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A bilayer cell-free protein synthesis system for high-throughput screening of gene products

Authors
Journal
FEBS Letters
0014-5793
Publisher
Wiley Blackwell (John Wiley & Sons)
Publication Date
Volume
514
Issue
1
Identifiers
DOI: 10.1016/s0014-5793(02)02329-3
Keywords
  • Cell-Free Protein Synthesis
  • Bilayer Reaction Method
  • Escherichia Colicoupled Transcription–Translation
  • Wheat Germ System
  • High-Throughput Screening
Disciplines
  • Biology

Abstract

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.

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