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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 李秀敏 | |
dc.contributor.author | Yu-Lun Tsai | en |
dc.contributor.author | 蔡雨倫 | zh_TW |
dc.date.accessioned | 2021-06-16T22:59:42Z | - |
dc.date.available | 2012-08-15 | |
dc.date.copyright | 2012-08-15 | |
dc.date.issued | 2012 | |
dc.date.submitted | 2012-08-07 | |
dc.identifier.citation | Alabadi D, Gil J, Blazquez MA, Garcia-Martinez JL (2004) Gibberellins repress photomorphogenesis in darkness. Plant Physiol 134: 1050-1057
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/64804 | - |
dc.description.abstract | 70 kD的熱休克蛋白 (heat shock protein 70 kD, Hsp70) 家族是細胞中普遍存在的伴護子 (chaperone),參與蛋白質轉位 (protein translocation)、摺疊 (folding) 及降解 (degradation) 的過程。植物葉綠體內的cpHsc70最近被發現參與在前驅蛋白 (precursor protein) 輸入葉綠體的轉位過程。因為Hsp70在細胞中參與的功能太多,需要J蛋白來協助Hsp70的功能專一性,但在葉綠體內的J蛋白至少有19個,所以我們實驗室欲找出幫助cpHsc70進行轉位作用的J蛋白。本篇文章裡研究的J25蛋白被預測具有穿膜區塊 (transmembrane domain),在本篇論文中,我發現J25不只位在類囊體膜 (thylakoid membrane)上,還有一小部分位在葉綠體內膜 (inner envelope membrane) 上。然而,從我的研究得知,j25突變株並不會造成蛋白質輸入葉綠體的過程受損。而此時,我們意外發現,在白光及紅光下生長的j25-1突變植株比起野生型有較長下胚軸 (hypocotyls) 的外表型,且j25與phyB (phytochrome B是最主要的紅光受器) 的雙突變株會有比兩個單突變株更長的下胚軸。另外,j25/phyB雙突變株種子的萌芽率也比兩個單突變株來的差。藉由j25突變株的特性分析,得知J25位在與紅光受器phyB不同的光訊息傳遞路徑上,讓我們得以將葉綠體的熱休克蛋白cpHsc70藉由J25與光傳導的訊息路徑做可能的連結。 | zh_TW |
dc.description.abstract | The 70 kD heat shock proteins (Hsp70s) are involved in protein translocation, folding and degradation. We have recently shown that chloroplast Hsp70s, cpHsc70s, are involved in protein import into chloroplasts. Since the J protein cochaperones are drivers that recruit Hsp70 for specific functions, we intend to identify the J protein assisting cpHsc70s in chloroplast protein import. There are more than one hundred J proteins in Arabidopsis, and at least 19 of them are localized in chloroplasts. Among them, J25 is predicted to localize in membranes. I found that J25 has a dual localization: the majority of J25 is localized in the thylakoid membrane and a small portion is localized in the inner envelope membrane. However, the j25 mutants show no defect in the import of a model chloroplast precursor protein. Nonetheless, I found another interesting phenotype of the j25 mutants: one of the alleles has long hypocotyls when grown under white and red light and double mutants of both alleles with a phyB (phytochrome B, the major red light sensor of plants) mutant have an additive long hypocotyl phenotype under white light and red light. In addition, the j25/phyB double mutants have an additive defect in seeds germination. These data suggest that J25 is involved in light sensing and that J25 and phyB are involved in independent red light signaling pathways. Characterizations of J25 provide a possible link between chloroplast Hsp70 and light-regulated signal transduction. | en |
