Peptide aptamers to investigate and disrupt protein function in plants

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PI: Stanton B. Gelvin (Purdue University)CoPI: Vitaly Citovsky (State University of New York, Stony Brook) Aptamers are small synthetic peptides of random amino acid sequence. When expressed in cells, aptamers can specifically bind to target proteins and inhibit function. They can thus be used to define the functional roles of proteins in various cellular processes, as well as defining specific domains within these proteins that are important for the proteins to mediate these roles. Furthermore, the aptamer sequences can be used as functional identification "marks" of plant proteins that naturally interact with the target proteins. An expression vector and libraries of millions of random aptamers will be developed for use in transient transformation experiments. The vector will allow expression and screening of aptamers first in yeast, and subsequently in plants. The constructions will incorporate bimolecular fluorescence complementation (BiFC) technology to detect aptamer-protein interactions, and Gateway technologies to facilitate mobilization of aptamers among various vectors. As proof of concept, tagged plant proteins important for development, and Agrobacterium virulence proteins important for Agrobacterium-mediated transformation will be probed with the aptamer library. This initial work will test whether random aptamers can be used to identify interacting proteins in two different organisms, yeast and plants. This project will provide research tools and resources for the plant science community to determine the functions of plant proteins, and protein-protein interactions in planta. Data will be released sequentially through a project website at Purdue University (accessible via http://bio.purdue.edu/people/faculty/index.php?refID=128) and the aptamer library will be provided to the ABRC for distribution.