Table 1.

PCR primers used in this study

Procedure5′ Primer3′ PrimeraCommentbAmplicon size (bp)
PCR
 Step 1 (Fig.1A)GAT/C GAT/C TTT/C TGT/C GCI CAA/G ATT/CA GCTGT/C TGI CCI AA/GI GTI GCI GAT/C TGI GCDerived from N terminus and internal peptide of mature protein180
 Step 2 (Fig.1B)TAA ATA AAA ATG TAA TAA TGAGT TGT GCT GCT TCT AATDerived from GenBank acc. no. 4098503691
 Step 3 (Fig. 1D)TGC AGC TAA TCA GGA GAC AAGA TAA ATA TAA CTG ACA CT5′ primer, derived from GenBank acc. no. 4098503; 3′ primer, from S. mutansdatabase765
 Step 4 (1st round) (Fig. 1E)CAA GCA GCA GCC GCA CAATAA TAC GAC TCA CTA TAG GG N NNN NNN NNN GAT ATC5′ primer, derived from 765-bp amplicon; 3′ primer, arbitrary primer (4)
 Step 4 (2nd round)
(Fig. 1E)
GCC GCA GCT GAT AAC ACATT ATG CTG AGT GAT ATC CC5′ primer, derived from 765-bp amplicon; 3′ primer, arbitrary primer (4)1,054
 Step 5 (Fig.1F)ATA TAC TGG CTT TTC TCT TCT TTCACT TTC TCT GCT AAT TCT TTG TTT5′ primer, derived from 100 bp upstream of gbpBpromoter; 3′ primer, derived from 100 bp downsteam of gbpBtermination codon1,644
RFLP analysis (Fig.5A)ATA TAC TGG GCT TTT CTC TTC TTT CGTC TTA GCC ACT TTC TCT GCDerived from UA159 sequence flanking ORF of GbpB (S. mutans genome database [http://www.genome.ou.edu/smutans.html])1,653
  • a I is inosine; N is A, T, G, or C.

  • b acc, accession.