TABLE 1.
Primers and oligonucleotides used in this study
| Primer or oligonucleotide | Sequence (5′–3′)a |
|---|---|
| Construction of SFV proteins | |
| pAc-v5/His-NS1-sense | GAATTCTATGGCCGCCAAAGTGCATGTTGATAT (EcoRI) |
| pAc-v5/His-NS1-anti | CTCGAGCGTGCACCTGCGTGATACTCTAGTTC (XhoI) |
| pAc-v5/His-NS2-sense | GCGGCCGCATGGGGGTCGTGGAAACACCTCGCA (NotI) |
| pAc-v5/His-NS2-anti | TCTAGACTACACCCGGCCGTGTGCATGGCTTCTC (XbalI) |
| pAc-v5/His-NS3-sense | GAATTCTATGGCACCATCCTACAGAGTTAAGAGAGC (EcoRI) |
| pAc-v5/His-NS3-anti | TCTAGACTTGCACCCGCGCGGCCTAGTCGCAGGAC (XbalI) |
| pAc-v5/His-NS4-sense | GAATTCTATGTATATTTTCTCCTCGGACACACTGGCA (EcoRI) |
| pAc-v5/His-NS4-anti | CTCGAGCGACGCACCAATCTAGGACCGCCGTAGAG (XhoI) |
| pAc-v5/His-capsid-sense | GCGGCCGCATGAATTACATCCCTACGCAAACGTTTT (NotI) |
| pAc-v5/His-capsid-anti | CTCGAGCGCCACTCTTCGGACCCCTCGGGGGTCAC (XhoI) |
| pAc-v5/His-E3-sense | GAATTCTATGTCCGCCCCGCTGATTACTGCCATGT (EcoRI) |
| pAc-v5/His-E3-anti | TCTAGAGCGCCGGTGTCTTGTTCCGTTTCGGCACG (XbalI) |
| pAc-v5/His-E2-sense | GCGGCCGCATGAGCGTGTCGCAACACTTCAACGTG (NotI) |
| pAc-v5/His-E2-anti | CTCGAGCGTGCGTGCGCCCGCGGGGCGCAGCAGAG (XhoI) |
| pAc-v5/His-6k-sense | GCGGCCGCCCATGGCTAGTGTGGCAGAGACTATGGC (NotI) |
| pAc-v5/His-6k-anti | CTCGAGCGAGCTCTGGCGGTTGCCCCGAGGCTCAGT (XhoI) |
| pAc-v5/His-E1-sense | GCGGCCGCATGTACGAACATTCGACAGTAATGCCGA (NotI) |
| pAc-v5/His-E1-anti | CTCGAGCGTCTGCGGAGCCCAATGCAAGTGACCACA (XhoI) |
| pRK-capsid-sense | GTCGACGATGAATTACATCCCTACGCAAACGTTTTAC (SalI) |
| pRK-capsid-anti | GCGGCCGCTTACCACTCTTCGGACCCCTCGGGGGTCA (NotI) |
| pMal-c2x-capsid-sense | GAATTCATGAATTACATCCCTACGCAAACGTTTTAC (EcoRI) |
| pMal-c2x-capsid-anti | GTCGACCCACTCTTCGGACCCCTCGGGGGTCACTC (SalI) |
| Construction of capsid mutants and SFV mutant | |
| K124A/K128A-sense | TCTTCGAAGTCGCACACGAAGGAGCGGTCACTGG (–) |
| K124A/K128A-anti | CCAGTGACCGCTCCTTCGTGTGCGACTTCGAAGA (–) |
| K139A/K142A-sense | GCCTGGTGGGCGACGCAGTCATGGCACCTGCCCA (–) |
| K139A/K142A-anti | TGGGCAGGTGCCATGACTGCGTCGCCCACCAGGC (–) |
| pMD18-T-K124A/K128A-sense | TCTAGAGACGGACATTGCATCATTC (XbalI) |
| pMD18-T-K124A/K128A-anti | CCTGCAGGAATTCACCCGGTGGGCA (SbfI) |
