ARABIDOPSIS 2010: Affinity Handle on Macromolecular Complexes

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Investigator: Nevan J. Krogan, PhD, MSc
Sponsor: University of Missouri System

Location(s): United States

Description

Networks of macromolecular interactions are fundamental to all biological processes. Until recently, the progress of functional networks studies in Arabidopsis has been propelled primarily by genetic-based approaches. On the other hand, the enormous potential of high-throughput studies of defined macromolecular entities has not been fully realized, largely because reliable affinity tag-enabled fractionation tools remain unavailable to most Arabidopsis researchers. To facilitate these studies, this project will develop and deliver a suite of high throughput cloning-adapted vectors incorporating quantitatively characterized tandem affinity tags, for rapid proteomic and RNomic analyses of Arabidopsis macromolecular complexes. In addition, the project will generate a set of robust purification protocols tailored to the specific affinity tags. These tangible deliverables will be disseminated to the Arabidopsis research community via the Arabidopsis Biological Resource Center (for binary vectors and plant lines), professional publications, TAIR as well as the project website (the purification protocols). This contribution will enable systems-level studies in a variety of areas of plant biology by many groups presently prevented from doing so by prohibitive cost of conducting the optimization studies in-house.