원문정보
초록
영어
Despite the promising potential of cell-free protein synthesis, the high cost of the required reagents has plagued its practical application. In particular, most of the reagent cost is attributed to the expenses for conventional energy sources such as phophsoenol pyruvate (PEP) and
creatine phopshate (CP). In this study, for the preparative expression of recombinant proteins at a reduced energy cost, we attempted to use the glycolytic intermediates to drive the cell-free protein synthesis in a continuous-exchange cell-free protein synthesis (CECF) reaction. Among
the glycolytic intermediates examined, use of fructose 1,6-bisphosphate (FBP) gave the highest yield of protein synthesis, producing approximately 7 mg/ml of a chloramphenicol acetyl transferase (CAT) protein. We also found that protein synthesis in the CECF reaction was able to be directed by PCR-amplified linear DNA templates by employing the measures to stabilize mRNA in the reaction mixture.