Exploring strategies for protein trapping in drosophila

Ana T. Quiñones-Coello, Lisa N. Petrella, Kathleen Ayers, Anthony Melillo, Stacy Mazzalupo, Andrew M. Hudson, Shu Wang, Claudia Castiblanco, Michael Buszczak, Roger A. Hoskins, Lynn Cooley

Research output: Contribution to journalArticle

120 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)1089-1104
Number of pages16
JournalGenetics
Volume175
Issue number3
DOIs
StatePublished - Mar 2007

ASJC Scopus subject areas

  • Genetics

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    Quiñones-Coello, A. T., Petrella, L. N., Ayers, K., Melillo, A., Mazzalupo, S., Hudson, A. M., Wang, S., Castiblanco, C., Buszczak, M., Hoskins, R. A., & Cooley, L. (2007). Exploring strategies for protein trapping in drosophila. Genetics, 175(3), 1089-1104. https://doi.org/10.1534/genetics.106.065995