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- Title
Stringent Analysis of Gene Function and Protein--Protein Interactions Using Fluorescently Tagged Genes.
- Authors
Neumüller, Ralph A.; Wirtz-Peitz, Frederik; Lee, Stella; Kwon, Young; Buckner, Michael; Hoskins, Roger A.; Venken, Koen J. T.; Bellen, Hugo J.; Mohr, Stephanie E.; Perrimon, Norbert
- Abstract
In Drosophila collections of green fluorescent protein (GFP) trap lines have been used to probe the endogenous expression patterns of trapped genes or the subcellular localization of their protein products. Here, we describe a method, based on nonoverlapping, highly specific, shRNA transgenes directed against GFP, that extends the utility of these collections to loss-of-function studies. Furthermore, we used a MiMIC transposon to generate GFP traps in Drosophila cell lines with distinct subcellular localization patterns, which will permit high-throughput screens using fluorescently tagged proteins. Finally, we show that fluorescent traps, paired with recombinant nanobodies and mass spectrometry, allow the study of endogenous protein complexes in Drosophila.
- Subjects
GENES; PROTEIN-protein interactions; MOLECULAR association; GREEN fluorescent protein; FLUORESCENT polymers
- Publication
Genetics, 2012, Vol 190, Issue 3, p931
- ISSN
0016-6731
- Publication type
Article
- DOI
10.1534/genetics.111.136465