Table 1.
Bacterial strains, plasmids, and primers
| Reagent | Description or sequence | Source or reference |
|---|---|---|
| Bacterial strains | ||
| V. cholerae O395 | O1 classical Ogawa, wild type, Smr | 53 |
| E. coli S17 λpir | thi pro recAhsdR17(rK− mK+)[RP4-2-Tc::Mu::Kmr Tn7] λpir; Tpr Smr | 13 |
| V. cholerae SJK70 | O395; ΔtcpA; Smr | R. K. Taylor |
| V. cholerae SI003 | SJK70, pMin1 | This study |
| V. cholerae JR003 | O395, ΔtcpAcofA | This study |
| ETEC 31-10 | Wild-type ETEC, carries 55-kb virulence plasmid with cof operon encoding CFA/III | 24 |
| ETEC 31-10p | ETEC 31-10, spontaneous CFA/III-deficient strain lacking 55-kb virulence plasmid | 51 |
| E. coli Origami(DE3) | Δ(ara-leu)7697 ΔlacX74 ΔphoAPvuII phoR araD139 ahpC galE galK rpsL F′[lac+lacIqpro] (DE3) gor522::Tn10trxB; Kmr Smr Tcr | Novagen |
| Plasmids | ||
| pTK1 | pKAS32::tcpACl-rplL; Apr | 26 |
| pTK1-ΔBbsI | pTK1, BbsI site deleted | This study |
| pJR2 | pTK1::cofA within tcpACl genomic flanking sequences | This study |
| pMin1 | pACYC184; Gmr Cmr | 38 |
| pMT5 | toxT; Apr | 14 |
| Primers | ||
| Ec-cofA-fpcr | GGAATTCCATATGGACTCACGTACTGTTTCTGAATTGGTC | This study |
| Ec-cofA-rpcr | CGGGATCCTTAACGGCTCGCCAAAGTAATAGAGTTA | This study |
| oOMP-fB2 | CATGGAAGACCGTTCAAATAAGTTTGTTTAACTTAATCTTAACG | This study |
| oOMP-rB1 | CATGGAAGACCGACCCTCTTGACCGGTTTTC | This study |
| oOMP-fA1 | CATGGAAGACCGGGGTATGAGCCTTCTGGAAGTCATCATC | This study |
| oOMP-rA2 | CATGGAAGACCGTGAATTAACGGCTCGCCAAAGTAATAG | This study |
| Vc-tcpA-f1.36 | GCAATGATCATCATGACAGATCTGC | This study |
| Vc-tcpA-flank-F2 | GTAAAATGGTGGAGTTACATAAATATGC | This study |
| Vc-tcpA-flank-R2 | GATACCACATTGAATTTAATAAGTTGC | This study |