In Vitro Synthesis of Proteins in Bacterial Extracts

This protocol describes the methods used to generate protein in a cell-free system derived from E. coli. The in vitro synthesis of protein has been used in studying many ribosome-based gene regulation steps (Gong and Yanofsky, 2001). Such techniques have also been used to study protein-protein, protein-DNA, and protein-RNA interactions, and to produce radiolabeled protein species. More recently, such approaches have been utilized to produce large quantities of toxic proteins for proteomic and structural studies (Yokoyama et al., 2000).