Generator

Part:BBa_K2971010:Design

Designed by: Simen Hermansen   Group: iGEM19_UiOslo_Norway   (2019-10-21)
Revision as of 22:50, 21 October 2019 by Simen (Talk | contribs) (→‎Design Notes)

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crtE and crtB in a single open reading frame


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1602
    Illegal BamHI site found at 841
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 43


Design Notes

Codon optimized for expression in Escherichia coli and iGEM registry compatibility.

The slightly unusual and simplified design, without an individual RBS for each gene, was to create a system where we could, with relative ease, produce several composite parts that produce various carotenoids. By combining crtEB (BBa_K2971004) with a phytoene desaturase of our choice, we could control what carotenoid that would be produced. When combined with CrtI from D. radiodurans(BBa K2971001) lycopene was produced in cultures expressing the composite part. If, for example, it was combined with CrtI from Rhodobacter capsulatus neurosporene would be produced instead. Xu et al., 2018 showed that this design was viable for producing plasmids intended for lycopene production.


References

Xu, X., Tian, L., Xu, J., Xie, C., Jiang, L., & Huang, H. (2018). Analysis and expression of the carotenoid biosynthesis genes from Deinococcus wulumuqiensis R12 in engineered Escherichia coli. AMB Express, 8(1), 94-94. doi:10.1186/s13568-018-0624-1

Source

Deinococcus radiodurans

References