A bilayer cell-free protein synthesis system for high-throughput screening of gene products.
A high-throughput cell-free protein synthesis method has been described. The methodology is based on a bilayer diffusion system that enables the continuous supply of substrates, together with the continuous removal of small byproducts, through a phase between the translation mixture and substrate mixture. With the use of a multititer plate the system was functional for a prolonged time, and as a consequence yielded more than 10 times that of the similar batch-mode reaction. Combining this method with a wheat germ cell-free translation system developed by us, the system could produce a large amount of protein sufficient for carrying out functional analyses. This novel bilayer-based cell-free protein synthesis system with its simplicity, minimum time and low cost may be useful practical methodology in the post-genome era.
Sawasaki
Tatsuya
T
Department of Applied Chemistry, Faculty of Engineering, The Venture Business Laboratory, Ehime University, 790-8577, Matsuyama, Japan.
Hasegawa
Yoshinori
Y
Tsuchimochi
Masateru
M
Kamura
Nami
N
Ogasawara
Tomio
T
Kuroita
Toshihiro
T
Endo
Yaeta
Y
D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D002474:Cell-Free System; D018076:DNA, Complementary / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005821:Genetic Techniques; D008051:Lipid Bilayers; D014176:Protein Biosynthesis
D001426:Bacterial Proteins; D018076:DNA, Complementary; D008051:Lipid Bilayers