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Plant Physiology 133:462-469 (2003)
© 2003 American Society of Plant Biologists

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BREAKTHROUGH TECHNOLOGIES

A Gateway Cloning Vector Set for High-Throughput Functional Analysis of Genes in Planta[w]

Mark D. Curtis* and Ueli Grossniklaus

Institute of Plant Biology and Zürich-Basel Plant Science Centre, University of Zürich, Zollikerstrasse 107, CH-8008 Zürich, Switzerland

The current challenge, now that two plant genomes have been sequenced, is to assign a function to the increasing number of predicted genes. In Arabidopsis, approximately 55% of genes can be assigned a putative function, however, less than 8% of these have been assigned a function by direct experimental evidence. To identify these functions, many genes will have to undergo comprehensive analyses, which will include the production of chimeric transgenes for constitutive or inducible ectopic expression, for antisense or dominant negative expression, for subcellular localization studies, for promoter analysis, and for gene complementation studies. The production of such transgenes is often hampered by laborious conventional cloning technology that relies on restriction digestion and ligation. With the aim of providing tools for high throughput gene analysis, we have produced a Gateway-compatible Agrobacterium sp. binary vector system that facilitates fast and reliable DNA cloning. This collection of vectors is freely available, for noncommercial purposes, and can be used for the ectopic expression of genes either constitutively or inducibly. The vectors can be used for the expression of protein fusions to the Aequorea victoria green fluorescent protein and to the {beta}-glucuronidase protein so that the subcellular localization of a protein can be identified. They can also be used to generate promoter-reporter constructs and to facilitate efficient cloning of genomic DNA fragments for complementation experiments. All vectors were derived from pCambia T-DNA cloning vectors, with the exception of a chemically inducible vector, for Agrobacterium sp.-mediated transformation of a wide range of plant species.

www.plantphysiol.org/cgi/doi/10.1104/pp.103.027979.

[w] The online version of this article contains Web-only data.

* Corresponding author; e-mail mcurtis{at}botinst.unizh.ch; fax 4116348204.

Received June 2, 2003; returned for revision June 17, 2003; accepted June 25, 2003.




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Recombinational Cloning with Plant Gateway Vectors
Plant Physiology, December 1, 2007; 145(4): 1144 - 1154.
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 J Exp BotHome page
M. Latijnhouwers, T. Gillespie, P. Boevink, V. Kriechbaumer, C. Hawes, and C. M. Carvalho
Localization and domain characterization of Arabidopsis golgin candidates
J. Exp. Bot., December 1, 2007; 58(15-16): 4373 - 4386.
[Abstract] [Full Text] [PDF]

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 Plant Physiol.Home page
M. Karimi, A. Bleys, R. Vanderhaeghen, and P. Hilson
Building Blocks for Plant Gene Assembly
Plant Physiology, December 1, 2007; 145(4): 1183 - 1191.
[Abstract] [Full Text] [PDF]

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 Plant Physiol.Home page
A. Himmelbach, U. Zierold, G. Hensel, J. Riechen, D. Douchkov, P. Schweizer, and J. Kumlehn
A Set of Modular Binary Vectors for Transformation of Cereals
Plant Physiology, December 1, 2007; 145(4): 1192 - 1200.
[Abstract] [Full Text] [PDF]

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 Plant CellHome page
A. Para, E. M. Farre, T. Imaizumi, J. L. Pruneda-Paz, F. G. Harmon, and S. A. Kay
PRR3 Is a Vascular Regulator of TOC1 Stability in the Arabidopsis Circadian Clock
PLANT CELL, November 1, 2007; 19(11): 3462 - 3473.
[Abstract] [Full Text] [PDF]

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 Proc. Natl. Acad. Sci. USAHome page
N. Uchida, B. Townsley, K.-H. Chung, and N. Sinha
Regulation of SHOOT MERISTEMLESS genes via an upstream-conserved noncoding sequence coordinates leaf development
PNAS, October 2, 2007; 104(40): 15953 - 15958.
[Abstract] [Full Text] [PDF]

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 Plant CellHome page
N. Winter, G. Kollwig, S. Zhang, and F. Kragler
MPB2C, a Microtubule-Associated Protein, Regulates Non-Cell-Autonomy of the Homeodomain Protein KNOTTED1
PLANT CELL, October 1, 2007; 19(10): 3001 - 3018.
[Abstract] [Full Text] [PDF]

