| Project/Area Number |
11660060
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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 | Niigata University |
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Principal Investigator |
SUEYOSHI Kuni Faculty of Agriculture, Niigata University, Associate Professor, 農学部, 助教授 (10216278)
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| Co-Investigator(Kenkyū-buntansha) |
OHYAMA Takuji Faculty of Agriculture, Niigata University, Professor, 農学部, 教授 (30152268)
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| Project Period (FY) |
1999 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2000: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1999: ¥2,500,000 (Direct Cost: ¥2,500,000)
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| Keywords | 14-3-3 protein / nitrate reductase / Immunohistology / Barley |
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| Research Abstract |
Nitrate reductase (NR) activity was markedly decreased by light to dark transition of barley plants. This inactivation was due to phosphorylation of NR protein and subsequent binding of 14-3-3 proteins to phosphorylated NR.In this study, we investigated tissue distribution and cellular localization of 14-3-3 proteins using immunological techniques in barley plants. The results are the following.
1.A cDNA for barley 14-3-3 protein were cloned by RT-PCR method using total RNA prepared from barley leaf as template. The plasmid that allows production of recombinant 14-3-3 protein was constructed and transformed into E.coli. The recombinant 14-3-3 protein expressed in E.coli was extract and purified. Specific antibody for 14-3-3 proteins was prepared by immunizing the recombinant 14-3-3 proteins.
2. Tissue distribution of 14-3-3 proteins was investigated. The protein extracts were prepared from various barley tissues including embryo and endosperm of germinating seeds, fully expanded and unexpanded leaf brades and mature roots and subjected to immunoblot analysis. The 14-3-3 proteins were present in all tissues with similar amount.
3. The sections of leaf and root tissues were prepared for immunohistological analysis of 14-3-3 proteins. It was suggested that 14-3-3 proteins located in vascular bundle sheath cells of leaf and in outer layer's cells of root cortex and stele.
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