TABLE 1.
PCR primers
| Method | Targeted site | Primer sequence (5′–3′) |
|
|---|---|---|---|
| Forward | Reverse | ||
| Mutagenesis | Nsp1-F44A | TTTATGCAATGCCGTgcCGTGTCCTTCGATCT | AGATCGAAGGACACGgcACGGCATTGCATAAA |
| Nsp15-H226A | GATTACGGCTTTGAGgcCGTTGTGTATGGTGAT | ATCACCATACACAACGgcCTCAAAGCCGTAATC | |
| Nsp15-H241A | ACCCTTGGTGGTTTGgccCTACTAATTTCGCAG | CTGCGAAATTAGTAGggcCAAACCACCAAGGGT | |
| Nsp16-D129A | GATTTAGTTATATCTGcaATGTATGATGGTAAG | CTTACCATCATACATtgCAGATATAACTAAATC | |
| Mutation stability RT-PCR | Nsp1: PEDV-210F/-627R | AGACAAACAGCCTTCCTCCG | ATGTTACCACCACGACGACC |
| Nsp15: PEDV-19071F/-19731R | CGACTTTGAGGGTGACGTCT | CCACAACATCCACCTCCACA | |
| Nsp15: PEDV-18319F/-19712R | CCAGAATTTTCTGTGGTCTGTCG | GTCTGGAGTTTATGATCCTTACACCAC | |
| Nsp16: PEDV-19923F/-19712R | TGCGTGTGCTACATCTTGGT | GTCAATCACAGCGCCACTTG | |
| Mutation sequencing | Nsp1: PEDV-235F/-549R | GTCTGGGGGTTGTGTGGATA | CAAATGTTTTTGGGGCGGCT |
| Nsp15: PEDV-19218F/-19637R | TGGTGTCCCAGTTAACACACA | TCGTCAAGCAGGAGATCCAT | |
| Nsp16: PEDV-20028F/-20345R | GTGTGGATTACGTTAGCGATGC | AGAATGCCTCTGACGATGACG | |
a
Underlined sequence characters represent the codon of mutated amino acid; lowercase sequence characters represent the mutated nucleotides.