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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 楊健志(Chien-Chih Yang) | |
| dc.contributor.author | Yi-Ting Shih | en |
| dc.contributor.author | 石依婷 | zh_TW |
| dc.date.accessioned | 2021-06-08T07:14:30Z | - |
| dc.date.copyright | 2008-08-06 | |
| dc.date.issued | 2008 | |
| dc.date.submitted | 2008-07-28 | |
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Arabinose-Induced Binding of AraC Protein to araIR2 RActivates the araBAD Operon Promoter. Proc. Natl. Acad. Sci. USA 84: 8814-8818 Leon-Kloosterziel, K. M., Verhagen, B. W., Keurentjes, J. J., VanPelt, J. A., Rep, M.,VanLoon, L. C., Pieterse, C. M. (2005). Colonization of the Arabidopsis rhizosphere by fluorescent Pseudomonas spp. activates a root-specific, ethylene- responsive PR-5 gene in the vascular bundle. Plant molecular biology 57(5): 731-748 Liu D, Rhodes D, D'Urzo MP, Xu Y, Narasimhan M.L., Hasegawa P.M., Bressan R.A., Abad L. (1996) In vivo and in vitro activity of truncated osmotin that is secreted into the extracellular matrix. Plant Science 121:123–131. Menu-Bouaouiche, Laurence, Vriet, Christelle, Peumans, Willy J., Barre, Annick, Van Damme, Els J. M., Pierre Rougé (2003). A molecular basis for the endo-β1,3- glucanase activity of the thaumatin-like proteins from edible fruits. Elsevier Science 85(1): 123-131 Narasimhan, M.L., Lee, H., Damsz, B., Singh, N.K., Ibeas, J.L., Mat sumoto, T.K., Woloshuk, C.P., Bressan, R.A. (2003). Overexpres sion of a cell wall glycolprotein in Fusarium oxysporum increases virulence and resistance to a plant PR-5 protein. Plant Jurnal 36: 390–400. Narasimhan, M. L., Coca, M. A., Jin, J., Yamauchi, T., Ito, Y., Kadowaki, T., Kim, K. K., Pardo, J. M., Damsz, B., Hasegawa, P. M., Yun, D. J., Bressan, R. A. (2005). Osmotin is a homolog of mammalian adiponectin and controls apoptosis in yeast through a homolog of mammalian adiponectin receptor. Molecular Cell 17(2): 171-80. Osmond, R. I., Hrmova, M., Fontaine, F., Imberty, A., Fincher, G. B. (2001). Binding interactions between barley thaumatin-like proteins and (1,3)- beta- D-glucans. Kinetics, specificity, structural analysis and biological implications. European journal of biochemistry / FEBS 268(15): 4190-4199. Palm G.J., Wlodower, A. (1999) Spectral variants of green fluorescent protein. Methods in enzymology 302: 378-393 Rakwal, R., Agrawal, G. K., Yonekura, M. (1999). Separation of proteins from stressed rice (Oryza sativa L.) leaf tissues by two-dimensional polyacrylamide gel electrophoresis: induction of pathogenesis-related and cellular protectant proteins by jasmonic acid, UV irradiation and copper chloride. Electrophoresis 20(17): 3472-3478. Salzman, R. A., Koiwa, H., Ibeas, J. I., Pardo, J. M., Hasegawa, P. M., Bressan, R. A. (2004). Inorganic cations mediate plant PR5 protein antifungal activity through fungal Mnn1- and Mnn4-regulated cell surface glycans. Molecular plant-microbe interactions : MPMI 17(7): 780-788. Schwartz, L. M., Smith, S. W., Jones, M. E., Osborne, B. A. (1993). Do all programmed cell deaths occur via apoptosis? Proc. Natl. Acad. Sci. USA 90(3): 980- 984. Sheen, J., Hwang, S., Niwa, Y., Kobayashi, H., Galbraith, D.W. (l995) Green fluorescent protein as a new vital marker in plant cells. Plant Jurnal 8: 777-784. Shigekazu NagataTT (2000). TTApoptotic DNA fragmentationT. Experimental Cell Research T256 (1): 12–18. Solomon M, Belenghi B, Delledonne M, Menachem E, Levine A. (1999). The involvement of cysteine proteases and protease inhibitor genes in the regulation of programmed cell death in plants.T TPlant Cell 11(3): 431-444. Song, W.Y., Sohn, E.J., Martinoia, E., Lee, Y.J., Yang, Y.Y., Jasinski, M., Forestier, C., Hwang, I., Lee, Y. (2003). Engineering tolerance and accumulation of lead and cadmium in transgenic plants. Nature biotechnology 21(8): 914-919. Taylor, J. A., West, D. W. (1980). The Use of Evan's Blue Stain to Test the Survival of Plant Cells after Exposure to High Salt and High Osmotic Pressure. Journal of experimental botany 31(2): 571-576. van Loon, L. C., Rep, M., Pieterse, C. M. (2006). Significance of inducible defense- related proteins in infected plants. 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Stress proteins on the yeast cell surface determine resistance to osmotin, a plant antifungal protein. Proc. Natl. Acad. Sci. USA 94: 7082–7087 | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/26544 | - |
| dc.description.abstract | Osmotin from tobacco can induce program cell death (PCD) of yeast by binding to a yeast membrane protein, YOL002c, which is a homolog of mammalian adiponectin receptor . Tobacco osmotin shares a similar structure fold with mammalian adiponectin with no sequence similarity.
