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The Wee-P Protein Trap:
Tagging Proteins With GFP at Their Native Genomic Loci

I. Introduction
II. Methods
III. Using Wee-Ps in P-element replacement experiments
IV. Table of available fusion lines
V. Table of expression patterns and images for Wee-P lines whose gene has not been determined

III. Using Wee-Ps in P-element replacement experiments

For some experiments it might be advantageous to have a particular gene tagged with a Wee-P element. If the gene of interest already has a P-element in the vicinity (preferably in an intron that is flanked by coding sequence), it should be possible to replace that P-element with a Wee-P element. The technique of P-element replacement has been described (Sepp KJ, Auld VJ. Conversion of lacZ enhancer trap lines to GAL4 lines using targeted transposition in Drosophila melanogaster. Genetics. 1999 Mar; 151(3):1093-101), and in the Davis lab such P-element replacements with Wee-Ps have been successfully made. We replace a previously existing P-element with a w+ starter Wee-P (making sure to use the correct phase), and then flip out the w+.

The details of how to do such P-element replacement crosses are highly dependent on the markers in the P-element that is to be replaced. We recommend using the crossing schemes in the Sepp and Auld paper (Sepp KJ, Auld VJ. Conversion of lacZ enhancer trap lines to GAL4 lines using targeted transposition in Drosophila melanogaster. Genetics. 1999 Mar;151(3):1093-101) as a starting point in designing a P-element replacement. In our hands, the rates of each of the steps in the crosses for P-element replacement were similar to the values reported in the Sepp and Auld paper.

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