Table II.
Primers for wild-type and mutant-type MRE in porcine IFN-β 3′UTR.
| Primera | Primer sequence (5′→3′)b |
|---|---|
| let-7b MRE-F | AAACTAGCGGCCGCTAGTATGTATTTAATTTTTTACCTTGT |
| let-7b MRE-R | CTAGACAAGGTAAAAAATTAAATACATACTAGCGGCCGCTAGTTT |
| let-7b Mut-F | AAACTAGCGGCCGCTAGTATGTATTTAATTTTGATGGAGGT |
| let-7b Mut-R | CTAGACCTCCATCAAAATTAAATACATACTAGCGGCCGCTAGTTT |
| miR-26a MRE-F | AAACTAGCGGCCGCTAGTCCCCTGTGGCTCTGGGAATTGACT |
| miR-26a MRE-R | CTAGAGTCAATTCCCAGAGCCACAGGGGACTAGCGGCCGCTAGTTT |
| miR-26a Mut-F | AAACTAGCGGCCGCTAGTCCCCTGTGGCTCTGGGTGAACTCT |
| miR-26a Mut-R | CTAGAGAGTTCACCCAGAGCCACAGGGGACTAGCGGCCGCTAGTTT |
| miR-34a MRE-F | AAACTAGCGGCCGCTAGTTTGACCATGTTGGCAATGATGT |
| miR-34a MRE-R | CTAGACATCATTGCCAACATGGTCAAACTAGCGGCCGCTAGTTT |
| miR-34a Mut-F | AAACTAGCGGCCGCTAGTTTGACCATGTTGGGTGACGGGT |
| miR-34a Mut-R | CTAGACCCGTCACCCAACATGGTCAAACTAGCGGCCGCTAGTTT |
| miR-145 MRE-F | AAACTAGCGGCCGCTAGTCTGTGGCTCTGGGAATTGACCT |
| miR-145 MRE-R | CTAGAGGTCAATTCCCAGAGCCACAGACTAGCGGCCGCTAGTTT |
| miR-145 Mut-F | AAACTAGCGGCCGCTAGTCTGTGGCTCTGGGTTGACCTCT |
| miR-145 Mut-R | CTAGAGAGGTCAACCCAGAGCCACAGACTAGCGGCCGCTAGTTT |
a
F and R, primer pairs were annealed and inserted into pmirGLO dual-luciferase miRNA target expression vector digested with PmeI and XbaI
b
NotI restriction endonuclease site is underlined; MRE, miRNA recognition elements; IFN-β, interferon-β; 3′UTR, 3′ untranslated region.