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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 莊榮輝(Rong-Huay Juang) | |
| dc.contributor.author | Tai-Tse Wu | en |
| dc.contributor.author | 吳岱澤 | zh_TW |
| dc.date.accessioned | 2021-06-13T02:22:45Z | - |
| dc.date.available | 2008-02-01 | |
| dc.date.copyright | 2007-02-01 | |
| dc.date.issued | 2007 | |
| dc.date.submitted | 2007-01-29 | |
| dc.identifier.citation | 吳 裕 仁 (2006) 綠竹筍生長過程蛋白質體變化及其抗體庫之建立
林 維 治 (1994) 林維治先生竹類論文集。林業叢刊第 69 號，張添榮主編，台灣省林業試驗所 Arnon DI, Rosenberg LL, Whatley FR (1954). A new glyceraldehydes phosphate dehydrogenase from photosynthetic tissues. Nature 173:1132–1134 Bustos DM, Iglesias AA (2002) Non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase is post-translationally phosphorylated in heterotrophic cells of wheat (Triticum aestivum). FEBS Lett 530:169–173 Bustos DM, Iglesias AA (2003) Phosphorylated non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase from heterotrophic cells of wheat interacts with 14-3-3 proteins. Plant Physiol 133:2081–2088 Cerff R (1995) Origin and evolution of phosphorylating and non-phosphorylating glyceraldehydes-3-phosphate dehydrogenase. In: Mathis P (ed) Photosynthesis: from light to biosphere. Kluwer Academic Publishers, Dordrecht, 1:933–938 D.Y. Arutyunov and V.I. Muronetz (2003) The activation of glycolysis performed by the nonphosphorylating glyceraldehyde-3-phosphate dehydrogenase in the model system. Biochemical and Biophysical Research Communications 300:149–154 Farres, J., Wang, T.T.Y., Cunningham, S.J., Weiner, H. (1995). Investigation of the active site cysteine residue of rat liver mitochondrial aldehyde dehydrogenase by site-directed mutagenesis. Biochemistry 34, 2592–2598. Go´mez-Casati DF, Sesma JI, Iglesias AA (2000) Structural and kinetic characterization of NADP-dependent, non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase from celery leaves. Plant Sci 154:107–115 Habenicht A (1997) The non-phosphorylating glyceraldehydes-3-phosphate dehydrogenase: Biochemistry, structure, occurrence and evolution. Biol Chem 378:1413–1419 Habenicht, A., Hellman, U., Cerff, R. (1994). Non-phosphorylating GAPDH of higher plants is a member of the aldehyde dehydrogenase superfamily with no sequence homology to the phosphorylating GAPDH. J. Mol. Biol. 237, 165–171. Iglesias AA, Serrano A, Guerrero MG, Losada M (1987) Purification and properties of NADP-dependent glyceraldehydes-3-phosphate dehydrogenase from green alga Chlamydomonas reinhardtii. Biochim Biophys Acta 925:1–10 Kelly GJ, Gibbs M (1973) Non-reversible D-glyceraldehyde -3-phosphate dehydrogenase of plant tissues. Plant Physiol 52:111–118 M. Arumugam Pillai • Z. Lihuang • T. Akiyama (2002) Molecular cloning, characterization, expression and chromosomal location of OsGAPDH, a submergence responsive gene in rice (Oryza sativa L.). Theor Appl Genet, 105:34–42 P. V. Dan’shina, E. V. Schmalhausen, D. Yu. Arutiunov, A. P. Pleten’, and V. I. Muronetz (2003) Acceleration of Glycolysis in the Presence of the non-phosphorylating and the oxidized phosphorylating glyceraldehyde-3-phosphate dehydrogenases. Biochemistry (Moscow), 68, 5: 593-600. Sachs MM, Subbiah CC, Saab IN (1996) Anaerobic gene expression and flooding tolerance in maize. J Exp Bot 47:1–15 Rosenberg LL, Arnon DI (1955) The preparation and properties of a new glycerladehyde-3-phosphate dehydrogenase from photosynthetic tissues. J Biol Chem 217:361–371 Rumpho ME, Edwards GE, Loescher WH (1983) A pathway for photosynthetic carbon flow to mannitol in celery leaves. Activity and localization of key enzymes. Plant Physiol 73:869–873 Umeda M, Uchimiya H (1994) Differential transcript levels of genes associated with glycolysis and alcohol fermentation in rice plants (Oryza sativa L) under submergence stress. Plant Physiol 106:1015–1022 | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/30958 | - |
