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First published online April 23, 2004; 10.1104/pp.104.040139

Plant Physiology 135:25-38 (2004)
© 2004 American Society of Plant Biologists

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BREAKTHROUGH TECHNOLOGIES

High-Throughput Fluorescent Tagging of Full-Length Arabidopsis Gene Products in Planta1

Guo-Wei Tian2, Amitabh Mohanty2, S. Narasimha Chary2, Shijun Li2, Brigitte Paap2, Georgia Drakakaki2, Charles D. Kopec, Jianxiong Li, David Ehrhardt, David Jackson*, Seung Y. Rhee, Natasha V. Raikhel and Vitaly Citovsky

Department of Biochemistry and Cell Biology, State University of New York, Stony Brook, New York 11794–5215 (G.-W.T., B.P., C.D.K., J.L., V.C.); Watson School of Biological Sciences (C.D.K.) and Cold Spring Harbor Laboratory (A.M., D.J.), Cold Spring Harbor, New York 11724; Center for Plant Cell Biology and Department of Botany and Plant Sciences, University of California, Riverside, California 92521–0124 (N.C., G.D., N.V.R.); and The Arabidopsis Information Resource (S.L., S.Y.R.) and Carnegie Institution of Washington (S.L., D.E., S.Y.R), Stanford, California 94305

We developed a high-throughput methodology, termed fluorescent tagging of full-length proteins (FTFLP), to analyze expression patterns and subcellular localization of Arabidopsis gene products in planta. Determination of these parameters is a logical first step in functional characterization of the approximately one-third of all known Arabidopsis genes that encode novel proteins of unknown function. Our FTFLP-based approach offers two significant advantages: first, it produces internally-tagged full-length proteins that are likely to exhibit native intracellular localization, and second, it yields information about the tissue specificity of gene expression by the use of native promoters. To demonstrate how FTFLP may be used for characterization of the Arabidopsis proteome, we tagged a series of known proteins with diverse subcellular targeting patterns as well as several proteins with unknown function and unassigned subcellular localization.

1 This work was supported by a grant from the 2010 Project of the National Science Foundation to V.C., D.E., D.J., N.R., and S.R.

2 These authors contributed equally to the paper.

www.plantphysiol.org/cgi/doi/10.1104/pp.104.040139.

* Corresponding author; e-mail jacksond{at}cshl.org; fax 516–367–8369.

Received February 3, 2004; returned for revision March 9, 2004; accepted March 12, 2004.




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