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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 張英? | |
| dc.contributor.author | Chia-Lun Chang | en |
| dc.contributor.author | 張家綸 | zh_TW |
| dc.date.accessioned | 2021-06-15T06:23:35Z | - |
| dc.date.available | 2010-08-12 | |
| dc.date.copyright | 2010-08-12 | |
| dc.date.issued | 2010 | |
| dc.date.submitted | 2010-08-09 | |
| dc.identifier.citation | Acaster MA, Kende H (1983) Properties and Partial Purification of 1-Aminocyclopropane-1-carboxylate Synthase. Plant Physiol 72: 139-145
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/47870 | - |
| dc.description.abstract | 鈣離子依存性激酶 (Calcium-dependent protein kinase; CDPK) 是植物細胞內十分重要的鈣離子感應器，它在接收到鈣離子訊號後會對下游的基質進行磷酸化，將鈣離子訊號傳遞下去。根據阿拉伯芥 (Arabidopsis thaliana) CDPK與CRK (CDPK-related kinase) 的胺基酸序列所做的親緣分析 (phylogenetic analysis)，34個CDPK可分為四子群 (subgroup)，且第四子群與CRK的親緣關係較為接近；而這樣的現象在匯入其他物種的CDPK與CRK同樣保持，且發現若進行區域 (domain) 的獨立分析，也觀察到相同現象。阿拉伯芥AtDi19 (drought-induced protein 19) 蛋白是近幾年被發現新的CDPK基質，AtDi19蛋白在阿拉伯芥中有七個家族成員，分別為AtDi19-1~7，具有兩個可能會與DNA結合的類鋅手指結構區域 (zinc-finger like motif)，目前發現AtDi19-2可能與高鹽逆境有關。前人發現AtDi19-2上找到一個磷酸化位點在Ser109，並命名為AtDi19-2-2。且AtDi19-2-2具有基質專一性 (substrate specificity)，只會被AtCPK16所磷酸化而不被其他CDPK磷酸化，在本研究中利用含有磷酸化位點的融合胜肽 (fusion peptide) 之定點突變 (site-directed mutagenesis) 株發現Di19-2-2上Ser109前後-1~+2對於AtCPK16的辨認十分重要。另外在本研究中，找到兩個尚未發表的阿拉伯芥CDPK的基質，分別為阿拉伯芥ACC合成酶5和7 (AtACS5, AtACS7)。 | zh_TW |
| dc.description.abstract | Calcium-dependent protein kinases (CDPKs) play important roles in the Ca2+ signal transduction in plants. CDPKs activated by Ca2+ signal phosphorylate proteins involved in the Ca2+ signal transduction. According to the phylogenetic analysis of 34 CDPKs and 8 CRKs (CDPK-related kinase) in Arabidopsis thaliana, CDPKs were divided into 4 subgroups, and there is the similarity among the CDPKs of subgroup IV and CRKs. This study includes the CDPKs and CRKs of other species, and showed the similar result. Moreover, the phylogenetic analysis of the domains also divided into 4 subgroups. The result of grouping is consistent in planta. In a previous study, the Arabidopsis thaliana AtDi19 (drought-induced protein 19) protein was found to be the substrate of CDPK. The AtDi19 gene family encodes 7 proteins, AtDi19-1~7, which contain two zinc-finger like motifs. AtDi19-2 could be involved in the salt stress response. Based on another research, Ser109 is the phosphorylation site of the AtDi19-2, namely AtDi19-2-2. In particular, AtDi19-2-2 was only phosphorylated by AtCPK16 but not by other CDPKs. The result showed the substrate specificity of CDPKs. In order to identify which amino acids are important for the recognition of AtCPK16, site-directed mutagenesis of fusion peptide containing the phosphorylation site revealed that -1~+2 amino acids next to Ser109 are important for the recognition. In addition, we discovered two additional substrates of Arabidopsis CDPKs, Arabidopsis ACC synthase 5 and 7 (AtACS5, AtACS7). | en |
| dc.description.provenance | Made available in DSpace on 2021-06-15T06:23:35Z (GMT). No. of bitstreams: 1 ntu-99-R96B42016-1.pdf: 2026182 bytes, checksum: aa9ea3ad0ddbcba63b40e5125f309688 (MD5) Previous issue date: 2010 | en |
| dc.description.tableofcontents | 中文摘要 I
Abstract II 縮寫與全名對照表 III 第一章 前言 1 一、鈣離子依存性激酶 (calcium-dependent protein kinase ; CDPK) 1 1. CDPK的基因家族 1 2. CDPK的一級結構 2 3. CDPK的生理功能 2 二、CDPK的基質 (substrate) 3 三、CDPK基質的磷酸化位點 4 四、Di19蛋白 (drought-induced protein 19) 4 五、AtDi19是CDPK的基質 5 六、研究目標 6 第二章 材料與方法 7 一、實驗材料 7 二、實驗方法 7 1. 大腸桿菌勝任細胞 (competent cell) 之製備 7 2. 少量質粒DNA之抽取 7 3. DNA洋菜膠體電泳 8 4. 細菌轉型作用 (transformation) 8 5. 變性聚丙烯胺電泳 (SDS-PAGE) 9 6. 蛋白質定量 9 7. 穀胱甘肽硫轉移酶 (glutathione S-transferase ; GST) 重組蛋白之純化 10 8. GST-6His tag重組蛋白之純化 11 9. 重組蛋白之定點突變 (site-directed mutagenesis) 11 11. 32p放射線訊號的標準化 (normalization) 13 第三章 結果 14 一、植物CDPK與CRK蛋白的親緣分析 14 二、GST-AtDi19-2與融合蛋白GST-CDPK-6H受激酶磷酸化之分析 15 三、GST-AtDi19-2的Ser109受激酶磷酸化之分析 15 四、GST-Di19-2的磷酸化位點之定點突變群的受激酶磷酸化之分析 16 五、番茄ACC合成酶融合蛋白受激酶磷酸化之分析 17 六、阿拉伯芥ACC合成酶融合蛋白之激酶反應 18 第四章 討論 19 參考文獻 23 圖表 44 附錄 57 | |
| dc.language.iso | zh-TW | |
| dc.subject | ACC合成酶 | zh_TW |
| dc.subject | 鈣離子依存性激酶 | zh_TW |
| dc.subject | 親緣分析 | zh_TW |
| dc.subject | AtDi19蛋白 | zh_TW |
| dc.subject | 基質專一性 | zh_TW |
| dc.subject | 定點突變 | zh_TW |
| dc.subject | ACC synthase | en |
| dc.subject | calcium-dependent protein kinases | en |
| dc.subject | phylogenetic analysis | en |
| dc.subject | AtDi19 protein | en |
| dc.subject | substrate specificity | en |
| dc.subject | site-direct mutagenesis | en |
| dc.title | 利用阿拉伯芥Di19-2蛋白研究阿拉伯芥鈣離子依存性激酶之基質專一性 | zh_TW |
| dc.title | Study of Substrate Specificity of Arabidopsis Calcium-Dependent Protein Kinases Using Arabidopsis Drought-Induced Protein 19 as a Substrate | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 98-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 林讚標,謝旭亮,謝明勳,鄭貽生 | |
| dc.subject.keyword | 鈣離子依存性激酶,親緣分析,AtDi19蛋白,基質專一性,定點突變,ACC合成酶, | zh_TW |
| dc.subject.keyword | calcium-dependent protein kinases,phylogenetic analysis,AtDi19 protein,substrate specificity,site-direct mutagenesis,ACC synthase, | en |
| dc.relation.page | 75 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2010-08-09 | |
| 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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