Difference between revisions of "Part:BBa K4343081"

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===Characterisation===
 
===Characterisation===
 
We integrated EhElo9 controlled by the TEF promoter to the SCP2 site of the genome of Y. lipolytica through this segment. Then we detected the proportion of C18:2 and C20:2 by gas chromatography (GC) and test the activity of ∆-9 elongase according to the results.<br>
 
We integrated EhElo9 controlled by the TEF promoter to the SCP2 site of the genome of Y. lipolytica through this segment. Then we detected the proportion of C18:2 and C20:2 by gas chromatography (GC) and test the activity of ∆-9 elongase according to the results.<br>
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<center>https://static.igem.wiki/teams/4343/wiki/puc-huh-ehelo9-map.png</center>
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<center>https://static.igem.wiki/teams/4343/wiki/puc-huh-scp2-ehelo9-map.png</center>
  
 
===Result===
 
===Result===

Revision as of 10:57, 9 October 2022


pUC-HUH-Scp2-EhElo9

EhElo9 gene encodes ∆-9 elongase from Emiliania huxleyi. This enzyme can catalyze the elongation of C18:2 to C20:2 in Y. lipolytica.

Characterisation

We integrated EhElo9 controlled by the TEF promoter to the SCP2 site of the genome of Y. lipolytica through this segment. Then we detected the proportion of C18:2 and C20:2 by gas chromatography (GC) and test the activity of ∆-9 elongase according to the results.

puc-huh-scp2-ehelo9-map.png

Result

C18:2 produced by them decreased from 62.7% to 32.8%, and the conversion of C18:2 to C20:2 reached 56.1%.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 1431
    Illegal XhoI site found at 3465
    Illegal XhoI site found at 3498
    Illegal XhoI site found at 7826
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 3653
    Illegal NgoMIV site found at 6886
    Illegal AgeI site found at 889
    Illegal AgeI site found at 1543
    Illegal AgeI site found at 1642
    Illegal AgeI site found at 2003
    Illegal AgeI site found at 4706
    Illegal AgeI site found at 5067
    Illegal AgeI site found at 6209
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1
    Illegal BsaI site found at 1546
    Illegal BsaI site found at 2183
    Illegal BsaI site found at 5247
    Illegal BsaI site found at 5784
    Illegal BsaI site found at 6212
    Illegal BsaI site found at 7035
    Illegal BsaI.rc site found at 401
    Illegal BsaI.rc site found at 1540
    Illegal BsaI.rc site found at 6206
    Illegal BsaI.rc site found at 7029
    Illegal BsaI.rc site found at 7299
    Illegal BsaI.rc site found at 8461
    Illegal BsaI.rc site found at 8845
    Illegal SapI.rc site found at 1908
    Illegal SapI.rc site found at 4972