Table 1. Primers used for the PCR amplification of Campylobacter genes encoding new vaccine candidates.
Restriction sites inserted in each primer (BamHI: GGATCC, NotI: GCGGCCGC, NcoI: CCATGG, SacI: GAGCTC) are displayed in bold type.
| Primer | Sequence | Cycling conditions | Polymerase |
|---|---|---|---|
| pcDNA3 cloning | |||
| YP437_F1 | 5'-GCCGCCGGATCCATGAGAAAAATTTTAGTTGTTTTAG-3' | 95°, 30 s/ 52°C, 30 s/ 72°C, 2 min | Vent |
| YP437_R1 | 5'-CCGCCGGCGGCCGCTTAATATTCTTTATTTTCATACC-3' | ||
| YP562_F1 | 5'-GCCGCCGGATCCATGAGAATTACAAATAAACTTAAC-3' | 95°, 30 s/ 55°C, 30 s/ 72°C, 2 min 30 s | Vent |
| YP562_R1 | 5'-CCGCCGGCGGCCGCTTACATATAATTTAATAAGCTAAG-3' | ||
| YP1115_F1 | 5'-GCCGCCGGATCCATGGGTATTTTTGGAACATTATAC-3' | 95°, 30 s/ 50°C, 30 s/ 72°C, 2 min | Vent |
| YP1115_R1 | 5'-CGGCGGGCGGCCGCTTAAGATTTAAGCCCTAGTAAGG-3' | ||
| YP9769_F1 | 5'-GCCGCCGGATCCATGATGAATTCATTTTATAATGGG-3' | 95°, 30 s/ 50°C, 30 s/ 72°C, 2 min | Vent |
| YP9769_R1 | 5'-CGGCGGGCGGCCGCTTATCTTTTCATATTAATGGC-3' | ||
| YP9817_F1 | 5'-GCCGCCGGATCCATGAAAATAATTAAAATTCTTTTTTTAGG-3' | 95°, 30 s/ 57°C, 30 s/ 68°C, 1 min 30 s | Taq HF |
| YP9817_R1 | 5'-CCGCCGGCGGCCGCTTAAAAGCCTAGATTTACTCCGCC-3' | ||
| YP9838_F1 | 5'-GCCGCCGGATCCATGAAAAAAATATTCACAGTAGC-3' | 95°, 30 s/ 50°C, 30 s/ 68°C, 1 min 30 s | Taq HF |
| YP9838_R1 | 5'-CCGCCGGCGGCCGCTTATTTTCTATTAGGTGAAGC-3' | ||
| pQE-trisystem cloning | |||
| YP437_F2 | 5'-GCCGCCCCATGGGCAGAAAAATTTTAGTTGTTTTAG-3' | 95°, 30 s/ 52°C, 30 s/ 72°C, 2 min | Vent |
| YP437_R2 | 5'-CCGCCGGCGGCCGCATATTCTTTATTTTCATACC-3' | ||
| YP562_F2 | 5'-GCCGCCCCATGGGCAGAATTACAAATAAACTTAAC-3' | 95°, 30 s/ 53°C, 30 s/ 68°C, 2 min 30 s | Taq HF |
| YP562_R2 | 5'-CCGCCGGCGGCCGCCATATAATTTAATAAGCTAAG-3' | ||
| YP1115_F2 | 5'-GCCGCCGAGCTCGGTATTTTTGGAACATTATAC-3' | 95°, 30 s/ 60°C, 30 s/ 72°C, 2 min | Vent |
| YP1115_R2 | 5'-CGGCGGGCGGCCGCAGATTTAAGCCCTAGTAAGG-3' | ||
| YP9769_F2 | 5'-CGGCCGCCATGGCCATGAATTCATTTTATAATGGG-3' | 95°, 30 s/ 50°C, 30 s/ 72°C, 2 min | Vent |
| YP9769_R2 | 5'-CCGCCGGCGGCCGCTCTTTTCATATTAATGGC-3' | ||
| YP9817_F2 | 5'-GCCGCCCCATGGGCAAAATAATTAAAATTCTTTTTTTAGG-3' | 95°, 30 s/ 57°C, 30 s/ 68°C, 1 min 30 s | Taq HF |
| YP9817_R2 | 5'-CCGCCGGCGGCCGCAAAGCCTAGATTTACTCCGCC-3' | ||
| YP9838_F2 | 5'-GCCGCCCCATGGGCAAAAAAATATTCACAGTAGC-3' | 95°, 30 s/ 51°C, 30 s/ 72°C, 1 min 30 s | Vent |
| YP9838_R2 | 5'-CCGCCGGCGGCCGCTTTTCTATTAGGTGAAGCTTTTG-3' | ||