| dc.description.abstract | 高等植物細胞質中Non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase (GAPN) 是一個具有高序列保守性蛋白質，負責催化重要能量及還原物質生成反應。在光合細胞中，此酵素參與了將葉綠體內NADPH運送至細胞質中的機制。為了清楚GAPN在綠竹筍快速生長中所扮演的角色，我們利用蛋白質純化方法，將綠竹筍中GAPN分離出來，並製備專一性的單株抗體。藉由二維電泳及免疫染色方法，我們發現綠竹筍在快速生長過程中，會表現出兩種不同的GAPN (GAPN-A與GAPN-B)，這兩種GAPN存在於在不同的組織中，並具有不同的pI值。這些結果暗示著GAPN-A與GAPN-B在竹筍的快速生長過程中扮演著全然不同的角色。 | zh_TW |
| dc.description.abstract | 英文摘要
Non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase (GAPN) is a conserved cytosolic protein and catalyze key steps in energy and reducing power partitioning in cells of higher plants. In photosynthetic cells, the enzyme is involved in a shuttle transfer mechanism to export NADPH from the chloroplast to the cytosol. To investigate the role of this enzyme in the rapid-growing green bamboo shoots, we purified and characterized GAPN from green bamboo shoots, and the GAPN specific monoclonal antibody was prepared by using purified GAPN as the antigen. We were identified two kinds of GAPN (GAPN-A and GAPN-B) in the rapid-growing green bamboo shoots by 2-DE and immunostaining. The two kinds of GAPN were found in different tissues during the rapid growth of green bamboo shoots and with distinct pI value. The results suggest that the GAPN-A and GAPN-B may have separate roles in the rapid-growing green bamboo shoots. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-13T02:22:45Z (GMT). No. of bitstreams: 1 ntu-96-R93442025-1.pdf: 2412825 bytes, checksum: 4a2e05493ac5047cebe4efbf051ee1de (MD5) Previous issue date: 2007 | en |
| dc.description.tableofcontents | 目錄
第一章 緒論 1 1.1 竹 1 1.1.1 分布 1 1.1.2 型態 1 1.1.3 獨特的生理現象 2 1.1.4 應用與經濟價值 2 1.2 Glyceraldehy-3-phosphate dehydrogenase (GAPDH) 3 1.2.1 Cytosolic phosphorylating GAPDH (GAPC) 4 1.2.2 Chloroplast phosphorylating GAPDH (GAPA/B) 4 1.2.3 Cytocolic non-phosphorylating GAPDH (GAPN) 6 1.3 研究動機及目的 8 第二章 材料與方法 9 2.1 一般分析法 9 2.1.1 蛋白質定量法 9 2.1.2 綠竹筍GAPN活性分析法 10 2.2 一般電泳檢定法 11 2.2.1 原態膠體電泳 11 2.2.2 SDS膠體電泳 14 2.2.3 Native-PAGE/SDS-PAGE膠體電泳 16 2.2.4 膠體染色法：Coomassie Brilliant Blue R-250 (CBR) 蛋白質染色法 17 2.2.5 綠竹筍GAPN 電泳膠片活性染色方法 18 2.2.6 綠竹GAPC電泳膠片活性染色方法 19 2.2.7 綠竹筍Alcohol dehydrogenase電泳膠片活性染色方法 20 2.2.8 膠片乾燥法及護貝 20 2.2.9蛋白質電泳轉印法 21 2.3綠竹筍GAPN抽取法 23 2.3.1 酵素粗抽取及硫酸銨分劃 23 2.3.2 管柱色層分析法之基本操作 24 2.3.3 離子交換法 26 2.3.4 膠體過濾法 27 2.3.5 疏水性層析法 28 2.3.6 Hydroxylapatite 29 2.3.7 製備式電泳與電泳溶離 30 2.4單株抗體之製備 32 2.4.1 抗原製備 32 2.4.2 小白鼠免疫 33 2.4.3細胞融合 34 2.4.4 細胞保存法 38 2.4.5 單株抗體的生產 39 2.5 免疫學檢定法 40 2.5.1 酵素免疫分析法 40 2.5.2 酵素免疫染色法 41 2.6 二維電泳相關實驗 44 2.6.1 二維電泳樣本蛋白質萃取法 44 2.6.2 蛋白質樣本溶解方法 45 2.6.3 二維電泳之蛋白質定量方法 45 2.6.4 膠體內蛋白脢水解 46 2.6.5 二維電泳 47 第三章 結果 49 3.1 未出土綠竹筍GAPN的純化與分離 49 3.1.1 硫酸銨分劃 49 3.1.2 DEAE Sephacel 49 3.1.3 Hydrophobic Interaction Chromatography (HIC) 50 3.1.4. Hydroxylapatite 50 3.1.5. 製備式電泳溶離 50 3.1.6. 純化表 51 3.1.7. 最適pH值測試 51 3.2綠竹筍GAPN之單株抗體製備 59 3.2.1 抗原製備 59 3.2.2 抗體製備 59 3.2.3 抗體效價與專一性測定 59 3.3 綠竹筍不同生長時期之GAPN之探討 62 3.3.1 不同生長時期二維電泳免疫染色圖譜比較 62 3.3.2 綠竹筍不同生長時期GAPN活性染色比較 71 3.3.3 綠竹筍不同生長時期GAPC活性染色比較 71 3.3.4 綠竹筍GAPN其他生化性質探討 71 第四章 結論與展望 77 4.1 綠竹筍中含有兩種形式的GAPN 77 4.2 綠竹筍中GAPN的生理功能探討 77 | |
| dc.language.iso | zh-TW | |
| dc.title | 綠竹筍不同生長時期non-phosphorylating glyceraldehyde-
3-phosphate dehydrogenase之純化與生化性質探討 | zh_TW |
| dc.title | Purification and characterization of non-phosphorylating
glyceraldehyde-3-phosphate dehydrogenase from green bamboo | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 95-1 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 楊健志,陳翰民,吳裕仁 | |
| dc.subject.keyword | 綠竹筍,純化, | zh_TW |
| dc.subject.keyword | Bambusa oldhamii, | en |
| dc.relation.page | 82 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2007-01-30 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 微生物與生化學研究所 | zh_TW |
| 顯示於系所單位： | 微生物學科所 | |
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