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Genetics. 2012 Mar;190(3):931-40. doi: 10.1534/genetics.111.136465. Epub 2011 Dec 14.

Stringent analysis of gene function and protein-protein interactions using fluorescently tagged genes.

Author information

1
Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115, USA.

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.

PMID:
22174071
PMCID:
PMC3296255
DOI:
10.1534/genetics.111.136465
[Indexed for MEDLINE]
Free PMC Article

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