Difference between revisions of "Part:BBa K4343092"
(→‎Characterisation)
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===Characterisation===
 
===Characterisation===
 
We integrated EhElo9 controlled by the FBAin 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. And the EhElo9 controlled by the TEF promoter (BBa_K4343081) was used as the control group.
 
We integrated EhElo9 controlled by the FBAin 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. And the EhElo9 controlled by the TEF promoter (BBa_K4343081) was used as the control group.
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<center>https://static.igem.wiki/teams/4343/wiki/puc-huh-scp2-ehelo9-pfabin-map.png</center>
  
 
===Result===
 
===Result===

Revision as of 14:17, 9 October 2022


pUC-HUH-Scp2-EhElo9(PFABin)

EhElo9 encodes ∆-9 elongase from Emiliania huxleyi which functions in the Δ-9 elongase/desaturase pathway and could convert C18:2 to C20:2.

Characterisation

We integrated EhElo9 controlled by the FBAin 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. And the EhElo9 controlled by the TEF promoter (BBa_K4343081) was used as the control group.

puc-huh-scp2-ehelo9-pfabin-map.png

Result

It was found that there was no significant change in EPA yield under the FBAin promoter (Fig. ), which demonstrates that the promoter optimization did not have a significant effect on EPA yield.

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 8261
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 3653
    Illegal NgoMIV site found at 7321
    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 6644
  • 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 6647
    Illegal BsaI site found at 7470
    Illegal BsaI.rc site found at 401
    Illegal BsaI.rc site found at 1540
    Illegal BsaI.rc site found at 6641
    Illegal BsaI.rc site found at 7464
    Illegal BsaI.rc site found at 7734
    Illegal BsaI.rc site found at 8896
    Illegal BsaI.rc site found at 9280
    Illegal SapI.rc site found at 1908
    Illegal SapI.rc site found at 4972