| Project/Area Number |
07555258
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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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 | NARA INSTITUTE OF SCIENCE AND TECHNOLOGY |
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Principal Investigator |
SHINMYO Atsuhiko NAIST,BIOLOGICAL SCIENCES,PROFESSOR, バイオサイエンス研究科, 教授 (30029235)
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| Co-Investigator(Kenkyū-buntansha) |
TAKIHARA Takanobu ITOEN Ltd.CENTRAL RESEARCH INSTITUTE,RESEARCHER, 中央研究所, 研究主事
KAKUDA Takami ITOEN Ltd.CENTRAL RESEARCH INSTITUTE,SENIOR SCIENTIST, 中央研究所, 第一研究室長
SEKINE Masami NAIST,BIOLOGICAL SCIENCES,ASSISTANT, バイオサイエンス研究科, 助手 (70226653)
YOSHIDA Kazuya NAIST,BIOLOGICAL SCIENCES,ASSISTANT PROF., バイオサイエンス研究科, 助教授 (50252622)
瀧原 隆宣 (株)伊藤園, 中央研究所, 研究主事
角田 隆巳 (株)伊藤園, 中央研究所, 第一研究室長
瀧原 孝宣 株式会社 伊藤園, 中央研究所, 研究主事
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| Project Period (FY) |
1995 – 1997
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| Project Status |
Completed (Fiscal Year 1997)
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| Budget Amount *help |
¥16,900,000 (Direct Cost: ¥16,900,000)
Fiscal Year 1997: ¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1996: ¥4,700,000 (Direct Cost: ¥4,700,000)
Fiscal Year 1995: ¥10,000,000 (Direct Cost: ¥10,000,000)
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| Keywords | TABACCO CULTURED CELLS / PROMOTER / ARABIDOPSIS THALIANA / HEAT-SHOCK PROTEIN / LOG PHASE CELLS / STATIONARY GROWTH PHASE / DIFFERENTIAL SCREENING / PECTIN ESTERASE / 制御シス配列 / 増殖停止期 / シロイヌナズナ / 転写因子 / タバコ細胞 / 熱ショックプロモーター / テアニン |
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| Research Abstract |
Construction of a gene expression system in tobacco cultured cells (BY2) was studied. A 925 bp promoter fragment of a heat-shock protein Nicotiana tabacum gene (HSP18.2) of Arabidopsis thaliana showed clear heat-shock response of expression of the beta-glucuronidase (GUS) reporter gene in BY2 cells. Similar results were observed in a 500ml flask and 3-l jar fermentor.
Isolation of strong promoters in BY2 cells was tried. cDNA clones of which mRNA level is high in log phase cells and copy number in genome is low were isolated. These clones showed high homology with F1-ATPase (mitochondria type), elongation factor 1-alpha, and a gene with unknown function of A.thaliana (clone 27), respectively. A5'-flanking region of clone 27 showed 6.2 times of promoter activity of the CaMV35S promoter in BY2 cells.
Three cDNA clones which are expressed in stationary growth phase of BY2 cells were isolated by a differential screening. There clones showed high sequence homologies to alcohol dehydrogenase, pectin esterase and extensin. Promoters of these genes will be useful in gene expression in high cell density culture.
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