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 J. Biol. Chem.Home page
Y. Tsegaye, C. G. Richardson, J. E. Bravo, B. J. Mulcahy, D. V. Lynch, J. E. Markham, J. G. Jaworski, M. Chen, E. B. Cahoon, and T. M. Dunn
Arabidopsis Mutants Lacking Long Chain Base Phosphate Lyase Are Fumonisin-sensitive and Accumulate Trihydroxy-18:1 Long Chain Base Phosphate
J. Biol. Chem., September 21, 2007; 282(38): 28195 - 28206.
[Abstract] [Full Text] [PDF]

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 Plant Physiol.Home page
F. Li, T. Asami, X. Wu, E. W.T. Tsang, and A. J. Cutler
A Putative Hydroxysteroid Dehydrogenase Involved in Regulating Plant Growth and Development
Plant Physiology, September 1, 2007; 145(1): 87 - 97.
[Abstract] [Full Text] [PDF]

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 Plant Physiol.Home page
S. A. Saracco, M. J. Miller, J. Kurepa, and R. D. Vierstra
Genetic Analysis of SUMOylation in Arabidopsis: Conjugation of SUMO1 and SUMO2 to Nuclear Proteins Is Essential
Plant Physiology, September 1, 2007; 145(1): 119 - 134.
[Abstract] [Full Text] [PDF]

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 Plant CellHome page
Y. Nakagawa, H. Hanaoka, M. Kobayashi, K. Miyoshi, K. Miwa, and T. Fujiwara
Cell-Type Specificity of the Expression of Os BOR1, a Rice Efflux Boron Transporter Gene, Is Regulated in Response to Boron Availability for Efficient Boron Uptake and Xylem Loading
PLANT CELL, August 1, 2007; 19(8): 2624 - 2635.
[Abstract] [Full Text] [PDF]

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 Plant CellHome page
Y. Yan, S. Stolz, A. Chetelat, P. Reymond, M. Pagni, L. Dubugnon, and E. E. Farmer
A Downstream Mediator in the Growth Repression Limb of the Jasmonate Pathway
PLANT CELL, August 1, 2007; 19(8): 2470 - 2483.
[Abstract] [Full Text] [PDF]

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 Plant Physiol.Home page
M. N. Martin, P. H. Saladores, E. Lambert, A. O. Hudson, and T. Leustek
Localization of Members of the {gamma}-Glutamyl Transpeptidase Family Identifies Sites of Glutathione and Glutathione S-Conjugate Hydrolysis
Plant Physiology, August 1, 2007; 144(4): 1715 - 1732.
[Abstract] [Full Text] [PDF]

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 Plant Physiol.Home page
C. Lunde, D. P. Drew, A. K. Jacobs, and M. Tester
Exclusion of Na+ via Sodium ATPase (PpENA1) Ensures Normal Growth of Physcomitrella patens under Moderate Salt Stress
Plant Physiology, August 1, 2007; 144(4): 1786 - 1796.
[Abstract] [Full Text] [PDF]

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 Plant CellHome page
K. Kobayashi, M. S. Otegui, S. Krishnakumar, M. Mindrinos, and P. Zambryski
INCREASED SIZE EXCLUSION LIMIT2 Encodes a Putative DEVH Box RNA Helicase Involved in Plasmodesmata Function during Arabidopsis Embryogenesis
PLANT CELL, June 1, 2007; 19(6): 1885 - 1897.
[Abstract] [Full Text] [PDF]

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 Plant CellHome page
K. Marinova, L. Pourcel, B. Weder, M. Schwarz, D. Barron, J.-M. Routaboul, I. Debeaujon, and M. Klein
The Arabidopsis MATE Transporter TT12 Acts as a Vacuolar Flavonoid/H+-Antiporter Active in Proanthocyanidin-Accumulating Cells of the Seed Coat
PLANT CELL, June 1, 2007; 19(6): 2023 - 2038.
[Abstract] [Full Text] [PDF]

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 GeneticsHome page
P. A. Reeves, Y. He, R. J. Schmitz, R. M. Amasino, L. W. Panella, and C. M. Richards
Evolutionary Conservation of the FLOWERING LOCUS C-Mediated Vernalization Response: Evidence From the Sugar Beet (Beta vulgaris)
Genetics, May 1, 2007; 176(1): 295 - 307.
[Abstract] [Full Text] [PDF]

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 Plant CellHome page
F. Chopin, M. Orsel, M.-F. Dorbe, F. Chardon, H.-N. Truong, A. J. Miller, A. Krapp, and F. Daniel-Vedele
The Arabidopsis ATNRT2.7 Nitrate Transporter Controls Nitrate Content in Seeds
PLANT CELL, May 1, 2007; 19(5): 1590 - 1602.
[Abstract] [Full Text] [PDF]

