MLST of Streptococcus zooepidemicus

Genes

Internal fragments of the following genes are used:

  • carbamate kinase (arcC)
  • ribonucleoside-diphosphate reductase (nrdE)
  • prolyl-tRNA synthetase (proS)
  • signal peptidase I (spi)
  • thymidylate kinase (tdk)
  • triosephosphate isomerase (tpi)
  • acetyl-CoA acetyltransferase (yqiL)

Amplification

Genes were amplified by PCR using the following primer pairs:

Locus Forward Reverse
arcC arcC1, 5'-AGC CAT CTA CCG ACT AAC AC-3' arcC4, 5'-TCT GAA AGG GTT TGG CTA GC-3'
nrdE nrdE1, 5'-TTC TCC TTC AGG TGA CAG ATG-3' nrdE4, 5'-AGA CTA GGC GTT TGA ACC TG-3'
proS proS1, 5'-TTG GTT GGA AAT GAC CAG ATC-3' proS4, 5'-CCT GAT CCT TGA CAT TAA CGG-3'
spi spi1, 5'-CTT AGA GCA TTG CGC TAA GC-3' spi4, 5'-TTG CCT GCT ATC TAG GGA AG-3'
tdk tdk1, 5'-GAA TTC AGG AAA GAC CAT TG-3' tdk4, 5'-TAA TGC TTG CGA CAA ACT GG-3'
tpi tpi1, 5'-GGC AGT AGT AAG CAA ATT ACC-3' tpi4, 5'-AGC AAG GCA AGG AAG CTA TC-3'
yqiL yqiL1, 5'-CCA CAT GGG AAT TAC AGC AG-3' yqiL4, 5'-TCA ATA CAG ACG TCC CTT GAC C-3'

 

Sequencing

Sequencing was performed using the following primer pairs:

Locus Forward Reverse
arcC arcC2, 5'-TGT CGC ACT TGG AGG AAA TG-3' arcC3, 5'-CAC CAC AAC ACC AGA ATC AAC-3'
nrdE nrdE2, 5'-CGC TCT ATT AAT TCT GCC TTG C-3' nrdE3, 5'-GCA TAG GTT GCT CAT GAT GAT G-3'
proS proS2, 5'-GAC TTC TTA GAG CCA GCT AG-3' proS3, 5'-AAC GGA GCT AGC TCT TTA GG-3'
spi spi2, 5'-CCT ACT TTT TGG ACT CTC ACG-3' spi3, 5'-GAT TTT ATT GAG TGG CCA AAA GCG-3'
tdk tdk2, 5'-CTT GAA GGT AGC ACA CAA TTA TG-3' tdk3, 5'-GGT CTC ATT GCC ACC AAT TTG-3'
tpi tpi2, 5'-GGT CAC TAC AAT TGA GGC TG-3' tpi3, 5'-TTC AGG CTT CAC AGA ACC AC-3'
yqiL yqiL2, 5'-AGT ACC GCA ACG TAA AGG TG-3' yqiL3, 5'-GCC TGA CGC TTT TCC ATC TC-3'