Part:BBa_K4343081

pUC-HUH-Scp2-EhElo9

EhElo9 gene encodes ∆-9 elongase from Emiliania huxleyi. This enzyme can catalyze the elongation of C18:2 to C20:2 in Y. lipolytica.

Characterisation

We integrated EhElo9 controlled by the TEF 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.

Result

C18:2 produced by them decreased from 62.7% to 32.8%, and the conversion of C18:2 to C20:2 reached 56.1%. (Fig: Po1f-8)

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 7826
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 3653
  Illegal NgoMIV site found at 6886
  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
  
• 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 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