Table 1.
Primers for MiΔ9FAD cloning and heterologous expression.
| Primer | Sequence 5′-3′ | Annealing Temperature/°C | Size of Amplificant/bp |
|---|---|---|---|
| RACE for cDNA cloning | |||
| acp-1 Forward | CTGTCGGCCCACCAGTTAGA | – | – |
| acp-1 Reverse | GCCGTAGATCCAGGAGAAGG | 60.9 | 1002 |
| acp-5-gsp1 | TCCTGGAAGGAGGTGTAGATGAAGC | 66 | – |
| acp-5-gsp2 | GGCAGCACCTGGTCCGTAACATA | 68 | 501 |
| acp-3-gsp1 | CCGCAACCTGTTTTCAGACTTCTCC | 65 | 1230 |
| DNA cloning | |||
| D1-Forward | ACGCGGGGAGTGACAACACCAGCTGT | 69.6 | 812 |
| D1-Reverse | CTCTGAGTTGCCGCTCCTTGTGGGGG | – | – |
| D2-Forward | ATGAACCCGAGTGGGCTCACGATG | 66.3 | 1542 |
| D2-Reverse | GTCCATGCCAGAGCCAATCAGGTT | – | – |
| D3-Forward | ACAGCCGAGGAGAACCGTCATGGT | 68.1 | 1376 |
| D3-Reverse | CTAGCAGTTGACGGTGCGGCCGTAGAT | – | – |
| D4-Forward | GGACGAGACGCCCTTCTCCTGGAT | 64.9 | 1005 |
| D4-Reverse | GCCCTGCAGCGTTTTACAGCGC | – | – |
| Heterologous expression in yeast | |||
| pY-Forward | cgGAATTCATGGCCGCAGAGGCCCCAT 1,2 | 68.5 | 1138 |
| pY-Reverse | gcTCTAGACATCTAGCAGTTGACGGTGC 1,2 | – | – |
1 Lower case letters: base pairs added on the restriction enzyme recognition site; 2 Underlined letters: restriction sites.