Team:Bielefeld-CeBiTec/Results/toolbox/labeling

Labeling

Short summary

For the labeling of a target protein in vivo a fluorescent amino acid should be incorporated through the amber stop codon. Our aim is to provide a (tRNA/aminacyl-synthetase) tRNA/aaRS to the iGEM community to easily incorporate the fluorescent amino acid L‑(7‑hydroxycoumarin‑4‑yl) ethylglycin (CouAA) during translation in response to the amber stop codon.
To demonstrate the advantages of this tool, we want to colocalize the ribulose 1,5‑bisphosphat carboxylase oxygenase (RuBisCo) (RuBisCo) from Halobacillus neaplitanus and the carboxysome in E. coli cells. The carboxysome is labeled with the green fluorescent protein (GFP) and the RuBisCo should be labeled with the fluorescent amino acid at different positions and with the red fluorescent protein (RFP) to compare the advantages and disadvantages of both labeling strategies.