dc.description.provenance | Made available in DSpace on 2021-06-16T22:59:42Z (GMT). No. of bitstreams: 1 ntu-101-R99B43032-1.pdf: 3044594 bytes, checksum: 51a52d55ad3844b493ccac2205af3484 (MD5) Previous issue date: 2012 | en |
dc.description.tableofcontents | 壹、 口試委員會審定書
貳、 誌謝………………………………………………………I 參、 中文摘要…………………………………………………II 肆、 英文摘要…………………………………………………III 伍、 目錄……………………………………………………IV-VI 陸、 前言…………………………………………………………1 柒、 材料與方法…………………………………………………5 1. 植物材料 2. 植物的種植與生長狀況 3. 引子 4. 檢驗阿拉伯芥突變株的T-DNA插入情形 5. 阿拉伯芥突變株的基因表現量分析 6. 阿拉伯芥突變株的蛋白質表現量分析 7. 阿拉伯芥葉綠體製備 8. 豌豆葉綠體製備 9. 抽取中量質體DNA 10. 活體外轉錄與轉譯 11. 活體外前驅蛋白質輸入葉綠體 12. 蛋白質濃度測定 13. 硫酸十二脂聚丙烯醯胺膠片的電泳分析 14. 西方墨點法 15. J25的定位分析 16. J25的抗體製備 17. 阿拉伯芥雙突變株的篩選 18. 阿拉伯芥幼苗的胚軸長度測量 19. 阿拉伯芥種子萌芽實驗 捌、 結果………………………………………………………25 1. 以J25-Ag1蛋白質做為抗原所製備的抗體可以成功辨認阿拉伯芥及豌豆的J25蛋白 2. J25為葉綠體內的穿膜蛋白 (integral membrane protein) 3. 內源性J25蛋白坐落在類囊體膜系 (thylakoid membrane) 及內膜 (inner envelope membrane) 上 4. j25-1及j25-2突變株中皆無可偵測到的J25蛋白,但j25-2仍有少部分的J25 mRNA存在 5. 南方墨點法 (Southern blot) 顯示j25-2為單個T-DNA插入的突變株,但j25-1很有可能含有其他的T-DNA插入位置 6. j25突變株不會造成蛋白質轉位的缺陷 7. j25/phyB 雙突變株在紅光及白光下生長的胚軸比j25及phyB單突變株的胚軸長 8. j25/phyB 雙突變株的萌芽率比j25及phyB單突變株還低 玖、 討論………………………………………………………31 壹拾、 參考文獻……………………………………………………...35 壹拾壹、 圖表………………………………………………………41 圖一、 葉綠體上的運輸機組示意圖 圖二、 電子傳遞鏈示意圖 圖三、 光敏素合成及特性示意圖 圖四、 J25被預測為是坐落在膜系上的葉綠體蛋白質 圖五、 純化J25-Ag1蛋白質做為製備J25抗體的抗原 圖六、 J25抗體可以成功辨認豌豆及阿拉伯芥葉綠體內的J25蛋白 圖七、 J25為葉綠體內的穿膜蛋白 圖八、 內源性J25可以抵抗嗜熱菌酶及胰蛋白酶 圖九、 J25的定位分析 圖十、 j25-1及j25-2突變株的特性分析 圖十一、 南方墨點法檢驗j25-1及j25-2的T-DNA插入情形 圖十二、 j25 突變株不會影響前驅蛋白輸入葉綠體的量 圖十三、 j25/phyB 雙突變株在白光下的下胚軸比單突變株長 圖十四、 j25/phyB 雙突變株在不同色光下的胚軸生長情形 圖十五、 j25/phyB雙突變株在黑暗中的萌芽率比野生型低 | |
dc.language.iso | zh-TW | |
dc.title | 阿拉伯芥葉綠體蛋白J25的功能與特性分析 | zh_TW |
dc.title | Characterizations of Arabidopsis Chloroplast J Protein J25 | en |
dc.type | Thesis | |
dc.date.schoolyear | 100-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 涂世隆,蘇百祥,吳素幸 | |
dc.subject.keyword | 葉綠體,熱休克蛋白,光,種子萌芽, | zh_TW |
dc.subject.keyword | chloroplast,heat shock protein,light,germination, | en |
dc.relation.page | 55 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2012-08-08 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 分子與細胞生物學研究所 | zh_TW |
顯示於系所單位: | 分子與細胞生物學研究所 |
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