Genetics, Vol. 165, 1433-1441, November 2003, Copyright © 2003

Green Fluorescent Protein Tagging Drosophila Proteins at Their Native Genomic Loci With Small P Elements

Peter J. Clynea, Jennie S. Brotmana, Sean T. Sweeneya, and Graeme Davisa
a Department of Biochemistry and Biophysics, University of California, San Francisco, California 94143-0448

Corresponding author: Graeme Davis, University of California, 513 Parnassus Ave., HSE 901, San Francisco, CA 94143-0448., gdavis{at}biochem.ucsf.edu (E-mail)

Communicating editor: K. ANDERSON

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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