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          Institute: MPI für molekulare Pflanzenphysiologie     Collection: Publikationen Pflanzenphysiologie     Display Documents

ID: 230012.0, MPI für molekulare Pflanzenphysiologie / Publikationen Pflanzenphysiologie
Versatile gene-specific sequence tags for Arabidopsis functional genomics: Trancript profiling and reverse genetics applications
Authors:Hilson, P.; Allemeersch, J.; Altmann, T; Aubourg, S; Avon, A.; Beynon, J.; Bhalerao, R. P.; Bitton, F.; Caboche, M.; Cannoot, B.; Chardakov, V.; Cognet-Holliger, C.; Colot, V.; Crowe, M.; Darimont, C.; Durinck, S.; Eickhoff, H.; de Longevialle, A. F.; Farmer, E. E.; Grant, M.; Kuiper, M. T. R.; Lehrach, H.; Leon, C.; Leyva, A.; Lundeberg, J.; Lurin, C.; Moreau, Y.; Nietfeld, W.; Paz-Ares, J.; Reymond, P.; Rouze, P.; Sandberg, G.; Segura, M. D.; Serizet, C.; Tabrett, A.; Taconnat, L.; Thareau, V.; Van Hummelen, P.; Vercruysse, S.; Vuylsteke, M.; Weingartner, M.; Weisbeek, P. J.; Wirta, V.; Wittink, F. R. A.; Zabeau, M.; Small, I.
Date of Publication (YYYY-MM-DD):2004-10
Title of Journal:Genome Research
Journal Abbrev.:Genome Res
Issue / Number:10B
Start Page:2176
End Page:2189
Review Status:not specified
Audience:Not Specified
Abstract / Description:Microarray transcript profiling and RNA interference are two new technologies crucial for large-scale gene function studies in multicellular eukaryotes. Both rely on sequence-specific hybridization between complementary nucleic acid strands, inciting us to create a collection of gene-specific sequence tags (GSTs) representing at least 21,500 Arabidopsis genes and which are compatible with both approaches. The GSTs were carefully selected to ensure that each of them shared no significant similarity with any other region in the Arabidopsis genome. They were synthesized by PCR amplification from genomic DNA. Spotted microarrays fabricated from the GSTs show good dynamic range, specificity, and sensitivity in transcript profiling experiments. The GSTs have also been transferred to bacterial plasmid vectors via recombinational cloning protocols. These cloned GSTs constitute the ideal starting point for a variety of functional approaches, including reverse genetics. We have subcloned GSTs on a large scale into vectors designed for gene silencing in plant cells. We show that in planta expression of GST hairpin RNA results in the expected phenotypes in silenced Arabidopsis lines. These versatile GST resources provide novel and powerful tools for functional genomics.
Free Keywords:repeat-containing gene
; secondary cell-wall
; vacuolar h+-atpase
; microarray analysis
; escherichia-coli
; wide expression
; restorer gene
; plant-growth
; design
; thaliana
External Publication Status:published
Document Type:Article
Communicated by:N. N.
Affiliations:MPI für molekulare Pflanzenphysiologie/Universitätsgastgruppen/AG Altmann
External Affiliations:State Univ Ghent, Flanders Interuniv Inst Biotechnol, Dept Plant Syst Biol, B-9052 Ghent, Belgium
State Univ Ghent, Flanders Interuniv Inst Biotechnol, Dept Plant Syst Biol, B-9052 Ghent, Belgium
UEVE, CNRS, INRA, Unite Rech Genom Vegetale, F-91057 Evry, France
Katholieke Univ Leuven, Fac Engn, Dept Elect Engn, ESAT, B-3001 Heverlee, Belgium
Univ Potsdam, Inst Biochem & Biol Genet, Max Planck Inst Mol Pflanzenphysiol, D-14476 Golm, Germany
Hort Res Int, Warwick CV35 9EF, England
Swedish Univ Agr Sci, Dept Forest Genet & Plant Physiol, S-90183 Umea, Sweden
Univ London Imperial Coll Sci Technol & Med, Dept Agr Sci, Ashford TN25 5AH, Kent, England
INRA, Genet & Ameliorat Plantes Stn, F-78026 Versailles, France
John Innes Ctr, CH-1015 Lausanne, Switzerland
Univ Lausanne, Dept Plant Mol Biol, Gene Express Lab, CH-1015 Lausanne, Switzerland
Max Planck Inst Mol Genet, Dept Vertebrate Genom, D-14195 Berlin, Germany
CSIC, Ctr Nacl Biotecnol, Dept Plant Mol Genet, E-28049 Madrid, Spain
AlbaNova Univ Ctr, Royal Inst Technol, KTH, Dept Biotechnol, S-10691 Stockholm, Sweden
UZ Gasthuisberg, VIB MicroArray Facil, B-3000 Louvain, Belgium
Univ Utrecht, Dept Mol Genet, NL-3584 CH Utrecht, Netherlands
Identifiers:ISI:000224514000024 [ID No:1]
ISI:000224514000024 [ID No:2]
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