Development and implementation of split-GFP-based bimolecular fluorescence complementation (BiFC) assays in yeast

Emma Barnard, Neil V. McFerran, Alan Trudgett, John Nelson, David J. Timson

    Research output: Contribution to journalArticlepeer-review

    Abstract

    BiFC (bimolecular fluorescence complementation) is a tool for investigating interactions between proteins. Non-fluorescent fragments of, for example, GFP (green fluorescent protein) are fused to the interacting partners. The interaction brings the fragments together, which then fold, reassemble and fluoresce. This process can be carried out in living cells and provides information both on the interaction and its subcellular location. We have developed a split-GFP-based BiFC assay for use in the budding yeast Saccharomyces cerevisiae in which the modifications are carried out at the genomic level, thus resulting in the tagged yeast proteins being expressed at wild-type levels. The system is capable of detecting interactions in all subcellular compartments tested (the cytoplasm, mitochondria and nucleus) and makes a valuable addition to techniques for the investigation of protein-protein interactions in this model organism.

    Original languageEnglish
    Pages (from-to)479-482
    Number of pages4
    JournalBiochemical Society Transactions
    Volume36
    Issue number3
    DOIs
    Publication statusPublished - 1 Jun 2008

    Keywords

    • Bimolecular fluorescence complementation
    • Green fluorescent protein (GFP)
    • Protein-fragment complementation assay
    • Protein-protein interaction
    • Saccharomyces cerevisiae
    • Yeast

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