A high-yield cell-free system of protein synthesis of mouse liver

1. A fractionated cell-free system of protein synthesis has been developed from mouse liver. It is composed of polysomes, "pH 5" fraction, Mg2+, K+, ATP and a ATP generating system. 2. It operates optimally at 30-37 degrees C, in the presence of 4 mM MgCl2 and 90 mM KCl. 3. Spermine is highly inhibitory, while spermidine shows a bimodal action, in that submillimolar concentrations stimulate, while millimolar concentrations inhibit protein synthesis. 4. Both spermine and spermidine show an interesting selectivity, in that, even though they inhibit incorporation of amino acids into most proteins, they stimulate incorporation into a few proteins. 5. The system can be rendered mRNA-dependent, either by preincubation or by treatment with micrococcal nuclease. In both cases globin mRNA as well as TMV RNA are faithfully translated. 6. Compared to other published mammalian fractionated cell-free systems, the mouse liver system is more efficient by approximately one order of magnitude, since the rate of incorporation of leucine per min is 30 pmol/mg protein or 435 pmol/mg RNA or 1 mol/mol ribosomes.