METHODS OF SYNTHESIZING CELL-FREE PROTEIN

摘要

A batch-type method of synthesizing a cell-free protein which is characterized by comprising, in a means of a reaction for synthesizing a cell-free protein, (1) bringing a synthetic reaction solution (a reaction phase) containing a biological extract into contact directly with a substrate and an energy source supplying solution (a supplying phase) without separating by a semi-permeable membrane, thus continuously supplying the substrate and the energy source molecules of the supplying phase into the reaction phase by free diffusion via the contact interface and, at the same time, taking off the by-products formed in the reaction phase into the supplying phase to thereby prolong the reaction time, and thus elevating the reaction efficiency; (2) pre-incubating (pre-heating) a reaction solution, containing a wheat germ extract, then diluting the synthesis reaction solution by adding a diluent solution to thereby prolong the synthetic reaction time, and thus elevating the synthesis efficiency; (3) re-supplying a substrate required in the synthesis of the protein (amino acid, ATP, GTP, creatine phosphate, etc.) and an energy source to the reaction mixture with the use of a gel filtration column or a semi-permeable membrane after ceasing the synthesis reaction and, at the same time, discontinuously taking off the by-products formed by the reaction to thereby elevate the efficiency of the reaction.