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Published on August 17, 2007; 10.1104/pp.107.105973

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Received July 26, 2007
Accepted August 13, 2007

Yeast-Plant Coupled Vector System for Identification of Nuclear Proteins

Adi Zaltsman , Bu-Young Yi , Alexander Krichevsky , Yedidya Gafni , and Vitaly Citovsky *

Department of Biochemistry and Cell Biology, State University of New York, Stony Brook, NY 11794-5215; Department of Environmental Horticulture, University of Seoul, Tongdaemoon, 130-743, Korea; Department of Genetics, Agricultural Research Organization, Volcani Center, P.O. Box 6, Bet Dagan 50250, Israel

* Corresponding author; email: vitaly.citovsky{at}stonybrook.edu.

Nuclear proteins are involved in many critical biological processes within plant cells, and, therefore, are in the focus of studies which 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 which 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 co-expression with any other proteins.




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T. Tzfira, S. V. Kozlovsky, and V. Citovsky
Advanced Expression Vector Systems: New Weapons for Plant Research and Biotechnology
Plant Physiology, December 1, 2007; 145(4): 1087 - 1089.
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