High throughput protein-protein interaction screening in yeast liquid culture
AssigneeUniversity of Washington
InventorsDavid BAKER, David YOUNGER, Eric KLAVINS
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Abstract
The present invention relates to methods and compositions for the high throughput screening of protein-protein interactions in yeast liquid culture. Protein fusions non-native to yeast may be expressed to replace endogenous sexual agglutination proteins and mediate library-by-library interrogation of protein interactions. The methods and compositions of the invention can be utilized for the characterization of protein interaction networks in high throughput for both binding affinity and specificity, which is crucial for understanding cellular functions, screening therapeutic candidates, and evaluating engineered protein networks.
Claims8
A recombinant haploid yeast cell comprising:
- one or more exogenous synthetic adhesion proteins, wherein each protein of the one or more exogenous synthetic adhesion proteins is expressed and displayed on the surface of the recombinant haploid yeast cell, each synthetic adhesion protein being (1) a fusion protein bound to a cell wall glycosylphosphatidylinositol (GPI) anchored protein residing on the surface of the recombinant haploid yeast cell or (2) a glycosylphosphatidylinositol (GPI) anchored fusion protein residing on the surface of the recombinant haploid yeast cell;
- and
- a nucleic acid construct which encodes the one or more exogenous synthetic adhesion proteins, the nucleic acid construct comprising:
- a primer binding site,
- a recombination site, and
- a oligonucleotide molecular barcode;
- wherein the recombinant haploid yeast cell is incapable of mating according to any native sexual agglutination process.
Dependencies are inferred from the claim text.