1997 Fiscal Year Final Research Report Summary
Construction of externally controllable gene promoters in plants
| Project/Area Number |
07554044
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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 |
植物生理
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| Research Institution | The University of Tokyo |
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Principal Investigator |
WATANABE Akira The University of Tokyo, Graduate School of Science, Professor, 大学院・理学系研究科, 教授 (70023471)
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| Co-Investigator(Kenkyū-buntansha) |
ITO Masaki The University of Tokyo, Graduate School of Science, Research Associate, 大学院・理学系研究科, 助手 (10242851)
FUJIMURA Tatsuhito Tsukuba University, Graduate School of Biosystem, Professor, 農林工学系バイオシステム研究科, 教授
MATSUOKA Makoto Nagoya University, BioScience Center, Professor, 生物分子応答研究センター, 教授 (00270992)
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| Project Period (FY) |
1995 – 1997
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| Keywords | Plant genes / Gene promoter / Gene expression / Arabidopsis thaliana / Sugar starvation / Tobacco BY-2 cells / Rice plant / Thiamin |
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| Research Abstract |
This research has been conducted to obtain a good externally controllable promoter to be used in the analysis of gene functions in plants. To do this we first isolated cDNA clones for genes that are expressed in plants under very specific conditions. These conditions included those deprivation of thiamine from culture media for tobacco BY-2 cells, dark treatment of Arabidopsis leaves and application of salicylic acid to tobacco plants. We could isolate several clones by a direct-display PCR procedure, but the expression of the genes complimentary to those cDNA sequences were not very specific, ie.the transcripts were found even in the presence of the vitamin. We were successful to find several genes that are expressed by a manner very specific to the second conditions. Three out of more than a dozen genes found to be dark-inducible were selected for rapid and specific expression after careful northern hybridization analysis. Their promoter regions were isolated and fused with a reporter gene (firefly luciferase). The fused genes were introduced into cultured BY-2 cells and looked at the expression after deprivation of sugar from the culture media, because we noticed in the northern analysis that the dark-induced expression was abolished if the leaves were fed with sucrose at 10mM.The transformed cells were very sensitive to sugar starvation in respect to the expression of the reporter gene. As the expression is very rapid, strong and reversible, and the control the promoter can be done by a very mild and naturally occurring stimuli, it appears promising as a tool for the analysis of functions of genes not favorable for normal growth, and for agricultural use for crop improvement as well.
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Research Products
(24 results)
