Table 2.

Plasmids and oligonucleotide primers used in mutagenesis and complementation

Plasmid or primerRelevant characteristics or sequenceaSource or reference
Plasmid or primerRelevant characteristics or sequenceaSource or reference
Plasmids:
    pKD3Plasmid utilized in λ phage recombinase mutagenesis method; Cmr41
    pKD46Expresses recombinases red, β, λ, and exo for construction of deletion mutants; Ampr41
    pMP2444pBBR1MCS-5 backbone; gfp expressed from lac promoter; Gmr41
    pJMK2pBBR1MCS-3 backbone; hofC gene inserted as KpnI-SacI fragment; TcrThis study
    pJMK3pBBR1MCS-3 backbone; hof gene cluster inserted as KpnI-SacI fragment; TcrThis study
    pJMK4pBBR1MCS-3 backbone; fimD gene inserted as KpnI-SacI fragment; TcrThis study
    pJMK5pBBR1MCS-3 backbone; fim gene cluster inserted as KpnI-SacI fragment; TcrThis study
    pJMK6pBBR1MCS-3 backbone; crl gene inserted as KpnI-SacI fragment; TcrThis study
    pJMK7pBBR1MCS-3 backbone; eae gene inserted as KpnI-SacI fragment; TcrThis study
Primers
    Ea1189ΔhofCF: 5′-ATGGGTGAACGCTTACTTTTCCGCTGGCAGGCTATTGACGATAGTGGGCAGTGTAGGCTGGAGCTGCTTC-3′This study
R: 5′-TTACCCAAGCGCATCTCCCAGCCTGAATACGGGCAAATACATGGCCACCACATATGAATATCCTCCTTA-3′
    Ea1189ΔhofF: 5′-ATGGGTGAACGCTTACTTTTCCGCTGGCAGGCTATTGACGATAGTGGGCAGTGTAGGCTGGAGCTGCTTC-3′This study
R: 5′-TCATGGCAACTGCTCCTCATCAAAACGGAACATATCCAGGCAAGCGTCCTCATATGAATATCCTCCTTA-3′
    Ea1189ΔfimDF: 5′-ATGACGCCTAAGGTGAAGAGGTATGTGCTATTTGACGAAGCTTTCTGCCGGTGTAGGCTGGAGCTGCTTC-3′This study
R: 5′-AATAAGTGTGCAARAGAGGACATTGAAAAGACAGAGAACGTCGAAGCGACCATATGAATATCCTCCTTA-3′
    Ea1189ΔfimF: 5′-ATGAAAAAAGTAATCAACTTTATTTTCCTGCTGCTGGCAGGTGCGGGTGAGTGTAGGCTGGAGCTGCTTC-3′This study
R: 5′-GATGACTATGAGTTATGACTGGCCGTCACTGTGCACGGTCGCGGCCCCTTCATATGAATATCCTCCTTA-3′
    Ea1189Δflg 3F: 5′-ATGATTATGAAAATAAAGCTTTTCCTGTTGGGCTGTGGACTTGGACTCGGGTGTAGGCTGGAGCTGCTTC-3′This study
R: 5′-TCAGAAAGACAAATCGACGGTCACCGTATCGCTATAGGACCCCGCCACCGCATATGAATATCCTCCTTA-3′
    Ea1189Δflg 4F: 5′-GGCAGCGGGGTGAAGTGATACAACAGATCGGCTTAGAATCAGGCTTTACGGTGTAGGCTGGAGCTGCTTC-3′This study
R: 5′-TTACGAATTGAGACTAAACAGTGACAGCTTCGACATCTGCTGGAATACGGCATATGAATATCCTCCTTA-3′
    Ea1189ΔcrlF: 5′-ATGACGTTACCGAGTGGACATCCTAAGAGTCGAATAATTAAGCGCTTTCAGTGTAGGCTGGAGCTGCTTC-3′This study
R: 5′-TCAGGCGGTCAGCTTCACCGGCTGGTCAGCGAAATCCGTTGCCGGTATCACATATGAATATCCTCCTTA-3′
    Ea1189ΔeaeF: 5′-ATGCAGGGGGGTAAAGCGGCTCCTCCTGCAATAATCTGGGACAAAGATGAGTGTAGGCTGGAGCTGCTTC-3′This study
R: 5′-TTATTTTATAATATCAACATAAGGCTTGTTTATTGCGCTGCCGCTGCCAACATATGAATATCCTCCTTA-3′
    ΔhofC compF: 5′-GGTACCATGGGTGAACGCTTACTTTTCC-3′This study
R: 5′-GAGCTCTTACCCAAGCGCATCTCC-3′
    Δhof compF: 5′-GGTACCATGGGTGAACGCTTACTTTTCC-3′This study
R: 5′-GAGCTCTCATGGCAACTGCTCCTCAT-3′
    ΔfimD compF: 5′-GGTACCATGACGCCTAAGGTGAAGAGG-3′This study
R: 5′-GAGCTCTTATTCACACGTTATCTCCTGTAACTT-3′
    Δfim compF: 5′-GGTACCTGAAATTATTAACGAATGCCTGC-3′This study
R: 5′-GAGCTCATGATAAAACTTGCGGATAAATATCG-3′
    Δcrl compF: 5′-GGTACCATGACGTTACCGAGTGGACAT-3′This study
R: 5′-GAGCTCTCAGGCGGTCAGCTTCAC-3′
    Δeae compF: 5′-GGTACCGGTAAAGCGGCTCCTCCT-3′This study
R: 5′-GAGCTCAATATCAACATAAGGCTTGTTTATTGC-3′
  • a KpnI (GGTACC) and SacI (GAGCTC) restriction sites are underlined. Abbreviations: F, forward; R, reverse; Amp, ampicillin; Cm, chloramphenicol; Gm, gentamicin; Tc, tetracycline.