Part:BBa_K4343066

pUC-HUH-EgElo9

The EgElo9 gene from Euglena gracilis encodes a ∆-9 elongase that catalyzes the conversion of linoleic acid (LA, C18:2) to eicosadienoic acid (EDA; C20: 2n-6)

Usage and Biology

We constructed a plasmid containing promoter TEF, EgElo9, and terminator Cyc1t through the T4 ligase. The plasmid was linearized by enzyme cleavage and then transformed into Yarrowia lipolytica and randomly integrated into its genome. EgElo9 gene was expressed under the action of promoter TEF, which can catalyze linoleic acid (LA, C18:2) to eicosadienoic acid (EDA; C20: 2n-6). Subsequently, the activity of EgElo9 was examined by gas chromatography.

Result

We integrate one copy of codon-optimized EgElo9 gene from Euglena gracilis under the control of strong promoter PTEF into the genome to obtain the engineered strain Po1f-1 by non-homologous End Joining (NHEJ). The percentage of eicosadienoic acid (EDA; C20: 2n-6) were 31.8%

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NotI site found at 5470
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal XhoI site found at 1920
  Illegal XhoI site found at 1953
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 2108
  Illegal AgeI site found at 97
  Illegal AgeI site found at 458
  Illegal AgeI site found at 3161
  Illegal AgeI site found at 3522
  Illegal AgeI site found at 4664
  Illegal AgeI site found at 5673
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 1
  Illegal BsaI site found at 638
  Illegal BsaI site found at 3702
  Illegal BsaI site found at 4239
  Illegal BsaI site found at 4667
  Illegal BsaI site found at 5194
  Illegal BsaI site found at 5676
  Illegal BsaI.rc site found at 4661
  Illegal BsaI.rc site found at 4723
  Illegal BsaI.rc site found at 5670
  Illegal BsaI.rc site found at 5940
  Illegal SapI.rc site found at 363
  Illegal SapI.rc site found at 3427