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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 楊健志 | |
| dc.contributor.author | Huei-Jyun Hong | en |
| dc.contributor.author | 洪蕙君 | zh_TW |
| dc.date.accessioned | 2021-06-07T23:44:57Z | - |
| dc.date.copyright | 2014-08-11 | |
| dc.date.issued | 2013 | |
| dc.date.submitted | 2014-07-03 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/16745 | - |
| dc.description.abstract | 阿拉伯芥 HHP1 (Heptahelical transmembrane protein) 是一個具有7個穿膜區塊的膜蛋白質,先前研究發現 HHP1 基因會受離層酸 (ABA)、滲透壓逆境及蔗糖誘導表現,其功能可能是 ABA 及滲透壓逆境訊息傳導的負向調控子。目前研究指出,葡萄糖的訊息傳導與 ABA 訊息傳導途徑有密切關係。本研究對 pHHP1::GUS 轉殖株進行葡萄糖處理,發現 HHP1 表現量會隨著葡萄糖濃度增加而上升,並且主要表現在未展開的子葉及下胚軸基部,顯示 HHP1 可能與植物對糖反應有關。在 55 及 167 mM 葡萄糖環境中,添加 ABA 會使 HHP1 表現量比以葡萄糖處理時些微增加,顯示葡萄糖和 ABA 共同影響 HHP1 表現可能不是加成作用,兩者可能透過同一訊息傳導路徑調控 HHP1 表現。ABI4 是參與糖及 ABA 訊息傳導之轉錄因子,在對 ABA 與糖不敏感的突變株 abi4-1 中,HHP1 表現量受葡萄糖誘導的程度明顯比 WT 低,表示 HHP1 受糖誘導的過程中可能需要 ABI4參與。綜合本研究之結果,HHP1 受葡萄糖誘導的機制可能與 ABI4 調控的 ABA 訊息傳導路徑有關,且 HHP1 可能作用於 ABI4 下游,並參與植物對糖的反應。 | zh_TW |
| dc.description.abstract | HHP1 (heptahelical protein 1) is a membrane protein with 7 transmembrane domains structure. Previous study has shown that HHP1 is induced by abscisic acid (ABA), osmotic stress and sucrose. Furthermore, HHP1 might act as a negative regulator in ABA and osmotic stress signaling. It is known that ABA signaling components are also involved in glucose signaling. As revealed by histochemical and quantitative analyses of GUS activity of pHHP1::GUS transgenic plants, the expression of HHP1 was increased with glucose concentration, and predominantly shown in the unexpanded cotyledon and the base of hypocotyl, indicating that HHP1 might participate in the response to glucose. Under 55 and 167 mM glucose condition, the glucose-induced HHP1 expression was slightly increased in the presence of ABA. Therefore, glucose and ABA might not act synergistically on the induction of HHP1 expression, suggesting they might act in the same signaling pathway to regulate HHP1 expression. ABI4 (ABA insensitive 4) is a transcription factor which participate in ABA and sugar signaling. In the abi4-1 mutant, which is insensitive to ABA and glucose, the glucose-induced HHP1 expression was significantly reduced than in WT. This result indicated that ABI4 was required for glucose-induced HHP1 expression. Taken together, these data suggested that HHP1 might act downstream of ABI4 and be regulated by ABA-mediated glucose response. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-07T23:44:57Z (GMT). No. of bitstreams: 1 ntu-102-R00b22004-1.pdf: 2552795 bytes, checksum: 347a054e65bbc38fd9161fd2f19106d8 (MD5) Previous issue date: 2013 | en |
| dc.description.tableofcontents | 目錄 I
