Part:BBa_K4343068
pUC-HUH-EgDes5
The EgDes5 gene encodes a ∆-5 desaturase from Euglena gracilis that catalyzes the conversion of dihomo-γ-linolenic acid (DGLA; C20: 3n-6) to arachidonic acid (ARA; C20:4 n-6).
Characterisation
We constructed a plasmid containing promoter TEF, EgDes5, and terminator Cyc1t through the T4 ligase. The plasmid was linearized by enzyme cleavage and then transformed into engineered Yarrowia lipolytica Po1f-1 and randomly integrated into its genome. EgDes5 gene was expressed under control of promoter TEF, which could catalyze the formation of arachidonic acid (ARA; C20:4 n-6) from eicosadienoic acid (EDA; C20:2n-6). The activity of EgDes5 was examined by gas chromatography.
Result
We integrate one copy of codon-optimized EgDes5 gene from Euglena gracilis under the control of strong promoter PTEF into the genome to obtain the engineered strain Po1f-1 by non-homologous End Joining (NHEJ) The percentage of arachidonic acid (ARA; C20:4 n-6) were 19.4%.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 1920
Illegal XhoI site found at 1953
Illegal XhoI site found at 4708 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 2108
Illegal AgeI site found at 97
Illegal AgeI site found at 458
Illegal AgeI site found at 3161
Illegal AgeI site found at 3522
Illegal AgeI site found at 4664
Illegal AgeI site found at 5629
Illegal AgeI site found at 6036 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1
Illegal BsaI site found at 638
Illegal BsaI site found at 3702
Illegal BsaI site found at 4239
Illegal BsaI site found at 4667
Illegal BsaI site found at 6039
Illegal BsaI.rc site found at 4661
Illegal BsaI.rc site found at 6033
Illegal BsaI.rc site found at 6303
Illegal SapI.rc site found at 363
Illegal SapI.rc site found at 3427
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