Difference between revisions of "Part:BBa K4343075"

(Characterisation)
(Result)
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===Result===
 
===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. ).  
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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 ).  
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<center>https://static.igem.wiki/teams/4343/wiki/part-6.jpg</center>
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

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.

puc-huh-igase2-map.png

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 ).

part-6.jpg

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 1920
    Illegal XhoI site found at 1953
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE 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
  • 1000
    INCOMPATIBLE 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