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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 葉開溫 | |
| dc.contributor.author | Jia-jyun Chen | en |
| dc.contributor.author | 陳佳君 | zh_TW |
| dc.date.accessioned | 2021-06-16T09:50:36Z | - |
| dc.date.available | 2022-02-16 | |
| dc.date.copyright | 2017-02-16 | |
| dc.date.issued | 2017 | |
| dc.date.submitted | 2017-01-17 | |
| dc.identifier.citation | 杜建勳。 2015。文心蘭SPLs (SQUAMOSAPROMOTER BINDING PROTEIN- LIKE 參與高室溫誘導開花之調控機制。國立台灣大學植物科學研究所碩士論文。
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/60013 | - |
| dc.description.abstract | 文心蘭的開花調控機制受到維他命C (ascorbate) 及穀胱甘肽 (glutathione) 的氧化還原態 (redox state) 所調控，然而氧化態的GSSG (oxidized form) 如何扮演訊號因子去啟動開花調控基因是一個未知的問題。本實驗利用抗穀胱甘肽抗體 (anti-GSH antiserum) 進行開花假球莖總蛋白質之篩選偵測，得到了一個約20 kDa的蛋白質分子，並證明此蛋白質為組蛋白H2B。試管內反應試驗結果，證實H2B重組蛋白分子可與GSSG進行共價結合，利用液相層析雙質譜儀分析結果也證實H2BCys40可與穀胱甘肽結合。若以1 mM GSSG溶液噴灑文心蘭幼苗期之假球莖，發現可縮短文心蘭抽花梗之時間，且H2B的穀胱甘肽化現象 (glutathionylation) 亦很明顯；利用yeast one-hybrid方法分析，初步顯示穀胱甘肽化的H2B可以和SPL10-2啟動子有親合能力。圓二色光譜 (circular dichroism) 分析也顯示穀胱甘肽化的H2B之構造受到改變。綜合上述，文心蘭的開花機制中有可能是染色質的H2B經過穀胱甘肽化後，影響到開花基因的表現所致。 | zh_TW |
| dc.description.abstract | The flowering regulation of Oncidesa is coordinately regulated by ascorbate and glutathione redox states. However, the role of glutathione disulfide (GSSG；oxidized form) as a signal factor to trigger flowering regulatory genes is still unknown. In this study, the glutathionylated proteins approximate 20 kDa were detected by anti-GSH antiserium, and the histone protein H2B was identified as the glutathionylated protein by LC-MS-MS analysis. The H2B-recombinant protein covalently bound with GSSG in vitro. By LC-MS-MS analysis, we found that the glutathionylation site of H2B was identified at cys40. Oncidesa plants in unsheath stage were treated with 1 mM GSSG. The result demonstrated that the early flowering of Oncidesa and the level of H2B glutathionylation were obviously detected. The glutathionyltion of H2B had affinity on SPL10-2 promoter in yeast one-hybrid assay. Further, the circular dichroism (CD) analysis suggested that glutathionylation of H2B affects its secondary structure. Collectively, we assumed that the glutathionylation of H2B alters the chromatin structure, leading to activation of flowering genes in Oncidesa. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-16T09:50:36Z (GMT). No. of bitstreams: 1 ntu-106-R03b42006-1.pdf: 3824252 bytes, checksum: 095fb53ad88d79d646d89d0c01af12b3 (MD5) Previous issue date: 2017 | en |
| dc.description.tableofcontents | 致謝 ii
中文摘要 iii Abstract iv 檢索表 v 目錄 vii 第一章 前言 1 第一節 文心蘭概論 1 第二節 植物開花生理 4 第三節 穀胱甘肽 (GSH) 與植物氧化逆境之關聯 9 第四節 蛋白質穀胱甘肽化與植物生理之關聯 13 第五節 文心蘭開花調控機制 16 第六節 研究目的 18 第二章 材料與方法 19 一、實驗材料 19 二、實驗方法 19 第一節 氧化態之穀胱甘肽 (GSSG) 處理文心蘭 19 第二節 文心蘭溫度處理 19 第三節 基因釣取 19 第四節 基因表現量測定 20 第五節 西方墨點法 (Western Blot) 24 第六節 載體構築 28 第七節 酵母菌轉型 32 第八節 純化候選基因之重組蛋白進行穀胱甘肽化分析 34 第九節 純化OgH2B重組蛋白進行圓二色光譜 (circular dichroism；CD) 36 第十節 OgH2B modeling 39 第三章 結果 40 第一節 文心蘭不同生長期之蛋白質穀胱甘肽化之挑選 40 第二節 文心蘭OgH2A與OgH2B之ORF (open reading frame) 釣取 41 第三節 OgH2A與OgH2B進行穀胱甘肽化之驗證 41 第四節 文心蘭OgH2B穀胱甘肽化之驗證 43 第五節 文心蘭不同生長期OgH2B調控機制之探討 43 第六節 較高的環境溫度 (30℃) 的開花誘導機制與OgH2B穀胱甘肽化之關係 44 第七節 OgH2B、OgH2BC40A與文心蘭開花基因啟動子交互作用 45 第八節 OgH2BC40A-C及OgH2BC40A-N與 OgSPL10-2啟動子交互作用. .46 第九節 OgH2B重組蛋白穀胱甘肽化對其結構影響之探討 47 第四章 討論 49 第一節 文心蘭不同生長期與蛋白質穀胱甘肽化之關係 49 第二節 OgH2B穀胱甘肽化 50 第三節 抽花芽期與30℃環境中穀胱甘肽化程度與OgH2B蛋白質累積量皆增加 52 第四節 OgH2B和OgH2BC40A與OgSPL10-2啟動子有交互作用之探討 53 第五節 OgH2B穀胱甘肽化對其結構與穩定性之探討 54 第六節 未來展望 56 參考文獻 57 | |
| dc.language.iso | zh-TW | |
| dc.title | 組蛋白H2B的穀胱甘肽化調控文心蘭開花之研究 | zh_TW |
| dc.title | Glutathionylation of H2B regulates flowering initiation
in Oncidesa orchid | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 105-1 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 謝旭亮,王淑珍,葉國禎,徐駿森 | |
| dc.subject.keyword | 文心蘭；組蛋白H2B；開花；穀胱甘?化, | zh_TW |
| dc.subject.keyword | flowering, glutathionylation； H2B； histone modification；Oncidesa, | en |
| dc.relation.page | 94 | |
| dc.identifier.doi | 10.6342/NTU201700119 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2017-01-18 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 植物科學研究所 | zh_TW |
| 顯示於系所單位： | 植物科學研究所 | |
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