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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/69924完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 李昆達(Kung-Ta Lee) | |
| dc.contributor.author | Chen-Lun Chen | en |
| dc.contributor.author | 陳振倫 | zh_TW |
| dc.date.accessioned | 2021-06-17T03:34:20Z | - |
| dc.date.available | 2021-03-02 | |
| dc.date.copyright | 2018-03-02 | |
| dc.date.issued | 2018 | |
| dc.date.submitted | 2018-02-12 | |
| dc.identifier.citation | 1 Weller, C. V. & Riker, A. D. Rhinosporidium Seeberi: Pathological Histology and Report of the Third Case from the United States. Am J Pathol 6, 721-732 711 (1930).
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Arabidopsis LTPG Is a Glycosylphosphatidylinositol-Anchored Lipid Transfer Protein Required for Export of Lipids to the Plant Surface. Plant Cell 21, 1230-1238, doi:10.1105/tpc.108.064451 (2009). 37 Edstam, M. M. & Edqvist, J. Involvement of GPI-anchored lipid transfer proteins in the development of seed coats and pollen in Arabidopsis thaliana. Physiol Plant 152, 32-42, doi:10.1111/ppl.12156 (2014). 38 Lee, S. B. et al. Disruption of Glycosylphosphatidylinositol-Anchored Lipid Transfer Protein Gene Altered Cuticular Lipid Composition, Increased Plastoglobules, and Enhanced Susceptibility to Infection by the Fungal Pathogen Alternaria brassicicola. Plant Physiol 150, 42-54, doi:10.1104/pp.109.137745 (2009). 39 Maldonado, A. M., Doerner, P., Dixon, R. A., Lamb, C. J. & Cameron, R. K. A putative lipid transfer protein involved in systemic resistance signalling in Arabidopsis. Nature 419, 399-403, doi:10.1038/nature00962 (2002). 40 Regente, M. C., Giudici, A. M., Villalain, J. & de la Canal, L. The cytotoxic properties of a plant lipid transfer protein involve membrane permeabilization of target cells. Lett Appl Microbiol 40, 183-189, doi:10.1111/j.1472-765X.2004.01647.x (2005). 41 Pagnussat, L., Burbach, C., Baluška, F. & de la Canal, L. An extracellular lipid transfer protein is relocalized intracellularly during seed germination. J Exp Bot 63, 6555-6563, doi:10.1093/jxb/ers311 (2012). 42 Carvalho, A. D. & Gomes, V. M. Role of plant lipid transfer proteins in plant cell physiology - A concise review. Peptides 28, 1144-1153, doi:10.1016/j.peptides.2007.03.004 (2007). 43 Howe, G. A. & Schilmiller, A. L. Oxylipin metabolism in response to stress. Curr Opin Plant Biol 5, 230-236 (2002). 44 Liavonchanka, A. & Feussner, I. Lipoxygenases: occurrence, functions and catalysis. J Plant Physiol 163, 348-357, doi:10.1016/j.jplph.2005.11.006 (2006). 45 Halim, V. A., Vess, A., Scheel, D. & Rosahl, S. 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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/69924 | - |
| dc.description.abstract | 當根毛農桿菌感染植物傷口組織，根毛農桿菌Ri質體上的T-DNA會插入植物的染色體中產生一癌化組織-毛狀根。我們實驗室多年來致力於研究毛狀根的誘導機制，並發現rolB或rolC缺失對於毛狀根的發育和側根發育有很大的影響，過去在ΔrolB、ΔrolC毛狀根的轉錄體分析中發現，RolB/RolC對於菸草脂質轉移蛋白 (lipid transfer protein, LTP) 的表現可能具有正向調控。因此，本研究以釀酒酵母菌/大腸桿菌表現系統對特定菸草脂質轉移蛋白，包含AB25、AF54、BQ56、DV77、DW88、EB85和FG54，分別進行異源表現，並進一步對個別的結合基質進行篩選。本研究首先建構表現質體，在釀酒酵母菌中僅能表現EB85與FG54，但後續難以純化。在重組LTP的N端加入融合蛋白Nus (N utilization substance protein A) 後，可使LTP成功表現在大腸桿菌中，經過純化後利用結合位競爭抑制方法，分析可與脂質轉移蛋白結合之脂質基質。BQ56、DV77、EB85、FG54可與C12~C18飽和脂肪酸、C16~C18不飽和脂肪酸、茉莉酸結合，其中BQ56對palmitoleic acid、DV77對linolenic acid可能有較高的結合能力，FG54會隨著脂質的碳鏈長度或雙鍵數增加而促進結合能力的趨勢，DW88則可能只能與油酸結合。上述結果顯示BQ56、DV77、DW88、EB85、FG54能夠在大腸桿菌中進行異源表現，菸草LTP可以廣泛地結合不同種脂質，其中包含被認為會影響根部發育的茉莉酸和羥脂前驅物亞麻油酸與次亞麻油酸。且對於脂質會有非專一性的結合。 | zh_TW |
