3. Associomics of membrane proteins in two model organisms, yeast and arabidopsis (ASSOCIOPORT)

Funding code: QLG2-CT-2001-01297

The EU project Associoport embraces the challenge of establishing a "large scale" procedure for systematic identification of protein-protein interactions with the specific focus on membrane proteins on the basis of the split-ubiquitin system. Saccharonyces cerevisiae and Arabidopsis thaliana serve as model organisms.

Project partners:

Zentrum für Molekularbiologie der Pflanzen (ZMBP)
Eberhard-Karls-Universität Tübingen

Lab. De Physiologie Cellulaire
Université libre de Bruxelles

Institut für Mikrobiologie
Heinrich-Heine Universität Düsseldorf

Departamento Microbiol. y Genetica
Universidad de Salamanca

Unité de Bioinformatique
Université libre de Bruxelles

Sympore GmbH, Tübingen

GenExpress GmbH, Berlin

As industrial partner GenExpress is responsible for providing suitable PCR products for in vivo recombination of these fragments in the split-ubiquitin system (SUS) as well as the establishment of a DNA chip for the yeast/arabidopsis protein expression monitoring.

For the generation of the PCR products GenExpress established a standardized procedure for optimal amplification of all relevant genes from arabidopsis and yeast. To this end, clusters were selected based on the length of the fragments (300bp-1000bp, 1000bp 3000bp and >3000bp). The variation of the PCR parameters allowed the high-throughput amplification of 419 arabidopsis and 281 yeast genes on a quantitative scale.

During the course of the project the question was raised whether it was possible to adjust the SUS via gateway technology. In this context, gene expression vectors were developed on the basis of the gateway system.



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