Development of a rapid and productive cell-free protein synthesis system

Prolonged cell‐free protein synthesis using dual energy sources: Combined use of creatine phosphate and glucose for the efficient supply of ATP and retarded accumulation of phosphate

“…A stable supply of ATP is one of key factors affecting the efficiency of cell-free protein synthesis (Kim and Swartz, 1999;Kim et al, 2006a). Since the translation of genetic information into protein molecules involves many steps that consume ATP or its equivalents, an adequate and steady supply of ATP is essential for the prolonged synthesis of proteins in a cell-free synthesis system.…”

A simple and cost-effective method to prepare template DNAs for cell-free protein synthesis

“…Depending on the experiment, 0.1 µg plasmid was used as the template to direct the protein synthesis. The synthesis reactions were conducted in a water bath at 37 o C for 2 h. The synthesized protein was quantified by measuring the TCA-precipitable radioactivity as described previously [10][11][12][13]. The molecular weight of the expressed protein was confirmed with a 13% Tricine-SDS-polyacrylamide gel [15] and Western blot analysis.…”

A highly efficient and economical cell-free protein synthesis system using the S12 extract of Escherichia coli

“…The use of glucose not only reduces the cost of the energy source, but also enables a steady supply of ATP over extended reaction periods. Moreover, unlike the energy sources that have been previously used such as phosphoenolpyruvate (PEP) and creatine phosphate (CP), the use of glucose does not result in the accumulation of inorganic phosphate, which can potentially inhibit cell-free protein synthesis by sequestrating magnesium ions [6]. However, the current protocols that involve glucose also have significant drawbacks.…”