TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Relevant genotype and phenotypea | Source or reference |
|---|---|---|
| Strains | ||
| E. coli | ||
| CC118λpir | Δ(ara-leu) araD ΔlacX74 galE galK phoA20 thi-1 rpsE rpoB argE(Am) recA1 λpir | 21 |
| S17-1λpir | recA thi pro rK− mK+ RP4:2-Tc::MuKm Tn7 Tpr Smr λpir | 13 |
| DH10B | F−mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 recA1 endA1 araΔ139 Δ(ara leu)7697 galU galK λ−rpsL (Strr) nupG | Invitrogen |
| V. cholerae | ||
| Fy_Vc_1 | V. cholerae O1 El Tor A1552, smooth variant, Rifr | 57 |
| Fy_Vc_2 | V. cholerae O1 El Tor A1552, rugose variant, Rifr | 57 |
| Fy_Vc_3 | Fy_Vc_1 ΔlacZ, Rifr | 8 |
| Fy_Vc_4 | Fy_Vc_2 ΔlacZ, Rifr | 8 |
| Fy_Vc_237 | Fy_Vc_1 mTn7-GFP, Rifr Gmr | 5 |
| Fy_Vc_240 | Fy_Vc_2 mTn7-GFP, Rifr Gmr | 32 |
| Fy_Vc_231 | Fy_Vc_1 Δvps-I operon, Rifr | 5 |
| Fy_Vc_234 | Fy_Vc_2 Δvps-I operon, Rifr | 6 |
| Fy_Vc_337 | Fy_Vc_1 ΔflaA, Rifr | This study |
| Fy_Vc_339 | Fy_Vc_2 ΔflaA, Rifr | This study |
| Fy_Vc_970 | Fy_Vc_1 ΔVC0900, Rifr | This study |
| Fy_Vc_3074 | Fy_Vc_1 ΔVC0900 ΔlacZ, Rifr | This study |
| Fy_Vc_3083 | Fy_Vc_1 ΔVC0900 mTn7-GFP, Rifr Gmr | This study |
| Fy_Vc_917 | Fy_Vc_2 ΔVC0900, Rifr | This study |
| Fy_Vc_3071 | Fy_Vc_2 ΔVC0900 ΔlacZ, Rifr | This study |
| Fy_Vc_3080 | Fy_Vc_2 ΔVC0900 mTn7-GFP, Rifr Gmr | This study |
| Fy_Vc_1592 | Fy_Vc_1 ΔVC1067, Rifr | This study |
| Fy_Vc_3077 | Fy_Vc_1 ΔVC1067 ΔlacZ, Rifr | This study |
| Fy_Vc_2089 | Fy_Vc_1 ΔVC1067 mTn7-GFP, Rifr Gmr | This study |
| Fy_Vc_861 | Fy_Vc_2 ΔVC1067, Rifr | This study |
| Fy_Vc_1139 | Fy_Vc_2 ΔVC1067 ΔlacZ, Rifr | This study |
| Fy_Vc_1232 | Fy_Vc_2 ΔVC1067 mTn7-GFP, Rifr Gmr | This study |
| Fy_Vc_3211 | Fy_Vc_1 ΔVC0900 ΔVC1067, Rifr | This study |
| Fy_Vc_3217 | Fy_Vc_1 ΔVC0900 ΔVC1067 ΔlacZ, Rifr | This study |
| Fy_Vc_3208 | Fy_Vc_2 ΔVC0900 ΔVC1067, Rifr | This study |
| Fy_Vc_3214 | Fy_Vc_2 ΔVC0900 ΔVC1067 ΔlacZ, Rifr | This study |
| Fy_Vc_1745 | Fy_Vc_2 ΔvpvC, Rifr | 6 |
| Fy_Vc_3584 | Fy_Vc_2 ΔvpvC ΔVC1067, Rifr | This study |
| Fy_Vc_183 | Fy_Vc_1 ΔhapR, Rifr | 4 |
| Fy_Vc_344 | Fy_Vc_1 ΔcdgA, Rifr | 32 |
| Fy_Vc_1675 | Fy_Vc_1 ΔhapR ΔcdgA, Rifr | 4 |
| Fy_Vc_3635 | Fy_Vc_1 ΔhapR ΔVC1067, Rifr | This study |
| Fy_Vc_3638 | Fy_Vc_1 ΔcdgA ΔVC1067, Rifr | This study |
| Fy_Vc_3643 | Fy_Vc_1 ΔhapR ΔcdgA ΔVC1067, Rifr | This study |
| Fy_Vc_346 | Fy_Vc_1 ΔcdgC, Rifr | 32 |
| Fy_Vc_354 | Fy_Vc_1 ΔrocS, Rifr | 32 |
| Fy_Vc_356 | Fy_Vc_1 ΔmbaA, Rifr | 32 |
| Fy_Vc_3599 | Fy_Vc_1 ΔrocS ΔcdgC, Rifr | This study |
| Fy_Vc_3611 | Fy_Vc_1 ΔrocS ΔmbaA, Rifr | This study |
