TABLE 1.
Oligonucleotide primers used in this study
| Primer | Nucleotide Sequence (5′ to 3′ Direction) | Purposea |
| Preparation of whole and partial regions of desaturases | ||
| FLAGf | GCAAAGCTTAAGATGGACTATAAGGATGATGATGAC | FLAG tag |
| FLAGr | CGTGGTACCCTTGTCATCATCATCCTTATAG | FLAG tag |
| 24aF+ | ACAGGTACCATGGGGAAGGGAGGTAACCAG | D6d |
| 24R+ | GTCTCTAGATTCATTTGTGGAGGTAGGCATCC | D6d |
| D5df | CCCGGTACCATGGCTCCCGACCCGGTGCAGACCC | D5d |
| D5dr | CCCCTGCAGCTATTGGTGAAGGTAAGCATCCAGCC | D5d |
| D6d-cytr | GGGCCGCGGAAGTACGAGAGGATGAACC | N-terminal region of D6d |
| D6d-cytf | CCCTTCCGCGGCAATGGCTGGATTCCC | Middle and C-terminal regions of D6d |
| D5d-cytr | GGGTTCCGCGGAAGATCCAAAGAGTGAGC | N-terminal region of D5d |
| D5d-cytf | CCCTTCCGCGGAACTTCCTTGGTGCCC | Middle and C-terminal regions of D5d |
| Amino acid substitution of D6d with D5d | ||
| d6d5-1 | ACCGTCATCACGGCCGTTCTGCTTGCTACCTCCC | F166V, V167L |
| d6d5-2 | ACGGCCTTTGTCCTTTCTACCGTCCAGGCCCAAGCTGGA | A169S, S171V |
| d6d5-3 | GGCTACAACATGATTTTGGCCACCTTTCTGT | Y182F |
| d6d5-4 | GCCACCTTTCTGTCTTTAGCACCTCCATATGGAAC | Y188F, K189S, K190T |
| d6d5-5 | TTTCTGTCTATAAGAAATCCACATGGAACCACATTGTC | I192T |
| d6d5-6 | TCCATATGGAACCACCTTGTCCACCATTTTGTCATTGGCCACTT | I196L, K199H |
| d6d5-7 | CACTTAAAGGGTGCCCCCGCCAGCTGGTGGAACCATCG | S209P, N211S |
| d6d5-8 | AACTGGTGGAACCATATGCATTTCCAGCACCAT | R216M |
| d6d5-9 | CATGCGAAGCCCAACTGCTTCCGCAAGGACCCCGACAT | I226C, H228R |
| d6d5-10 | GGACCCCGACATAAACATGCACGTGTTTGTCC | K234N, S235M, L236Δ |
| d6d5-11 | ATAAAGAGCCTGCACCCATTGGTGTTTGTCCTTGGA | Δ238P, Δ239L |
| d6d5-12 | ATAAAGAGCCTGCACTTCTTTGCCCTTGGAGAGTGGCA | V238F, V240A |
| d6d5-13 | GTGTTTGTCCTTGGAAAGGTGCTGCCCCTCGAGTATGG | E243K, W244V, Q245L |
| d6d5-14 | CTTGGAGAGTGGCAGTCCGTCGAGCTTGGCAAGAAGAAGCTG | P246S, L247V, Y249L |
| d6d5-15 | CTCGAGTATGGCAAGGAGAAGAAGAAATATCTGCCCTA | K252E, L254K |
| d6d5-16 | AAGAAGAAGCTGAAACATATGCCCTACAACCACC | Y256H, L257M |
| d6d5-17 | TACAACCACCAGCATAAATACTTCTTCCTGA | E264K |
| d6d5-18 | ATCTTGGGAGCCCTGTGTCTTTTCAACTTTATCAGGT | V321C, F322L, L323F |
| d6d5-19 | GCCCTGGTTTTCCTCTTCATTGTCAGGTTCCTGGAGA | N324F, F325I, I326V |
| d6d5-20 | AGGTTCCTGGAGAGCAACTGGTTTGTGTGGG | H332N |
| d6d5-21 | CAGATGAACCACATTCCCATGCACATTGATCTTGATCAC | V344P, E346H |
| d6d5-22 | TCATGGAGATTGATCATGATCGCTACCGGGACTGGTTCA | L349H, H351R |
| d6d5-23 | ATTGATCTTGATCACAACGTGGACTGGTTCAGCAGC | Y352N, R353V |
| d6d5-24 | CACTACCGGGACTGGGTCAGCACCCAGCTGGCAGCCAC | F356V, S358T |
| d6d5-25 | TTCAGCAGCCAGCTGCAAGCCACCTGCAATGT | A361Q |
| d6d5-26 | GCCACCTGCAATGTGCACCAGTCCTTCTTCA | E367H |
| d6d5-27 | AATGTGGAGCAGTCCGCCTTCAATAACTGGTTCAGCGGGC | F370A, D373N |
