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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
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dc.contributor.advisor | 李昆達(Kung-Ta Lee) | |
dc.contributor.author | Ke-Jin Lin | en |
dc.contributor.author | 林科錦 | zh_TW |
dc.date.accessioned | 2021-06-17T06:34:09Z | - |
dc.date.available | 2023-08-21 | |
dc.date.copyright | 2018-08-21 | |
dc.date.issued | 2018 | |
dc.date.submitted | 2018-08-16 | |
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Arabidopsis B-cell lymphoma2 (Bcl-2)-associated athanogene 7 (BAG7)-mediated heat tolerance requires translocation, sumoylation and binding to WRKY29. New Phytol., 214(2), 695-705. Lin, H.-w., Kwok, K. H., & Doran, P. M. (2003). Development of Linum flavum hairy root cultures for production of coniferin. Biotechnol. Lett., 25(7), 521-525. Liu, Z. Q., Gao, J., Dong, A. W., & Shen, W. H. (2009). A truncated Arabidopsis NUCLEOSOME ASSEMBLY PROTEIN 1, AtNAP1;3T, alters plant growth responses to abscisic acid and salt in the Atnap1;3-2 mutant. Mol Plant, 2(4), 688-699. Marhavý, P., Bielach, A., Abas, L., Abuzeineh, A., Duclercq, J., Tanaka, H., . . . Kleine-Vehn, J. (2011). Cytokinin modulates endocytic trafficking of PIN1 auxin efflux carrier to control plant organogenesis. Dev. Cell, 21(4), 796-804. Maurel, C., Barbier-Brygoo, H., Spena, A., Tempe, J., & Guern, J. (1991). Single rol genes from the Agrobacterium rhizogenes TL-DNA alter some of the cellular responses to auxin in Nicotiana tabacum. Plant Physiol., 97(1), 212-216. Maurel, C., Brevet, J., Barbier-Brygoo, H., Guern, J., & Tempé, J. (1990). Auxin regulates the promoter of the root-inducing rolB gene of Agrobacterium rhizogenes in transgenic tobacco. Molecular and General Genetics MGG, 223(1), 58-64. Maurel, C., Leblanc, N., Barbier-Brygoo, H., Perrot-Rechenmann, C., Bouvier-Durand, M., & Guern, J. (1994). Alterations of auxin perception in rolB-transformed tobacco protoplasts (time course of rolB mRNA expression and increase in auxin sensitivity reveal multiple control by auxin). Plant Physiol., 105(4), 1209-1215. Nilsson, O., Moritz, T., Imbault, N., Sandberg, G., & Olsson, O. (1993). Hormonal characterization of transgenic tobacco plants expressing the rolC gene of Agrobacterium rhizogenes TL-DNA. Plant Physiol., 102(2), 363-371. Palazón, J., Cusidó, R., Roig, C., & Pinol, M. (1998). Expression of the rolC gene and nicotine production in transgenic roots and their regenerated plants. Plant Cell Rep., 17(5), 384-390. Pistelli, L., Giovannini, A., Ruffoni, B., Bertoli, A., & Pistelli, L. (2010). Hairy root cultures for secondary metabolites production. In Bio-Farms for Nutraceuticals (pp. 167-184): Springer. Rogner, U. C., Spyropoulos, D. D., Le Novere, N., Changeux, J. P., & Avner, P. (2000). Control of neurulation by the nucleosome assembly protein-1-like 2. Nat. Genet., 25(4), 431-435. Schmülling, T., Schell, J., & Spena, A. (1988). Single genes from Agrobacterium rhizogenes influence plant development. The EMBO journal, 7(9), 2621-2629. Schmülling, T., Fladung, M., Grossmann, K., & Schell, J. (1993). Hormonal content and sensitivity of transgenic tobacco and potato plants expressing single rol genes of Agrobacterium rhizogenes T-DNA. The Plant Journal, 3(3), 371-382. Shanks, J. V., & Morgan, J. (1999). Plant ‘hairy root’culture. Curr. Opin. 