COOPERATIVITY OF STABILIZED MESSENGER-RNA AND ENHANCED TRANSLATION ACTIVITY IN THE CELL-FREE SYSTEM

Detailed analysis of cell-free translation, coupled transcription-tran slation in static conditions and a continuous how system based on E. c oli S30 extracts was performed. Degradation of template mRNA was the p redominant trigger to terminate the protein synthesis. In a coupled sy stem, mRNA was preserved by repeated transcription whereas the starvat ion of nucleotide triphosphates led to the termination of protein synt hesis in less than 1 h. In the CFCF system, NTP was held at the level of initial concentration and therefore did not arrest the translation for 15 h. The accurate coupling of transcriptional rate and translatio nal rate was also crucial to enhance the efficiency of protein synthes is.