TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Relevant genotypea | Reference or source |
|---|---|---|
| Strains | ||
| V. cholerae | ||
| O395 | Classical O1 biotype, Smr | Laboratory collection |
| Mut3 | O395 epsE::lacZ kan Smr Kanr | 77 |
| JJM43 | O395 ΔtoxR43 ΔctxA1 Smr | 27 |
| Mut4 | O395 ΔtoxR43 ΔctxA1 epsE::lacZ kan Smr | 77 |
| O395ΔrpoE | O395ΔrpoE Smr | 49 |
| TRH7000 | El Tor O1 biotype, thy Hgr Δ(ctxA-ctxB) | 29 |
| Mut1 | TRH7000 epsE::lacZ kan Kanr | 75 |
| Mut6 | TRH7000 epsF::kan Kanr | 75 |
| Mut5 | TRH7000 epsG::kan Kanr | 77 |
| Mut 8 | TRH7000 epsL::kan Kanr | 75 |
| PU3 | TRH7000 epsM::Tn5 Kanr | 62 |
| TRHΔepsC | TRH7000 ΔepsC Kanr | S. R. Lybarger et al. (unpublished data) |
| TRHΔepsL | TRH7000 ΔepsL Kanr | This study |
| PBAD::eps | TRH7000 PBAD::eps | This study |
| TRHΔeps | TRH7000 ΔepsC-N Cmr | This study |
| N16961 | Wild-type El Tor O1 biotype, Smr | Laboratory collection |
| NΔeps | N16961 ΔepsC-N Cmr | This study |
| 569B | Classical O1 biotype Smr | Laboratory collection |
| M14 | 569B epsE-A321V | 32 |
| Bah2 | El Tor O1 biotype, E7946, ΔattRS | 87 |
| Bah2 epsD::kan | Bah2 epsD::kan | 14, 15 |
| E. coli | ||
| MC1061 | F−araD139 Δ(ara-leu)7697 Δ(lac)X74 rpsL hsdR2 mcrA mcrB1 | 11 |
| S17.1 | F− recA pro hsdR RP4-2 Tcr::Mu Tnr::Tn7 | 81 |
| SY327 λpir | Δ(lac-pro) argE(Am) recA56 rpoB λpir Rifr | 55 |
| MM294 (pRK2013) | Donor of transfer function for triparental conjugation | 52 |
| Plasmids | ||
| pPCR-Script | Cloning vector, Ampr | Stratagene |
| pBAD33 | Cloning vector, araC-PBAD Cmr | 25 |
| pJN105 | Cloning vector, ori pBBR1 araC-PBAD mob Gmr | 59 |
| pK18mobsacB | ori-pMB1 oriT (RP4) sacB lacZα Kanr | 79 |
| pMMB-GFP | pMMB66-gfp coding sequence, Ampr | 80 |
| pMMB68 | etxB under PTAC control in pMMB66EH, Ampr | 76 |
| pMMB190 | Cloning vector, pTACplacUV5 Ampr | 57 |
| pCH40 | pbla::phoA Tetr | 31 |
| pMS43 | epsE-K270A under PTAC control in pMMB207, Cmr | 14, 75 |
| pCVD442 | ori R6K mobRP4 sacB Ampr | 17 |
| pUC18K | Cloning vector, aphA-3 Ampr Kanr | 51 |
| pBBRlux | ori pBBR1, promoterless luxCDABE Cmr | 46 |
| pAES100 | 1,078-bp EcoRI-XbaI fragment upstream of epsC in pK18mobsacB, Kanr | This study |
| pAES101 | 1,075-bp fragment downstream of epsN cloned into XbaI- and SphI-digested pAES100, Kanr | This study |
| pAES102 | 923-bp fragment containing Cmr cassette cloned into XbaI site of pAES101, Kanr Cmr | This study |
| pAES103 | 1,070-bp fragment upstream of epsC cloned into SphI-ClaI site of pJN105, Gmr | This study |
| pAES104 | 1,079-bp fragment of full-length, promoterless epsC cloned into EcoRI- and XbaI-cut pAES103, Gmr | This study |
| pAES105 | SphI-XbaI 3,479-bp fragment containing upstream region of epsC, araC, and pBAD::epsC from pAES104 in pK18mobsacB, Kanr | This study |
| pAES106 | rpoEP2 cloned into pBBRlux, Cmr | This study |
| pMMB131 | pMMB68 gene fusion between etxB signal sequence and phoA under PTAC control, Ampr | This study |
| pEps | 15-kb XbaI-SphI fragment containing entire eps operon (from epsC to epsN) in pMMB190, Ampr | This study |
a
Ampr, ampicillin resistance; Cmr, chloramphenicol resistance; Gmr, gentamicin resistance; Hgr, mercury resistance; Kanr, kanamycin resistance; Rifr, rifampin resistance; Tetr, tetracycline resistance.