Table 2.
Site-directed mutagenesis primers.
| Primers | Sequences (5′ to 3′) | Digestion site |
|---|---|---|
| SSA1 promoter-f | aag-CTGCAG-agtattgtagaataaattattcagc | PstI (CTGCAG) |
| SSA1 promoter-r | aag-GTCGAC-aatttaattttttgtttaattgtgtttt | SalI (GTCGAC) |
| SSA2 CDS + 582 bp UTR-f | aag-GTCGAC-atgtctaaagctgttggtattga | SalI (GTCGAC) |
| SSA2 CDS + 582 bp UTR-r | aag-GGATCC-atccaatcataatccagtaaattg | BamHI (GGATCC) |
| Ssa2128−132A3 | gatcttgggtaaaatgaaggaagccgCTGCAGgtgccttgggtaccac | PstI (CTGCAG)* |
| Ssa2131−135A3 | aaatgaaggaaaccgctgAAGCTTtcgcgggtgccactgttaaagatgctgttgtc | HindIII (AAGCTT)* |
| Ssa2150−156A4 | actgtcccagcttacttcaatgcttccgcaagagCAGCTGccaaagatgctggtaccattgc | PvuII (CAGCTG)* |
| Ssa2334−338A3 | caaatctaaagttgatgaaattgCCTTGGctggtgcttctaccagaattccaaaggttc | StyI (CCTTGG)* |
Sequences of sense mutagenesis primers for each construct are shown. Each PstI digestion site (CTGCAG), SalI digestion site (GTCGAC) and BamHI digestion site (GGATCC) in the template primers are capitalized. All mutated nucleotides are indicated by bold underline. New digestion sites (*) generated by mutation and used for rapid colony selection are indicated by capitals and italics. UTR, 3′ untranslated regions.