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Title: Exploring strategies for protein trapping in Drosophila

Abstract

The use of fluorescent protein tags has had a huge impact oncell biological studies in virtually every experimental system.Incorporation of coding sequence for fluorescent proteins such as greenfluorescent protein (GFP) into genes at their endogenous chromosomalposition is especially useful for generating GFP-fusion proteins thatprovide accurate cellular and subcellular expression data. We testedmodifications of a transposon-based protein trap screening procedure inDrosophila to optimize the rate of recovering useful protein traps andtheir analysis. Transposons carrying the GFP-coding sequence flanked bysplice acceptor and donor sequences were mobilized, and new insertionsthat resulted in production of GFP were captured using an automatedembryo sorter. Individual stocks were established, GFP expression wasanalyzed during oogenesis, and insertion sites were determined bysequencing genomic DNA flanking the insertions. The resulting collectionincludes lines with protein traps in which GFP was spliced into mRNAs andembedded within endogenous proteins or enhancer traps in which GFPexpression depended on splicing into transposon-derived RNA. We report atotal of 335 genes associated with protein or enhancer traps and aweb-accessible database for viewing molecular information and expressiondata for these genes.

Authors:
; ; ; ; ; ; ; ; ; ;
Publication Date:
Research Org.:
COLLABORATION - Yale U. School ofMedicine
OSTI Identifier:
923654
Report Number(s):
LBNL-62587
Journal ID: ISSN 0016-6731; GENTAE; R&D Project: L0111; BnR: 600305000; TRN: US200804%%1379
DOE Contract Number:  
DE-AC02-05CH11231
Resource Type:
Journal Article
Resource Relation:
Journal Name: Genetics; Journal Volume: 175; Journal Issue: 3; Related Information: Journal Publication Date: March 2007
Country of Publication:
United States
Language:
English
Subject:
59; DNA; DROSOPHILA; EMBRYOS; GENES; MODIFICATIONS; OOGENESIS; PRODUCTION; PROTEINS; RNA; SPLICING; TRANSPOSONS; TRAPPING

Citation Formats

Quinones-Coello, Ana T., Petrella, Lisa N., Ayers, Kathleen, Melillo, Anthony, Mazzalupo, Stacy, Hudson, Andrew M., Wang, Shu, Castiblanco, Claudia, Buszczak, Michael, Hoskins, Roger A., and Cooley, Lynn. Exploring strategies for protein trapping in Drosophila. United States: N. p., 2006. Web. doi:10.1534/genetics.106.065995.
Quinones-Coello, Ana T., Petrella, Lisa N., Ayers, Kathleen, Melillo, Anthony, Mazzalupo, Stacy, Hudson, Andrew M., Wang, Shu, Castiblanco, Claudia, Buszczak, Michael, Hoskins, Roger A., & Cooley, Lynn. Exploring strategies for protein trapping in Drosophila. United States. doi:10.1534/genetics.106.065995.
Quinones-Coello, Ana T., Petrella, Lisa N., Ayers, Kathleen, Melillo, Anthony, Mazzalupo, Stacy, Hudson, Andrew M., Wang, Shu, Castiblanco, Claudia, Buszczak, Michael, Hoskins, Roger A., and Cooley, Lynn. Mon . "Exploring strategies for protein trapping in Drosophila". United States. doi:10.1534/genetics.106.065995.
@article{osti_923654,
title = {Exploring strategies for protein trapping in Drosophila},
author = {Quinones-Coello, Ana T. and Petrella, Lisa N. and Ayers, Kathleen and Melillo, Anthony and Mazzalupo, Stacy and Hudson, Andrew M. and Wang, Shu and Castiblanco, Claudia and Buszczak, Michael and Hoskins, Roger A. and Cooley, Lynn},
abstractNote = {The use of fluorescent protein tags has had a huge impact oncell biological studies in virtually every experimental system.Incorporation of coding sequence for fluorescent proteins such as greenfluorescent protein (GFP) into genes at their endogenous chromosomalposition is especially useful for generating GFP-fusion proteins thatprovide accurate cellular and subcellular expression data. We testedmodifications of a transposon-based protein trap screening procedure inDrosophila to optimize the rate of recovering useful protein traps andtheir analysis. Transposons carrying the GFP-coding sequence flanked bysplice acceptor and donor sequences were mobilized, and new insertionsthat resulted in production of GFP were captured using an automatedembryo sorter. Individual stocks were established, GFP expression wasanalyzed during oogenesis, and insertion sites were determined bysequencing genomic DNA flanking the insertions. The resulting collectionincludes lines with protein traps in which GFP was spliced into mRNAs andembedded within endogenous proteins or enhancer traps in which GFPexpression depended on splicing into transposon-derived RNA. We report atotal of 335 genes associated with protein or enhancer traps and aweb-accessible database for viewing molecular information and expressiondata for these genes.},
doi = {10.1534/genetics.106.065995},
journal = {Genetics},
number = 3,
volume = 175,
place = {United States},
year = {Mon Dec 18 00:00:00 EST 2006},
month = {Mon Dec 18 00:00:00 EST 2006}
}