Evaluation of cell-free protein synthesis using PDMS-based microreactor arrays.
A living cell has numerous proteins, only a few thousand of which have been identified to date. Cell-free protein synthesis is a useful and promising technique to discover and produce various proteins that might be beneficial for biotechnological, pharmaceutical, and medical applications. For this study, we evaluated the performance and the general applicability of our previously developed microreactor array chip to cell-free protein synthesis by comparisons with a commercially available system. The microreactor array chip comprises a temperature control chip made of glass and a disposable reaction chamber chip made of polydimethylsiloxane (PDMS). For evaluation of the microreactor array chip, rat adipose-type fatty acid binding protein, glyceraldehyde-3-phosphate dehydrogenase, cyclophilin, and firefly luciferase were synthesized from their respective DNA templates using a cell-free extract prepared from Escherichia coli. All these proteins were synthesized in the microreactor array chip, and their respective amounts and yields were investigated quantitatively.
Yamamoto
Takatoki
T
Center for International Research on MicroMechatronics, Institute of Industrial Science, The University of Tokyo, Meguro, Tokyo, Japan. takatoki@iis.u-tokyo.ac.jp
Hino
Mami
M
Kakuhata
Rei
R
Nojima
Takahiko
T
Shinohara
Yasuo
Y
Baba
Yoshinobu
Y
Fujii
Teruo
T
D000818:Animals; D002474:Cell-Free System / Q000737:chemistry; D021983:Cyclophilins / Q000138:chemical synthesis* / Q000737:chemistry; D004129:Dimethylpolysiloxanes / Q000737:chemistry*; D050556:Fatty Acid-Binding Proteins / Q000138:chemical synthesis* / Q000737:chemistry; D050536:Glycerol-3-Phosphate Dehydrogenase (NAD+) / Q000138:chemical synthesis* / Q000737:chemistry; D008156:Luciferases / Q000138:chemical synthesis* / Q000737:chemistry; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods; D051381:Rats; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012828:Silicones / Q000737:chemistry*
D004129:Dimethylpolysiloxanes; D050556:Fatty Acid-Binding Proteins; D012828:Silicones; C013830:baysilon; D050536:Glycerol-3-Phosphate Dehydrogenase (NAD+); D008156:Luciferases; D021983:Cyclophilins