| Project/Area Number |
03454007
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
植物生理学
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| Research Institution | Kokkaido Tokai Univ. |
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Principal Investigator |
YAMADA Mitsuhiro Department of Biosclence and Technology, Hokkaido Tokai University Prof., 工学部, 教授 (50012266)
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| Co-Investigator(Kenkyū-buntansha) |
ARIGA Sanae Dept.Medical Technol.Hokkaido Univ., Ass.Prof., 大学部, 助教授 (90184283)
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| Project Period (FY) |
1991 – 1993
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| Project Status |
Completed (Fiscal Year 1993)
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| Budget Amount *help |
¥6,500,000 (Direct Cost: ¥6,500,000)
Fiscal Year 1993: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1992: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1991: ¥4,600,000 (Direct Cost: ¥4,600,000)
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| Keywords | Castor bean / Cell wall / fatty acid-binding protein / modified LTP / LTP / High density vesicle / LTP gene / Wax / ヒマ種子 / 果皮 / 脂質転移タンパク質 / 脂質転多タンパク質遺伝子 / 修飾脂質転移タンパク質 / クチクラ / 植物 / 登熟種子 / ゲノムDNA / イントロン / グリオキシソ-ム / 器官特異性 / イソ型タンパク質 |
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| Research Abstract |
1. Function of lipid transfer protein (LTP) and its homologus proteins
LPT was isolated as a protein which transfers polar lipids between biomembranes in vitro, but it is unlikely that LTP functions in vivo for intracellular lipid trasfer. LTPs did not occur in cytosol but in density vesicles in cells and cell walls. The expression of LTP genes and the accumulation of LTPs were observed in the surface area of developing organs such as embryos, endosperms and primordia of leaves and floweres. In addition, LTPs have the capacities of binding fatty acids and acylCoAs. From these results. LTP seems to functions in vivo as a carrier of wax or cutin monomers. This assumption was supported by evidence that wax synthesis induced by UV irradiation in cucmber leaves was followed by the accumulation of LTPs in the cuticles. On the other hand , the localization of LTPs in density vesicles in castor bean cotyledon cells and assembly of these vesicles into a vacuoles remains unsolved.
2. Cloning of LTP genes and its homologus protein genes
There are organ-specific isoforms of LTP, A - D, in castor bean seedlings. We cloned the isoform C gene by PCR method using the isoform C _cDNA.Intron sturucture of the isoform C gene has been determined, but its promoter structure has not, because of unsuccessfulness in isolating the upstream region of LTP gene. Several clones have been isolated from genomic library prepared from castor bean cotyledons and the analysis of nucleotides is in progress.
Eight cysteines in LTP sequence were conservative. To examine the role of these cysteines for binding of LTP to fatty acid, the isoform C _cDNA was modified to yield the isoform C with serines instead the cysteines and the modified LTP is now to be expressed.
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