Development of cell-free gene translator as an equipment to support researches in bioscience and biotechnology

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Wth aims of developing a laboratory instrument which enables to obtain several mg of foreign active protein in one or two days, and of establishing method of preparing large amount of Escherichia coli S30 extract which is used in the system, the following have been achieved.1. Achievement of Nagoya UniversityA cell-discruption equipment(trade name : beads schocker) was found to be useful to get large amount of S30 extract. Maximally about 50mL of the extract could be obtained. Optimal operational conditions of this equipment was established. A reaction condition to synthesize active protein having correct disulfide bridge was also elucidated.2. Achievement of Tomy Manufacturing Co., Ltd.A small closed dialyzer having semipermiable membrane with working volume of 0. 1〜 0. 3mL was developed. Using this dialyzer, ca. 1. 2mg/mL of chlaramphenicol acetyltransferase could be synthesized by cell-free transcription/ translation system containing E. coli S30 extract in 12 hours.3. Achievement of Nippon Flour Mills Co., Ltd.Long-term stability of S30 extract in its storage in liquid nitrogen was tested. The extract was found to be stably stored for about one year.