Difference between revisions of "Part:BBa K4343076"

 
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<partinfo>BBa_K4343076 short</partinfo>
 
<partinfo>BBa_K4343076 short</partinfo>
  
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The PaDes17 gene encodes a &#8710;-17 elongase from Pythium aphanidermatum that catalyzes the conversion of arachidonic acid (ARA; C20:4 n-6) to Eicosapentaenoic acid (EPA), an &#969; 3 polyunsaturated fatty acid (PUFA).  
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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).
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===Characterisation===
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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.
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===Result===
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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. ).
  
 
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Revision as of 14:13, 9 October 2022


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. ).

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE 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
  • 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 5766
  • 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 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