Table 1 Details of primers and restriction enzymes used for multilocus restriction typing (MLRT) of Y. enterocolitica biovar 1A
Target gene | Primer | Position* | Sequence (5'-3') | Annealing temperature | Amplicon size (bp) | Restriction enzyme | Restriction fragments (bp)†|
|---|---|---|---|---|---|---|---|
mdh (malate dehydrogenase) | Mdh1 Mdh2 | 484705...484726 485301...485280 | TAT ATG ACA TCG CGC CAG TGA C CAG CTT GCC CCA TAG ACA GAG T | 61°C | 597 | Hae III Rsa I | 102, 164, 331 179, 191, 227 |
cya (adenylate cyclase) | AdC1 AdC2 | 224199...224222 225200...225181 | AAC CGC CTG CAA AAG AAA TGT AGT CCA GCC CGG ACG GTT AGC AC | 66°C | 1,002 | Hae III Sau96 I | 22, 157, 346, 477 24, 128, 216, 634 |
glnA (glutamine synthetase) | GN1 GN2 | 36808...36830 37528...37506 | TTC CGG TGG CAA GTC ATA CAG GT CAA ATA CGA AGG CGG CAA CAA AG | 65°C | 721 | Bgl I Sau96 I | 70, 651 39, 85, 237, 360 |
zwf (glucose-6-phosphate dehydrogenase) | G6P1 G6P2 | 2570039...2570061 2570679...2570659 | CCT GAA TAC CGC GCA TCG TCT CT AGG GCG CTG GGG CTA TTT TGA | 65°C | 641 | Rsa I BstN I | 32, 62, 109, 189, 249 128, 243, 376 |
icdA (isocitrate dehydrogenase) | IDH1 IDH2 | 1923868...1923889 1925035...1925014 | GCG CTG AAG GAG AGG TTG ATG G CGC CTT CGG TGC CTT TGA TAA T | 57°C | 1,168 | Hae III Rsa I | 136, 185, 365, 480 125, 127, 221, 304, 391 |
gdhA (glutamate dehydrogenase) | GmD1 GmD2 | 4416077...4416094 4416600...4416579 | GGG CAA AGG CGG CTC TGA TAC GTT CGC GGC ATA ATC TTC | 66°C | 524 | Hae III Mse I | 11, 42, 141, 320 21, 50, 121, 432 |