TABLE 1.
Bacterial strains and plasmids used
| Strain or plasmid | Relevant characteristics | Reference or source |
|---|---|---|
| Vibrio sp. | ||
| 569B | Classical biotype, wild type | 23 |
| 569Bstp10rif | Spontaneous Strr Rifr mutant of 569B | This study |
| M14 | epsE mutant of 569B | 23 |
| 0395 | Classical biotype | 59 |
| U1 | El Tor biotype | 40 |
| JBK70 | El Tor biotype | 31 |
| 569B(chiA::Kanr) | Derivative of 569B with a Kanr gene inserted into chiA | This study |
| A. hydrophila | Undescribed isolate | R. K. Holmes |
| E. coli | ||
| DH5α | F− φ80dlacZΔM15 Δ(lacZYA-argF)U169 deoR recA1 endA1 phoA hsdR17 (rK− mK+) supE44 λ− thi-1 gyrA96 relA1 | Life Technologies |
| DH5αF′tet | DH5α transconjugant with [F′ proAB lacIqZΔM15 Tn10(Tetr)] | Life Technologies and Stratagene |
| LE392 | F− e14− (McrA−) hsdR514 (rK− mK+) supE44 supF58 lacY1 galK2 galT22 metB1 trpR55 | 35 |
| S17λpir | Tpr SmrrecA thi pro hsdRM+ [RP4-1-Tc::Mu:KmrTn7] (λpir) | 36 |
| Plasmids | ||
| pCOS5 | Cosmid cloning vector, Ampr Chlr | 10 |
| pBluescriptKS+ | Phagemid cloning vector, Ampr | Stratagene |
| pBluescriptSKII+ | Phagemid cloning vector, Ampr | Stratagene |
| pUCK4 | Plasmid encoding a Kanr gene cassette | Pharmacia Biotech |
| pKAS32 | Suicide vector, Ampr Stps | 55 |
| pRS415 | Promoter probe vector, Ampr | 53 |
| pmal-p2 | Expression vector for engineering translational fusions to MalE, Ampr | New England Biolabs |
| pTDCC1 | Cosmid clone of V. cholerae 569B, chiA+ | This study |
| pTDCC2 | 2.9-kbp Sau3AI fragment from pTDCC1 into the BamHI site of pBluescriptKS− | This study |
| pTDCC2.3 | Frameshift mutation at the NheI site of the insert in pTDCC2 | This study |
| pTDCC2.5 | BamHI/EcoRI fragment of pTDCC2.4 into BamHI/EcoRI site of pBluescriptSKII+ | This study |
| pTDCC2.6 | E-tag epitope fused to the 3′ end of chiA cloned in pBluescriptSKII+ | This study |
| pJL1 | chiA/E-tag hybrid gene of pTDCC2.6 fused to the 3′ end of malE in pmal-p2 | This study |
| pZ1 | Nucleotides 31–465 of the insert of pTDCC2 cloned into pRS415 | This study |
| pZ3 | Nucleotides 99–465 of the insert of pTDCC2 cloned into pRS415 | This study |
| pZ13 | Nucleotides 18–465 of the insert of pTDCC2 cloned into pRS415 | This study |
| pZ56 | Nucleotides 157–465 of the insert of pTDCC2 cloned into pRS415 | This study |
| pAKT-1 | Kanr gene of pUCK4 inserted into the unique HpaI site of pTDCC2 | This study |
| pJPF3 | KpnI/SstI fragment of pAKT-1 into the same sites of pKAS32 | This study |
| pTDCepsE | Wild-type copy of epsE gene of 569B | 11 |