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Curr Protoc Neurosci. 2007 Oct;Chapter 5:Unit 5.27. doi: 10.1002/0471142301.ns0527s41.

Split-ubiquitin system for identifying protein-protein interactions in membrane and full-length proteins.

Author information

1
Universität Tübingen, Tübingen, Germany.

Abstract

Protein-protein interactions play a fundamental role in the regulation of almost all cellular processes. Thus, the identification of interacting proteins can help to elucidate their function. The mating-based split-ubiquitin system (mbSUS) uses yeast as a test organism to identify potential interactions between full-length membrane proteins or between a full-length membrane protein and a soluble protein. The mbSUS can also be used to provide further evidence for protein-protein interactions detected with other methods and to map the interaction domains of selected proteins. The mbSUS is optimized for systematic screening approaches employing a mating-based approach, as typically used to determine protein interactions on a genomic scale. Construction of bait and prey fusions is simplified by adapting two different cloning procedures: (i) in vivo cloning in yeast, and (ii) Gateway cloning in E. coli. Protocols for small-scale interaction tests, as well as systematic approaches using sorted bait and prey arrays, are described.

PMID:
18428659
DOI:
10.1002/0471142301.ns0527s41
[Indexed for MEDLINE]

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