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Zaltsman, A., Department of Biochemistry and Cell Biology, State University of New York, Stony Brook, NY 11794-5215, United States
Yi, B.-Y., Department of Biochemistry and Cell Biology, State University of New York, Stony Brook, NY 11794-5215, United States, Department of Environmental Horticulture, University of Seoul, Tongdaemoon, Seoul 130-743, South Korea
Krichevsky, A., Department of Biochemistry and Cell Biology, State University of New York, Stony Brook, NY 11794-5215, United States
Gafni, Y., Department of Genetics, Agricultural Research Organization, Volcani Center, Bet Dagan 50250, Israel
Citovsky, V., Department of Biochemistry and Cell Biology, State University of New York, Stony Brook, NY 11794-5215, United States
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. © 2007 American Society of Plant Biologists.
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Yeast-plant coupled vector system for identification of nuclear proteins
145
Zaltsman, A., Department of Biochemistry and Cell Biology, State University of New York, Stony Brook, NY 11794-5215, United States
Yi, B.-Y., Department of Biochemistry and Cell Biology, State University of New York, Stony Brook, NY 11794-5215, United States, Department of Environmental Horticulture, University of Seoul, Tongdaemoon, Seoul 130-743, South Korea
Krichevsky, A., Department of Biochemistry and Cell Biology, State University of New York, Stony Brook, NY 11794-5215, United States
Gafni, Y., Department of Genetics, Agricultural Research Organization, Volcani Center, Bet Dagan 50250, Israel
Citovsky, V., Department of Biochemistry and Cell Biology, State University of New York, Stony Brook, NY 11794-5215, United States
Yeast-plant coupled vector system for identification of nuclear proteins
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. © 2007 American Society of Plant Biologists.
Scientific Publication
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