Table 1.
Oligonucleotides used for cloning and mutagenesis
| Primer | Direction | Mutation | Sequencea, b |
|---|---|---|---|
| 1 | Forward | N/A | 5'-TATTCTTATCATATGAAAAAGTTTCCCGAAGGGTTC |
| 2 | Reverse | N/A | 5'-TATTCTTATCTCGAGATCTGTTAGTCCGTTGTTTTTG |
| 3 | Forward | N/A | 5'-AATACGACTCACTATAGG |
| 4 | Reverse | N/A | 5'-CTAGTTATTGCTCAGCGG |
| 5 | Forward | F219L | 5'GACGGAAAGATAGGGATTGTTTTAAACAACGGATACTTCGA |
| 6 | Forward | N221S | 5'-GGGATTGTTTTCAACAGCGGATACTTCGAACCTGC |
| 7 | Forward | G222M | 5'-GATTGTTTTCAACAACATGTACTTCGAACCTGCAAG |
| 8 | Forward | G222Q | 5'- GGATTGTTTTCAACAACCAATACTTCGAACCTGCAAGTGAGAGAG |
| 9 | Reverse | F219L | 5'- GACGGAAAGATAGGGATTGTTTTAAACAACGGATACTTCGA |
| 10 | Reverse | N221S | 5'- GCAGGTTCGAAGTATCCGCTGTTGAAAACAATCCC |
| 11 | Reverse | G222M | 5'-CTCTCTCACTTGCAGTACCGAAGTATTGGTTGTTGAA |
| 12 | Reverse | G222Q | 5'- CTCTCTCACTTGCAGGTTCGAAGTATTGGTTGTTGAAAACAATCC |
aThe restriction sites for cloning are underlined. bInduced changes are shown in bold.