Table 1.
Oligonucleotide primers used for desaturase gene characterization and functional expression
| Degenerated | Primer sequencea |
|---|---|
| PR3 | 5'-GGYATYACVGCHGGNGCWCA-3' |
| PR4 | 5'-TGRTARTTRTGGAABSCYTCNCC-3' |
| Gene specificb | Primer sequence |
| Lc1–5a | 5'-CAGGCCAGCGGCATCAGCAATAAGTA-3' |
| Lc1–5b | 5'-CGAGGATTAATCTCAGCGGCCACT-3' |
| Lc1–3a | 5'-TACTTATTGCTGATGCCGCTGGCCTGC-3' |
| Lc1–3b | 5'-TCCTGGAAAAACGCGCTCTTCGTAGCTGCAATG-3' |
| Lc2–5a | 5'-TCCTTTGGCCTTGATTTCGGGGTGTTTC-3' |
| Lc2–5b | 5'-CCATGAGCCAACCCATGTGGGAGAAGAA-3' |
| Lc2–3a | 5'-GGTGAAGAAACACCCCGAAATCAAGGCC-3' |
| Lc3–5a | 5'-TCGCTCATGTCAATGCTGGCTCCCTTCT-3' |
| Lc3–5b | 5'-ACGTTTGGCGTTGTGAGGATCAGCGTC-3' |
| Lc3–3a | 5'-TTGAGGCGGACCCCATCGTCATGTTT-3' |
| Lc3–3b | 5'-TGGTACGTGGCTACAATCCTGCGGTTCA-3' |
| Lc4–5a | 5'-GTCAACAGCCAACCGACATGCGAA-3' |
| Lc4–5b | 5'-GAGTCGGGATAACGAAGCAAACCA-3' |
| Lc4–3a | 5'-TACGCAATTCCCTTCGTCGGATTG-3' |
| Lc4–3b | 5'-CTTATGCCGATGTACTTCTGGAACG-3' |
| Full-length amplificationb | Primer sequence |
| Lc1-fls | 5'-GATTCATAGATTCGTGTTCGGTGA-3' |
| Lc1-flas | 5'-CAGGGACCTCGAAGTGACCTTT-3' |
| Lc2-fls | 5'-GCAGTGATTGGTGTCGTGCGGA-3' |
| Lc2-flas | 5'-AACAATAAAATATTTATTTACATTAATTTGG-3' |
| Lc3-fls | 5'-TAAAGCAAGTTACTGTCGCCG-3' |
| Lc3-flas | 5'-GAAATACGTTTATTATTGGAATAAATAA-3' |
| Lc4-fls | 5'-GAGTCACAAGTTGTTCAGTCGTAA-3' |
| Lc4-flas | 5'-TCCATAAGATGGGTACGTATATTGT-3' |
| Expression YEpOLEX | Primer sequencec |
| Lc1ORFs | 5'-gctctagaATGCCGCCGAACGTGACAG-3' |
| Lc1ORFas | 5'-cgagctcgCTAATCATCTTTACGGTTAATG-3' |
| Lc2ORFs | 5'-gctctagaATGCCGCCCGAGGGTCTAAT-3' |
| Lc2ORFas | 5'-cgagctcgTCAAAAAGATTTCTCCGGAT-3' |
| Lc3ORFs | 5'-gctctagaATGGCTCCAAACATCCTGGG-3' |
| Lc3ORFas | 5'-cgagctcgTCACTGTTCTTTAGGATGTGC-3' |
| Expression pYEX | Primer sequencec |
| Lc4ORFs | 5'-ggatccATAATGGCGCCATATCCTGAAG-3' |
| Lc4ORFas | 5'-gaattcCGATTTCATATTAGTTCATTTTAATC-3' |
a [21]; Y = C or T; V = A, G or C; H = A, C or T; N = A, G, C or T; W = A or T; R = A or G; B = G, T or C; S = G or C.
b Lc1= Lca-QPAQ, Lc2 = Lca-SPVE, Lc3 = Lca-GATD, Lc4= Lca-KPVQ
c Restriction sites are indicated in lower case letters, start and stop codons are marked in bold.