[Animal and viral mRNA translation in a mRNA-dependent cell-free system and rabbit reticulocytes]

Abstract

A system of cell-free synthesis from rabbit reticulocytes is described. A method of repeated bloodlettings for induction of reticulocytosis was used. The lysate of washed reticulocytes was treated with micrococcus nuclease enabling effective inhibition of synthesis in endogenous templates. The translating activity is manifested most completely in the presence of the following compounds: ATP, GTP, ATP-regenerating system, SH-compounds, polyamines, tRNA. This system must be optimized in K+ and Mg2+ concentrations for different templates. The system is tolerant to high concentrations of rRNA which allows the utilization of total RNA preparations in the experiments. This system may be satisfactorily used for translation of mRNA of different origins.