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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 王愛玉 | |
| dc.contributor.author | Shu-Ping Yang | en |
| dc.contributor.author | 楊書萍 | zh_TW |
| dc.date.accessioned | 2021-06-08T04:48:04Z | - |
| dc.date.copyright | 2009-07-31 | |
| dc.date.issued | 2009 | |
| dc.date.submitted | 2009-07-29 | |
| dc.identifier.citation | 莊榮輝 (1985) 水稻蔗糖合成酶之研究。，博士論文，國立台灣大學農業化學研究所。
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Mol Gen Genet 252: 303-310 Chourey PS, Taliercio EW, Carlson SJ, Ruan YL (1998) Genetic evidence that the two isozymes of sucrose synthase present in developing maize endosperm are critical, one for cell wall integrity and the other for starch biosynthesis. Mol Gen Genet 259: 88-96 Delmer DP, Amor Y (1995) Cellulose biosynthesis. Plant Cell 7: 987-1000 Ding X, Richter T, Chen M, Fujii H, Seo YS, Xie M, Zheng X, Kanrar S, Stevenson RA, Dardick C, Li Y, Jiang H, Zhang Y, Yu F, Bartley LE, Chern M, Bart R, Chen X, Zhu L, Farmerie WG, Gribskov M, Zhu JK, Fromm ME, Ronald PC, Song WY (2009) A rice kinase-protein interaction map. Plant Physiol 149: 1478-1492 Duncan KA, Hardin SC, Huber SC (2006) The three maize sucrose synthase isoforms differ in distribution, localization, and phosphorylation. Plant Cell Physiol 47: 959-971 Fujikawa Y, Kato N (2007) Split luciferase complementation assay to study protein-protein interactions in Arabidopsis protoplasts. 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Plant Physiol 134: 1427-1438 Hennen-Bierwagen TA, Lin Q, Grimaud F, Planchot V, Keeling PL, James MG, Myers AM (2009) Proteins from multiple metabolic pathways associate with starch biosynthetic enzymes in high molecular weight complexes: a model for regulation of carbon allocation in maize amyloplasts. Plant Physiol 149: 1541-1559 Huang JW, Chen JT, Yu WP, Shyur LF, Wang AY, Sung HY, Lee PD, Su JC (1996) Complete structures of three rice sucrose synthase isogenes and differential regulation of their expressions. Biosci Biotechnol Biochem 60: 233-239 Jackson V (1978) Studies on histone organization in the nucleosome using formaldehyde as a reversible cross-linking agent. Cell 15: 945-954 Jiang CJ, Imamoto N, Matsuki R, Yoneda Y, Yamamoto N (1998) In vitro characterization of rice importin beta1: molecular interaction with nuclear transport factors and mediation of nuclear protein import. FEBS Lett 437: 127-130 Kim R, Yokota H, Kim SH (2000) Electrophoresis of proteins and protein-protein complexes in a native agarose gel. Anal Biochem 282: 147-149 Kluger R, Alagic A (2004) Chemical cross-linking and protein-protein interactions-a review with illustrative protocols. Bioorg Chem 32: 451-472 Kobayashi T, Hearing VJ (2007) Direct interaction of tyrosinase with Tyrp1 to form heterodimeric complexes in vivo. J Cell Sci 120: 4261-4268 Koch K (2004) Sucrose metabolism: regulatory mechanisms and pivotal roles in sugar sensing and plant development. Curr Opin Plant Biol 7: 235-246 Lakey JH, Raggett EM (1998) Measuring protein-protein interactions. Curr Opin Struct Biol 8: 119-123 Liao YC, Wang AY (2003) Sugar-modulated gene expression of sucrose synthase in suspension-cultured cells of rice. Physiologia Plantarum 118: 319-327. Matic S, Akerlund HE, Everitt E, Widell S (2004) Sucrose synthase isoforms in cultured tobacco cells. Plant Physiol Biochem 42: 299-306 Melcher K, Chen HT (2007) Identification and analysis of multiprotein complexes through chemical crosslinking. Curr Protoc Cell Biol Chapter 17: