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PMID: 17194782 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The carnegie protein trap library: a versatile tool for Drosophila developmental studies.

Genetics ·Vol. 175 ·No. 3 ·2007-03-00 ·Pages 1505-31

Buszczak M, Paterno S, Lighthouse D, Bachman J, Planck J, Owen S, Skora AD, Nystul TG, Ohlstein B, Allen A, Wilhelm JE, Murphy TD, Levis RW, Matunis E, Srivali N, Hoskins RA, Spradling AC

Abstract

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.

MeSH Terms
Animals Base Sequence DNA Primers DNA Transposable Elements/genetics Drosophila/genetics Drosophila Proteins/genetics,metabolism Female Gene Expression Profiling/methods Gene Expression Regulation, Developmental Gene Library Genes, Insect/genetics Green Fluorescent Proteins/metabolism Molecular Sequence Data Mutagenesis Ovary/metabolism Reverse Transcriptase Polymerase Chain Reaction Salivary Glands/metabolism Sequence Analysis, DNA
Chemicals
DNA Primers DNA Transposable Elements Drosophila Proteins Green Fluorescent Proteins
Authors & Affiliations
17 authors, click to expand affiliations / ORCID
Buszczak Michael
Howard Hughes Medical Institute Research Laboratories, Department of Embryology, Carnegie Institution of Washington, Baltimore, Maryland 21218, USA.
Paterno Shelley
Lighthouse Daniel
Bachman Julia
Planck Jamie
Owen Stephenie
Skora Andrew D
Nystul Todd G
Ohlstein Benjamin
Allen Anna
Wilhelm James E
Murphy Terence D
Levis Robert W
Matunis Erika
Srivali Nahathai
Hoskins Roger A
Spradling Allan C
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Article Info
Journal
Genetics
Abbr.
Genetics
ISSN
0016-6731
Published
2007-03-00
Epub
2006-00-28
Pages
1505-31
Language
English
Region
United States
NLM ID
0374636
PMCID
PMC1840051
Subset
IM
Grants
Howard Hughes Medical Institute · United States
NICHD NIH HHS · R01 HD040307 · United States
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