Difference between revisions of "Part:BBa K4239001:Design"

(→‎Design Notes)
(→‎Design Notes)
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===Design Notes===
 
===Design Notes===
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Every Igem restriction sites (EcoR1, Xba1, Spe1 and Pst1) were removed from the part.
+
The Igem restriction site Pst1 was removed into the iluxC part, to create fiatluxC. This mutation does not change the protein sequence.
 +
 
 +
iluxC had mutations from luxC (N10T, N59D, E74D, S256P, M355T, N360D). These mutations were errore-prone PCR induced according to Gregor et al.’s study in 2018. They change the protein sequence, in order to create a system that produced more light.
  
 
===Source===
 
===Source===

Revision as of 12:48, 8 October 2022


Enhanced luciferase subunits fiatluxC


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1311


Design Notes

The Igem restriction site Pst1 was removed into the iluxC part, to create fiatluxC. This mutation does not change the protein sequence.

iluxC had mutations from luxC (N10T, N59D, E74D, S256P, M355T, N360D). These mutations were errore-prone PCR induced according to Gregor et al.’s study in 2018. They change the protein sequence, in order to create a system that produced more light.

Source

fiatluxCD is made up of fiatluxC and fiatluxD.

References

Gregor C, Gwosch KC, Sahl SJ, Hell SW. Strongly enhanced bacterial bioluminescence with the ilux operon for single-cell imaging. Proc Natl Acad Sci U S A. 2018 Jan 30;115(5):962-967. doi: 10.1073/pnas.1715946115. Epub 2018 Jan 16. PMID: 29339494; PMCID: PMC5798359.