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Originally published as Genetics Published Articles Ahead of Print on December 18, 2006.

Genetics, Vol. 175, 1089-1104, March 2007, Copyright © 2007
doi:10.1534/genetics.106.065995

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Exploring Strategies for Protein Trapping in Drosophila

Ana T. Quiñones-Coello*,1, Lisa N. Petrella*,1, Kathleen Ayers*,1, Anthony Melillo*, Stacy Mazzalupo*,2, Andrew M. Hudson*, Shu Wang*,3, Claudia Castiblanco*, Michael Buszczak{dagger},{ddagger}, Roger A. Hoskins§ and Lynn Cooley*,{dagger},**,4

* Department of Genetics, ** Department of Cell Biology and {dagger} Department of Molecular, Cellular and Developmental Biology, Yale University School of Medicine, New Haven, Connecticut 06520, {ddagger} Department of Embryology, Carnegie Institution of Washington, Baltimore, Maryland 21218 and § Department of Genome Biology, Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720

4 Corresponding author: Department of Genetics, Yale University School of Medicine, P.O. Box 208005, 333 Cedar St., New Haven, CT 06520-8005.
E-mail: lynn.cooley{at}yale.edu

The use of fluorescent protein tags has had a huge impact on cell biological studies in virtually every experimental system. Incorporation of coding sequence for fluorescent proteins such as green fluorescent protein (GFP) into genes at their endogenous chromosomal position is especially useful for generating GFP-fusion proteins that provide accurate cellular and subcellular expression data. We tested modifications of a transposon-based protein trap screening procedure in Drosophila to optimize the rate of recovering useful protein traps and their analysis. Transposons carrying the GFP-coding sequence flanked by splice acceptor and donor sequences were mobilized, and new insertions that resulted in production of GFP were captured using an automated embryo sorter. Individual stocks were established, GFP expression was analyzed during oogenesis, and insertion sites were determined by sequencing genomic DNA flanking the insertions. The resulting collection includes lines with protein traps in which GFP was spliced into mRNAs and embedded within endogenous proteins or enhancer traps in which GFP expression depended on splicing into transposon-derived RNA. We report a total of 335 genes associated with protein or enhancer traps and a web-accessible database for viewing molecular information and expression data for these genes.


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Genetics 2007 175: NP. [Full Text]  



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Copyright © 2007 by the Genetics Society of America.