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Use of roots transformed by Agrobacterium rhizogenes in rhizosphere research: applications in studies of cadmium assimilation from sewage sludges. Plant Mol. Biol., 13(3), 295-302. Vanneste, S., De Rybel, B., Beemster, G. T., Ljung, K., De Smet, I., Van Isterdael, G., . . . Tasaka, M. (2005). Cell cycle progression in the pericycle is not sufficient for SOLITARY ROOT/IAA14-mediated lateral root initiation in Arabidopsis thaliana. The Plant Cell, 17(11), 3035-3050. Veena, V., & Taylor, C. G. (2007). Agrobacterium rhizogenes: recent developments and promising applications. In Vitro Cellular & Developmental Biology-Plant, 43(5), 383-403. Wang, J.-H., Lin, H.-H., Liu, C.-T., Lin, T.-C., Liu, L.-y. D., & Lee, K.-T. (2014). Transcriptomic analysis reveals that reactive oxygen species and genes encoding lipid transfer protein are associated with tobacco hairy root growth and branch development. Mol. Plant-Microbe Interact., 27(7), 678-687. White, F., Taylor, B., Huffman, G., Gordon, M., & Nester, E. (1985). Molecular and genetic analysis of the transferred DNA regions of the root-inducing plasmid of Agrobacterium rhizogenes. J. Bacteriol., 164(1), 33-44. Xu, H., Xu, W., Xi, H., Ma, W., He, Z., & Ma, M. (2013). The ER luminal binding protein (BiP) alleviates Cd(2+)-induced programmed cell death through endoplasmic reticulum stress-cell death signaling pathway in tobacco cells. J. Plant Physiol., 170(16), 1434-1441. Zhao, D., Fu, C., Chen, Y., & Ma, F. (2004). Transformation of Saussurea medusa for hairy roots and jaceosidin production. Plant Cell Rep., 23(7), 468-474. 易松輝. (2014). 菸草毛狀根中Root Locus B蛋白質交互作用之研究. 臺灣大學, Available from Airiti AiritiLibrary database. (2014年) 林大中. (2015). 利用雙分子螢光互補法研究與 RolB 有交互作用之蛋白. 臺灣大學, Available from Airiti AiritiLibrary database. (2015年) | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/72300 | - |
dc.description.abstract | 毛狀根為植物受到根毛農桿菌感染後,農桿菌將其Ri質體上的T-DNA隨機插入植物染色體產生的根狀組織,具有快速生長、基因組成穩定、能在不含植物賀爾蒙培養基中長期繼代培養、累積高量植物次級代謝物的特性。因為有這些特性,毛狀根被廣泛應用在高價值植物藥與重組蛋白質生產當中。然而,並不是所有的植物都能被根毛農桿菌感染產生毛狀根,因此了解毛狀根分化生成的分子作用機制將能大幅提升毛狀根的應用潛力。在T-DNA當中,rolB與rolC基因被認為在誘導毛狀根扮演重要關鍵的角色。本實驗室先前以酵母菌雙雜合法,發現ORF13a、PHI-2與NtbZIP和RolB蛋白質有交互作用。但以同樣的方法尋找與RolC有交互作用的蛋白質時,發現單獨表現GAL4 DNA-BD/RolC融合蛋白的酵母菌會自動活化下游的報導基因,造成篩選結果幾乎為偽陽性。本研究透過隨機突變rolC基因降低RolC的自活化能力,再進行酵母菌雙雜合實驗,找出在菸草毛狀根當中可能和RolC蛋白質有交互作用的蛋白質。此外,舊有的酵母菌接合轉形方法在本研究中存在接合轉形效率 (mating efficiency) 過低的問題,故本研究改良了接合轉形方法,進一步提升酵母菌的接合轉形效率,並以此新的方法,發現46個可能與RolC蛋白質有交互作用的蛋白質。 | zh_TW |