Unit 17 10 Miernyk JA, Thelen JJ (2008) Biochemical approaches for discovering protein-protein interactions. Plant J 53: 597-609 Nguyen-Quoc B, Krivitzky M, Huber SC, Lecharny A (1990) Sucrose Synthase in Developing Maize Leaves: Regulation of Activity by Protein Level during the Import to Export Transition. Plant Physiol 94: 516-523 O'Reilly G, Clarke F (1993) Identification of an actin binding region in aldolase. FEBS Lett 321: 69-72 Pozueta-Romero J, Ardila F, Akazawa T (1991) ADP-Glucose Transport by the Chloroplast Adenylate Translocator Is Linked to Starch Biosynthesis. Plant Physiol 97: 1565-1572 Pozueta-Romero J, Yamaguchi J, Akazawa T (1991) ADPG formation by the ADP-specific cleavage of sucrose-reassessment of sucrose synthase. 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Anal Chem 78: 8059-8068 Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A 76: 4350-4354 Wang AY, Kao MH, Yang WH, Sayion Y, Liu LF, Lee PD, Su JC (1999) Differentially and developmentally regulated expression of three rice sucrose synthase genes. Plant Cell Physiol 40: 800-807 Wang AY, Yu WP, Juang RH, Huang JW, Sung HY, Su JC (1992) Presence of three rice sucrose synthase genes as revealed by cloning and sequencing of cDNA. Plant Mol Biol 18: 1191-1194 Winter H, Huber JL, Huber SC (1998) Identification of sucrose synthase as an actin-binding protein. FEBS Lett 430: 205-208 Winter H, Huber SC (2000) Regulation of sucrose metabolism in higher plants: localization and regulation of activity of key enzymes. Crit Rev Biochem Mol Biol 35: 253-289 Yu WP, Wang AY, Juang RH, Sung HY, Su JC (1992) Isolation and sequences of rice sucrose synthase cDNA and genomic DNA. Plant Mol Biol 18: 139-142 | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/23218 | - |
| dc.description.abstract | 蔗糖合成酶 (sucrose synthase, SuS) 在植物體中催化蔗糖和 UDP 形成果糖和 UDP-glucose 之可逆反應，此酵素對於在多醣類合成時的基質供應，以及在植物發育上都扮演重要角色。本論文利用化學聯結劑disuccinimidyl tartarate (DST)，探討水稻懸浮培養細胞 (Oryza sativa L. cv. Tainung 67) 中與蔗糖合成酶 (rice sucrose synthase, RSuS) 具交互作用之蛋白質。以 anti-RSuS 抗體進行西方點墨法的結果顯示，有數個與 RSuS 具聯結之蛋白質複合體存在，進一步以膠體過濾層析純化，並以 anti-RSuS 抗體進行免疫共沉澱後，所得之蛋白質進行 MALDI-TOF-MS 分析。在 in vitro 化學聯結實驗中觀察到的一些蛋白質複合體，在 in vivo 化學聯結實驗中存在類似結果，顯示在 in vivo 狀態下，RSuS 與其它蛋白質具交互作用。 | zh_TW |
| dc.description.abstract | Sucrose synthase (SuS) catalyzes the reversible interconversion of sucrose and UDP into fructose and UDP-glucose in plants. The enzyme plays an important role in providing substrates for polysaccharide synthesis and is crucial for plant development. To investigate whether SuS interact with other proteins, the chemical cross-linker, disuccinimidyl tartarate (DST), was used to covalently cross-link the interacted proteins in rice suspension-cultured cells of rice (Oryza sativa L. cv. Tainung 67). Western analysis with anti-RSuS antibody showed the presence of several cross-linked protein complexes containing RSuS. The RSuS-containing protein complexes were partially purified by gel filtration chromatography and immunoprecipitated with anti-RSuS antibody, and then subjected to MALDI-TOF-MS analysis. The in vivo cross-linking experiment revealed that some RSuS-containing different protein complexes observed in the in vitro cross-linking experiment were also present in the in vivo cross-linked proteins of rice suspension-cultured cells, suggesting that the interactions between RSuS and other proteins occur in vivo. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-08T04:48:04Z (GMT). No. of bitstreams: 1 ntu-98-R96b47211-1.pdf: 4301408 bytes, checksum: f839bacba1322c2619c471bca61e8dbb (MD5) Previous issue date: 2009 | en |
