RT Journal Article
SR Electronic
T1 Exploring Strategies for Protein Trapping in Drosophila
JF Genetics
JO Genetics
FD Genetics Society of America
SP 1089
OP 1104
DO 10.1534/genetics.106.065995
VO 175
IS 3
A1 Quiñones-Coello, Ana T.
A1 Petrella, Lisa N.
A1 Ayers, Kathleen
A1 Melillo, Anthony
A1 Mazzalupo, Stacy
A1 Hudson, Andrew M.
A1 Wang, Shu
A1 Castiblanco, Claudia
A1 Buszczak, Michael
A1 Hoskins, Roger A.
A1 Cooley, Lynn
YR 2007
UL http://www.genetics.org/content/175/3/1089.abstract
AB 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.