TY - JOUR
T1 - Development and implementation of split-GFP-based bimolecular fluorescence complementation (BiFC) assays in yeast
AU - Barnard, Emma
AU - McFerran, Neil V.
AU - Trudgett, Alan
AU - Nelson, John
AU - Timson, David J.
PY - 2008/6/1
Y1 - 2008/6/1
N2 - 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.
AB - 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.
KW - Bimolecular fluorescence complementation
KW - Green fluorescent protein (GFP)
KW - Protein-fragment complementation assay
KW - Protein-protein interaction
KW - Saccharomyces cerevisiae
KW - Yeast
UR - http://www.scopus.com/inward/record.url?scp=45249092170&partnerID=8YFLogxK
U2 - 10.1042/BST0360479
DO - 10.1042/BST0360479
M3 - Article
C2 - 18481985
AN - SCOPUS:45249092170
SN - 0300-5127
VL - 36
SP - 479
EP - 482
JO - Biochemical Society Transactions
JF - Biochemical Society Transactions
IS - 3
ER -