TABLE 1.
Primers used in this study
| Primer | Nucleotide sequence (5′-3′) (restriction enzyme)a | Use |
|---|---|---|
| For cDNA cloning | ||
| A2 | GGTCACATAACGCCCCTATAGCCAT | RT of segment 7R or 7RΔ |
| 7ribozS | ACGGTGGCGGCCGCTAATACGACTCACTATAGGCTTTTAA (NotI) | Full-length PCR of segment 7R or 7RΔ |
| 7ribozR | GACGTCACCGGTCACATAACGCCCCTATAGCCATTTAGGT (AgeI) | Full-length PCR of segment 7R or 7RΔ |
| For site-directed mutagenesis | ||
| 7-mut-Bst-S | AGCTCTATTATTAATACTTCTTTCGAATCTGCAGTCGTTGCTGC (BstBI) | Creation of BstBI site in segment 7R (change T to C) |
| 7-mut-Eco-S | GAATGATATTGAACAGCAGTTGAATTCAATTGATTTAATTAATCCC (EcoRI) | Creation of EcoRI site in segment 7RΔ (change A to G) |
| 7-del-stop-S | AGCAATGCAACTATGAATATGCATATGATTATGCTTTTCAGTGGTTG | Deletion of stop codon in segment 7R |
| For specific detection of rearranged segments 7 | ||
| RPZJ3 | CTAAGTTTATTCACGTCTTCATC | RT and PCR |
| DPZJ3 | GAGTGGTATCTAAGGTCTATGG | PCR |
a
Restriction enzyme sites are underlined. Letters in italics indicate the T7 RNA polymerase promoter sequence, and letters in bold indicate the nucleotides substituted in the gene 7 sequence.