Table 3. Primers used for RT-PCR amplification and sequencing of the M RNA of the TSWV isolates
|
Primer name |
Forward or reverse |
Sequence |
Annealing temperature, ° C |
|
1 |
F |
AGAGCAATCAGTGCATCAGAAATATACCTATTATACA |
55 |
|
1 |
R |
CACTACCAAAAGAAACCCC |
|
|
2 |
F |
GAAGATGAACAACACCCC |
40 |
|
3 |
F |
TCAGTTGAAGAGGAAGA |
41 |
|
3 |
R |
TATGTTAATGAAAGATACAA |
|
|
4 |
F |
TGAGAGAAATCCATAGG |
55 |
|
4 |
R |
GGTTTAGAGCAAATATCAG |
|
|
5 |
F |
CCGCATAGAAGACAGCC |
55 |
|
5 |
R |
TACAGGAAACTGCGACAC |
|
|
6 |
F |
AGAAGTTGGTGTTACA |
46 |
|
6 |
R |
GATGCTTTTATGTTCCAGG |
|
|
7 |
F |
GTGCCAAAGATACTCTCTATG |
46 |
|
7 |
R |
CTGAGGAAATGTTGGATGG |
|
|
8 |
F |
GCAATCTCTGACTCTTT |
52 |
|
8 |
R |
ATGATGATTCTGCTGAG |
|
|
9 |
F |
GTATCTGACGGGTTCCAGG |
55 |
|
9 |
R |
AGAGCAATCAGTGCAAACAAAAACCTTAATCC |
The PCR primers were chosen from a published sequence of Tomato spotted wilt virus (TSWV) isolate BR-01 (1), except primer 3 (2). Amplification of IGR (primer 3) was conducted as described (2).
1. Kormelink, R., De Haan, P., Meurs, C., Peters, D. & Goldbach, R. (1992) J. Gen. Virol. 73, 2795-2804.
2. Bhat, A. I., Pappu, S. S., Pappu, H. R., Deom, C. R. & Culbreath, A. K. (1999) Virus Res. 61, 161-170.