Difference between revisions of "Part:BBa K4239003:Design"

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===Design Notes===
 
===Design Notes===
  
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The Igem restriction site EcoRI is removed from iluxA.
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Some mutations were realize in luxA K22E, T119A, S178A
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<p><i>fiatluxA</i> did not have any mutation compare to <i>iluxA</i>, because no igem restriction site (EcoR1, Xba1, Spe1 and Pst1) were into the gene.
  
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<p><i>iluxA</i> had mutations from <i>luxA</i> (K22E, T119A, S178A). They changed the protein sequence, in order to create a system that produced more light. The first and the last letters correspond to the Amino Acid before and after mutation, and the number indicates its position in the protein. These mutations were error-prone PCR induced according to Gregor et al.’s study in 2018.</p>
  
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Fiatlux genes come from ilux genes (C, D, A, B, E). They were modified to remove every Igem restriction site (EcoR1, Xba1, Spe1 and Pst1) included in genes. They were also adapted to include the biobrick format.
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===Source===
 
===Source===
  
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azaze
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The source of fiatluxA
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===References===
 
===References===
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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.
 
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.
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Revision as of 20:28, 9 October 2022


Enhanced luciferase substrate forming units fiatluxA


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 504
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 1023


Design Notes

fiatluxA did not have any mutation compare to iluxA, because no igem restriction site (EcoR1, Xba1, Spe1 and Pst1) were into the gene.

iluxA had mutations from luxA (K22E, T119A, S178A). They changed the protein sequence, in order to create a system that produced more light. The first and the last letters correspond to the Amino Acid before and after mutation, and the number indicates its position in the protein. These mutations were error-prone PCR induced according to Gregor et al.’s study in 2018.

Source

The source of fiatluxA

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.