HHP1 (UhUeptaUhUelical UpUrotein 1) is a homolog of adiponectin receptor in Arabidopsis with 51% sequence similarity. It is interesting to find out if there is any physiological or biochemical interaction between AtOSM34 and HHP1 of Arabidopsis. AtOSM34 is a homolog of tobacco osmotin in Arabidopsis. Heterologous protein expression of AtOSM34 in E.coli was established. Cellular localization of AtOSM34 was studied using transient expression of AtOSM34-mGFP5 fusion protein in onion epidermal cells and Aradidopsis protoplasts. The plasmid used for AtOSM34-mGFP5 fusion protein expression was constructed using pCAMBIA1302 vector. It is likely that AtOSM34 is located in the cytosol or other subcellular organelle. We also tried to treat Arabidopsis protoplasts using recombinant AtOSM34. The possibility of a putative receptor for AtOSM34 is discussed. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-08T07:14:30Z (GMT). No. of bitstreams: 1 ntu-97-R95b47211-1.pdf: 1927572 bytes, checksum: 6557e3520bedbd39313adb17c19c2cb4 (MD5) Previous issue date: 2008 | en |
| dc.description.tableofcontents | 目錄………………………………………………………………………I
縮寫表…………………………………………………………………IV 摘要……………………………………………………………………VI Abstract………………………………………………………………VII 第一章 緒論 11 PR5 家族蛋白質………………………………………1 12 Osmotin 相關功能的研究………………………………3 13 程序性細胞死亡 (Programmed Cell Death)……………7 131 細胞凋亡 (Apoptosis)…………………………………7 14 模式植物：阿拉伯芥………………………………………9 15 組織定位中常用的報導基因………………………………10 151 glucuronidase (GUS)……………………………………10 152 綠色螢光蛋白 GFP (Green fluorescent protein)……10 16 Binary vector 介紹………………………………………12 17 研究動機、目的與方法……………………………………13 第二章 材料與方法 21 實驗材料…………………………………………………………14 211 植物材料………………………………………………14 212 載體 (vectors)…………………………………14 213 大腸桿菌菌株…………………………………………16 214 農桿菌菌株……………………………………………16 22 實驗藥品………………………………………………………17 221 一般化學藥劑…………………………………………17 222 酵素…………………………………………………17 223 培養基…………………………………………………17 23 儀器設備…………………………………………………………18 24 實驗方法……………………………………………………20 241 阿拉伯芥種植…………………………………………20 2411 培養基配製…………………………………21 2412 種子之表面消毒與低溫處理………………22 2413 土壤培養……………………………………23 2414 收集種子……………………………………24 242 DNA 之抽取與分析方法………………………………24 2421 質體 DNA 之抽取與分析……………………24 2422 DNA 限制酶切割反應分析…………………27 2423 洋菜膠體電泳………………………………27 2424 DNA 片段的分離及純化……………………28 2425 DNA 之去磷酸化反應………………………29 2426 DNA 之磷酸化反應…………………………29 2427 DNA 定量……………………………………29 243 AtOSM34、AtOLP 基因序列選殖………………………30 2431 專一性引子設計……………………………30 2432 聚合酶鏈鎖反應 (Polymerase chain reaction, PCR)………………………………………………………31 2433 T-A cloning…………………………………31 2434 DNA 接合反應………………………………32 2435 大腸桿菌之質體轉形………………………32 2436 重組質體的檢定……………………………34 