Cell-Free Protein Production System
Cell-free protein production system consists of RNA polymerase, transfer RNAs (tRNAs), aminoacyl-tRNA synthetases, translation factors, ribosomes, and DNA for the template of a target protein. It is reconstituted by E. coli extract and chemical compounds, e.g. aminoacids, NTP (nucleotide 5'-triphosphates: ATP, CTP, GTP and UTP), and so on.
Our cell-free system uses E. coli BL21 strain and produces 1 mg of GFP (Green Fluorescence Protein) per mL of a reaction. Both plasmid and linear DNA are usable for the template. A quick screening for protein production by SDS-PAGE can be achieved by using PCR products. In cooperation with Nonnatural Amino Acid Technology Team, we develop an advanced cell-free system through the site-specific incorporation of modified aminoacids into a target protein. Since it makes possible introductions of chemical labeling without damage for protein function, it will contribute to developing novel techniques for life science research, e.g. molecular imaging and probing of partner biomolecules.