Part:BBa_K4343082

pUC-HUH-Scp2-IgASE1

IgASE1 gene encodes ∆-9 elongase from Isochrysis galbana. This enzyme can catalyze the elongation of C18:2 to C20:2 in Y. lipolytica.

Characterisation

We integrated IgASE1 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

The conversion of C18:2 to C20:2 under the catalytic action of IgASE1 was around 26.3%, with only 16.5% of C20:2 produced.(fig:Po1f-9)

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 7817
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE 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
  
• 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 7026
  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 7020
  Illegal BsaI.rc site found at 7290
  Illegal BsaI.rc site found at 8452
  Illegal BsaI.rc site found at 8836
  Illegal SapI site found at 6501
  Illegal SapI.rc site found at 1908
  Illegal SapI.rc site found at 4972
  Illegal SapI.rc site found at 6984