Part:BBa_K4239007

Enhanced units fiatluxABE

Description

fiatluxA (BBa_K239003) and fiatluxB (BBa_K239004) are made to be used together. They code for the luciferase protein.

Luciferase has as substrats FMNH₂, O₂ and Fatty aldehydes, and produces H₂0, Fatty Acids and FMN and emits luminescence.

The systeme fiatluxA/fiatluxB is made to be used with fiatluxC, fiatluxD and fiatluxE (BBa_K239006) , gathered in the fiatluxCDABE operon.

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.

The ilux operon was born from a mutated natural luminescence operon present in the bacteria P.luminescens: the lux operon. These mutations were error-prone PCR induced according to Gregor et al.’s study in 2018 (Gregor et al. 2018). The aim was to create a system of genes that produced more light.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 2853
  
• 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
  

Construction

resultats de l'overlap PCR , j'ai pas trop compris ce qui a était fait...!!!!!!!!!!!!!!!!!

Characterization

caractérisation dans E.coli DH5alpha...!!!!!!!!!!!!!!

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.