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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 楊健志(Chien-Chih Yang) | |
| dc.contributor.author | Shih-Yu Kuo | en |
| dc.contributor.author | 郭時鈺 | zh_TW |
| dc.date.accessioned | 2021-06-16T23:33:25Z | - |
| dc.date.available | 2018-07-26 | |
| dc.date.copyright | 2012-08-01 | |
| dc.date.issued | 2012 | |
| dc.date.submitted | 2012-07-27 | |
| dc.identifier.citation | Acevedo-Hernandez GJ, Leon P, Herrera-Estrella LR (2005) Sugar and ABA responsiveness of a minimal RBCS light-responsive unit is mediated by direct binding of ABI4. Plant J 43: 506-519
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Plant Cell Environ 34: 1031-1042 Zhu Y, Bond J, Thomas P (2003a) Identification, classification, and partial characterization of genes in humans and other vertebrates homologous to a fish membrane progestin receptor. Proc Natl Acad Sci U S A 100: 2237-2242 Zhu Y, Rice CD, Pang Y, Pace M, Thomas P (2003b) Cloning, expression, and characterization of a membrane progestin receptor and evidence it is an intermediary in meiotic maturation of fish oocytes. Proc Natl Acad Sci U S A 100: 2231-2236 高艾玲 (2010). 阿拉伯芥 AtMAPRs 之分子特性研究. 博士論文. 國立臺灣大學微生物與生化學研究所. | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/65262 | - |
| dc.description.abstract | 豬的 MAPR (membrane associated progesterone receptor component 1) 與黃體激素及血基質結合有關，從阿拉伯芥中發現四個與 MAPR 相似度為30-40 % 之同源蛋白質，將其命名為 AtMAPR2、AtMAPR3、AtMAPR4 與 AtMAPR5。這四種蛋白質皆具有高度保守性的 cytochrome b5 like heme/steroid binding domain。(高艾玲，2010)
本論文針對 AtMAPR3 與血基質結合特性，探討 AtMAPR3 的表現量是否會受到血基質影響。以血基質合成前驅物 δ-aminolevulinic acid (ALA) 處理11天大阿拉伯芥幼苗，發現在正常光照下 AtMAPR3 表現量上升 3.5倍，完全黑暗下基因表現也上升2.5倍，而其他 AtMAPRs 卻不受影響。處理 ALA 後再加入 dipyridyl (DPD) 來抑制 Fe-chelatase 使得血基質無法合成，AtMAPR3 表現量上升情形即消失。AtMAPR3 可能與過氧化物的代謝有關，分別以 H2O2 及 H2O2 類似物 tert-butyl hydroxide (tBHP) 處理11天大幼苗後，AtMAPR3 表現量約上升6倍及4倍。先前文獻指出，植物對於 jasmonic acid (JA) 產生的反應可能是利用 H2O2 作為訊息分子啟動一些防禦基因之表現，以不同濃度 JA 處理 (50-100 μM)，會使 AtMAPR3 表現量上升4倍。實驗室已有 AtMAPR3 基因剔除突變株 (AtMAPR3-KO)，需要建構 AtMAPR3 基因過量表現突變株 (AtMAPR3-OX)，以瞭解 AtMAPR3 所參與的生理功能。將此兩種突變株及 WT 進行 H2O2、tBHP 及 methyl viologen (MeV) 氧化逆境處理，發現 AtMAPR-KO 存活率比 WT 略低，但無顯著差異，此外也無觀察到 AtMAPR-OX 有存活率較高現象。綜合這些實驗結果推論，阿拉伯芥細胞中 ALA 下游產物量，如血基質或 Mg-protoporphyrin IX，可能是影響 AtMAPR3 表現量的訊息分子之一。而AtMAPR3 在 ROS 所引發反應中扮演何種生理角色，需要進一步實驗探討。 | zh_TW |
| dc.description.abstract | Four functional-unknown proteins, AtMAPR2, AtMAPR3, AtMAPR4 and AtMAPR5, possessing sequence 30-40% similarity to MAPR (membrane associated progesterone receptor) are studied in this group. The cytochrome b5 like heme/steroid binding domain are highly conserved in these proteins, and they bind progesterone and heme in animals. The UV-visible absorption spectra showed AtMAPRs are hemoproteins. (Kao, 2010, unpublished)
To study the physiological role of heme-binding ability of AtMAPR3, we analyzed whether or not the expression of AtMAPR3 are subjected to the heme concentration. In this study, 10-day-old seedlings were treated with a precursor of heme, δ-aminolevulinic acid (ALA). The results showed AtMAPR3 expression was three-fold-increased by ALA treatment under 16 h light / 8 h dark, and two-fold-increased under 24 h dark; however, when adding dipyridyl (DPD), the inhibitor of Fe-chelatase, the induction was blocked. The expression of