Table 1.

Bacterial strains and plasmids used

Bacterial strain or plasmidRelevant phenotype and/or genotypeaReference or source
Strain
A. naeslundii
  T14Vb Wild-type strain, expresses type 1 and 2 fimbriae, SmrKms 8
  5951Spontaneous mutant of strain T14V, lacks type 1 fimbriae, expresses type 2 fimbriae, Smr Kms 8
  147Spontaneous mutant of strain 5951, lacks type 1 and 2 fimbriae, Smr Kms 8
  MYT2-DC7c Lacks type 2 fimbriae, ΔfimA::kan SmrKmr This study
  MYT2-SC8d Lacks type 2 fimbriae, ΔfimA::kan, contains a copy of pMY2201, Smr Kmr This study
  MYT2-SC3d Expresses FimA, contains a copy of pMY2201, ΔfimA::kanSmr Kmr This study
  MY2366-DC2c Expresses FimA, Δorf365::kan SmrKmr This study
  MY2366-SC1d Expresses type 2 fimbriae, Δorf365::kan, contains a copy of pMY2366, Smr Kmr This study
S. oralis 34Synthesizes cell wall polysaccharide that serves as the receptor for Actinomycestype 2 fimbriae 26, 27
E. coli
  DH5αAps Gibco-BRL
  TG1Aps Amersham
  JM109Aps Gibco-BRL
  AV3502TG1 carrying pGP1-2 and pAV3502, Apr Kmr This study
Plasmids
 pAV1402Contains a 48-kbA. naeslundii T14V chromosomal DNA subcloned onto pHC79 13
 pAV3022Contains a 9.0-kbHindIII DNA fragment from pAV1402 subcloned onto pUC13, Apr This study
 pAV3502Contains a 2.4-kbSmaI DNA fragment from pAV3022 subcloned onto pGEM3Z for high-level expression of FimA, Apr This study
 pAV2606Contains a 1.85-kb BamHI DNA fragment from pAV3022 that encoded a 22-kDa truncated amino-terminal portion of FimA, Apr This study
 pAV2621Contains a 4.4-kbBamHI DNA fragment from pAV3022 that encoded a 35-kDa truncated carboxyl-terminal portion of FimA, Apr This study
 pMY300Contains the kan gene from pJRD215 subcloned onto pGEM7Zf(+), Apr Kmr This study
 pMY221Contains a 2.4-kb SmaI DNA fragment from pAV3022 subcloned onto pUC13 that harbors A. naeslundiiT14V fimA, Apr This study
 pMY2201The 975-bp KpnI DNA fragment internal tofimA is deleted and replaced with the kan gene from pJRD215; transcription of kan is opposite that offimA; ΔfimA::kanApr Kmr This study
 pMY2366The 600-bp BstXI DNA fragment internal to orf365 is substituted with the kan gene; transcription ofkan is opposite that of orf365; Δorf365::kan AprKmr This study
 pJRD215KmrSmr 10
 pUC13Apr Gibco-BRL
 pGEM7Zf(+)Apr Promega
 pGEM3ZApr Promega
 pGP1-2Contains the T7 RNA polymerase gene, Kmr 40
  • a Km, kanamycin; Sm, streptomycin; Ap, ampicillin.

  • b Made resistant to streptomycin by selection of colonies from a growth medium containing the antibiotic.

  • c Mutant strain generated by a double-crossover recombinational event.

  • d Mutant strain generated by a single-crossover recombinational event predicted by the mechanism of Campbell.