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Preparation and Characterization of a Highly Active Cell-free Protein-synthesizing System from Dry Pea Primary Axes ¹

Laboratorium voor Plantenbiochemie, Katholieke Universiteit Leuven, Vaartstraat 24, B-3000 Leuven, Belgium

A highly active and reliable cell-free protein-synthesizing system from dry pea primary axes has been developed. The system has been optimized with respect to both overall activity and translational fidelity. Under optimal conditions, the system incorporates nearly 1,000 picomoles leucine per 50 microliters incubation mixture. At limiting tobacco mosaic virus RNA concentrations, its efficiency exceeds 1,000 picomoles leucine per picomole template molecules. The maximal size of the polypeptides coded for by tobacco mosaic virus RNA goes up to 180,000. A procedure has been worked out to reduce the endogenous template activity of the system. the dramatic increase of the activity of this pea cell-free system as compared to previous attempts can be explained by the use of Cl^–-free buffers.

¹ This work was supported in part by grants of the Belgian "National Fonds voor Wetenschappelijk Onderzoek" (N. F. W. O.).