TABLE 1.
Names, sequences, polarities, and restriction enzyme sites of primers used in this study to generate all baculovirus recombinants for expression in insect cells and the GFP constructs for expression in mammalian cells
| Name | Sequence (5′ to 3′)a | Sequence locationa | Sense | Restriction enzyme site | Constructs generated |
|---|---|---|---|---|---|
| NV132 | GGCCTGATCAATGATGATGGCGTCTAAGGAC | 5357-5385 | + | BclI | ORF2, ORF2+3, ORF2+3+3′UTR, ORF2+3′UTR |
| NV-3′UTR-Xho | CCCGCTCGAGAACATCAAATTAAACCTAAT | 7624-7637 | − | XhoI | ORF2+3+3′UTR, ORF3+3′UTR |
| NV133 | CCCGCTCGAGCATCGCCTATTATTTGCGAAT | 7687-7666 | − | XhoI | ORF2+3 |
| Luc-5′ | CCCGCTGGATCCATGACTTCGAAAGTTTATGATCCA | 1-24 | + | BamHI | Luc-Fast, Luc-3′UTR-Fast |
| Luc-3′ | CCCGCTCGAGGAATTATTGTTCATTTTTGAGAACTC | 745-775 | − | XhoI | Luc-Fast |
| NV131 | CCCGCTCGAGTTATCGGCGCAGACCAAGCCT | 6928-6950 | − | XhoI | ORF2 |
| ORF2+3′UTR | CCCGCTCGAGAACATCAAATTAAACCTAATTAAACCTAA TTAAATGAATATGATGCCCACATTTCATATTACAACATT ATCGGCGCAGACCAAGCCT | 6929-6952/7611-7654 | − | XhoI | ORF2+3′UTR |
| Luc-3′UTR NV | CCCGCTCGAGAACATCAAATTAAACCTAATTAAACCTAA TTAAATGAATATGATGCCCACATTTCATATTACAACAGA ATTATTGTTCATTTTTGAGAACTCGCT | 7611-7654/745-775 | − | XhoI | Luc-3′UTR-Fast |
| AUG-ORF3-5 | GGCTTGGTCTGCGCCGATAAGGATCCATGGCCCAAGCC ATAATTG | 6929-6970 | + | None | Site-directed mutagenesis-AUG-ORF3 |
| AUG-ORF3-3 | GCACCAATTATGGCTTGGGCCGTTATCGGCGCAGACCA AGCC | 6929-6973 | − | None | Site-directed mutagenesis-AUG-ORF3 |
| GFP-NV-5 | GGGCCTCGAGGCGCGATGATGATGGCGTCTAAGGAC | 5357-5385 | + | XhoI | GFP-ORF2+3+3′UTR NV |
| GFP-NV-3 | CCCGGGAACATCAAATTAAACCTAATT | 7637-7654 | − | XmaI | GFP-ORF2+3+3′UTR NV |
| ORF3-5-BamHI | GCGGGGGGATCCATGATGATGGCCCAAGCCATAA TTGGT | 6950-6973 | + | BamHI | ORF3+3′UTR |
a
Sequence locations for each primer on Norwalk sequence (gi9630803) and Renilla luciferase (shown in italics [gi28864243]).