Protein synthesizing system from wheat germ: efficient translation of synthetic and natural messages

Abstract

Optimum conditions for translation of eukaryotic, prokaryotic and synthetic templates in wheat germ cell-free extract were determined. 1. Translation of eukaryotic message (BMV RNA and TMV RNA) was at optimum at the same concentrations of K+, Mg(2+), HEPES and spermine. In optimal conditions the efficiency of translation was high, for BMV RNA being equal to 220 pmoles and for TMV RNA, to 280 pmoles of leucine incorporated per 1 microgram of template. Prokaryotic template (Qbeta RNA) was translated under different ionic conditions. 2. Translation of synthetic template [poly(U)] was at optimum at fairly higher concentrations of K+ and Mg(2+) than those optimal for natural template translation. 3. Efficient translation of natural and synthetic templates depends on complete removal of inhibitors found in the wheat germ cell-free extract. Action of these inhibitors could be mimicked by adenine nucleotides.