AtMAPR3 was also induced by H2O2 and tert-butyl hydroxide (tBHP) about six-fold. H2O2 could be used as a second messenger to express the defence-related genes by jasmonic acid (JA), and JA increased AtMAPR3 expression about four-fold. To further study the physiology roles of AtMAPR3, we need to analyze the phenotype of AtMAPR3-KO and other mutants. In this study, AtMAPR3-KO, AtMAPR3-OX and WT were treated with H2O2, tBHP and methyl viologen (MeV), and the survival rate of AtMAPR3-KO was lower than WT slightly. However, AtMAPR3-OX showed no differences with WT. The results revealed that the downstream products of ALA, such as Mg-protoporphyrin IX or heme, could be a signal molecule which involved in the expression of AtMAPR3. The expression of AtMAPR3 in response to ROS needs to be further investigated in physiological aspect. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-16T23:33:25Z (GMT). No. of bitstreams: 1 ntu-101-R99b22048-1.pdf: 3344077 bytes, checksum: af4e62117983864796f396e99e9ff406 (MD5) Previous issue date: 2012 | en |
| dc.description.tableofcontents | 目錄 I
圖目錄 IV 縮寫表 VI 摘要 IX Abstract X 第一章 緒論 1 1.1黃體激素 2 1.1.1 Nuclear Progesterone Receptor：PR-A 及 PR-B 3 1.1.2 Membrane Progestin Receptor：mPRα、mPRβ 及mPRγ 3 1.1.3 Membrane Associated Progesterone Receptor component 1：MAPR (PGRMC1 或 PGMRC1) 4 1.2 阿拉伯芥中 MAPRs (AtMAPRs) 6 1.2.1 阿拉伯芥 7 1.2.2 AtMAPRs 蛋白質序列特性 8 1.2.3 AtMAPRs 功能 8 1.2.4 本論文之研究主角：AtMAPR3 10 1.3 血基質 (heme) 11 1.4 光與光呼吸作用 (light and photorespiration) 17 1.5 氧化逆境 (oxidative stress) 20 1.6 研究方向及重點 21 第二章 材料與方法 23 2.1實驗材料 23 2.1.1 植物材料 23 2.1.2 載體 (vector) 23 2.1.3 菌種 23 2.2 實驗藥品 24 2.2.1 一般化學藥品 24 2.2.2 酵素 24 2.2.3 培養基 24 2.3 儀器設備 25 2.4 實驗方法 26 2.4.1 阿拉伯芥種植 27 2.4.2 AtMAPR3表現載體之建構 29 2.4.3 AtMAPR3轉殖株之建立 31 2.4.4 DNA抽取與相關分析 32 2.4.5 RNA之抽取與相關分析 37 2.4.6 AtMAPRs於不同處理下的基因表現 41 2.4.7 AtMAPR3 T-DNA突變株之鑑定與分析 47 2.4.8阿拉伯芥外表型 (phenotype) 之觀察 53 第三章 結果與討論 54 3.1 AtMAPR3 基因表現分析 54 3.1.1 AtMAPR3 於 ALA/DPD/Hemin 處理下之表現 54 3.1.2 AtMAPR3 於 H2O2/MeV/tBHP 處理下之表現 56 3.1.3 AtMAPR3 於 JA/SA處理下之表現情形 57 3.2 過量表現 AtMAPR3 (AtMAPR3-OX) 突變株篩選 57 3.3 AtMAPR3 突變株之外表型觀察 59 3.3.1 H2O2 處理 59 3.3.2 tBHP 處理 60 3.3.3 MeV 處理 60 3.4 結論 62 第四章 未來展望 64 4.1 AtMAPR3 突變株外表型之觀察 64 4.2 探討 AtMAPR3 與 ROS 之關係 64 4.3 探討 AtMAPR3 與 AGT1 之關係 65 4.4 探討 AtMAPR3 是否受 retrograde signaling 所調控 65 參考文獻 66 實驗結果圖 72 附錄一、pCAMBIA1302 載體之圖譜 94 附錄二、TA cloning 載體之圖譜及序列 95 附錄三、本論文所使用的引子序列 96 問答集 97 | |
| dc.language.iso | zh-TW | |
| dc.subject | 血基質 | zh_TW |
| dc.subject | 阿拉伯芥 | zh_TW |
| dc.subject | 逆境 | zh_TW |
| dc.subject | AtMAPR3 | en |
| dc.subject | d-amino levulinic acid | en |
| dc.subject | heme | en |
| dc.title | 阿拉伯芥中 AtMAPR3 表現受 d-amino levulinic acid 及逆境誘導之研究 | zh_TW |
| dc.title | Study of the AtMAPR3 expression induced by d-amino levulinic acid and stresses in Arabidopsis | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 100-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 李昆達(Kung-Ta Lee),李平篤(Ping-Du Lee),廖憶純(Yi-Chun Liao),黃楓婷(Feng-Ting Huang) | |
| dc.subject.keyword | 阿拉伯芥,逆境,血基質, | zh_TW |
| dc.subject.keyword | AtMAPR3,d-amino levulinic acid,heme, | en |
| dc.relation.page | 102 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2012-07-27 | |
| 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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