Engineering Enzymes Through Genetic Selection
Doyle, Donald F.
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The disclosure provide a system for the selection of variant nuclear receptor ligand binding domains, or for the selection of variant enzymes that have an enhanced ability to synthesize a nuclear receptor ligand or a precursor thereof. The disclosure provides yeast cells comprising: a yeast transcription modulating system comprising a nucleic acid expression system encoding a nuclear receptor ligand-binding domain linked to a DNA-binding domain, a nucleic acid expression system encoding a coactivator domain linked to a yeast transcriptional activator, a heterologous enzyme system for generating a nuclear receptor ligand, and a selective genetic locus expressed in the presence of the recombinant nuclear receptor polypeptide and a nuclear receptor ligand specifically bound to the recombinant nuclear receptor polypeptide. The disclosure also provides methods of using the yeast cell system for identifying variant nuclear receptor ligand binding domains or variant enzymes synthesizing a nuclear receptor ligand.
- Georgia Tech Patents