Table 1.

Strains and plasmids used in this study

Strain or plasmidGenotype or featuresSource or reference
Strains
 E2348/69Prototype O127:H6 EPEC strain 25
 UMD901E2348/69 bfpA C129S 50
 UMD928E2348/69bfpG::aphA3 This study
 UMD923E2348/69bfpB::aphA3 This study
 UMD924E2348/69bfpC::aphA3 This study
 UMD926E2348/69bfpD::aphA3 This study
 UMD932E2348/69bfpP::aphA3 This study
 UMD918E2348/69bfpH::aphA3 This study
 DH5α supE44 ΔlacU169(φ80 lacZΔM15) hsdR17 recA1 endA1 gyrA96 thi-1 relA1 36
 DH5αλpir DH5α(λpir) 27
Plasmids
 pBluescriptHigh-copy-number cloning vectorStratagene
 pMSD205 bfpA gene cloned into PCR1000 under control of an isopropyl-β-d-thiogalactopyranoside-inducible promoter 13
 pRPA1001-kbBamHI-HindIII fragment of the bfpcluster containing bfpA cloned into pWKS30This study
 pRPA1013-kb BamHI-BalI fragment of thebfp cluster containing bfpA-bfpB cloned into pWKS30This study
 pRPA1034.1-kbBamHI-XbaI fragment of the bfp cluster containing bfpA-bfpC cloned into pWKS30This study
 pRPA1042.3-kb BamHI-HindIII fragment of the bfp cluster containing bfpA-bfpG cloned into pWKS30This study
 pRPA1066.5-kbBamHI-KpnI fragment of the bfp cluster containing bfpA-bfpD cloned into pWKS30This study
 pRPA1071.3-kb PstI-BamHI fragment of the bfp cluster containing bfpP cloned into pWKS30This study
 pCVD442Positive-selection suicide vector 14
 pRK2073pRK2 derivative suitable for mobilization of plasmids 10
 pKDS302Entire bfp cluster cloned into pTRC99A under control of trc promoter 40
 pMSD230 bfpA cloned into pTRC99A under control oftrc promoterThis study
 pMSD2335.3-kbBamHI fragment of the bfp cluster containingbfpA-bfpU cloned into pWKS30This study
 pDN19PB bfpP cloned into pDN19 51
 pDN19PBΔ bfpP with a 93-bp deletion cloned into pDN19 51
 pUC18K aphA3 gene cloned into pUC18, creating a nonpolar kanamycin cassette 27
 pUC18K2pUC18K with 1 bp added after the ATG start codonK. Jarvis and J. Kaper
 pUC18K3pUC18K with 2 bp added after the ATG start codonK. Jarvis and J. Kaper