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. 2003 Nov;165(3):1433–1441. doi: 10.1093/genetics/165.3.1433

Green fluorescent protein tagging Drosophila proteins at their native genomic loci with small P elements.

Peter J Clyne 1, Jennie S Brotman 1, Sean T Sweeney 1, Graeme Davis 1
PMCID: PMC1462835  PMID: 14668392

Abstract

We describe a technique to tag Drosophila proteins with GFP at their native genomic loci. This technique uses a new, small P transposable element (the Wee-P) that is composed primarily of the green fluorescent protein (GFP) sequence flanked by consensus splice acceptor and splice donor sequences. We demonstrate that insertion of the Wee-P can generate GFP fusions with native proteins. We further demonstrate that GFP-tagged proteins have correct subcellular localization and can be expressed at near-normal levels. We have used the Wee-P to tag genes with a wide variety of functions, including transmembrane proteins. A genetic analysis of 12 representative fusion lines demonstrates that loss-of-function phenotypes are not caused by the Wee-P insertion. This technology allows the generation of GFP-tagged reagents on a genome-wide scale with diverse potential applications.

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Selected References

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