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Elife. 2016 Feb 20;5:e12068. doi: 10.7554/eLife.12068.

A genome-wide resource for the analysis of protein localisation in Drosophila.

Author information

1
Max Planck Institute of Cell Biology and Genetics, Dresden, Germany.
2
Muscle Dynamics Group, Max Planck Institute of Biochemistry, Martinsried, Germany.
3
Department of Proteomics and Signal Transduction, Max Planck Institute of Biochemistry, Martinsried, Germany.
4
Centre for Cellular and Molecular Platforms, National Centre for Biological Sciences, Tata Institute of Fundamental Research, Bangalore, India.
5
Heidelberg Institute of Theoretical Studies, Heidelberg, Germany.
6
Department of Molecular Cell and Developmental Biology, University of California, Los Angeles, Los Angeles, United States.
7
Institute of Neuroscience, Trinity College Dublin, Dublin, Ireland.

Abstract

The Drosophila genome contains >13000 protein-coding genes, the majority of which remain poorly investigated. Important reasons include the lack of antibodies or reporter constructs to visualise these proteins. Here, we present a genome-wide fosmid library of 10000 GFP-tagged clones, comprising tagged genes and most of their regulatory information. For 880 tagged proteins, we created transgenic lines, and for a total of 207 lines, we assessed protein expression and localisation in ovaries, embryos, pupae or adults by stainings and live imaging approaches. Importantly, we visualised many proteins at endogenous expression levels and found a large fraction of them localising to subcellular compartments. By applying genetic complementation tests, we estimate that about two-thirds of the tagged proteins are functional. Moreover, these tagged proteins enable interaction proteomics from developing pupae and adult flies. Taken together, this resource will boost systematic analysis of protein expression and localisation in various cellular and developmental contexts.

KEYWORDS:

<i>d. melanogaster</i>; developmental biology; drosophila; evolutionary biology; genetics; genomics; live imaging; protein tagging; recombineering; resource; stem cells

Comment in

PMID:
26896675
PMCID:
PMC4805545
DOI:
10.7554/eLife.12068
[Indexed for MEDLINE]
Free PMC Article
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