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Molecular & Cellular Proteomics 6:413-424, 2007.
© 2007 by The American Society for Biochemistry and Molecular Biology, Inc.
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From the
Alliance for Cellular Signaling,
Molecular Biology Laboratory, California Institute of Technology, Pasadena, California 91125, ¶ Microscopy Laboratory, Stanford University, Palo Alto, California 94305, and || Bioinformatics and Data Coordination Laboratory, University of California San Diego, La Jolla, California 92093
Cellular responses to inputs that vary both temporally and spatially are determined by complex relationships between the components of cell signaling networks. Analysis of these relationships requires access to a wide range of experimental reagents and techniques, including the ability to express the protein components of the model cells in a variety of contexts. As part of the Alliance for Cellular Signaling, we developed a robust method for cloning large numbers of signaling ORFs into Gateway® entry vectors, and we created a wide range of compatible expression platforms for proteomics applications. To date, we have generated over 3000 plasmids that are available to the scientific community via the American Type Culture Collection. We have established a website at www.signaling-gateway.org/data/plasmid/ that allows users to browse, search, and blast Alliance for Cellular Signaling plasmids. The collection primarily contains murine signaling ORFs with an emphasis on kinases and G protein signaling genes. Here we describe the cloning, databasing, and application of this proteomics resource for large scale subcellular localization screens in mammalian cell lines.
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