Part:BBa_K4239004:Design
Enhanced luciferase substrate forming unit fiatluxB
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
fiatluxB did not have any mutation compare to iluxB, because no igem restriction site (EcoRI, XbaI, SpeI and PstI) were into the gene.
iluxB had mutations from luxB (S13P, V121A, N259D). 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 was the ilux operon available in a pGEX plasmid. An overlap PCR was performed to reconstitute directly fiatluxABE fragments which had been cut by the restriction enzymes.
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.