| dc.description.abstract | When Agrobacterium rhizogenes infects the wound plant tissue, transfer DNA (T-DNA) of root-inducing plasmid (pRi) inserts into plant chromosome and develops the tumorized tissue – hairy root. Our previous studies suggested that lacking rolB/rolC gene cause impaired hair root syndrome and rolB/rolC gene plays an important role in root inducing capacity. Moreover, it also indicated that RolB/RolC may positively regulate the expression of proteins related lipid transport, called lipid transfer proteins, LTPs. In this study, we used Escherichia coli and Saccharomyces cerevisiae as host for heterologous expression of specific tobacco LTPs comprising of AB25, AF54, BQ56, DV77, DW88, EB85 and FG54. First, we tried to express LTPs in S. cerevisiae, but only EB85 and FG54 could be expressed. Afterwards, recombinant LTPs fused with N-terminal NusA (N utilization substance protein A) were successfully expressed in E. coli. Binding substrate analysis was used to screen what the possible substrates are. The result showed BQ56, DV77, EB85, FG54 had similar binding pattern which could bind to C12~18 saturated fatty acid, C16~18 unsaturated fatty acid harboring one to three double bond and jasmonate. In addition, BQ56 corresponding to palmitoleic acid and DV77 corresponding to linolenic acid might have higher binding ability. The binding ability of FG54 was enhanced when the length of carbon chain and number of double bond increased. DW88 merely bound to oleic acid. These results revealed that tobacco LTPs broadly bind to several kinds of lipid substrate including jasmonate, linoleic acid, and linolenic acid which are related to root growth. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-17T03:34:20Z (GMT). No. of bitstreams: 1 ntu-107-R04b22041-1.pdf: 3742780 bytes, checksum: 050034a3c9bdcfff374800a2c25a3d0b (MD5) Previous issue date: 2018 | en |
| dc.description.tableofcontents | 口試委員審定書 I
誌謝 II 中文摘要 III Abstract IV 圖目錄 VII 表目錄 IX 縮寫表 X 第一章 緒論 1 第一節 文獻回顧 1 第一項 毛狀根 1 第二項 植物非專一性脂質轉移蛋白質 2 第三項 羥脂 (Oxylipin) 4 第四項 異源表現LTP 5 第五項 TNS螢光猝滅 5 第二節 研究目的 7 第三節 研究架構 8 第二章 實驗材料與方法 9 第一節 材料 9 第一項 質體與菌株 9 第二項 引子 9 第二節 方法 10 第一項 目標基因增幅 10 第二項 建構嵌入菸草脂質轉移蛋白之載體 11 第三項 表現重組脂質轉移蛋白於釀酒酵母菌/大腸桿菌並純化之 13 第四項 分析重組LTP蛋白質 18 第五項 分析重組脂質轉移蛋白與不同疏水性物質之結合能力 19 第三章 結果與討論 21 第一節 菸草LTP基因序列分析 21 第二節 具有菸草LTP基因cDNA之質體製備 21 第三節 表現菌株之建立 22 第四節 以西方墨點法 (Western blot) 確認LTP蛋白之表現與純化 23 第五節 LTP蛋白之結合基質篩選 25 第六節 討論 26 第四章 結論 29 第五章 圖表 30 第六章 參考文獻 59 第七章 附錄 68 | |
| dc.language.iso | zh-TW | |
| dc.subject | 菸草 | zh_TW |
| dc.subject | 脂質轉移蛋白 | zh_TW |
| dc.subject | 毛狀根 | zh_TW |
| dc.subject | 異源表現 | zh_TW |
| dc.subject | 大腸桿菌 | zh_TW |
| dc.subject | 釀酒酵母菌 | zh_TW |
| dc.subject | Escherichia | en |
| dc.subject | lipid transfer protein | en |
| dc.subject | Nicotiana tabaccum | en |
| dc.subject | hairy root | en |
| dc.subject | Saccharomyces cerevisiae | en |
| dc.subject | heterologous expression | en |
| dc.title | 菸草毛狀根脂質轉移蛋白之異源表現 | zh_TW |
| dc.title | Heterogolous expression of Tobacco Hairy Root Lipid Transfer Proteins | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 106-1 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 楊健志(Chien-Chih Yang),張世宗(Shih-Chung Chang),劉?德(Chi-Te, Liu) | |
| dc.subject.keyword | 脂質轉移蛋白,菸草,毛狀根,異源表現,大腸桿菌,釀酒酵母菌, | zh_TW |
| dc.subject.keyword | lipid transfer protein,Nicotiana tabaccum,hairy root,heterologous expression,Escherichia,Saccharomyces cerevisiae, | en |
| dc.relation.page | 78 | |
| dc.identifier.doi | 10.6342/NTU201800264 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2018-02-13 | |
| 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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