| Fy_Vc_3602 | Fy_Vc_1 ΔmbaA ΔcdgC, Rifr | This study |
| Fy_Vc_3614 | Fy_Vc_1 ΔrocS ΔmbaA ΔcdgC, Rifr | This study |
| Fy_Vc_3590 | Fy_Vc_1 ΔrocS ΔVC0900, Rifr | This study |
| Fy_Vc_3593 | Fy_Vc_1 ΔmbaA ΔVC0900, Rifr | This study |
| Fy_Vc_3587 | Fy_Vc_1 ΔcdgC ΔVC0900, Rifr | This study |
| Fy_Vc_3617 | Fy_Vc_1 ΔrocS ΔmbaA ΔcdgC ΔVC0900, Rifr | This study |
| Fy_Vc_1643 | Fy_Vc_2 ΔplzA, Rifr | This study |
| Fy_Vc_1647 | Fy_Vc_2 ΔplzB, Rifr | This study |
| Fy_Vc_1651 | Fy_Vc_2 ΔplzC, Rifr | This study |
| Fy_Vc_1636 | Fy_Vc_2 ΔplzD, Rifr | This study |
| Fy_Vc_1640 | Fy_Vc_2 ΔplzE, Rifr | This study |
| Fy_Vc_2504 | Fy_Vc_2 ΔplzA ΔplzC ΔplzD ΔplzE, Rifr | This study |
| Fy_Vc_3655 | Fy_Vc_2 ΔplzA ΔplzB ΔplzC ΔplzD ΔplzE, Rifr | This study |
| Fy_Vc_3661 | Fy_Vc_2 ΔplzA ΔplzC ΔplzD ΔplzE mTn7-GFP, Rifr | This study |
| Fy_Vc_3781 | Fy_Vc_2 ΔvpvC ΔplzC, Rifr | This study |
| Fy_Vc_3784 | Fy_Vc_2 ΔVC1067 ΔplzC, Rifr | This study |
| Plasmids | ||
| pGP704-sac28 | pGP704 derivative; sacB, Apr | 8 |
| pFY-449 | pGP704-sac28:: ΔVC0900, Apr | This study |
| pFY-399 | pGP704-sac28:: ΔVC1067, Apr | This study |
| pCC2 | pGP704-sac28:: ΔlacZ, Apr | 8 |
| pSM1 | pWM91:: ΔflaA(VC2188) | 38 |
| pAJH5 | pWM91:: Δvps-I operon | 20 |
| pFY-367 | pGP704-sac28:: ΔvpvC, Apr | 6 |
| pFY-9 | pGP704-sac28:: ΔhapR, Apr | Gary Schoolnik |
| pFY-149 | pGP704-sac28:: ΔcdgA, Apr | 32 |
| pFY-151 | pGP704-sac28:: ΔcdgC, Apr | 32 |
| pFY-161 | pGP704-sac28:: ΔrocS, Apr | 32 |
| pFY-163 | pGP704-sac28:: ΔmbaA, Apr | 32 |
| pFY-640 | pGP704-sac28:: ΔplzA, Apr | This study |
| pFY-641 | pGP704-sac28:: ΔplzB, Apr | This study |
| pFY-642 | pGP704-sac28:: ΔplzC, Apr | This study |
| pFY-643 | pGP704-sac28:: ΔplzD, Apr | This study |
| pFY-644 | pGP704-sac28:: ΔplzE, Apr | This study |
| pMCM11 | pGP704::mTn7-GFP, Gmr Apr | 32 |
| pUX-BF13 | oriR6K helper plasmid, provides Tn7 transposition function in trans, Apr | 3 |
| pBAD/myc-His-B | Arabinose-inducible expression vector with C-terminal myc epitope and His6 tags | Invitrogen |
| pFY-573 | pBAD/myc-His-B::VC0900, Apr | This study |
| pFY-572 | pBAD/myc-His-B::VC1067, Apr | This study |
| pFY-590 | pBAD/myc-His-B::VC0900, E445A and E446A mutations, Apr | This study |
| pFY-645 | pBAD/myc-His-B::VC0900, R434A and D437A mutations, Apr | This study |
| pFY-589 | pBAD/myc-His-B::VC1067, G605A mutation, Apr | This study |
| pCC12 | pRS415 vpsL promoter, Apr | 8 |
| pCC10 | pRS415 vpsR promoter, Apr | 8 |
| pCC25 | pRS415 vpsT promoter, Apr | 8 |
a
Rifr, rifampin resistance; Apr, ampicillin resistance; Gmr, gentamicin resistance.