dc.description.abstract | Hairy roots are root-like tissues, which are resulted from the infection of Agrobacterium rhizogenes. During infection, Agrobacterium would transfer the T-DNA from its Ri plasmid to plant cell and randomly insert into plant chromosome. Hairy roots are characterized by high growth rate, genetic stability, and high accumulation of plant secondary metabolite, and they could be maintained in hormone-free medium. Due to these features, hairy roots are widely applied in production of high-value pharmaceutical molucules and recombinant proteins. However, still many plants are resistant to the infection of A. rhizogenes, the utilization of hairy root culture in these plants is limited. Understanding the molecular mechanism during hairy root formation increases applicability of medical herbs hairy root. Our previous study found that RolB protein can interact with ORF13a, NtbZIP, and PHI-2 by using yeast two-hybrid assay. But when applying the same method to screen the interacting proteins of RolC, we found that RolC would autoactivate the the reporter genes in Y2HGold, resulting too many false positive. In this study, we randomly mutated rolC gene by error-prone PCR to reduce autoactivity of RolC, and used these mutant RolC as bait to screen the interactors in tobacco hairy roots. Besides, we applied new mating method to increase the mating efficiency of yeasts. Using the new mating method, we found 46 potential interacting proteins of RolC. | en |
dc.description.provenance | Made available in DSpace on 2021-06-17T06:34:09Z (GMT). No. of bitstreams: 1 ntu-107-R03b22014-1.pdf: 1509467 bytes, checksum: b5978fa84d6679bba696644e40220171 (MD5) Previous issue date: 2018 | en |
dc.description.tableofcontents | 摘要 a
Abstract b 目錄 c 圖表目錄 e 前言 1 1.1 毛狀根 1 1.2 RolB以及RolC為影響毛狀根發育最關鍵的兩個因子 2 1.3 研究目的 3 第2章 材料與方法 5 2.1 生物材料與培養條件 5 2.1.1. Saccharomyces cerevisiae之培養條件 5 2.1.2. 菸草毛狀根之培養條件 6 2.2 酵母菌雙雜合 (Yeast two-hybrid) 6 2.2.1. 建立菸草毛狀根cDNA library 6 2.2.2. 酵母菌勝任細胞製作與轉形 7 2.2.3. 篩選低自活化的rolC突變序列 (rolC mutant with low autoactivation ) 7 2.2.4. Y2Gold [pGBKT7-mutant rolC] 存活性試驗 8 2.2.5. 以模擬酵母菌雙雜合接合轉形情境測試mutant RolC自活化能力 8 2.2.6. 酵母菌雙雜合接合轉形 (Clontech之方法) 9 2.2.7. 酵母菌雙雜合接合轉形 (新方法提升轉形接合效率) 10 2.2.8. 調整參數進一步提升新方法的轉形接合效率 10 2.2.9. 確認並定序篩選後的候選cDNA片段 11 2.2.10. 分析與RolC有交互作用的候選序列 11 第3章 結果 12 3.1 篩選低自活化的rolC突變序列 (rolC mutant with low autoactivation ) 12 3.2 Y2Gold [pGBKT7-mutant rolC] 存活性試驗 12 3.3 以模擬接合轉形的情境但不加入Y187 [pGADT7-cDNA library]轉形株,測試Y2HGold [pGBKT7-mutant rolC] 轉形株之自活化能力 13 3.4 酵母菌雙雜合篩選 13 第4章 討論 15 4.1 RolC自活化Y2HGold下游報導基因的能力無法被完全消除 15 4.2 接合轉形效率受到bait影響 15 4.3 使用突變rolC的方式可能無法找到RolC真正的交互作用者 15 4.4 BAG7以及BLP1和RolB的交互作用可能參與了內質網逆境反應的調控 15 4.5 NAP1作為histone chaperone,可能和RolC共同參與了植物賀爾蒙離層酸 (ABA) 反應的路徑調控以及調節植物的生長發育 16 4.6 Auxin efflux carrier和RolC的交互作用可能影響毛狀根的auxin濃度分布,進而促進側根生長 17 第5章 結論 18 表格 19 圖 30 參考文獻 35 附錄 39 | |
dc.language.iso | zh-TW | |
dc.title | 以酵母菌雙雜合系統篩選與RolC 具交互作用之蛋白質 | zh_TW |
dc.title | Screening of the Interaction Proteins of RolC by Use of Yeast Two-hybrid System | en |
dc.type | Thesis | |
dc.date.schoolyear | 106-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 楊健志,靳宗洛,張英? | |
dc.subject.keyword | 毛狀根,根毛農桿菌,rolC,蛋白質交互作用體學, | zh_TW |
dc.subject.keyword | A. rhizogenes A4,hairy roots,rolC,Interactomic, | en |
dc.relation.page | 39 | |
dc.identifier.doi | 10.6342/NTU201803764 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2018-08-16 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 生化科技學系 | zh_TW |
顯示於系所單位: | 生化科技學系 |
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