TABLE 1.
Primers used for PCR amplifications and mutagenesis
| Primer | Sequence (5′–3′)a | ORF1a positionb |
|---|---|---|
| Cloning primers | ||
| PLP NS | TCGATCGCATATGTCTATCTTGGATGAGCTTCAA | 3393–3413 |
| PLP CA | GAGTCTAGATAACTTTTCAGCTATAGCACCTGC | 4301–4281 |
| PLP-2 FCP | GAAGGAGATCGCGGATCCTTTGATGAACCACAACTGCTG | 5250–5270 |
| PLP-2 RCP | GCCCGCCTGCAGAAGCTTCTACGATAAATCTGGCTTATA | 6017–6000 |
| Mutagenesis primers | ||
| FMPH1272P | GATGTTAATGATTGTCCCTCTATGGCTGTAGTAGAGGG | 4008–4045 |
| RMPH1272P | CCCTCTACTACAGCCATAGAGGGACAATCATTAACATC | 4045–4008 |
| FMPH1873P | GGTGGTAGTGTGGGCCCTTACACGCATGTGAAATG | 5811–5843 |
| RMPH1873P | CATTTCACATGCGTGTAAGGGCCCACACTACCACC | 5843–5811 |
| FMPD904G | CCTTGTAAGGAGCATGGTGTGATAGGCACAAAAG | 2904–2937 |
| RMPD904G | CTTTTGTGCCTATCACACCATGCTCCTTACAAGG | 2937–2904 |
a
Restriction sites introduced into primers are shown in boldface; mutated codons are underlined.
b
Reverse position of ORF1a indicates a negative-strand primer.