Table 1.

Bacterial strains, plasmids, and oligonucleotides used in the study

Strain, plasmid, or oligonucleotideRelevant characteristic(s) or sequenceSource or reference
E. coli DH10BF mcrA Δ(mrr-hsdRMS-mcrBC) φ80dlacZΔM15 lacX74 deoR recA1 araD139 (ara leu)7697 galU galK I-rpsL endA1 nupGGibco/BRL
C. perfringens strains
    Strain 13Gangrene-associated strain20
    SM125pilT mutant39
    SM125(pSM271)pilT mutant complemented with wild-type pilT39
N. gonorrhoeae strains
    MS11Wild type, ΔpilE234
    MS11-307ΔpilE1::erm ΔpilE226
    MS11-310ΔpilE1::kan2
    KR106ΔpilE1::erm ΔpilE2, C. perfringens pilA2+This study
    KR107ΔpilE1::erm ΔpilE2, C. perfringens pilA1+This study
Plasmids
    pKH35E. coli origin of replication, lacIq, OPOPlac, three Neisseria uptake sequences, chloramphenicol resistance11
    pKR106pKH35 with C. perfringens pilA2 and E. coli ribosome binding siteThis study
    pKR107pKH35 with C. perfringens pilA1 and E. coli ribosome binding siteThis study
    pKR112PGem-T Easy with pilT from N. gonorrhoeae MS11This study
    pKR113pKR112, Kanr inserted into SacII site in pilTThis study
Oligonucleotides
    OKR0115′-CTTTAAAAAATAAAGCTTAGGAGGCATAAATGTTATTACTGAAAGC-3′
    OKR0135′-CATATTAAGCTTAGGAGGAAAACCAATGAATACAAAAAAAC-3′
    OKR0145′-CACCTTATTATTTTACTAATTCTAGAACTTTTAATACTATTGATTATTTC-3′
    OKR0195′-GCCCCTATGAAATCTAGATAATTAAAGTATATTTATACTATAGACACC-3′
    OKR0635′-CGAAACCATATGCTAAACAGAGCCGCA-3′
    OKR0645′-GCCTTGCCATGGCGCC-3′
    OKR0655′-CCGCGGCCGTCTGAACTCAAAATC-3′
    OKR0665′-CCGCGGTTGATGAGAGCTTTGTTGTAG-3′