Team:KUAS Korea/Parts
Parts
Through this project we have newly produced two parts; BBa_K3107000 and BBa_K3107001. Detailed characterization process of each part is described on the experiments and results page.
BBa_K3107000, nnLuz, is a fungal luciferase gene derived from Neonothopanus nambi, a type of luminescent mushroom. It oxidizes-react with oxygen-the fungal luciferin, 3-hydroxyhispidin, and convert it into caffeylpyruvate.[1] Bioluminescence is caused by the release of energy in the form of light during this catalytic process.
BBa_K3107001, nnh3h, is also a gene derived from N. Nambi, which uses NADPH to hydroxylate the hispidin and synthesize 3-hydroxyhispidin, a fungal luciferin. Working with the enzyme BBa_K3107000, it can elicit luminescence through two stages of reaction from the hispidin.
In making parts, we have optimized the sequence so that the genes can express properly in the E. coli. In addition, by replacing codons without changing the amino acid sequence, unnecessary restriction site were removed to meet the assembly standards. [2]
In particular, NgoMIV, Bsa1, EcoR1 restriction sites that occurred in BBa_K3107001 were removed. <groupparts>iGEM19 KUAS_Korea</groupparts>References
[1] Alexey A. Kotlobay et al., Genetically encodable bioluminescent system from fungi, PNAS, 2018.
[2]Paul M. Sharp et al., Codon usage patterns in Escherichia coli, Bacillus subtilis, Saccharomyces cerevisiae, Schizosaccharomyces pombe, Drosophila melanogaster and Homo sapiens; a review of the considerable within-species diversity, Nucleic Acids Research, 1988, p. 8207–8211.