title: A bilayer cell-free protein synthesis system for high-throughput screening of gene products.
abstract: 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.
authors.affiliation: Department of Applied Chemistry, Faculty of Engineering, The Venture Business Laboratory, Ehime University, 790-8577, Matsuyama, Japan.
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Sawasaki
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Hasegawa
authors.forename: Masateru
authors.initials: M
authors.lastname: Tsuchimochi
authors.forename: Nami
authors.initials: N
authors.lastname: Kamura
authors.forename: Tomio
authors.initials: T
authors.lastname: Ogasawara
authors.forename: Toshihiro
authors.initials: T
authors.lastname: Kuroita
authors.forename: Yaeta
authors.initials: Y
authors.lastname: Endo
mesh_terms: 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
chemical_list: D001426:Bacterial Proteins; D018076:DNA, Complementary; D008051:Lipid Bilayers