Difference between revisions of "Part:BBa K4343075"

Revision as of 14:30, 9 October 2022

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