Table 1 Overlapping primers used for generating the specified mutation sites in hRFVT-2
Construct | Forward and reverse primers (5’-3’) |
|---|---|
GFP-hRFVT-2[F119A] | TTAGCACTGGCCGCTGTGCTGGCACTG; CAGTGCCAGCACAGCGGCCAGTGCTAA |
GFP-hRFVT-2[V120A] | GCACTGGCCTTTGCGCTGGCACTGGCA; TGCCAGTGCCAGCGCAAAGGCCAGTGC |
GFP-hRFVT-2[L121A] | CTGGCCTTTGTGGCGGCACTGGCATGC; GCATGCCAGTGCCGCCACAAAGGCCAG |
GFP-hRFVT-2[L123P] | TTTGTGCTGGCACCGGCATGCTGTGCC; GGCACAGCATGCCGGTGCCAGCACAAA |
GFP-hRFVT-2[L339P] | TCCTTGGCAGGGCCGGGCGGCCTCTCT; AGAGAGGCCGCCCGGCCCTGCCAAGGA |
GFP-hRFVT-2[L342A] | GGGCTGGGCGGCGCCTCTCTGCTGGGC; GCCCAGCAGAGAGGCGCCGCCCAGCCC |
Real-Time PCR primers | Â |
hRFVT-2 | CCCTGGTCCAGACCCTA; ACACCCATGGCCAGGA |
β-Actin | AGCCAGACCGTCTCCTTGTA; TAGAGAGGGCCCACCACAC |