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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 羅凱尹 | |
dc.contributor.author | Ting-Jyun Lu | en |
dc.contributor.author | 呂庭郡 | zh_TW |
dc.date.accessioned | 2021-05-19T18:01:40Z | - |
dc.date.available | 2025-08-06 | |
dc.date.available | 2021-05-19T18:01:40Z | - |
dc.date.copyright | 2015-12-02 | |
dc.date.issued | 2015 | |
dc.date.submitted | 2015-08-06 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/7989 | - |
dc.description.abstract | 核醣體的功能為製造蛋白質,因此核醣體的缺失會對細胞的生理活性造成很大的影響。核醣體由核醣蛋白以及rRNA組成,而組裝的步驟需要兩百多個因子參與其中,這些因子的功能包含協助組裝、修飾以及運輸。對於細胞的生理意義而言,蛋白質轉譯作用極其重要,核醣體的生合成機制在各物種間具有高度的保守性,因此酵母菌經常作為探討核醣體生合成機制的模式生物。目前已知Bcp1參與60S核醣體的生合成:缺少Bcp1的時候會造成帶有綠色螢光標記的60S核醣體蛋白在細胞核累積,並且多核醣體圖譜中的60S核醣體的量有明顯的下降以及80S核醣體與多核醣體的部分有Halfmer產生。本實驗室前人透過高通量抑制子的篩選,發現BCP1與RPL23有所關聯,Rpl23是構成60S核醣體的其中一個核醣蛋白。本研究透過在酵母菌中測試Bcp1對於Rpl23蛋白質穩定性的影響,以及利用大腸桿菌系統測試Bcp1與Rpl23之間的結合,證明Bcp1為Rpl23的伴護子 (chaperone)。 | zh_TW |
dc.description.provenance | Made available in DSpace on 2021-05-19T18:01:40Z (GMT). No. of bitstreams: 1 ntu-104-R02623025-1.pdf: 2704874 bytes, checksum: e72895dabb1cb8e91dda25262e448584 (MD5) Previous issue date: 2015 | en |
dc.description.tableofcontents | 目錄
致謝 i 中文摘要 ii Abstract iii 目錄 iv 表目錄 vii 圖目錄 viii 附圖目錄 ix 壹、前言 1 一、 核醣體生合成 (Ribosome biogenesis) 1 二、 核醣體出核運輸 4 三、 Bcp1簡介 5 四、 研究動機 6 貳、材料與方法 7 一、 菌株與質體 7 二、 轉型作用 (transformation) 7 (一) 大腸桿菌轉型作用 (E. coli heat shock transformation): 7 (二) 快速酵母菌轉型作用 (Quick yeast transformation): 7 (三) 高效酵母菌轉型作用 (High-efficiency yeast transformation): 7 三、 生長測試 (Spot assay) 8 四、 酵母菌雙雜合系統 (Yeast two-hybrid system) 8 五、 多核醣體圖譜分析 (Polysome analysis) 8 六、 螢光顯微鏡觀察 9 七、 免疫沉澱 (Immunoprecipitation) 9 八、 In vitro interaction assay 9 九、 蛋白質染色 (Coomassie blue) 10 十、 bcp1突變基因的建構 10 参、結果 11 一、 BCP1參與60S核醣體生合成 11 二、 Bcp1會與Rpl23結合 13 三、 Bcp1為Rpl23的伴護子 (Chaperone) 14 四、 Bcp1具有全長以及剪切掉N端的兩種蛋白質型式 15 五、 Bcp1會與Rkm1結合 17 六、 bcp1突變株的建構 18 七、 Bcp1透過N端與Rkm1結合 20 八、 Bcp1、Rkm1的結合位不位於Rpl23的C端 22 九、 在酵母菌中探討Bcp1、Rpl23與Rkm1對彼此之間結合的影響 23 十、 探討Mss4對於Bcp1參與60S核醣體生合成的影響 24 肆、結論 26 伍、討論 27 一、 Bcp1、Rpl23以及Rkm1之間的結合 27 二、 Bcp1的兩種蛋白質形式 28 三、 探討Rkm1對於Bcp1與Rpl23結合的影響 29 四、 探討Bcp1與Mss4的結合區塊 (Domain) 30 五、 探討Mss4對於Bcp1參與60S核醣體生合成的影響 31 六、 Bcp1可能在兩條路徑之間扮演調控的角色 32 陸、參考文獻 33 表目錄 表 一 、本實驗所使用之酵母菌菌株 38 表 二、本實驗所使用之質體 40 表 三、各bcp1突變株的生長表現及螢光分布 42 表 四、bcp1突變株定序結果 45 圖目錄 圖 一、BCP1參與60S核醣體生合成 46 圖 二、Bcp1會與Rpl23結合 48 圖 三、Bcp1為Rpl23的伴護子 50 圖 四、Bcp1具有全長以及剪切掉N端的兩種蛋白質型式 52 圖 五、Bcp1會與Rkm1結合 54 圖 六、Bcp1藉由N端與Rkm1結合 56 圖 七、在酵母菌中探討Bcp1、Rpl23與Rkm1對彼此之間結合的影響 59 圖 八、探討Mss4對於Bcp1參與60S核醣體生合成的影響 61 | |
dc.language.iso | zh-TW | |
dc.title | Bcp1與Rpl23間交互作用之探討 | zh_TW |
dc.title | Functional characterization of the interaction between Bcp1 and Rpl23 | en |
dc.type | Thesis | |
dc.date.schoolyear | 103-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 陳美瑜,黃偉邦,冀宏源,徐駿森 | |
dc.subject.keyword | Bcp1,Rpl23,伴護子,Rkm1,Mss4,核醣體生合成, | zh_TW |
dc.subject.keyword | Bcp1,Rpl23,chaperone,Rkm1,Mss4,ribosome biognenesis, | en |
dc.relation.page | 61 | |
dc.rights.note | 同意授權(全球公開) | |
dc.date.accepted | 2015-08-06 | |
dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
dc.contributor.author-dept | 農業化學研究所 | zh_TW |
dc.date.embargo-lift | 2025-08-06 | - |
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