Isotope-Assisted Proteomic Analysis of Hormone Receptor Mutants

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There are two specific aims in this project, one focused on genomic technology development and the other, on identifying proteins involved in mediating the effect of a specific hormone, cytokinin, in plant cells. The technology to be developed involves using mass spectrometers and heavy isotopes to measure the concentration and modifications of as many of the 30,000 different plant proteins as possible. This new technology in the field commonly known as 'proteomics' is badly needed in order to convert genomic sequence data into knowledge that represents a real understanding of the underlying biological mechanisms. These measurements will be performed with specific 'knockout' mutant plants that are resistant to the effects of the plant hormone. By measuring how these mutations affect the presence of particular proteins, the handful of proteins that are specifically involved in mediating the effects of the hormone will be identified. Broader impacts of this project beyond the development of a much needed, generally applicable technology include organizing workshops for the research community in the methods that will be developed and the introduction of concepts in functional genomics into high school biology classrooms.