Table 1.
Oligonucleotides used in this study
| Oligonucleotidea | Polarity | Sequence (5′→3′) | Nt binding region |
|---|---|---|---|
| 3′UTRPSEU(+) | − | CTGCAAGTCATCCATTCTGATAGAGAGTG | nts 63–91 in BCoV 3′ UTRb |
| 5RAAP(−)c | (+) | GGCCACGCGTCGACTAGTACGGGGGGGGGG | 3′ poly(C) on cDNA |
| Ada3(+) | − | GGAATTCTCGAGCTCAAGCTTTTTTTTTTTTTTTT | 3′ poly(A) tail |
| DI3′(+) | − | CGGGATCCGTCGACACGCGTTTTTTTTTTTTTTTTTTTT | 3′ poly (A) tail |
| DI420(−) | + | GTTGTGTGCAGTCTAGCCTAATAC | nts 419–442 in pDrep1 |
| HpaI(+) | − | GGCTGAAAGCTGTTAACAGCAGAAATG | nts 140–166 in pDrep1 |
| Ia1096(+) | − | CGCACAACGTGCCATGCCAC | nts 1096–1215 in BCoV genomed |
| Leader(−) | + | GAGCGATTTGCGTGCGTGCATCCCGC | nts 7–32 in pDrep1 |
| NdeI(+) | − | CCTCCAAATCATATGGACGTGTATTC | nts 456–481 in pDrep1 |
| Pst(−) | + | GAGAGTTGACGCAGAGGAC | nts 1799–1818 in pDrep1 |
| TGEV8(+) | − | CATGGCACCATCCTTGGCAACCCAGA | nts 1098–1123 in pDrep1 |
a
The positive and negative symbols in the oligonucleotide names indicate the polarity of the nucleic acid to which the oligonucleotide anneals.
b
The sequence in nts 63–91 in BCoV is common to all the group 2 coronaviruses examined in this study (note Fig. 3).
c
Supplied with the RACE kit (Invitrogen).
d
From BCoV-Mebus genome sequence as determined by K. Nixon and D. Brian (GenBank Accession No. U00735).