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

eLife

K VijayRaghavan¹  Aishwarya Krishnamoorthy¹  Vinay Vikas KJ¹  RT Krishnan¹  Volker Hartenstein²  Matthias Mann³  Marco Y Hein³  Siegfried Schloissnig⁴  Philipp Kämpfer⁴  Mani Ramaswami⁵  Stephan Janosch⁶  Dana Suchold⁶  Helena Jambor⁶  Mihail Sarov⁶  Susanne Hasse⁶  Radoslaw K Ejsmont⁶  Christopher Schmied⁶  Elisabeth Vinis⁶  Pavel Tomancak⁶  Elisabeth Knust⁶  Frank Schnorrer⁷  Bettina Stender⁷  Irene RS Ferreira⁷  Nicole Plewka⁷  Christiane Barz⁷  Katja Finkl⁷ 

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

关键词: drosophila;    resource;    protein tagging;    live imaging;    recombineering;    genetics;   

DOI  :  10.7554/eLife.12068

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown