2001 Fiscal Year Final Research Report Summary
Development of cell-free gene translator as an equipment to support researches in bioscience and biotechnology
| Project/Area Number |
10555287
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 展開研究 |
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| Research Field |
生物・生体工学
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| Research Institution | Nagoya University |
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Principal Investigator |
YAMANE Tsuneo Nagoya University, Graduate School of Bioagricultural Sciences, Professor, 大学院・生命農学研究科, 教授 (70026102)
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| Co-Investigator(Kenkyū-buntansha) |
SEKIGUCHI Satoshi Nippon Flours Mills Co. Ltd., Senior Researcher, 中央研究所, 主任研究員
TSUKUI Sadashi Tomy manufaturing Co.Ltd., Senior Researcher, 開発本部, 主任研究員
NAKANO Hideo Nagoya University, Graduate School of Bioagricultural Sciences, Associate Professor, 大学院・生命農学研究科, 助教授 (00237348)
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| Project Period (FY) |
1998 – 2001
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| Keywords | cell-free protein synthesis / transcription / translation / bioreactor / ATP / stabilization of protein / E. coli S30 extract / Wheat germ extract |
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| Research Abstract |
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 University
A 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.
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