Open Access
Table I.
Oligonucleotide primer sequences used in this study.
| Designation | Sequence (5′–3′). Lower case indicates base changes to introduce restriction sites | Application | Template | Annealing temp (°C) |
|---|---|---|---|---|
| p247 ISS1 fwd | GCTCTTCGGATTTTCGGTATC | ISS1 probe | pGh9::ISS1 | 58 |
| p250 ISS1 rev | CATTTTCCACGAATAGAAGGACTGTC | ISS1 probe | pGh9::ISS1 | 61 |
| p261 | TGGTTGAAGCAGAAGCTGAA | Screening for ISS1 within srtA ORF vs p247 | pGh9::ISS1 | 55 |
| p616 | CTAAGAAAATCACAGTTGACATGC | Screening for ISS1 within sub0826 ORF vs p247/p250 | pGh9::ISS1 | 59 |
| p621 | CAACGAATCAACAAACTGAAAGC | Screening for ISS1 within sub1370 ORF vs p247/p250 | pGh9::ISS1 | 59 |
| p628 | CATGTCTTTCGGTACCGTGGTTTTCTCG | Amplification of srtA ORF | S. uberis 0140J genomic DNA | 62.5 |
| p633 | ACAGTATTAAAGGCACAAATGGAAG | Amplification of srtA ORF | S. uberis 0140J genomic DNA | 60 |
| p641 | CAAATGCTAAAGTTGGCTTAAAAAC | Screening for ISS1 within sub0135 ORF vs p247/p250 | pGh9::ISS1 | 60 |
| p642 | CAATAATCAACAAATAAAGCGCAAT | Screening for ISS1 within sub0135 ORF vs p247/p250 | pGh9::ISS1 | 60 |
| p645 | ATCAAACTATTGTACCCTTTGGTGA | Screening for ISS1 within sub1095 ORF vs p247/p250 | pGh9::ISS1 | 60 |
| p646 | TGTAATGAATCTAGGAGGGGTAGGT | Screening for ISS1 within sub1095 ORF vs p247/p250 | pGh9::ISS1 | 60 |
| p649 | ATTAAAGAAGGTGATGTGGTTCAAA | Screening for ISS1 within sub1730 ORF vs p247/p250 | pGh9::ISS1 | 60 |
| p650 | GTCATTTAGCTTCTCCCATGTAAAA | Screening for ISS1 within sub1730 ORF vs p247/p250 | pGh9::ISS1 | 60 |
| p653 | CTTTATGAAAATAGCCAAGCTGAAA | Screening for ISS1 within sub0888 ORF vs p247/p250 | pGh9::ISS1 | 60 |
| p654 | CTTAATTGTTGCTGGTCTTTCATTT | Screening for ISS1 within sub0888 ORF vs p247/p250 | pGh9::ISS1 | 60 |
| p657 | TTTTCTTCTTAATTGCACACCTGA | Screening for ISS1 within sub0207 ORF vs p247/p250 | pGh9::ISS1 | 60 |
| p658 | AATCCAAGCTGTTGAACTATCCTC | Screening for ISS1 within sub0207 ORF vs p247/p250 | pGh9::ISS1 | 60 |
| p667 | TGATCTTGTGCATATCAATCTGG | Screening for ISS1 within sub0145 ORF vs p247/p250 | pGh9::ISS1 | 59 |
| p668 | AAGGAACCTGAAAGAACCATCTC | Screening for ISS1 within sub0145 ORF vs p247/p250 | pGh9::ISS1 | 61 |
| p675 | ACGTTCCCTACAAAAGAAAAAGG | Screening for ISS1 within sub0241 ORF vs p247/p250 | pGh9::ISS1 | 59 |
| p676 | TTGCCACTTAGGCTGATAAAAAC | Screening for ISS1 within sub0241 ORF vs p247/p250 | pGh9::ISS1 | 59 |
| p689 | GTGATTAAGCgAAtTCAATAAGGAAATAATGG | Amplification of sub1154 gene fragment and upstream flanking sequence. Incorporating an EcoRI restriction site. | S. uberis 0140J genomic DNA | 62 |
| p690 | CTTGTCGTCGTCATCCTTGTAGTCATCCGACTGCTTTTCATTTTGCAA | Amplification of sub1154 gene fragment, incorporating FLAG-TAG | S. uberis 0140J genomic DNA | > 60 |
| p691 | GACTACAAGGATGACGACGACAAGTATCCGCAGTCAATAAGCCAAA | Amplification of sub1154 gene fragment, incorporating FLAG-TAG | S. uberis 0140J genomic DNA | > 60 |
| p692 | ACTTACTTGAAAGCtTCACAAGTGGTAAAGT | Amplification of sub1154 gene fragment and downstream flanking sequence. Incorporating a HindIII restriction site. | S. uberis 0140J genomic DNA | 63 |