TABLE 1.
Oligonucleotides used in the study
| Oligonucleotide | Positionsa | Restriction enzyme site | Sequenceb | Sensec |
|---|---|---|---|---|
| P50 | 4754-4776 | GGAACATTGCAGAAATGCAGCCC | + | |
| P51 | 1420-1443 | BamHI | GCGCCGGATCCAGCGGTGCGAAACTCCGTACTTAC | + |
| P73 | 4623-4646 | PstI | GCCGCCTGCAGCCAATTCATGCTGGCTGGTATTGC | − |
| P9 | 4757-4776 | SphI/SmaI | GGCGGCCCGCATCCCGGGCTGCATTTCTGCAATGTTCC | − |
| PF4 | 1285-1304 | XbaI | GCGCGTCTAGAAGAAACGGGAAGCTCGCTCG | + |
| PF6 | 4398-4417 | PstI | GCGCGCTGCAGAGCGAGTAAGACAGACTCTT | + |
| PF7 | 1-20 | SacI | GGCGGAGCTCTAATACGACTCACTATAGGAAATTCTCCAGGATTTCTC | + |
| PR21 | 4645-4668 | NsiI | CCGCGCATGCATATTCCTGTTTACGAAAGTTAGG | + |
| PR23 | 4398-4420 | PstI | CGCCGCTGCAGAGCGAGTAAGAGCGCCTCTTCAG | + |
| PR31 | 4398-4432 | GAGCGAGTAAGACAGACTCTTCAGTCTGAGTTTGTGCTGCA | + | |
| PR32 | 4398-4432 | GCACAAACTCAGACTGAAGAGTCTGTCTTACTCGCTCTGCA | − | |
| PR33 | 4413-4447 | GCTCTTCAGTCTGAGTTTGTGGAGATGAGTGTAAATCTGCA | + | |
| PR34 | 4413-4447 | GATTTACACTCATCTCCACAAACTCAGACTGAAGAGCTGCA | − | |
| PR35 | 4437-4467 | GGAGATGAGTGTAAATCTGGCATAGCATACAGGTTACTGCA | + | |
| PR36 | 4437-4467 | GTAACCTGTATGCTATGCCAGATTTACACTCATCTCCTGCA | − | |
| PR37 | 4686-4708 | GACCCAGACACGGTTGATCTCAC | + | |
| PR38 | 4727-4739 | GATCGCTGGAAGCACTACCGGAC | + | |
| PR41 | 4398-4432 | PstI | CCGGCCTGCAGAGCGAGTAAGAGCGCCTCTTCAGACGCACTTTGTGCTGCAG | + |
| PR44 | 4398-4435 | PstI | GCCGCCTGCAGAGCGAGTAAGAGCGCCTCTTCAGACGCAGTTTGTGGAG | + |
| PR45 | 4398-4432 | PstI | CCGGCCTGCAGAGCGAGTAAGAGCGCCTCTTCAGTCTGAGTTTGTGCTGCAG | + |
| PR46 | 4398-4432 | PstI | CGCGCCTGCAGAGCGAGTAAGACAGACTCTTCAGACGCACTTTGTGCTGCAG | + |
| PR47 | 4730-4749 | GTCCGGTAGTGCTCCAGCG | − | |
| PR48 | 4407-4426/4647-4668 | PstI | GGCGCCTGCAGGACAGACTCTTCAGTCTGAGATTCCTGTTTACGAAAGTTAGG | + |
| PR58 | 4398-4432/4647-4673 | PstI | CGGCGCTGCAGAGCGAGTAAGACAGACTCTTCAGTCTGAATTAGTGCTGCAGATTCCTG | + |
| PR60 | 4398-4432 | PstI | CCGGCCTGCAGAGCGAGTAAGATAGGCTCTTCAGTCTGAGTTTGTG | + |
| PR61 | 4407-4432/4647-4668 | PstI | CCGGCCTGCAGGACAGACTCTTCAGTCTGAGTTTGTGATTCCTGTTACGAAAGTTAGG | + |
| PR63 | 1426-1445 | GCGAAACTCCGTACTTACAC | + | |
| PR71 | 4386-4417 | BstBI | CGCATGCATCCTGCCTTCGAAAAAGAAAGCGAGTAAG | + |
| PR113 | 4398-4439/4647-4653 | PstI | CCGGCCTGCAGAGCGAGTAAGACAGATTCTTCAGTTTGAGTTTGTGCTGCAGATTCCTG | + |
| PR114 | 4398-4439/4647-4653 | PstI | CCGGCCTGCAGAGCGAGTAAGACAAACTCTTCAATCTGAGTTTGTGCTGCAGATTCCTG | + |
| PR95 | 4411-4432 | XbaI | GGCCGTCTAGACACAAACTCAGACTGAAGAGTC | − |
| PR96 | 4398-4422 | XbaI | GCGCCTCTAGAAGCGAGTAAGACAGACTCTTCAGTC | + |
| PR97 | 4398-4442/161-168 | XbaI | GCGCGTCTAGACACAAACTCAGACTGAAGAGGCGCTCTTACTCGCTTCTAGACATGTCGC | − |
| PR98 | 4399-4432 | XbaI | CCGGCTCTAGACACAAACTCGCGTCTGAAGAGTCTGTCTTACTCGC | − |
| PR99 | 4398-4432/161-168 | XbaI | CGCGGTCTAGACACAAACTCGCGTCTGAAGAGGCGCTCTTACTCGCTTCTAGACATGTCGC | − |
a
Coordinates correspond to those of the TBSV genome (15).
b
Viral sequences corresponding to the coordinates shown are underlined. Restriction enzyme sites are italicized, and individual mutations are in bold.
c
Refers to the sense of the oligonucleotide in reference to the plus-sense viral RNA.