Table 1.

Bacterial strains and plasmids used in this study

Strain or plasmidRelevant characteristicsaSource or reference
Strains
P. gingivalis
  33277Type strain from ATCCThis laboratory
  UPFDerivative of 33277,upf:lacZ gene, Emr This study
  MPF10Derivative of 33277, mpf10:lacZgene, Emr This study
  MPS10Derivative of 33277, mps10:lacZ gene, Emr This study
  MPF35Derivative of 33277,mpf35:lacZ gene, Emr This study
  MPS35Derivative of 33277, mps35:lacZgene, Emr This study
  MP58Derivative of 33277, mp58:lacZ gene, Emr This study
  MP150Derivative of 33277,mp150:lacZ gene, Emr This study
  MP60Derivative of 33277, mp60:lacZgene, Emr This study
  MP59Derivative of 33277, mp59:lacZ gene, Emr This study
E. coli DH5α endA1 hsdR17 supE44 thi-1 recA gyrA96 relA1 Δ(lacZYA-argF)U169λ-φ80 dlacZ ΔM15; recipient for recombinant plasmidsBRL
Cloning vectors
 pTZBg21.1Containing a 2.5-kb SacI DNA fragment with the fimA gene, Amr 32
 pUC19 E. coli cloning vector, Amr BRL
 pDN19lacContains 3.6-kbBamHI-SalI fragment containing promoterlesslacZ gene with ribosome binding site 35
 pJRD215Wide-host-range cosmid vector, Kmr Smr Mob+, unable to replicate in P. gingivalis 28
 pBF4Contains 3.8-kb EcoRI fragment in Tn4351 carrying two antibiotic resistance genes: Tcr expressed in E. coli and Emrexpressed in P. gingivalis 33
 R751IncP plasmid used to mobilize vectors from E. coli to Bacteroides recipient, TprTra+ 28
 pCR2.1-TOPOLinearized plasmid with single 3′ dT residues, Kmr Invitrogen
Recombinant plasmids
 pUPF1 P. gingivalis fimA promoter region (upf) containing sequence from −606 to +86 in plasmid pCR2.1-TOPO, Amr Kmr This study
 pUPF5 upf:lacZ gene in pJRD215 with a 3.8-kb EcoRI fragment from Tn4351, Tcr Kmr Smr This study
 pMPF101 upf:lacZ gene in pUC19 with 3-base change from TAT to CCG at positions −23 to −21 (mpf10:lacZ), Amr This study
 pMPF103 mpf10:lacZ gene in pJRD215 with a 3.8-kb EcoRI fragment from Tn4351, Tcr Kmr Smr This study
 pMPS101 upf:lacZ gene in pUC19 with 3-base change from TAA to GCC at positions −11 to −9 (mps10:lacZ), Amr This study
 pMPS103 mps10:lacZ gene in pJRD215 with 3.8-kb EcoRI fragment from Tn4351, Tcr Kmr Smr This study
 pMPF351 upf:lacZ gene in pUC19 with 3-base change from TTG to CCA at positions −46 to −44 (mpf35:lacZ), Amr This study
 pMPF353 mpf35:lacZ gene in pJRD215 with 3.8-kb EcoRI fragment from Tn4351, Tcr Kmr Smr This study
 pMPS351 upf:lacZ gene in pUC19 with 3-base change from TGG to CAC at positions −43 to −41 (mps35:lacZ), Amr This study
 pMPS353 mps35:lacZ gene in pJRD215 with 3.8-kb EcoRI fragment from Tn4351, Tcr Kmr Smr This study
 pMP1501 upf:lacZ gene in pCR2.1-TOPO with 150-bp deletion from −240 to −90 (mp150:lacZ), Amr This study
 pMP1504 mp150:lacZ, gene in pJRD215 with 3.8-kb EcoRI fragment from Tn4351, Tcr Kmr Smr This study
 pMP601 mp150:lacZ gene with 15-bp deletion from −85 to −71 (mp60:lacZ) in pUC19, Amr This study
 pMP603 mp60:lacZ gene in pJRD215 with 3.8-kb EcoRI fragment from Tn4351, Tcr Kmr Smr This study
 pMP591pMP601 with 17-bp deletion from −64 to −48 (mp59:lacZ), Amr This study
 pMP593 mp59:lacZ gene in pJRD215 with 3.8-kb EcoRI fragment from Tn4351, Tcr Kmr Smr This study
 pMP581 upf:lacZ gene in pUC19 with 16-bp deletion from −23 to −8 (mp58:lacZ), Amr This study
 pMP583 mp58:lacZ gene in pJRD215 with 3.8-kb EcoRI fragment from Tn4351, Tcr Kmr Smr This study
  • a Kmr, Smr, Tcr, Emr, Tpr, and Amr, resistance to kanamycin, streptomycin, tetracycline, erythromycin, trimethoprim, and ampicillin; Mob+, can be mobilized; Tra+, capable of self-transfer.