King, R.W., Lustig, K.D., Stukenberg, P.T., McGarry, T.J., Kirschner, M.W.
Notes: This Tech. Sight article describes the in vitro expression cloning technique (IVEC). The technique involves expression of plasmid pools (~50-100 clones/pool) in the TNT® Coupled Reticulocyte Lysate System, where approximately 30 proteins can be expressed in a single reaction. Each pool is screened for a certain biochemical activity, and positive pools are progressively subdivided until a single cDNA encoding the active protein is isolated. The technique is demonstrated diagrammatically with post-translational modifications (e.g., phosphorylation, degradation and cleavage of radiolabeled protein), but other activities could be assayed with this technique. (0901)