Objectif The objective of this proposal is to develop new tools for the functional identification of new plant genes. It aims at assigning genes to identified proteins In a first step, we propose to focus on a particular set of proteins and choose the plant plasma membrane proteins because (i) functional studies of this interface between the cell interior and external medium are at the forefront of plant cell biology, (ii) less than 5 % of the total number of plasma membrane proteins are represented within databases, and (iii) it will be a challenge to fill this gap in our knowledge by systematic generation of sequence information on plasma membrane proteins and respective DNA sequences. This information must be generated and stored in a centrally available database to provide an important resource for researchers from both academia and industry. We propose a coordinated effort to construct a fundamental database on Arabidopsis plasma membrane proteins, connecting protein expression and sequence information to available or new EST data. Our action will be based on: the construction of a comprehensive directory of plasma membrane proteins (by 2D PAGE) - the generation of partial amino acid sequence data - the development of new methodologies allowing both to identify rare plasma membrane proteins (by using an original expression cloning procedure) and to assign protein sequence data to partial DNA sequences available in EST databases (by using an innovative gene chips methodology) and the construction of a database accessible either directly or through cross referred existing databases. A first part of the work involving most partners will concern, beside an effort for the standardization of procedures, (i) the analysis of plasma membrane protein expression patterns in various Arabidopsis tissues and under various environmental conditions and (ii) additional characterization of given protein subsets A large part of the project, concerning again most partners, will be devoted to automated protein sequencing using a main preparative material that will be selected from the results obtained at the preceeding step. At the same time, a new procedure using ordered arrays of available ESTs on glass grids will be developped for Arabidopsis by one partner (already running this system for other organisms) to connect protein sequence data to cDNA sequences. In addition, rare plasma membrane proteins will be isolated by one partner using protein expression and insertion into the plasma membrane of mammalian cells according to a procedure this partner recently developped for plasma membrane proteins. Finally, during all the project, one partner will be specifically in charge to construct first the protein directory and then the database that will contain both General Image (GI) data sets and Quantitative and Qualitative Individual Spots (QQIS) data sets. The final plasma membrane database is expected to constitute a resource of vital importance as a basic directory that increases our knowledge of the Arabidopsis genome, and as a link between other coordinated initiatives from the gene to function. Champ scientifique natural sciencescomputer and information sciencesdatabasesnatural sciencesbiological sciencesgeneticsDNAnatural sciencesbiological sciencesbiochemistrybiomoleculesproteinsnatural sciencesbiological sciencescell biologynatural scienceschemical sciencesorganic chemistryamines Programme(s) FP4-BIOTECH 2 - Specific research, technological development and demonstration programme in the field of biotechnology, 1994-1998 Thème(s) 020204 - New methods for genome analysis Appel à propositions Data not available Régime de financement CSC - Cost-sharing contracts Coordinateur Institut National de la Recherche Agronomique Contribution de l’UE Aucune donnée Adresse 2,Place Pierre Viala 34060 Montpellier France Voir sur la carte Coût total Aucune donnée Participants (6) Trier par ordre alphabétique Trier par contribution de l’UE Tout développer Tout réduire INSTITUT NATIONAL DE LA RECHERCHE AGRONOMIQUE France Contribution de l’UE Aucune donnée Adresse Route de Saint-Cyr RD10 78026 VERSAILLES Voir sur la carte Coût total Aucune donnée MAX-PLANCK-GESELLSCHAFT ZUR FOERDERUNG DER WISSENSCHAFTEN E.V. Allemagne Contribution de l’UE Aucune donnée Adresse Carl-von-Linné-Weg 10 50829 KOELN Voir sur la carte Coût total Aucune donnée Stiftung Deutsches Krebsforschungszentrum Allemagne Contribution de l’UE Aucune donnée Adresse 280,Im Neuenheimer Feld 280 69120 Heidelberg Voir sur la carte Coût total Aucune donnée UNIVERSITE CATHOLIQUE DE LOUVAIN Belgique Contribution de l’UE Aucune donnée Adresse 2/20,Place Croix du Sud 2/20 1348 LOUVAIN-LA-NEUVE Voir sur la carte Coût total Aucune donnée Universiteit Gent Belgique Contribution de l’UE Aucune donnée Adresse 35,Ledeganckstraat 9000 Gent Voir sur la carte Coût total Aucune donnée University of Cambridge Royaume-Uni Contribution de l’UE Aucune donnée Adresse Downing Street CB2 3EA Cambridge Voir sur la carte Coût total Aucune donnée