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PMID:14668392
| Citation |
Clyne, PJ, Brotman, JS, Sweeney, ST and Davis, G (2003) Green fluorescent protein tagging Drosophila proteins at their native genomic loci with small P elements. Genetics 165:1433-41 |
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| 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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| Keywords |
Animals; Artificial Gene Fusion; Base Sequence; DNA Primers; DNA Transposable Elements; Drosophila/genetics; Drosophila Proteins/genetics; Genomics; Green Fluorescent Proteins; Luminescent Proteins/genetics; Recombinant Fusion Proteins/genetics; Subcellular Fractions/metabolism |
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Significance
Annotations
| Gene product | Qualifier | GO ID | GO term name | Evidence Code | with/from | Aspect | Notes | Status |
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