縮寫表 IV 中文摘要 VII Abstract VIII 圖目錄 IX 表目錄 X 第一章 緒論 1 1.1 離層酸 (Abscisic acid, ABA) 1 1.1.1 離層酸之生合成及代謝 1 1.1.2 離層酸之訊息傳導 4 1.2 糖訊息傳導 (sugar signaling) 6 1.2.1 葡萄糖之感知 (glucose sensing) 7 1.2.2 葡萄糖之訊息傳遞機制 (glucose signaling) 8 1.2.3 葡萄糖訊息傳導突變株 8 1.2.4 葡萄糖與 ABA 生合成反應之交互作用 10 1.2.5 葡萄糖與 ABA 訊息傳導之交互作用 11 1.3 糖與離層酸的轉錄調控 13 1.3.1 與糖相關之轉錄調控 13 1.3.2 與離層酸相關之轉錄調控 15 1.4 HHP 蛋白質家族 16 1.4.1 HHP1 17 1.5 本論文之研究緣起及動機 19 第二章 材料與方法 20 2.1 實驗材料 20 2.1.1 植物材料 20 2.1.2 載體 (vectors) 21 2.1.3 菌種 21 2.2 實驗藥品 21 2.2.1 一般化學藥劑 21 2.2.2 酵素 21 2.2.3 培養基 22 2.2.4 植物組織培養用藥品 22 2.3 儀器設備 23 2.4 實驗方法 24 2.4.1 阿拉伯芥之種植 24 2.4.1.1 種子表面消毒及低溫處理 24 2.4.1.2 種子之無菌培養 24 2.4.1.3 種子之土壤培養 24 2.4.1.4 種子之收集 25 2.4.2 GUS 活性分析實驗 25 2.4.2.1 GUS 活性染色分析 25 2.4.2.2 GUS 活性定量分析 26 2.4.3 RNA 之抽取與相關分析實驗 27 2.4.3.1 阿拉伯芥 total RNA 之抽取 27 2.4.3.2 甲醛瓊脂糖膠體電泳 27 2.4.3.3 核酸定量 28 2.4.3.4 DNase treatment 29 2.4.3.5 反轉錄反應 (Reverse Transcription) 30 2.4.4 HHP1 基因表現量分析 31 2.4.4.1 定量性即時聚合酶連鎖反應 31 2.4.4.2 阿拉伯芥不同糖處理之樣品收集 32 2.4.4.3 各處理樣品之定量性即時聚合酶連鎖反應 32 2.4.5 目標載體之建構 32 2.4.5.1 以 Phusion DNA polymerase 進行聚合酶連鎖反應(polymerase chain reaction, PCR) 33 2.4.5.2 DNA 瓊脂糖膠體電泳 (Agarose Gel Electrophoresis) 34 2.4.5.3 PCR 產物之分離與純化 34 2.4.5.4 DNA 之磷酸化反應 35 2.4.5.5 Sticky-end PCR 35 2.4.5.6 質體 DNA 小量製備 37 2.4.5.7 DNA 限制酶切割反應 (Restriction Enzyme Digestion) 37 2.4.5.8 DNA 之去磷酸化反應 37 2.4.5.9 接合反應 38 2.4.5.10 大腸桿菌熱衝擊法轉形 (Heat-shock Transformation) 38 2.4.5.11 菌落 PCR 檢定法 (colony PCR) 38 第三章 結果與討論 40 3.1. 以 pHHP1::GUS 轉殖株觀察 HHP1 於不同處理下之表現情形 40 3.1.1. 添加葡萄糖及蔗糖可誘導HHP1的表現 41 3.1.2. 葡萄糖和離層酸對 HHP1 表現的影響 42 3.1.3. 添加 fluridone 對 HHP1 表現的影響 43 3.2. 以 qRT-PCR 分析 HHP1 於不同處理下的基因表現量 45 3.3. 分析 ABA 相關突變株中 HHP1 的表現情形 46 3.3.1. HHP1 於 ahg1-3 中的基因表現情形 47 3.3.2. HHP1 於 abi4-1 中的基因表現情形 48 3.4. 分析 HHP1 上游序列中可能含有的糖相關之轉錄調控序列 49 3.5. 尋找 HHP1 上游序列中與 ABA 訊息傳導相關轉錄因子結合的轉錄調控序列 50 3.6. 結論 52 第四章 未來展望 54 4.1 觀察轉錄因子 ABI4 是否與 HHP1 上游序列結合並調控其基因表現 54 4.2 不同糖對 HHP1 基因表現之影響 54 4.3 探討葡萄糖誘導 HHP1 表現之機制 55 4.4 探討 HHP1 是否受 Retrograde signaling 調控 56 參考文獻 57 第五章 圖與表 68 附錄一、本論文所使用之引子序列 79 論文口試問答摘要 80 | |
| dc.language.iso | zh-TW | |
| dc.subject | ABI4 | zh_TW |
| dc.subject | HHP1 | zh_TW |
| dc.subject | 葡萄糖 | zh_TW |
| dc.subject | 離層酸 | zh_TW |
| dc.subject | glucose | en |
| dc.subject | ABA | en |
| dc.subject | ABI4 | en |
| dc.subject | HHP1 | en |
| dc.title | 阿拉伯芥 HHP1 基因受葡萄糖誘導與離層酸訊息傳導之關係 | zh_TW |
| dc.title | The relationship between abscisic acid signaling and glucose-induced HHP1 expression in Arabidopsis | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 102-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 王愛玉,陳佩燁,常怡雍,黃楓婷 | |
| dc.subject.keyword | HHP1,葡萄糖,離層酸,ABI4, | zh_TW |
| dc.subject.keyword | HHP1,glucose,ABA,ABI4, | en |
| dc.relation.page | 86 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2014-07-03 | |
| 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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