Difference between revisions of "Part:BBa K4343092"
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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. | ||
+ | <center>https://static.igem.wiki/teams/4343/wiki/puc-huh-scp2-ehelo9-pfabin-map.png</center> | ||
===Result=== | ===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. | + | 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. (Fig: PFBAin) |
− | + | <center>https://static.igem.wiki/teams/4343/wiki/promoter-change.png</center> | |
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===Usage and Biology=== | ===Usage and Biology=== |
Latest revision as of 13:10, 10 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.
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. (Fig: PFBAin)
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE 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 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE 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 - 1000INCOMPATIBLE 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