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doi:10.1534/genetics.106.065961
A more recent version of this article appeared on March 1, 2007.
REGULAR RESEARCH PAPERS |
The Carnegie protein trap library: a versatile tool for Drosophila developmental studies
Michael Buszczak 1, Shelley Paterno 1, Daniel Lighthouse 1, Julia Bachman 1, Jamie Plank 1, Stephanie Owen 1, Andrew Skora 1, Todd Nystul 1, Benjamin Ohlstein 1, Anna Allen 1, James Wilhelm 1, Terence Murphy 1, Bob Levis 1, Erika Matunis 2, Nahathai Srivali 1, Roger Hoskins 3 and Allan Spradling 1*
1 Carnegie Institution
2 Johns Hopkins Medical School
3 Lawrence Berkeley National Labs
* To whom correspondence should be addressed. E-mail: spradling{at}ciwemb.edu.
Submitted on September 18, 2006
Revised on November 4, 2006
Accepted on 18 December 2006
Metazoan physiology depends on intricate patterns of gene expression that remain poorly known. Using transposon mutagenesis in Drosophila, we constructed a library of 7,404 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 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 245 lines trapped different identifiable protein isoforms, while insertions likely to act as GFP-enhancer traps were found in 284 additional genes. At least seven 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.
Key Words: P-element, gene expression, oogenesis, piggyBac, protein fusion
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