | This study |
| FY_Vc_105 | RΔrbmA | This study |
| FY_Vc_222 | Rugose-gfp | This study |
| FY_Vc_224 | RΔrbmA-gfp | This study |
| E. coli | ||
| DH5α | F′ endA1 hsdR17 supE44 thi-1 recA1 gyrA96 relA1 Δ(argF-lacZYA) U169 (φ80dlacΔM15) | Promega |
| CC118 (λ pir) | D(ara-leu) araD DlacX74 galE galK phoA20 thi-1 rpsE rpoB argE(Am) recA1 λ pir | 21 |
| S17-1 (λ pir) | Tpr SmrrecA thi pro rK− mK+RP4:2-Tc:MuKm Tn7 λ pir | 15 |