Directed Evolution of
Tyrosyl-tRNA Synthetase
Mussel adhesive proteins introduction
Mussel adhesive proteins (MAPs) are a class of protein complexes secreted through mussel foot glands that react with the surface of the substrate to produce extremely strong adhesion. They are currently used as biomedical
adhesives because of their strong adhesion, degradability and excellent biocompatibility in the marine environment.
In
order to generate MAPs
in a large
scale by microbiology, we hope to obtain a kind of aminoacyl-tRNA synthetase
(aaRS) that allows L-DOPA to be site-specially incorporated into recombinant
MAPs by directed evolution.
We built mutation libraries by
error-prone PCR and site-directed mutagenesis.
Meanwhile, Phage-assisted Continuous Evolution (PACE) strategy was also
used for the directed evolution of tyrosyl-tRNA synthetase (TyrRS).
Meanwhile, Phage-assisted Continuous Evolution (PACE) strategy was also
used for the directed evolution of tyrosyl-tRNA synthetase (TyrRS).
Fluorescent screening strategy
We built mutation libraries by error-prone PCR and site-directed mutagenesis. In addition, a mutated sfGFP was applied to operate fluorescence screening. Mutants were then selected by fluorescence with plate reader.
PACE strategy
PACE (Phage-assisted continuous evolution), a advanced system used to evolve the aminoacyl-tRNA synthetase.
