14] Isolation and translation of mouse interferon mRNAs

Abstract

It has been observed that the wheat germ cell-free protein-synthesizing system translates mouse interferon mRNA to protein that was active in inhibiting viral multiplication in mouse cells. Easy availability of the wheat embryo and the simplicity of the procedure for the preparation of the extract make this material a good choice for studies of protein synthesis. Mouse interferon mRNAs isolated by the two methods as discussed in this chapter are translated in the wheat germ extract with high efficiency. The fidelity of the translation is reflected in the production of the biologically active product. A high efficiency of translation of mouse interferon mRNA to biologically active protein in the wheat germ ceil-free system needs the presence of polyamines. Translation of many plant viral RNAs in the wheat germ extract is stimulated by addition of polyamines such as spermine or spermidine. It is important to maximize the translation system for both the ionic requirements and the polyamine concentration. The product of translation of mouse mRNA in the wheat germ extract is characterized as mouse interferon by (a) its inhibition of virus multiplication in homologous cells, (b) its absence of activity in heterologous cells, (c) its neutralization by anti-mouse interferon antiserum, and (d) the absence of inhibitory effects on viral multiplication by the translation products obtained from mRNA isolated from non-induced cells.