244 表現載體之建構………………………………………36 2441 Binary vector 之建構……………………36 245 阿拉伯芥基因轉殖……………………………………37 2451 植物材料之準備……………………………37 2452 農桿菌轉形…………………………………38 2453 農桿菌感染 2454 轉植株之初步篩選………………………………………41 246 阿拉伯芥原生質體轉形………………………………42 2461 原生質體抽取方法…………………………42 2462 原生質體基因轉形方法……………………44 247 基因鎗 (bombardment) 轉殖………………………………44 2471 植物材料之準備……………………………………44 2472 DNA 包裹鎢粒子之處理…………………………45 2473 操作基因鎗………………………………………45 2474 螢光染劑 DAPI 之染色…………………………46 248 顯微鏡及影像分析…………………………………………47 249 重組蛋白質之表現與檢定…………………………………48 2491 重組蛋白質最佳誘導條件之探討……………48 2492 重組蛋白質之大量表現………………………48 2493 重組蛋白質之純化……………………………49 2494 電泳檢定法……………………………………50 2410 蛋白質免疫轉印法……………………………………52 24101 蛋白質電泳轉印法…………………………52 24102 酵素免疫染色法……………………………53 2411 外加蛋白質處理原生質體……………………………53 24111 蛋白質前處理………………………………54 24112計算存活率與觀察細胞凋亡情形……………54 第三章 結果與討論 31 建立 AtOSM34 過度表現之阿拉伯芥轉殖株…………………55 311 AtOSM34 基因序列的選殖……………………………55 312 建構表現載體…………………………………………55 313 阿拉伯芥之基因轉殖…………………………………57 32 AtOSM34 的定位研究……………………………………………59 321 AtOSM34 之序列比對與分析…………………………59 322 洋蔥表皮定位分析……………………………………59 323 阿拉伯芥原生質體的定位分析………………………60 33 AtOSM34 及 AtOLP 重組蛋白質之表現與分析………………61 331 AtOSM34、AtOLP 基因序列選殖與表現載體的建構…61 332 AtOSM34 及 AtOLP 重組蛋白質最佳表現條件之探討………………………………………………………………………62 333 重組蛋白質純化與純化條件之探討…………………………63 第四章 結論與未來展望……………………………………………65 41 結論………………………………………………………………65 42 未來展望…………………………………………………………65 參考文獻………………………………………………………………66 圖與表…………………………………………………………………71 附錄……………………………………………………………………81 | |
| dc.language.iso | zh-TW | |
| dc.subject | 細胞凋亡 | zh_TW |
| dc.subject | 奇異果甜蛋白質 | zh_TW |
| dc.subject | 抗黴菌 | zh_TW |
| dc.subject | 七個穿膜區域 | zh_TW |
| dc.subject | 螢光融合蛋白 | zh_TW |
| dc.subject | 蛋白質異源表現系統 | zh_TW |
| dc.subject | 冷適應 | zh_TW |
| dc.subject | HHP1 | en |
| dc.subject | TLP | en |
| dc.subject | PRs | en |
| dc.subject | OLP | en |
| dc.subject | OSM | en |
| dc.subject | CTPP | en |
| dc.subject | GFP | en |
| dc.title | 阿拉伯芥中 Osmotin 同源蛋白質的的選殖與生理研究 | zh_TW |
| dc.title | Molecular Cloning of Osmotin Homologs in Arabidopsis thaliana and Physiology Studies | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 96-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 蘇仲卿(Jong-Ching Su),常怡雍(Yee-yung Charng),章為皓(Wei-Hau Chang),李平篤(Ping-Du Lee),陳佩燁(Rita P.-Y. Chen) | |
| dc.subject.keyword | 奇異果甜蛋白質,抗黴菌,七個穿膜區域,螢光融合蛋白,蛋白質異源表現系統,冷適應,細胞凋亡, | zh_TW |
| dc.subject.keyword | OLP,OSM,CTPP,GFP,HHP1,PRs,TLP, | en |
| dc.relation.page | 69 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2008-07-30 | |
| 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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