TABLE 1.
Primers used in the study
| Primer name | Sequence (5′–3′)a | Restriction site (promoter or tag) | Purpose |
|---|---|---|---|
| 3ABC-Fwd | GCGAAGCTTCCACCATGGATTACAAGGATGACGACGATAAGATCTCAATTCCTTCCC | HindIII (FLAG) | FMDV 3ABC amplification |
| 3ABC-Rev | GCGGGTACCTATTAAGCGTAATCAGGAACGTCGTAAGGGTACTCGTGGTGTGGTTCG | KpnI (HA) | |
| 3A-Rev | GCGGGTACCTATTATTCAGCTTGTGGTTGTTC | KpnI | FMDV 3A amplification |
| 3B-Fwd | GCGAAGCTTCCACCATGGATTACAAGGATGACGACGATAAGGGACCCTACACCGGT | HindIII (FLAG) | FMDV 3B amplification |
| 3B-Rev | GCGGGTACCTATTACTCAGTGACAATCAAG | KpnI | |
| 3C-Fwd | GCGAAGCTTCCACCATGGCTAGTGGTGCTCCCCCGACT | HindIII | FMDV 3C amplification |
| 3C-H46Y-Fwd | TACCTTGTTCCTCGTTATCTTTTCGCAGAGAAG | Site 46 of 3C mutagenesis | |
| 3C-H46Y-Rev | CTTCTCTGCGAAAAGATAACGAGGAACAAGGTA | ||
| 3C-D84N-Fwd | CAGGACATGCTCTCAAACGCCGCGCTCATGGTG | Site 84 of 3C mutagenesis | |
| 3C-D84N-Rev | CACCATGAGCGCGGCGTTTGAGAGCATGTCCTG | ||
| 3C-C163G-Fwd | ACCAAGGCGGGTTACGGTGGAGGAGCCGTTCTT | Site 163 of 3C mutagenesis | |
| 3C-C163G-Rev | AAGAACGGCTCCTCCACCGTAACCCGCCTTGGT | ||
| 3C-H205R-Fwd | CTTAAAATGAAGGCACGCATCGATCCCGAACCA | Site 205 of 3C mutagenesis | |
| 3C-H205R-Fwd | TGGTTCGGGATCGATGCGTGCCTTCATTTTAAG | ||
| KPNA1-Fwd | GGCGGATCCACCATGGATTACAAGGATGACGACGATAAGATGACCACCCCAGGAAA | BamHI (FLAG) | KPNA1 amplification |
| KPNA1-Rev | GCGCTCGAGTCAAAGCTGGAAACCT | XhoI | |
| KPNA2-Fwd | GGCGGATCCACCATGGATTACAAGGATGACGACGATAAGATGTCCACCAACGAGA | BamHI (FLAG) | KPNA2 amplification |
| KPNA2-Rev | GCGCTCGAGCTAAAAGTTAAAGGT | XhoI | |
| KPNA3-Fwd | GGCGCGGCCGCCACCATGGATTACAAGGATGACGACGATAAGATGGCCGAGAACCCC | NotI (FLAG) | KPNA3 amplification |
| KPNA3-Rev | GCGCTCGAGTTAAAAATTAAATTC | XhoI | |
| KPNA4-Fwd | GGCGCGGCCGCCACCATGGATTACAAGGATGACGACGATAAGATGGCGGACAACGAG | NotI (FLAG) | KPNA4 amplification |
| KPNA4-Rev | GCGCTCGAGCTAAAACTGGAACCCTTCT | XhoI | |
| GAPDH-Fwd | ATGGCCTTCCGTGTCC | GAPDH amplification | |
| GAPDH-Rev | GTGGGTGTCGCTGTTGA | ||
| ISG15-Fwd | GACAAATGCGACGAACCTC | ISG15 amplification | |
| ISG15-Rev | CCGCTCACTTGCTGCTT | ||
| ISG56-Fwd | CCTCCTTGGGTTCGTCTACA | ISG56 amplification | |
| ISG56-Rev | GGCTGATATCTGGGTGCCTA | ||
| Mx1-Fwd | CCATCGGAATCTTGACG | Mx1 amplification | |
| Mx1-Rev | GCTTCGGACAGGCTCAG | ||
| OAS-Fwd | CTCAAGAGCCTCATCCGC | OAS amplification | |
| OAS-Rev | GACCGTCCGAAATCCCT | ||
| PKR-Fwd | TAACGAGAAGGCGGAGCG | PKR amplification | |
| PKR-Rev | CCATTTGGATGAAAAGGCACT | ||
| FMDV-A-Fwd | TATGGCGCGCCTAATACGACTCACTATAGGTTGAAAGGGGGCGTTAGGG | AscI (T7) | FMDV fragment A amplification |
| FMDV-A-Rev | TGAGCGGCCGCCTCAGGTGTTTTG | NotI | |
| FMDV-Am-Fwd | ACTGGCGAGTGCTAGC*AACAGCACTGTTGCT | NheI | NheI site mutagenesis |
| FMDV-Am-Rev | AGCAACAGTGCTGTTG*CTAGCACTCGCCAGT | NheI | |
| FMDV-B-Fwd | GAGGCGGCCGCTCACTGCATTCAT | NotI | FMDV fragment B amplification |
| FMDV-B-Rev | TGTCCCGGGAGAGTTTTCTCTCTGA | XmaI | |
| FMDV-C-Fwd | TCTCCCGGGACACAAGGCGAGTGAT | XmaI | FMDV fragment C amplification |
| FMDV-C-Rev | GCGGCGGCGTTAATTAATTTTTTTTTTTTTTT | PacI |
a
Restriction endonuclease sites are underlined. The Flag or HA tag or T7 promoter is in boldface. The mutated sites are in boldface and underlined. The asterisk refers to the mutated nucleotide of FMDV type O, strain Tibet/CHA/99, sequence.