Table 1.
Primer pairs for site-directed mutagenesis of N-Myc-AAH
| Primer | Sequence (5′→3′) | Primer | Sequence (5′→3′) |
|---|---|---|---|
| S9A (M1)-F | GCGTAAGAATGCCAAGGCC | S115A (M11)-F | CTTAAAGAGAGATCTACTGC |
| AGCGGCAACAGCAGCAG | AGAGCCAGCAGTCC | ||
| S9A (M1)-R | CTGCTGCTGTTGCCGCTGGC | S115A (M11)-R | GGACTGCTGGCTCTGCAGTAG |
| CTTGGCATTCTTACGC | ATCTCTCTTTAAG | ||
| S10A (M2)-F | GTAAGAATGCCAAGAGCGCC | S198A (M12)-F | GTAGATGATAGATTTGAGGCC |
| GGCAACAGCAGCAGCAG | CTGGAACCTGAAG | ||
| S10A (M2)-R | CTGCTGCTGCTGTTGCCGGC | S198A (M12)-R | CTTCAGGTTCCAGGGCCTC |
| GCTCTTGGCATTCTTAC | AAATCTATCATCTAC | ||
| S14A (M3)-F | GAGCAGCGGCAACAGCGCC | T340A (M13)-F | TAAATAAATTTGATAAGGC |
| AGCAGCGGCTCCGGCAG | TATTAAAGCTGAACTTG | ||
| S14A (M3)-R | CTGCCGGAGCCGCTGCTGGC | T340A (M13)-R | CAAGTTCAGCTTTAATAGCC |
| GCTGTTGCCGCTGCTC | TTATCAAATTTATTTA | ||
| S16A (M4)-F | GGCAACAGCAGCAGCGCCG | S374A (M14)-F | GCAAATACCCTCAGGCTCCAC |
| GCTCCGGCAGCGG | GAGCAAGATATG | ||
| S16A (M4)-R | CCGCTGCCGGAGCCGGCGCT | S374A (M14)-R | CATATCTTGCTCGTGGAGCCT |
| GCTGCTGTTGCC | GAGGGTATTTGC | ||
| S18A (M5)-F | CAGCAGCAGCGGCGCCGGC | S427A (M15)-F | GTTTGAAGCGTCGCGCAGACA |
| AGCGGTAGCAC | GGCAACAATTTC | ||
| S18A (M5)-R | GTGCTACCGCTGCCGGCGCC | S427A (M15)-R | GAAATTGTTGCCTGTCTGCG |
| GCTGCTGCTG | CGACGCTTCAAAC | ||
| S20A (M6)-F | GCAGCGGCTCCGGCGCCGGT | T484A (M16)-F | GAGGTGCTGAGTGTGGCACCT |
| AGCACGAGTGC | AATGATGGCTTTG | ||
| S20A (M6)-R | GCACTCGTGCTACCGGCGCC | T484A (M16)-R | CAAAGCCATCATTAGGTGCCA |
| GGAGCCGCTGC | CACTCAGCACCTC | ||
| S24A (M7)-F | GGCAGCGGTAGCGCGAGTG | T577A (M17)-F | CACAGCCTTGGTGGGCCCCAA |
| CGGGCAGCAG | AAGAAACGG | ||
| S24A (M7)-R | CTGCTGCCCGCACTCGCGCT | T577A (M17)-R | CCGTTTCTTTTGGGGCCCACC |
| ACCGCTGCC | AAGGCTGTG | ||
| T23A (M8)-F | GCAGCGGTAGCACGGCTGC | T748A (M18)-F | GCATCCGGAACTGGCACCACA |
| GGGCAGCAGCAG | GCAGAGAC | ||
| T23A (M8)-R | CTGCTGCTGCCCGCAGCCGT | T748A (M18)-R | GTCTCTGCTGTGGTGCCAGTT |
| GCTACCGCTGC | CCGGATGC | ||
| S29A (M9)-F | GGGCAGCAGCGCCCCCGGG | H675Q (M19)-F | CATGCACCCCGGGACTCAGGT |
| GCCCG | GTGGCCGCACACAGG | ||
| S29A (M9)-R | CGGGCCCCGGGGGCGCTGCT | H675Q (M19)-R | CCTGTGTGCGGCCACACCTGA |
| GCCC | GTCCCGGGGTGCATG | ||
| T36A (M10)-F | GCCCGGAGAGAGGCAAAGC | H675R (M20)-F | CATGCACCCCGGGACTCGCGT |
| ATGGAGGAC | GTGGCCGCACACAGG | ||
| T36A (M10)-R | GTCCTCCATGCTTTGCCTCTC | H675R (M20)-R | CCTGTGTGCGGCCACACGCGA |
| TCCGGGC | GTCCCGGGGTGCATG |
Ser/Thr residues on N-Myc-AAH predicted to be phosphorylated were mutated to Ala by site-directed mutagenesis using forward and reverse primer pairs containing a mutated nucleotide sequence (under-lined).