A STABILIZED REACTION MIXTURE FOR IN-VITRO TRANSLATION OF MESSENGER-RNA

Here we describe a method for obtaining a ready-to-use stabilized reac tion mixture for in vitro translation of mRNA. We also demonstrate the stabilization of a complete translation mixture containing wheat germ extract, amino acids, ATP, GTP, creatine phosphate, creatine kinase, and the reaction buffer by lyophilization in the presence of various s ugars. The greatest stabilizing effect is achieved by supplementing th e mixture with 10% (mass/volume) trehalose, which is also a unique tra nslation activator, enhancing the translation of various mRNAs. A lyop hilized complete translation mixture containing trehalose can be store d at 4-8 degrees C for several months without losing its activity. The mixture can be easily reconstituted by adding an aqueous mRNA solutio n and retains the potential for reproducible functioning. This allows the employment of such a cell-free translation system for analytical s creening of a broad spectrum of compounds inhibiting translation at va rious stages.