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Yeast-Plant Coupled Vector System for Identification of Nuclear Proteins^1,[OA]

Department of Biochemistry and Cell Biology, State University of New York, Stony Brook, New York 11794–5215 (A.Z., B.-Y.Y., A.K., V.C.); Department of Environmental Horticulture, University of Seoul, Tongdaemoon, Seoul 130–743, Korea (B.-Y.Y.); and Department of Genetics, Agricultural Research Organization, Volcani Center, Bet Dagan 50250, Israel (Y.G.)

Nuclear proteins are involved in many critical biological processes within plant cells and, therefore, are in the focus of studies that usually begin with demonstrating that the protein of interest indeed exhibits nuclear localization. Thus, studies of plant nuclear proteins would be facilitated by a convenient experimental system for identification of proteins that are actively imported into the cell nucleus and visualization of their nuclear accumulation in vivo. To this end, we developed a system of vectors that allows screening for cDNAs coding for nuclear proteins in a simple genetic assay in yeast cells, and verification of nuclear accumulation in planta following one-step transfer and autofluorescent tagging of the identified clones into a multiple cloning site-compatible and reading frame-compatible plant expression vector. In a recommended third experimental step, the plant expression cassette containing the identified clone can be transferred, also by a one-step cloning, into a binary multigene expression vector for transient or stable coexpression with any other proteins.

The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (www.plantphysiol.org) is: Vitaly Citovsky (vitaly.citovsky{at}stonybrook.edu).

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www.plantphysiol.org/cgi/doi/10.1104/pp.107.105973

^* Corresponding author; e-mail vitaly.citovsky{at}stonybrook.edu.

Received July 26, 2007; accepted August 13, 2007; published August 17, 2007.

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