Part:BBa_K4343075
pUC-HUH-IgASE2
IgASE2 encodes ∆-9 elongase from Isochrysis galbana which functions in the Δ-9 elongase/desaturase pathway and could convert C18:2 to C20:2.
Characterisation
Two copies of this gene, three copies of EhElo9 (BBa_K4343074), and two copies of EgDes8 (BBa_K4343067) were randomly integrated into the genome of the engineered strain Po1F-18 by transformation. The ∆ 9 elongase (EhElo9) gene was expressed under the action of promoter PTEF to generate ∆ 9 elongase, which catalyzes the formation of Eicosadienoic acid (EDA; C20:2n-6), the engineered strain Po1F-19 was obtained. After fermentation and culture, the effect of the increased copy number of these three genes on the increase of EPA content was tested by gas chromatography, and 16.3% of EPA in the engineered strain Po1F-18 was obtained.
Result
Furthermore, EhElog9, IgASE2, and EgDes8 were randomly integrated in the genome of Po1f-18 to obtain Po1f-19. It can be found that EPA Po1f-19 produced made up 16.3% of TFAs. The results of qPCR demonstrate that the genome of Po1f-19 was integrated of 3 copies of the EnElo9, 3 copies of the IgASE2, and 2 copies of the EgDes8. (Fig: Po1f-10 ).
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 1920
Illegal XhoI site found at 1953 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE 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 5472 - 1000INCOMPATIBLE 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 5475
Illegal BsaI.rc site found at 4661
Illegal BsaI.rc site found at 5469
Illegal BsaI.rc site found at 5739
Illegal SapI.rc site found at 363
Illegal SapI.rc site found at 3427
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