| Project/Area Number |
09680044
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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 |
家政学
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| Research Institution | Nagoya Women's University |
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Principal Investigator |
OBA Kazuko Nagoya Women's Uniersity, Department of Food Science and Nutrition, Professor, 家政学部, 教授 (80023480)
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| Co-Investigator(Kenkyū-buntansha) |
MATSUOKA Makoto Nagoya University, Bioscience center, Professor, 生物分子応答研究センター, 教授 (00270992)
YAMAMOTO Atsuko Nagoya Women's Uniersity, Department of Food Science and Nutrition, Research associate, 家政学部, 助手
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| Project Period (FY) |
1997 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 1999: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1998: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1997: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Keywords | ascorbic acid / ascorbate oxidase / L-galactonolactone dehydrogenase / L-gulonolactone oxidase / potato tuber / sweet potato / tomato / transgenic plants / センスコンストラクト / アンチセンスコンストラクト / cDMAクローニング / ゲノミックサザン / L-ガラクトノラクトンデヒドロゲナーテ / アミノ酸シークエンス |
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| Research Abstract |
3. cDNA sequence analysis of L-galactonolactone dehydrogenase (GLDHase) from sweet potato : N-terminal partial sequences of 4 internal polypeptides generated by partial digestion of GLDH-ase with V8 protease were determined. The clone encoded a polypeptide of 581 amino acid residues with a molecular mass of 66kDa. The deduced amino acid sequence showed 77% identity with that of cauliflower GLDHase, and significant homology to that of GLO (22% identity) from rat.
1. Analysis of trangenic potato and tomato with the gene for L-gulonolactone oxidase (GLO) : L-gulonolactone oxidase (GLO) which catalyzes the last step of ascorbic acid (AsA) biosynthesis in animals oxidizes L-galactonolactone, a precursor of AsA in plants and yeast. We introduced a rat GLO gene to potato and tomato under the control of CaMV 35S promoter to elucidate whether the expression of GLO gene was confirmed by PCR using specific primers. GLO activity was detected in several transformants of potato and tobacco. Some of t
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ransgenic potato and tomato with GLO activity contained higher levels of AsA than that in wild type.
2. Sense and antisense expression of ascorbate oxidase (AAO) gene in tobacco plants. Effect on ascorbate levels in tobacco leaves : We introduced tobacco genes with a sense and an antisense construct of the AAO gene under control of the constitutive CaMV 35S promoter to elucidate the function of AAO gene under control of the constitutive CaMV 35S promoter to elucidate the function of AAO. All of sense transformant (TィイD2oィエD2) had higher levels of AAO-mRNA and lower levels of AsA than those in wild type. Three antisense TィイD2oィエD2 plants had high levels of AsA, although AAO-mRNA levels in all antisense TィイD2oィエD2 plants were much low compared with those in wild type. All TィイD21ィエD2-progenies of self-polinated sense TィイD2oィエD2 plants gown in the presence of Kanamycin had high levels of AAO-mRNA and AAO-protein. AsA content in 5 antisense TィイD21ィエD2-progenies were higher than in wild type. Growth rate of several antisense TィイD21ィエD2-progenies with high AsA content was higher than that of sense TィイD21ィエD2-progenies. These results suggest that antisense expression of AAO in vegetables may be useful for breeding vegetables containing high levels of AsA.
Gnomic Southern analysis suggested that GLDHase gene exists as a single copy in the genome of sweet potato. Less
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