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          Institute: MPI für molekulare Zellbiologie und Genetik     Collection: Publikationen MPI-CBG 2008     Display Documents



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ID: 414325.0, MPI für molekulare Zellbiologie und Genetik / Publikationen MPI-CBG 2008
BAC TransgeneOmics: a high-throughput method for exploration of protein function in mammals
Authors:Poser, Ina; Sarov, Mihail; Hutchins, James R A; Heriche, Jean-Karim; Toyoda, Yusuke; Pozniakovsky, Andrei; Weigl, Daniela; Nitzsche, Anja; Hegemann, Bjorn; Bird, Alexander W; Pelletier, Laurence; Kittler, Ralf; Hua, Sujun; Naumann, Ronald; Augsburg, Martina; Sykora, Martina M; Hofemeister, Helmut; Zhang, Youming; Nasmyth, Kim; White, Kevin P; Dietzel, Steffen; Mechtler, Karl; Durbin, Richard; Stewart, A Francis; Peters, Jan-Michael; Buchholz, Frank; Hyman, Anthony A
Date of Publication (YYYY-MM-DD):2008
Title of Journal:Nature Methods
Volume:5
Issue / Number:5
Start Page:409
End Page:415
Copyright:not available
Audience:Experts Only
Intended Educational Use:No
Abstract / Description:The interpretation of genome sequences requires reliable and standardized methods to assess protein function at high throughput. Here we describe a fast and reliable pipeline to study protein function in mammalian cells based on protein tagging in bacterial artificial chromosomes (BACs). The large size of the BAC transgenes ensures the presence of most, if not all, regulatory elements and results in expression that closely matches that of the endogenous gene. We show that BAC transgenes can be rapidly and reliably generated using 96-well-format recombineering. After stable transfection of these transgenes into human tissue culture cells or mouse embryonic stem cells, the localization, protein-protein and/or protein-DNA interactions of the tagged protein are studied using generic, tag-based assays. The same high-throughput approach will be generally applicable to other model systems.
External Publication Status:published
Document Type:Article
Version Comment:Automatic journal name synchronization
Communicated by:n.n.
Affiliations:MPI f�r molekulare Zellbiologie und Genetik
Identifiers:LOCALID:987
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