Primer sets used for preparation of monkey and rabbit AOX1 mutant cDNA
| Mutant | Amino Acid Substitutions | Primer Sequences | |
|---|---|---|---|
| Monkey | |||
| 1 | K1004Q | Forward | 5′-cggagaattactggaagcagaaaggactggccatg-3′ |
| Reverse | 5′-catggccagtcctttctgcttccagtaattctccg-3′ | ||
| 2 | M1009I | Forward | 5′-aagaagaaaggactggccatagtccccctgaag-3′ |
| Reverse | 5′-cttcagggggactatggccagtcctttcttctt-3′ | ||
| 3 | A1023Y | Forward | 5′-ccttggctcacgcgcttatggtcaggctgc-3′ |
| Reverse | 5′-gcagcctgaccataagcgcgtgagccaagg-3′ | ||
| 4 | I1032V | Forward | 5′-ctgctgccttggttcacgtttatcttgacggctct-3′ |
| Reverse | 5′-agagccgtcaagataaacgtgaaccaaggcagcag-3′ | ||
| 5 | 1 + K1005R | Forward | 5′-cggagaattactggaagcagagaggactggccatggtcc-3′ |
| Reverse | 5′-ggaccatggccagtcctctctgcttccagtaattctccg-3′ | ||
| 6 | 5 + (M1009I, V1010I) | Forward | 5′-gaagcagagaggactggccataatccccctgaagt-3′ |
| Reverse | 5′-acttcagggggattatggccagtcctctctgcttc-3′ | ||
| 7 | 6 + (R1021V, A1023Y) | Forward | 5′-agtatcctgttggccttggctcagtcgcttatggtcaggctgc-3′ |
| Reverse | 5′-gcagcctgaccataagcgactgagccaaggccaacaggatact-3′ | ||
| 8 | 7 + I1032V | Forward | 5′-ctgctgccttggttcacgtttatcttgacggctct-3′ |
| Reverse | 5′-agagccgtcaagataaacgtgaaccaaggcagcag-3′ | ||
| 9 | 8 + (G1064K, I1067M) | Forward | 5′-caggtggtcagccgtgaattaaaaatgccaatgtcgaatgtccacct-3′ |
| Reverse | 5′-aggtggacattcgacattggcatttttaattcacggctgaccacctg-3′ | ||
| 10 | A1083T, V1085A | Forward | 5′-gaacaagcacagaaaccgtccctaatacgaatgcctctggaggttctg-3′ |
| Reverse | 5′-cagaacctccagaggcattcgtattagggagcgtttctgtgcttgttc-3′ | ||
| 11 | A1083T | Forward | 5′-cacagaaaccgtccctaatacgaatgtctctggaggttct-3′ |
| Reverse | 5′-agaacctccagagacattcgtattagggacggtttctgtg-3′ | ||
| 12 | V1085A | Forward | 5′-cgtccctaatgcaaatgcctctggaggttctgtgg-3′ |
| Reverse | 5′-ccacagaacctccagaggcatttgcattagggacg-3′ | ||
| Rabbit | |||
| 13 | A1081V | Forward | 5′-ctgtccctaatactaatgtctctggaggatctgtgg-3′ |
| Reverse | 5′-ccacagatcctccagagacattagtattagggacag-3′ |