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Cell ; 150(4): 855-66, 2012 Aug 17.
Artículo en Inglés | MEDLINE | ID: mdl-22901814

RESUMEN

Understanding the in vivo dynamics of protein localization and their physical interactions is important for many problems in biology. To enable systematic protein function interrogation in a multicellular context, we built a genome-scale transgenic platform for in vivo expression of fluorescent- and affinity-tagged proteins in Caenorhabditis elegans under endogenous cis regulatory control. The platform combines computer-assisted transgene design, massively parallel DNA engineering, and next-generation sequencing to generate a resource of 14,637 genomic DNA transgenes, which covers 73% of the proteome. The multipurpose tag used allows any protein of interest to be localized in vivo or affinity purified using standard tag-based assays. We illustrate the utility of the resource by systematic chromatin immunopurification and automated 4D imaging, which produced detailed DNA binding and cell/tissue distribution maps for key transcription factor proteins.


Asunto(s)
Animales Modificados Genéticamente , Proteínas de Caenorhabditis elegans/análisis , Caenorhabditis elegans/genética , Ingeniería Genética/métodos , Genoma de los Helmintos , Factores de Transcripción/análisis , Animales , Caenorhabditis elegans/química , Caenorhabditis elegans/metabolismo , Proteínas de Caenorhabditis elegans/genética , Factores de Transcripción/genética
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