Optimisation of an in vitro enzyme selection

Isolation of a catalyst for a given chemical reaction may be achieved by in vitro selection of enzymes from a protein library. Here, we investigate the polymerisation reaction on filamentous phage and the cross-linking of substrate on phage to optimise an in vitro selection for DNA polymerase activity. The efficiency of the optimised selection is measured by enrichment factors up to 3.8 x 10(3), the highest described so far for an in vitro selection of proteins for catalysis. It should be useful for directed polymerase evolution towards novel catalytic activities. (C) 2003 Academie des sciences. Publie par Editions scientitiques et medicales Elsevier SAS. Tons droits reserves.