Abstract
We describe a reliable GAL4-based yeast two-hybrid system for identifying and isolating clones encoding proteins interacting with a protein of interest. This two-hybrid system gives extremely low background and few false-positive clones, making it ideal for library screening purposes. We have successfully used it not only to isolate Arabidopsis NPR1-interactors from rice but also to pull out the rice NPR1 ortholog using one of the interactors as bait.
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References
Chevray, P. M. and Nathans, D. (1992) Protein interaction cloning in yeast: identification of mammalian proteins that react with the leucine zipper of jun. Proc. Natl. Acad. Sci. USA 89, 5789–5793.
Chern, M., Fitzgerald, H. A., Canlas, P. E., Navarre, D. A., and Ronald, P. C. (2005) Overexpression of a rice NPRI homolog leads to constitutive activation of defense response and hypersensitivity to light. Mol. Plant Microbe Interact. 18, 511–520.
Chern, M.-S., Fitzgerald, H. A., Yadav, R. C., Canlas, P. E., Dong, X., and Ronald, P. C. (2001). Evidence for a disease-resistance pathway in rice similar to the NPR1-mediated signaling pathway in Arabidopsis. Plant J. 27, 101–113.
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© 2007 Humana Press Inc.
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Chern, M., Richter, T., Ronald, P.C. (2007). Yeast Two-Hybrid Approaches to Dissecting the Plant Defense Response. In: Ronald, P.C. (eds) Plant-Pathogen Interactions. Methods in Molecular Biology, vol 354. Humana Press. https://doi.org/10.1385/1-59259-966-4:79
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DOI: https://doi.org/10.1385/1-59259-966-4:79
Publisher Name: Humana Press
Print ISBN: 978-1-58829-448-7
Online ISBN: 978-1-59259-966-0
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