Composite

Part:BBa_K4343091

Designed by: Chen Hao   Group: iGEM22_NNU-China   (2022-10-09)
Revision as of 14:34, 9 October 2022 by MINTKIM (Talk | contribs) (→‎Characterisation)


hisG-URA-hisG selection marker + EXP promoter + EhElo9 + Cyc1t terminator+ homologous arm SCP2-up

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 EXP 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-pexp-map.png

Result

it was found that there was no significant change in EPA yield under the EXP 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
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 6528
  • 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 8176
    Illegal XhoI site found at 9092
    Illegal XhoI site found at 9383
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 3653
    Illegal NgoMIV site found at 7575
    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 6898
  • 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 6901
    Illegal BsaI site found at 7724
    Illegal BsaI.rc site found at 401
    Illegal BsaI.rc site found at 1540
    Illegal BsaI.rc site found at 6895
    Illegal BsaI.rc site found at 7718
    Illegal BsaI.rc site found at 7988
    Illegal BsaI.rc site found at 9406
    Illegal SapI site found at 8904
    Illegal SapI.rc site found at 1908
    Illegal SapI.rc site found at 4972


[edit]
Categories
Parameters
None