Preparation and Optimization of a Cell-free Translation System from Vicia sativa Germ Lacking Ribosome-inactivating Protein Activity

A cell-free translation system from Vicia sativa with high activityfor reading endogenous messengers was prepared. The concentrationsof several of the components were optimized. The system washighly sensitive to cycloheximide, puromycin and fusidic acid,but was completely insensitive to chloramphenicol. The systemlacks apparent free endogenous ribosome-inactivating proteinactivity. It was highly sensitive to several well-known ribosome-inactivatingproteins, which makes it very suitable for studying the effectsand the mechanism of action of these plant toxins. Key words: Translation system, protein synthesis, cell-free extracts, ribosome-inactivating proteins, Vicia sativa