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Table 3 List of primers used for PCR amplification of protein-encoding genes from Treponema denticola strains

From: Multilocus sequence analysis of Treponema denticolastrains of diverse origin

Gene

Primer

Sequence(5′to 3′)

Strains amplified

dnaN

dnaNF

ATGAAAATAAGTTTTGACAGAGACAC

dnaF + dnaR: all strains (55-50°C)

 

dnaNR

TTACTCCGTCTGCATAGGC

 

recA

recAF1

GTGGCAAAAGCAAAAAAC

recAF1 + recAR1: most strains (55-47°C)

 

recAR1

TTAAAAAAGACTGTCGTCCG

recAF2 + recAR2: ATCC 700768, MS25 (54-47°C)

 

recAF2

TTCATATTGGCCGCATTTG

recAF1 + recArecAR2: ATCC 700771 (55-49°C)

 

recAR2

TTGTGTACTCATAATGCCGCTC

 
 

recAF

GTGGCAAAAGCAAAAAACGAAG

recAF + recAR: OMZ852, OMZ853, NY531, NY553 (58-53°C)

 

recAR

TTAAAAAAGACTGTCGTCCGCC

 

radC

radCF1

ATGATAGACTATAAAAATTCGTCCAATAC

radCF1 + radCR1: most strains (55-50°C)

 

radCR1

TTAAATATCAAACCTCGTTCCG

radCF1 + radCR2: MS25 (55-49°C)

 

radCF2

AACATGGCTTTCCGAAATC

radCF2 + radCR1: ATCC 700768 (55-49°C)

 

radCR2

GTGCAGCGGCTCTAAAAG

 

ppnK

TDE1591F1

ATATGGATCCCATATGAAAAAAG

TDE1591F1 + TDE1591R1: most strains (52-45°C)

 

TDE1591R1

AATTCTCGAGTCAATTCAGTTTGGG

TDE1591F2 + TDE1591R2: OKA3, MS25,GM1, ST10A,

 

TDE1591F2

AGCTACCCTGCCCTAATTTC

ATCC 700768, ATCC 700771 (57-52°C)

 

TDE1591R2

AACATCCTTAAAAAGCGGC

 

flaA

TDE1712F

ATGAAAAAAACATTTATACTTGTTG

TDE1712F + TDE1712R: all strains (52-46°C)

 

TDE1712R

TTATTGTTGGTTCTTTTCGG

 

era

eraF1

ATGAACAGCGGAGTTGTAAC

eraF1 + eraR1: most strains (55-50°C)

 

eraR1

TTAATACGAGATTTTTTTTATGATATTATC

 
 

eraF2

GGTACTTGTGCTTACCGAAAAC

eraF2 + eraR2: MS25 (54-47°C)

 

eraR2

CCGACACAATCGAGGAAG

 
 

eraF4

CGCTTAGAAGAAGGGGATGC

eraF4, eraR4 separately used for direct chromosome sequencing of ATCC 700768†

 

eraR4

CTTTTTCGACATAGAGGAAGGC

 

pyrH

pyrHF

ATGGTAACTGTTTTGTCGGT

pyrHF + pyrHR: all strains (54-47°C)

 

pyrHR

TTAGCCGATTACCGTTCCTT

 
  1. Genetic loci are based on the ATCC 35405 type strain of Treponema denticola.
  2. F: Forward primer; R: Reverse primer. Values in parenthesis indicate annealing temperatures used in ‘touchdown PCR’ procedures.
  3. †PCR amplification was unsuccessful; sequencing of chromosomal DNA employed.