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Analysis of the expression patterns, subcellular localisations and interaction partners of Drosophila proteins using a pigP protein trap library

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journal contribution
posted on 2016-02-22, 11:39 authored by N. Lowe, J. S. Rees, J. Roote, E. Ryder, I. M. Armean, G. Johnson, E. Drummond, H. Spriggs, J. Drummond, J. P. Magbanua, H. Naylor, B. Sanson, R. Bastock, S. Huelsmann, V. Trovisco, M. Landgraf, S. Knowles-Barley, J. D. Armstrong, H. White-Cooper, Celia Hansen, R. G. Phillips, K. S. Lilley, S. Russell, D. St Johnston
Although we now have a wealth of information on the transcription patterns of all the genes in the Drosophila genome, much less is known about the properties of the encoded proteins. To provide information on the expression patterns and subcellular localisations of many proteins in parallel, we have performed a large-scale protein trap screen using a hybrid piggyBac vector carrying an artificial exon encoding yellow fluorescent protein (YFP) and protein affinity tags. From screening 41 million embryos, we recovered 616 verified independent YFP-positive lines representing protein traps in 374 genes, two-thirds of which had not been tagged in previous P element protein trap screens. Over 20 different research groups then characterized the expression patterns of the tagged proteins in a variety of tissues and at several developmental stages. In parallel, we purified many of the tagged proteins from embryos using the affinity tags and identified co-purifying proteins by mass spectrometry. The fly stocks are publicly available through the Kyoto Drosophila Genetics Resource Center. All our data are available via an open access database (Flannotator), which provides comprehensive information on the expression patterns, subcellular localisations and in vivo interaction partners of the trapped proteins. Our resource substantially increases the number of available protein traps in Drosophila and identifies new markers for cellular organelles and structures.

Funding

This work was supported by a project grant from the Wellcome Trust [076739], by a Wellcome Trust Principal Research Fellowship to D.StJ. [049818 and 080007], and by core support from the Wellcome Trust [092096] and Cancer Research UK [A14492].

History

Citation

Development 2014 141: 3994-400

Version

  • VoR (Version of Record)

Published in

Development 2014 141: 3994-400

Publisher

Company of Biologists: Development

issn

0950-1991

eissn

1477-9129

Acceptance date

2014-08-13

Copyright date

2014

Available date

2016-02-22

Publisher version

http://dev.biologists.org/content/141/20/3994

Notes

Supplementary material available online at http://dev.biologists.org/lookup/suppl/doi:10.1242/dev.111054/-/DC1

Associated authors

The UK Drosophila Protein Trap Screening Consortium

Language

en

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