Genetic analyses of response mechanisms for aluminum stress in plant
| Project/Area Number |
16580046
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Plant nutrition/Soil science
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| Research Institution | Okayama University |
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Principal Investigator |
EZAKI Bunichi Okayama University, Research Institute for Bioresources, Associate Professor, 資源生物科学研究所, 助教授 (90243500)
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| Co-Investigator(Kenkyū-buntansha) |
NAKASHIMA Susumu Research Institute for Bioresources, Associate Professor, 資源生物科学研究所, 助教授 (60033122)
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| Project Period (FY) |
2004 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 2006: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2005: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 2004: ¥1,900,000 (Direct Cost: ¥1,900,000)
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| Keywords | aluminum (Al) stress / gene response mechanism / Al stress induced gene / transcription factor / glutathione S-transferase / oxidative stress / signal transduction / Arabidopsis thaliana / Ring Zinc finger protein / Homeobox-Leucine Zipper / 転写調節因子 / アルミニウム(Al)ストレス / ゲルシフトアッセイ / Bio-panning / AtGST11遺伝子 / 遺伝子発現応答機構 / aluminum(Al)stress / glutathione S-transferase (GST) |
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| Research Abstract |
1. Construction of cDNA library to isolate and characterize several candidate clones encoding transcription factor(s) which are related for the gene expression of the AtGST11 gene under aluminum (Al) stress.
Using mRNA molecules derived from Al treated Arabidopsis thaliana, cDNA library was constructed. The constracted cDNA library were applied for a 3 times repeated screening designated "Biopanning". Finally, we could isolate 50 candidates clones in the third screening.
2. Molecular genetical analysis of the isolated clones
Isolated 50 clones were applied to DNA sequencing and could be classified into 8 genes. Ring Zn finger protein (#13) and Homeobox-Leucine Zipper protein 6 (#43) which have been already reported as transcription factors were included in these clones. One clone (#4) which encodes an unknown protein was also included. Unfortunately, other 5 genes seemed not to be related to gene-expression of the AtGST11 gene.
3. Gel shift assay
To confirm whether the three clones (#4, 13 and 43) actually can bind to the promoter region of the AtGST11 gene, gel shift assay was performed using the purified three proteins. These proteins were extracted and purified by His-tag affinity column before the gel shift assay. The result suggested that all of these proteins can bind to the promoter.
4. New approach (One hybrid assay) for an isolation of transcription factors
In the last year of this project (2006), we also started a new approach to isolate transcription factor(s) which are related to the gene-expression of either the AtGST1 or AtGST11 gene under Al stress by "Yeast One Hybrid System"., We could isolate four new candidates from this screening.
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Report
(4 results)
Research Products
(21 results)
