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High-throughput identification of dominant negative polypeptides in yeast

Michael W. Dorrity, Christine Queitsch, Stanley Fields
doi: https://doi.org/10.1101/418608
Michael W. Dorrity
1Department of Genome Sciences, University of Washington, Seattle, WA 98195
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Christine Queitsch
1Department of Genome Sciences, University of Washington, Seattle, WA 98195
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Stanley Fields
1Department of Genome Sciences, University of Washington, Seattle, WA 98195
2Department of Medicine, University of Washington, Seattle, WA 98195
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Abstract

Dominant negative polypeptides can act as inhibitors by binding to the wild type protein or by titrating an essential ligand. Here, we use high-throughput sequencing of DNA libraries composed of fragments of yeast genes to identify dominant negative polypeptides based on their depletion during cell growth. The method uncovers numerous inhibitory polypeptides for a protein and thus is capable of defining interacting domains with exquisite resolution, even pinpointing individual residues that contact ligands.

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Posted September 15, 2018.
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High-throughput identification of dominant negative polypeptides in yeast
Michael W. Dorrity, Christine Queitsch, Stanley Fields
bioRxiv 418608; doi: https://doi.org/10.1101/418608
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High-throughput identification of dominant negative polypeptides in yeast
Michael W. Dorrity, Christine Queitsch, Stanley Fields
bioRxiv 418608; doi: https://doi.org/10.1101/418608

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