The carnegie protein trap library: a versatile tool for Drosophila developmental studies.
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.
Buszczak
Michael
M
Howard Hughes Medical Institute Research Laboratories, Department of Embryology, Carnegie Institution of Washington, Baltimore, Maryland 21218, USA.
Paterno
Shelley
S
Lighthouse
Daniel
D
Bachman
Julia
J
Planck
Jamie
J
Owen
Stephenie
S
Skora
Andrew D
AD
Nystul
Todd G
TG
Ohlstein
Benjamin
B
Allen
Anna
A
Wilhelm
James E
JE
Murphy
Terence D
TD
Levis
Robert W
RW
Matunis
Erika
E
Srivali
Nahathai
N
Hoskins
Roger A
RA
Spradling
Allan C
AC
D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D004251:DNA Transposable Elements / Q000235:genetics; D004330:Drosophila / Q000235:genetics*; D029721:Drosophila Proteins / Q000235:genetics* / Q000378:metabolism; D005260:Female; D020869:Gene Expression Profiling / Q000379:methods; D018507:Gene Expression Regulation, Developmental; D015723:Gene Library; D017344:Genes, Insect / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D008969:Molecular Sequence Data; D016296:Mutagenesis; D010053:Ovary / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012469:Salivary Glands / Q000378:metabolism; D017422:Sequence Analysis, DNA
D017931:DNA Primers; D004251:DNA Transposable Elements; D029721:Drosophila Proteins; D049452:Green Fluorescent Proteins