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        Français

        PSB

        Carl-Ivar Br?ndén building (CIBB)
        71, Avenue des Martyrs
        CS 90181
        F-38042 Grenoble
        Intranet

        ESPRIT

        Purpose of the platform (operated by ISBG-IBS)

        Proprietary library-based screening technology to identify solubly expressing constructs of problematic proteins


        Expression of soluble protein by random incremental truncation

        - The ESPRIT construct screening platform has been developed at EMBL to identify soluble constructs of "difficult-to-express" protein targets that resist the classical approach of bioinformatics and PCR cloning.

        - It employs the principles of directed evolution whereby a diverse random library of DNA constructs is generated and screened to identify rare clones of interest (soluble expressers).

        - All unidirectional truncations of the target gene, both 5’ and 3’, are synthesised by exonuclease degradation to generate potential expression constructs.

        - Additionally, we have developed a "scanning" version to identify internal domains. In each experiment, 30,000 individual clones are assayed in parallel for yield and solubility using a highly automated colony array format.


        Responsible

        Darren Hart
        Tel.: 04 57 42 85 86


        Staff in charge of the platform

        Philippe Mas tel: 04 76 20 94 81


        In practice

        High workload of a library construction and screen means that collaborating scientist should help in preparation of experiments. Experiment is approximately 1 month in duration.


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