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 DevelopmentHome page
J. F. Uhrig, M. Mutondo, I. Zimmermann, M. J. Deeks, L. M. Machesky, P. Thomas, S. Uhrig, C. Rambke, P. J. Hussey, and M. Hulskamp
The role of Arabidopsis SCAR genes in ARP2-ARP3-dependent cell morphogenesis
Development, March 1, 2007; 134(5): 967 - 977.
[Abstract] [Full Text] [PDF]

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 Plant CellHome page
S. Yehudai-Resheff, S. L. Zimmer, Y. Komine, and D. B. Stern
Integration of Chloroplast Nucleic Acid Metabolism into the Phosphate Deprivation Response in Chlamydomonas reinhardtii
PLANT CELL, March 1, 2007; 19(3): 1023 - 1038.
[Abstract] [Full Text] [PDF]

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 Plant Physiol.Home page
Q. Zeng, X. Wang, and M. P. Running
Dual Lipid Modification of Arabidopsis G{gamma}-Subunits Is Required for Efficient Plasma Membrane Targeting
Plant Physiology, March 1, 2007; 143(3): 1119 - 1131.
[Abstract] [Full Text] [PDF]

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 Plant Physiol.Home page
A. B. Sivitz, A. Reinders, M. E. Johnson, A. D. Krentz, C. P.L. Grof, J. M. Perroux, and J. M. Ward
Arabidopsis Sucrose Transporter AtSUC9. High-Affinity Transport Activity, Intragenic Control of Expression, and Early Flowering Mutant Phenotype
Plant Physiology, January 1, 2007; 143(1): 188 - 198.
[Abstract] [Full Text] [PDF]

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 Plant CellHome page
M. Chen, G. Han, C. R. Dietrich, T. M. Dunn, and E. B. Cahoon
The Essential Nature of Sphingolipids in Plants as Revealed by the Functional Identification and Characterization of the Arabidopsis LCB1 Subunit of Serine Palmitoyltransferase
PLANT CELL, December 1, 2006; 18(12): 3576 - 3593.
[Abstract] [Full Text] [PDF]

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 J. Biol. Chem.Home page
S. Goepfert, J. K. Hiltunen, and Y. Poirier
Identification and Functional Characterization of a Monofunctional Peroxisomal Enoyl-CoA Hydratase 2 That Participates in the Degradation of Even cis-Unsaturated Fatty Acids in Arabidopsis thaliana
J. Biol. Chem., November 24, 2006; 281(47): 35894 - 35903.
[Abstract] [Full Text] [PDF]

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 J Exp BotHome page
M. E. Salvucci, B. P. DeRidder, and A. R. Portis Jr
Effect of activase level and isoform on the thermotolerance of photosynthesis in Arabidopsis
J. Exp. Bot., November 1, 2006; 57(14): 3793 - 3799.
[Abstract] [Full Text] [PDF]

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 CSH ProtocolsHome page
N. Basherudin and M. D. Curtis
Identification of Positive GATEWAY Expression Clones When Both the pENTRY and pDEST Vectors Contain the Same Marker for Bacterial Selection
CSH Protocols, November 1, 2006; 2006(29): pdb.prot4647 - pdb.prot4647.
[Abstract] [Full Text]

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 Plant CellHome page
K. Ohashi-Ito and D. C. Bergmann
Arabidopsis FAMA Controls the Final Proliferation/Differentiation Switch during Stomatal Development
PLANT CELL, October 1, 2006; 18(10): 2493 - 2505.
[Abstract] [Full Text] [PDF]

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 Proc. Natl. Acad. Sci. USAHome page
Y.-S. Wang, R. Shrestha, A. Kilaru, W. Wiant, B. J. Venables, K. D. Chapman, and E. B. Blancaflor
Manipulation of Arabidopsis fatty acid amide hydrolase expression modifies plant growth and sensitivity to N-acylethanolamines
PNAS, August 8, 2006; 103(32): 12197 - 12202.
[Abstract] [Full Text] [PDF]

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 Plant Physiol.Home page
L. Brand, M. Horler, E. Nuesch, S. Vassalli, P. Barrell, W. Yang, R. A. Jefferson, U. Grossniklaus, and M. D. Curtis
A Versatile and Reliable Two-Component System for Tissue-Specific Gene Induction in Arabidopsis
Plant Physiology, August 1, 2006; 141(4): 1194 - 1204.
[Abstract] [Full Text] [PDF]

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 Plant Physiol.Home page
R. Bari, B. Datt Pant, M. Stitt, and W.-R. Scheible
PHO2, MicroRNA399, and PHR1 Define a Phosphate-Signaling Pathway in Plants
Plant Physiology, July 1, 2006; 141(3): 988 - 999.
[Abstract] [Full Text] [PDF]


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