Part:BBa_K4343076
pUC-HUH-PaDes17
The PaDes17 gene encodes a ∆-17 elongase from Pythium aphanidermatum that catalyzes the conversion of arachidonic acid (ARA; C20:4 n-6) to Eicosapentaenoic acid (EPA), an ω 3 polyunsaturated fatty acid (PUFA).
Characterisation
Two copies of this gene and two copies of ∆-5 desaturase (EgDes5) From Euglena gracilis were randomly integrated into the genome of the Engineered strain Po1F-19 by transformation. The ∆-17 desaturase (PaDes17) gene was expressed under the action of promoter TEF to generate ∆-17 elongase, which can catalyze Arachidonic acid (ARA; C20:4n-6) generated Eicosapentaenoic acid (EPA), the engineered strain Po1F-20 was obtained. After fermentation and culture, the effect of the increase of the copy number of ∆-17 elongase and ∆-5 desaturase on the increase of EPA content was tested by gas chromatography.
Result
Po1f-20 was further randomly integrated with 2 copies of EgDes5 and 2 copies of PaDes17 in the genome of the engineered strain Po1f-19, and EPA proportion reached 22.4% of the TFAs. (Fig: Po1f-2 )
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 5099
Illegal XhoI site found at 1920
Illegal XhoI site found at 1953
Illegal XhoI site found at 4841 - 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 5766 - 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 5769
Illegal BsaI.rc site found at 4661
Illegal BsaI.rc site found at 5763
Illegal BsaI.rc site found at 6033
Illegal SapI.rc site found at 363
Illegal SapI.rc site found at 3427
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