| d6d5-28 | TGCCAAGACACAACTACCACAAGGTTGCCCCACTGGTGA | L396Y, I399V |
| d6d5-29 | AAGATTGCCCCACTGGTGCAGTCTCTCTGCGCCA | K404Q |
| d6d5-30 | TCTCTCTGCGCCAAGTATGGCATTAAATACCAAGAGAAGC | H410Y, E413K |
| d6d5-31 | CATGGCATTGAATACGAATCGAAGCCGCTGCTGAG | Q415E, E416S |
| d6d5-32 | AGAAGCCGCTGCTGACGGCCTTCGCCGACATTGTGAGTTC | R421T, L423F, L424A |
| d6d5-33 | CTGCTCGACATTGTGTATTCACTGAAGAAGTC | S428Y |
| d6d5-34 | GTGAGTTCACTGAAGGAGTCTGGGCAGCTGTGGCTGGATG | K432E, E435Q |
| d6d5-35 | GATGCCTACCTCCACCAATGAATCTAGTGAA | K444Q |
| Amino acid substitution of D5d with D6d | ||
| d6d5-36 | CACTTAAAGGGTGCCTCCGCCAGCTGGTGGAAC | P209S |
| d6d5-37 | AGGGTGCCCCCGCCAACTGGTGGAACCAT | S211N |
| d6d5-38 | CTGGTGGAACCATCGACATTTCCAGCACCAT | M216R |
| d6d5-39 | GGACCCCGACATAAAGATGCACCCATTGGTGT | N234K |
| d6d5-40 | ACCCCGACATAAACAGCCACCCATTGGTGTTTG | M235S |
| d6d5-41 | CCGACATAAACATGCTGCACCCATTGGTGTTTG | Δ236L |
| d6d5-42 | ATAAACATGCACTTGGTGTTTGTCCTTGGA | P238Δ |
| d6d5-43 | ATAAACATGCACCCAGTGTTTGTCCTTGGA | L239Δ |
| d6d5-44 | GTGTTTGTCCTTGGAGAGGTGCTGCCCCTCGA | K243E |
| d6d5-45 | TTTGTCCTTGGAAAGTGGCTGCCCCTCGAGTA | V244W |
| d6d5-46 | GTCCTTGGAAAGGTGCAGCCCCTCGAGTATGG | L245Q |
| d6d5-47 | CAGATGAACCACATTGTCATGCACATTGATCTT | P344V |
| d6d5-48 | AACCACATTCCCATGGAGATTGATCTTGATCAC | H346E |
| Amino acid substitution of D6d with zD5/6d | ||
| d6zebd5-1 | TCGTACTTCGGCACTGGCTGGATTCCC | N156T |
| d6zebd5-2 | ACCGTCATCACGGCCGTTGTCCTTGCTACCTCC | F166V |
| d6zebd5-3 | TGGCTACAACATGATTTCGGCCACCTTTCTGTC | Y182F |
| d6zebd5-4 | CACCTTTCTGTCTTCAAGACCTCCATATGGAAC | Y188F, K190T |
| d6zebd5-5 | TCCATATGGAACCACCTCGTCCACAAGTTTGTC | I195L |
| d6zebd5-6 | GGACCCCGACATAAATATGCTGCACGTGTTTG | K234N, S235M |
| d6zebd5-7 | CTTGGAGAGGTCCAGCCCGTCGAGTATGGC | W245V, L248V |
| d6zebd5-8 | AAGAAGAAGCTGAAACACCTGCCCTACAACCAC | Y257H |
| d6zebd5-9 | TACAACCACCAGCATAAGTACTTCTTCCTGATC | E265K |
| d6zebd5-10 | TCCAGTACCAGATCTTCATGACCATGATCAG | I284F |
| d6zebd5-11 | GCCATCAGCTACTATGTTCGTTTCTTCTACACC | A305V |
| d6zebd5-12 | TTGGGAGCCCTGGTTCTCTTCAACTTTATCAGGTTC | F322L, L323F |
| d6zebd5-13 | GTTTTCCTCAACTTTGTCAGGTTCCTGGAGAGC | I326V |
| d6zebd5-14 | CAGATGAACCACATTCCCATGGAGATTGATCTTG | V344P |
| d6zebd5-15 | ATTGATCTTGATCACAACCGGGACTGGTTCAGCAG | Y352N |
| d6zebd5-16 | AATGTGGAGCAGTCCGCCTTCAATGACTGGTTC | F370A |
| d6zebd5-17 | TGCCAAGACACAACTATCACAAGATTGCCCC | L396Y |
| d6zebd5-18 | CTCTGCGCCAAGTACGGCATTAAGTACCAAGAGAAG | H410Y |
| d6zebd5-19 | GCCAAGTACGGCATTAAATACCAAGAGAAGCCG | E413K |
| d6zebd5-20 | CGCTGCTGAGGGCCTTCGCTGACATTGTGAGTTC | L423F, L424A |
a
Amino acids are indicated by single characters. Δ indicates a gap in the amino acid sequence alignment.