Table 3. Sequences of the primers and vectors used for RHDV1 type-specific MAb epitope identification.
| Fragment | Primer sequences (5′–3′)A | Position In VP60 geneB | Vector | |
|---|---|---|---|---|
| Sense | Negative –sense | |||
| pR1-VP60 | GGAATTCATGGAGGGCAAAACCCGCAC | CCCAAGCTTGTCAGACATAAGAAAAGCCATTGGT | 1–1740 | pET-32a |
| eR1-VP60 | CCCAAGCTTATGGAGGGCAAAACCCGCAC | GGAATTCTCAGACATA AGA AAAGCCATTGGT | 1–1740 | pcDNA3.1(+) |
| R1-VP60-1 | GGAATTCATGGAGGGCAAAACCCGCAC |  |
1–342 | pGEX-6p-1 |
| R1-VP60-2 | GGAATTCCCATTCACAGCCGTGCTGAG | CCCAAGCTTGTAGTGTGGGGACAAGGCCAG | 301–570 | pET-32a |
| R1-VP60-3 | GGAATTCGACTTGCGTCCCAACAT | CCCAAGCTTGTGCGGGTGAGATTGAGT | 514–720 | pET-32a |
| R1-VP60-4 | GGAATTCAGAGCCCCCTCCAGCAAGAC | CCCAAGCTTGGCCTCTTCGATGGTCAATGTC | 679–900 | pET-32a |
| R1-VP60-5 | GGAATTCTGGTCAAGCCCTCGGTTTGC | CCCAAGCTTGAGCAGGAGTGCCGGGTGTAG | 859–1353 | pET-32a |
| R1-VP60-6 | GGAATTCGTCACATACACGCCCCAAC | CCCAAGCTTGTCAGACATAAGAAAAGCCATTGGT | 1300–1740 | pET-32a |
| R1-VP60-4.1 | GGAATTC CCCGCAGGCCTCCTCACGAC | CCCAAGCTTGTTGCAGTCCCACTATTTGGC | 715–795 | pET-32a |
| R1-VP60-4.2 | GGAATTCATAGTGGGACTGCAACCAGT | CCCAAGCTTGCCGAGGGCTTGACCAGCCAT | 781–873 | pET-32a |
| R1-VP60-4.3 | GGAATTCCCCGCAGGCCTCCTCACGAC | CCCAAGCTTGGGAAAACCCCCCAGGTACTG | 715–816 | pET-32a |
| R1-VP60-4.4 | GGAATTCAGGTGGAACGGCCAAATAGT | CCCAAGCTTGCCGAGGGCTTGACCAGCCAT | 766–873 | pET-32a |
| 1D6-1 | GGAATTCTGGAACGGCCAAATAGTGGGACTGCAA | CCCAAGCTTGTTGCAGTCCCACTATTTGGCCGTTCCA | 769–795 | pET-32a |
| 1D6-2 | GGAATTCAACGGCCAAATAGTGGGACTGCAA | CCCAAGCTTGTTGCAGTCCCACTATTTGGCCGTT | 771–795 | pET-32a |
| 1D6-3 | GGAATTCAGGTGGAACGGCCAAATAGTGGGACTG | CCCAAGCTTGCAGTCCCACTATTTGGCCGTTCCACCT | 766–792 | pET-32a |
| 1D6-4 | GGAATTCAGGTGGAACGGCCAAATAGTGGGA | CCCAAGCTTGTCCCACTATTTGGCCGTTCCACCT | 766–789 | pET-32a |
| 1D6-5 | GGAATTCAGGTGGAACGGCCAAATAGTG | CCCAAGCTTGCACTATTTGGCCGTTCCACCT | 766–786 | pET-32a |
| 1D6-6 | GGAATTCAGGTGGAACGGCCAAATA | CCCAAGCTTGTATTTGGCCGTTCCACCT | 766–783 | pET-32a |
| 1D6-7 | GGAATTCAGGTGGAACGGCCAA | CCCAAGCTTGTTGGCCGTTCCACCT | 766–780 | pET-32a |
| 1D6-8 | GGAATTCAGGTGGAACGGC | CCCAAGCTTGGCCGTTCCACCT | 766–777 | pET-32a |
| R1-A1-1 | GGAATTCCGAAGAGGCAGTGCAAGTTATTCTGGGAACAACTCC | CCCAAGCTTGGGAGTTGTTCCCAGAATAACTTGCACTGCCTCTTCG | 892–927 | pET-32a |
| R1-A1-2 | GGAATTCTCTGGGAACAACTCCACCAACGTGCTCCAG | CCCAAGCTTGCTGGAGCACGTTGGTGGAGTTGTTCCCAGA | 913–942 | pET-32a |
| R1-A1-3 | GGAATTCACCAACGTGCTCCAGTTTTGGTACGCTAAT | CCCAAGCTTGATTAGCGTACCAAAACTGGAGCACGTTGGT | 928–957 | pET-32a |
| 1H2-1 | GGAATTCAACGTGCTCCAGTTTTGGTACGCTAAT | CCCAAGCTTGATTAGCGTACCAAAACTGGAGCACGTT | 931–957 | pET-32a |
| 1H2-2 | GGAATTCGTGCTCCAGTTTTGGTACGCTAAT | CCCAAGCTTGATTAGCGTACCAAAACTGGAGCAC | 934–957 | pET-32a |
| 1H2-3 | GGAATTCCTCCAGTTTTGGTACGCTAAT | CCCAAGCTTGATTAGCGTACCAAAACTGGAG | 937–957 | pET-32a |
| 1H2-4 | GGAATTCCAGTTTTGGTACGCTAAT | CCCAAGCTTGATTAGCGTACCAAAACTG | 940–957 | pET-32a |
| 1H2-5 | GGAATTCACCAACGTGCTCCAGTTTTGGTACGCT | CCCAAGCTTGAGCGTACCAAAACTGGAGCACGTTGGT | 928–954 | pET-32a |
| 1H2-6 | GGAATTCACCAACGTGCTCCAGTTTTGGTAC | CCCAAGCTTGGTACCAAAACTGGAGCACGTTGGT | 928–951 | pET-32a |
| 1H2-7 | GGAATTCACCAACGTGCTCCAGTTTTGG | CCCAAGCTTGCCAAAACTGGAGCACGTTGGT | 928–948 | pET-32a |
| 1H2-8 | GGAATTCACCAACGTGCTCCAGTTT | CCCAAGCTTGAAACTGGAGCACGTTGGT | 928–945 | pET-32a |
| 1H2-9 | GGAATTCACCAACGTGCTCCAG | CCCAAGCTTGCTGGAGCACGTTGGT | 928–942 | pET-32a |
AUnderlined letters indicate additional restriction enzyme sites: EcoRI and Hind III for the vector pET-32a, EcoRI and SalI for the vector pGEX-6P-1, and HindIII and EcoRI for the vector pcDNA3.1(+) were introduced into each primer. The boxed TTAs are the stop codon additions introduced into the antisense primers of the VP60 fragments for use with pGEX-6P-1. BNucleotide positions correspond to those in the sequence of the RHDV1 VP60 gene (GenBank accession no.: AF453761.1).