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Candidate enzymes for screening are most often selected by sequence homology (even those with weak homology, to augment diversity) with reference proteins (known proteins with a specific activity).
The candidate genes are amplified from DNA from a collection of strains from Genoscope and the “Cloaca maxima” metagenome, then cloned in 96-well microtiter plates by a cloning method which employs restriction and ligation enzymes.
The cloned genes are then over-expressed in E. coli BL21 in 96-well microtiter plates followed by the preparation of cellular lysates. Enzymatic screening of these plates is carried out in 96 or 384-well format.
This platform, which has been functioning for more than a year, has a current output of about 5000 clonings per year.