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Originally published as Genetics Published Articles Ahead of Print on December 28, 2006.

Genetics, Vol. 175, 1505-1531, March 2007, Copyright © 2007
doi:10.1534/genetics.106.065961

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The Carnegie Protein Trap Library: A Versatile Tool for Drosophila Developmental Studies

Michael Buszczak*, Shelley Paterno*, Daniel Lighthouse*, Julia Bachman*, Jamie Planck*, Stephenie Owen*, Andrew D. Skora*, Todd G. Nystul*, Benjamin Ohlstein*, Anna Allen*, James E. Wilhelm*, Terence D. Murphy*, Robert W. Levis*, Erika Matunis{dagger}, Nahathai Srivali*, Roger A. Hoskins{ddagger} and Allan C. Spradling*,1

* Howard Hughes Medical Institute Research Laboratories, Department of Embryology, Carnegie Institution of Washington, Baltimore, Maryland 21218, {dagger} Department of Cell Biology, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205 and {ddagger} Department of Genome Biology, Life Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, California 94720

1 Corresponding author: Carnegie Institution, 3520 San Martin Dr., Baltimore, MD 21218.
E-mail: spradling{at}ciwemb.edu

Metazoan physiology depends on intricate patterns of gene expression that remain poorly known. Using transposon mutagenesis in Drosophila, we constructed a library of 7404 protein trap and enhancer trap lines, the Carnegie collection, to facilitate gene expression mapping at single-cell resolution. By sequencing the genomic insertion sites, determining splicing patterns downstream of the enhanced green fluorescent protein (EGFP) exon, and analyzing expression patterns in the ovary and salivary gland, we found that 600–900 different genes are trapped in our collection. A core set of 244 lines trapped different identifiable protein isoforms, while insertions likely to act as GFP-enhancer traps were found in 256 additional genes. At least 8 novel genes were also identified. Our results demonstrate that the Carnegie collection will be useful as a discovery tool in diverse areas of cell and developmental biology and suggest new strategies for greatly increasing the coverage of the Drosophila proteome with protein trap insertions.


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