Difference between revisions of "Part:BBa K4343074"

Revision as of 14:37, 9 October 2022

pUC-HUH-EhElo9

EhElo9 encodes ∆-9 elongase from Emiliania huxleyi which functions in the Δ-9 elongase/desaturase pathway and could convert C18:2 to C20:2.

Characterisation

Three copies of this gene, two copies of IgASE2 (BBa_K4343075), and two copies of EgDes8 (BBa_K4343067) were randomly integrated into the genome of the engineered strain Po1F-18. The ∆-9 elongase (EhElo9) gene was expressed under the control of promoter TEF to generate ∆-9 elongase, which catalyzes the formation of Eicosadienoic acid (EDA; C20:2n-6), the engineered strain Po1F-19 was obtained. After fermentation, 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, EhElo9, 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-8).

Sequence and Features