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Methods Mol Biol. 2018;1672:599-612. doi: 10.1007/978-1-4939-7306-4_39.

Rewiring the Budding Yeast Proteome using Synthetic Physical Interactions.

Author information

1
Mitotic Control Laboratory, The Francis Crick Institute, 1 Midland Road, London, NW1 1AT, UK.
2
Mitotic Control Laboratory, The Francis Crick Institute, 1 Midland Road, London, NW1 1AT, UK. Peter.Thorpe@crick.ac.uk.

Abstract

Artificially tethering two proteins or protein fragments together is a powerful method to query molecular mechanisms. However, this approach typically relies upon a prior understanding of which two proteins, when fused, are most likely to provide a specific function and is therefore not readily amenable to large-scale screening. Here, we describe the Synthetic Physical Interaction (SPI) method to create proteome-wide forced protein associations in the budding yeast Saccharomyces cerevisiae. This method allows thousands of protein-protein associations to be screened for those that affect either normal growth or sensitivity to drugs or specific conditions. The method is amenable to proteins, protein domains, or any genetically encoded peptide sequence.

KEYWORDS:

Chromobody; GFP-binding protein (GBP); Green fluorescent protein (GFP); High-throughput screen; Nanobody; Protein-protein interactions (PPI); Selective ploidy ablation

PMID:
29043650
DOI:
10.1007/978-1-4939-7306-4_39
[Indexed for MEDLINE]

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