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PSB

Carl-Ivar Brändén 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.