| qRT-PCR | |
| EGFP-sense | GACAAGCAGAAGAACGGCATC |
| EGFP-anti | CGGACTGGGTGCTCAGGTA |
| SFV-sense | CCGGAGGACGCACAGAAGTTG |
| SFV-anti | TGCGACGGCCACAATCGGAAG |
| Human actin-sense | AGAGCTACGAGCTGCCTGAC |
| Human actin-anti | AGCACTGTGTTGGCGTACAG |
| Aedes aegypti Rp49-sense | GCTATGACAAGCTTGCCCCCA |
| Aedes aegypti Rp49-anti | TCATCAGCACCTCCAGCT |
| Amplification of templates for in vitro transcription dsRNAs and ssRNAs | |
| EGFP-ds400-sense | TAATACGACTCACTATAGATGGTGAGCTAGGGCGAGGA |
| EGFP-ds400-anti | TAATACGACTCACTATAGGCTTGTGCCCCAGGATGTTGC |
| EGFP-ds200-sense | TAATACGACTCACTATAGATGGTGAGCTAGGGCGAGGA |
| EGFP-ds200-anti | TAATACGACTCACTATAGGCGGCTGAAGCACTGCACGC |
| dsAgo2–sense | TAATACGACTCACTATAGATGGGAAAAAAAGATAAGAACAAGC |
| dsAgo2–anti | TAATACGACTCACTATAGCACCTTGTTGACCTTGTTTAGTCCAC |
| EGFP-ss200-sense | TAATACGACTCACTATAGATGGTGAGCTAGGGCGAGGA |
| EGFP-ss200-anti | CTATAGTGAGTCGTATTAGCGGCTGAAGCACTGCACGC |
| Amplification of templates for in vitro transcription RNA probes | |
| EGFP-probe-sense | GATCCGCCACAACATCGAGGACGGC |
| EGFP-probe-anti | TAATACGACTCACTATAGTTACTTGTACAGCTCGTCCATGCC |
| S2-Rp49-sense | CCGTGAATACTGTGGTGAAATTGCC |
| S2-Rp49-anti | TAATACGACTCACTATAGGTTTTTTTTTCACTTTTAACGTTTCA |
| GAPDH-probe-sense | GGCGTGATGGCCGCGGGGCTCTCC |
| GAPDH-probe-anti | TAATACGACTCACTATAGGCAAAGGTGGAGGAGTGGGTGTCGC |
| SFV-probe-sense | GCTGCCAAATCAAAACGAACCCTGTCAG |
| SFV-probe-anti | TAATACGACTCACTATAGGTCTGCGGAGCCCAATGCAAGTGACC |
| FHV-probe-sense | ATGCCAAGCAAACTCGCGCTAATCCAGGAACTTCCCG |
| FHV-probe-anti | TAATACGACTCACTATAGGCCTCTAGGTATGCCACCACGCTGGGTTTCTC |
| siEGFP-sense | GCUGACCCUGAAGUUCAUCUU |
| siEGFP-anti | GAUGAACUUCAGGGUCAGCUU |
| SFV-vsiRNA-probe-sense | TAATACGACTCACTATAGATGGCGGATGTGTGACATACACGACGCCAAAAGATTTTGTTCCAGCTCCT |
| SFV-vsiRNA-probe-anti | AGGAGCTGGAACAAAATCTTTTGGCGTCGTGTATGTCACACATCCGCCATCTATAGTGAGTCGTATTA |
| EGFP-siRNA Probe | DIG-GCAAGCUGACCCUGAAGUUCAUC-DIG |
| U6-Probe | DIG-GCAGGGGCCAUGCUAAUCUUCU-DIG |
a
Restriction enzymes are indicated in parentheses where applicable. –, No restriction enzyme. DIG, digoxigenin labeled.