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Fig. 1 | BMC Biology

Fig. 1

From: A straightforward approach for bioorthogonal labeling of proteins and organelles in live mammalian cells, using a short peptide tag

Fig. 1

Using a GCE-tag for fluorescence labeling of proteins via genetic code expansion (GCE) and bioorthogonal chemistry. a Schematic representation of the labeling approach. A GCE-tag (blue) is attached to the N-terminal of the protein (gray). Binding of the Fl-dye (green) to the GCE-tag leads to labeling of the protein. b Graphical illustration of the experimental design. The stop codon, UAG, and nucleotides encoding a short polypeptide tag are added upstream to the coding sequence of a protein. During ribosomal translation, the ncAA BCN-Lys (red) is incorporated into the protein in response to the in-frame UAG codon using a specific, orthogonal pair of tRNA (dark blue)/tRNA-Synth (purple). Finally, a tetrazine-conjugated Fl-dye (green) is covalently attached to BCN-Lys via a bioorthogonal reaction. As a result, the protein is directly labeled with Fl-dye via a small polypeptide tag.

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