Composite

Part:BBa_K4343071

Designed by: Xiaolu Sun   Group: iGEM22_NNU-China   (2022-10-09)
Revision as of 13:53, 10 October 2022 by Milulu (Talk | contribs) (→‎Result)

(diff) ← Older revision | Latest revision (diff) | Newer revision → (diff)


Sc4Des

The Sc4Des gene encodes a ∆-4 desaturase from Schizochytrium sp. that catalyzes the conversion of Docosapentaenoic acid (DPA; C22:5n-3) to Docosahexaenoic acid (DHA; C22:6n- 3).

Characterisation

We constructed a plasmid containing promoter TEF, Sc4Des, and terminator Cyc1t through the T4 ligase. The plasmid was linearized by enzyme cleavage and then transformed into engineered strain with one copy of TsElo5. The Sc4Des gene was expressed in the promoter TEF, which catalyzes the conversion of Docosapentaenoic acid (DPA; C22:5n-3) to Docosahexaenoic acid (DHA; C22:6n-3). The activity of Sc4Des was examined by gas chromatography.

puc-huh-sc4des-map.png

Result

Δ-4 desaturase was identified to convert DPA to DHA. The engineered strain Po1f-20 contained TsElo5. Thus, in order to obtain an engineered strain for co-production of DHA and EPA, Sc4Des gene from Schizochytrium sp. were randomly integrated in the genome of engineered strain Po1f-20 contained TsElo5 to obtain strain Po1f-21. The GC analysis indicated that the percentage of EPA and DHA in the engineered strain were 2.5% and 18.9% of TFAs, respectively, indicating successful heterologous synthesis of DHA and EPA in the Yarrowia lipolytica.

favorite.png

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 5022
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 1920
    Illegal XhoI site found at 1953
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE 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 5970
  • 1000
    INCOMPATIBLE 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 5330
    Illegal BsaI site found at 5479
    Illegal BsaI.rc site found at 4661
    Illegal BsaI.rc site found at 5967
    Illegal BsaI.rc site found at 6237
    Illegal SapI.rc site found at 363
    Illegal SapI.rc site found at 3427


[edit]
Categories
Parameters
None