Table 1.
Primers used for 3′-, 5′-RACE, ORF cloning, and plasmid construction
| Gene | Direction | Primer sequences |
|---|---|---|
| 3′-RACE | ||
| Universal | ||
| RT primer | Reverse | 5′-GCTGTCAACGATACGCTACGTAACGGCATGACAGTG(T)18-3′ |
| 1st PCR | Reverse | 5′-GCTGTCAACGATACGCTACGTAACG-3′ |
| 2nd PCR | Reverse | 5′-CGCTACGTAACGGCATGACAGTG-3′ |
| MT1 | ||
| 1st PCR | Forward | 5′-AAACCGGACAGCAAACCCAAACT-3′ |
| 2nd PCR | Forward | 5′-GGGCCCCACTCAACTTCATAGGTC-3′ |
| MT2 | ||
| 1st PCR | Forward | 5′-CCGAAGGAAGGCAAAGGCTGAGAG-3′ |
| 2nd PCR | Forward | 5′-CCAGAAGGGCTTTTTGTCACCAG-3′ |
| 5′-RACE | ||
| MT1 | ||
| RT primer | Reverse | 5′-pAGTAACTAGCCACGAAGA-3′ |
| 1st PCR | Forward | 5′-AAACCGGACAGCAAACCCAAACT-3′ |
| Reverse | 5′-GCCCAGGATGTCCACCACGATAGT-3′ | |
| 2nd PCR | Forward | 5′-GGGCCCCACTCAACTTCATAGGTC-3′ |
| Reverse | 5′-ACGACGGCCGCGATCTTATTTC-3′ | |
| MT2 | ||
| RT primer | Reverse | 5′-pATTAAGGCAGCTGTTGAA-3′ |
| 1st PCR | Forward | 5′-CCGAAGGAAGGCAAAGGCTGAGAG-3′ |
| Reverse | 5′-GGTGCTGGCTGTCTGGATGAAGGT-3′ | |
| 2nd PCR | Forward | 5′-CCAGAAGGGCTTTTTGTCACCAG-3′ |
| Reverse | 5′-TGACTGCAGGCTCGGTGGTAGGT-3′ | |
| ORF cloning | ||
| MT1 | Forward | 5′-GCGCGGGGCTACAGGATGATG-3′ |
| Reverse | 5′-CAGGAGGTCCGCTCCAACACTATG-3′ | |
| MT2 | Forward | 5′-GGAGCGCCCCCAAGCAGT-3′ |
| Reverse | 5′-AGGTCAAGGTGGCAGGGAAAATG-3′ | |
| Plasmid construction | ||
| MT1 | Forward | 5′-CACCATGAAGGGCAATGTCAGC-3′ |
| Reverse | 5′-TTAAACAGAGTCCACCTTTATTAAATT-3′ | |
| MT2 | Forward | 5′-CACCATGCCTGACAACAGCTCCAT-3′ |
| Reverse | 5′-CTAGAGAGCACCTTCCTGGACAG-3′ | |