Overview
Toolbox
Figure 1: Overview of the different functions provided by specific ncAAs
We want to integrate plasmids with the sequences of specific synthetases into our toolbox and provide them for the iGEM as well as the synthetic biology community.
- Analysing: Two different ncAAs are incorporated which are labeled with fluorophores in a chemically reaction. With the help of Foerster Resonance Energy Transfer (FRET) the distance between the amino acids could be measured.
- Photoswitching: A photoisomerisable amino acid could be incorporated which changes its conformation when it is irradiated with light of different wavelengths. In one conformation it inhibits the function of the target protein. With this photoswitch, reactions could be switched on and off on the protein level only by light exposure.
- Labeling: A fluorescent amino acid could be used to label the target protein in vivo. The advantages compared to fluorescent proteins lay in the smaller size of the fluorescent amino acid.
- Photolysis: Photolysis amino acids break the peptide backbone at the position they were incorporated. This could be used to activate or deactivate proteins by cleaving them.
- Fusing: Two ncAAs which could form a specific covalent bond to each other could be used to fuse proteins together or immobilize the target protein not only at the C- or N-termini.
We hope that our toolkit will be used frequently by future iGEM-teams and help them at their projects. We also wish that they will add new synthetases for other ncAAs and thus expand the possibilities of further applications in protein design.
Two different ways on how to incorporate ncAAs
Figure 2: Expanded code sun
Portrayal of the code sun after the incorporation of one new unnatural base to expand the genetic code and create new blank codons (purple) that can be used to evolve aaRS to make many ncAAs available for advanced protein design.