Part:BBa_K4343087
pUC-HUH-Scp2-MaElo16
MaElo16 gene encodes a C16/18 elongase from Misgurnus anguillicaudatus, which can catalyze C16:0 to C18:0. This gene is expressed in Y. lipolytica.
Characterisation
We constructed a plasmid containing promoter TEF, MaElo16, terminator Cyc1t, and homologous arm through the T4 ligase. The plasmid was linearized by enzyme cleavage and then was integrated into the Scp2 site of strain Po1f-3. MaElo16 gene was expressed under the control of TEF promoter, which can catalyze the elongation of C16:0 to C18:0, it also has catalytic activity for C18:1. Then the activity of MaElo16 was tested by gas chromatography.
Result
The C16:0 content was reduced to 15.5% of TFAs and the percentage of C18:2 they produced were almost doubled. (Fig:Po1f-14)
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 6782
Illegal XhoI site found at 1431
Illegal XhoI site found at 3465
Illegal XhoI site found at 3498
Illegal XhoI site found at 7826 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 3653
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 - 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 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
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