| dc.description.tableofcontents | 目錄………………………………………………………………………………….Ⅰ
縮寫表……………………………………………………………………………….Ⅴ 摘要………………………………………………………………………….………Ⅶ Abstract……………………………………………………………………………..Ⅷ 第一章 研究背景…………………………………………………………..…………1 第一節 蛋白質間交互作用..…………………………….……………..……..1 1.1 蛋白質間交互作用之重要性 1 1.2 蛋白質間交互作用的形式與作用力 1 1.3 常見之分析方法 2 第二節 化學聯結法 …………………………………………………………3 2.1 化學聯結劑之官能基 3 2.1.1 與胺基反應之聯結劑官能基 3 2.1.2 與硫氫基反應之聯結劑官能基 3 2.1.3 光活化官能基 4 2.1.4 其他形式之聯結劑官能基 4 2.2 化學聯結劑的種類 4 2.2.1 雙同官能基聯結劑 4 2.2.2 雙異官能基聯結劑 5 2.2.3 三官能基聯結劑 5 2.2.4 Zero-length cross-linkers 5 2.3 如何選擇化學聯結劑 5 第三節 蔗糖合成酶…………………………………………………………...7 3.1 蔗糖合成酶與其異構酶 7 3.2 蔗糖合成酶的生理特性 8 3.3 蔗糖合成酶與其他蛋白質交互作用之可能性 8 3.3.1 參與細胞壁纖維素合成 8 3.3.2 參與澱粉合成 9 3.3.3 與肌動蛋白質 (actin) 結合 9 第四節 研究目的……………………………………………………………..9 第二章 材料與方法 ………………………………………………………………….11 第一節 實驗材料…………………………………….………...…………...11 1.1 水稻懸浮培養細胞 11 1.2 藥品 11 1.2.1 一般化學藥品 11 1.2.2 化學聯結劑 11 第二節 實驗設備…………………………………………………………..14 2.1 化學聯結蛋白質樣本製備及純化設備 14 2.2 蛋白質樣本分析設備 14 2.3 其他設備 14 第三節 實驗方法…………………………………..………………………14 3.1 水稻懸浮培養細胞培養 14 3.2 化學聯結法 15 3.2.1 In vitro 化學聯結法 15 3.2.1 In vivo 化學聯結法 15 3.3 快速蛋白質液相層析 (FPLC) - 膠體過濾法 16 3.3.1 管柱流洗 16 3.3.2 膠體過濾法分離蛋白質 16 3.3.3 以膠體過濾法測定原態分子量 16 3.4 免疫共沉澱法 17 3.4.1 配製 protein A Sepharose 溶液 17 3.4.2 免疫共沉澱 17 3.5 蛋白質定量與分析 18 3.5.1 蛋白質定量 18 3.5.2 蛋白質電泳 18 3.5.3 製備式電泳 19 3.5.4 二維電泳 19 3.5.5原態瓊脂糖膠體電泳 21 3.6 膠體染色法 22 3.6.1 Coomassie Brilliant Blue R (CBR) 染色法 22 3.6.2 硝酸銀染色法 22 3.6.3 負染色法 22 3.7 西方點墨法 23 3.7.1 蛋白質轉印 23 3.7.2 免疫染色 23 3.8 蛋白質身分鑑定 24 3.9 抗體製備 24 3.9.1 抗原製備 24 3.9.2 免疫 24 3.9.3 採血 25 3.9.4 效價測試 25 第三章 結果與討論……………………………………………..…………..………26 第一節 Anti-RSuS3 多株抗體的製備………………………………………26 第二節 化學聯結劑的選擇…………………………………………………..26 2.1 不同化學聯結劑之聯結效率 26 2.2 討論 27 第三節 試管中 (in vitro) 聯結反應條件探討………………………………28 3.1 DST 最適濃度探討 28 3.2 討論 29 3.2.1高濃度 DST 處理的影響 29 3.2.2 DST 反應的最佳條件 29 3.2.3 與 RSuS 聯結之蛋白質分子量推測 30 第四節 RSuS 聯結蛋白質複合體之分離與純化…………………………...30 4.1 二維電泳 30 4.2原態瓊脂糖膠體電泳 31 4.3膠體過濾法與免疫共沉澱法 31 4.4 討論 32 4.4.1分離 RSuS 聯結蛋白質複合體方法探討 32 4.4.2 NaIO4 的影響 33 第五節 聯結蛋白質身分鑑定………………………………………….……34 5.1 MALDI-TOF-MS分析結果 34 5.2 討論 34 第六節 In vivo 化學聯結法…………………………………………………35 6.1 In vivo DST 化學聯結結果 35 6.2 討論 35 第四章 結論與未來展望……………………………………………………………37 第一節 結論…………………………………………………………………..37 第二節 未來展望……………………………………………………………...37 2.1確認與 RSuS 聯結之蛋白質的身分 37 2.2 利用 in vivo 化學聯結法確認與 RSuS 有交互作用的蛋白質 38 參考文獻……………………………………………………………………………..39 圖表集………………………………………………………………………………..45 | |
| dc.language.iso | zh-TW | |
| dc.subject | 水稻懸浮培養細胞 | zh_TW |
| dc.subject | 蔗糖合成酶 | zh_TW |
| dc.subject | 化學聯結法 | zh_TW |
| dc.subject | 蛋白質交互作用 | zh_TW |
| dc.subject | protein-protein interactions | en |
| dc.subject | rice suspension-cultured cell | en |
| dc.subject | chemical cross-linking | en |
| dc.subject | Sucrose synthase | en |
| dc.title | 利用化學聯結法探討水稻懸浮培養細胞中與蔗糖合成酶具交互作用之蛋白質 | zh_TW |
| dc.title | Studies on Sucrose Synthase Interacted Proteins in Rice Suspension-Cultured Cells by Chemical Cross-Linking | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 97-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 宋賢一,張珍田,林忠亮,楊健志 | |
| dc.subject.keyword | 蔗糖合成酶,化學聯結法,蛋白質交互作用,水稻懸浮培養細胞, | zh_TW |
| dc.subject.keyword | Sucrose synthase,chemical cross-linking,protein-protein interactions,rice suspension-cultured cell, | en |
| dc.relation.page | 65 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2009-07-29 | |
| 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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