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A bilayer cell‐free protein synthesis system for high‐throughput screening of gene products
Author(s) -
Sawasaki Tatsuya,
Hasegawa Yoshinori,
Tsuchimochi Masateru,
Kamura Nami,
Ogasawara Tomio,
Kuroita Toshihiro,
Endo Yaeta
Publication year - 2002
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(02)02329-3
Subject(s) - bilayer , cell free protein synthesis , cell free system , translation (biology) , throughput , chemistry , substrate (aquarium) , cell , diffusion , biophysics , nanotechnology , biological system , computer science , protein biosynthesis , gene , biochemistry , materials science , membrane , biology , messenger rna , enzyme , physics , telecommunications , ecology , wireless , thermodynamics
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.