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http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/47426完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 張鑫 | |
| dc.contributor.author | Pei-Chun Wu | en |
| dc.contributor.author | 吳佩純 | zh_TW |
| dc.date.accessioned | 2021-06-15T05:59:18Z | - |
| dc.date.available | 2015-09-09 | |
| dc.date.copyright | 2010-09-09 | |
| dc.date.issued | 2010 | |
| dc.date.submitted | 2010-08-17 | |
| dc.identifier.citation | Bartenschlager, R., Frese, M., and Pietschmann, T. (2004). Novel insights into hepatitis C virus replication and persistence. Adv Virus Res 63, 71-180.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/47426 | - |
| dc.description.abstract | C型肝炎病毒為單股正向RNA病毒，其正股基因體可轉譯成一多蛋白質前驅物，經由細胞內部及病毒本身的蛋白酶切割成結構性和非結構性蛋白質。非結構性蛋白質NS3由631個胺基酸組成，具有多功能性；其N端部分為serine protease，C端部分為RNA helicase。先前實驗室已證實，NS3蛋白質在NS4A蛋白質的輔助之下，可進行病毒多蛋白質切割和內部截切的功能。內部截切產生主要的截切產物NS3(1-402)，相較於其野生型具有更高度的細胞轉型能力。先前研究亦指出，NS3能使細胞在沒有貼附及低血清的情況下穩定生長。一般的細胞藉由貼附細胞外基質活化焦點黏著 (focal adhesion) 訊息傳遞路徑，調節細胞的增生、遷移、生長和細胞凋亡等功能。為了探究NS3蛋白質造成細胞轉型的機制，是否透過對focal adhesion訊息傳遞的影響，本研究首先利用共同免疫沉澱和GST pull down進行分析，結果偵測到focal adhesion細胞因子integrin β1能和NS3形成複合物。當NS3和integrin β1同時存在時，integrin β1蛋白質的表現量較單獨表現時會些微下降，且細胞週期相關蛋白質CDK inhibitor p21和p27會因NS3的同時表現有些微改變，推測 NS3的存在會干擾integrin β1活化的訊息傳遞。根據細胞附著實驗的結果顯示，細胞的貼附能力會受到NS3和integrin調節。MTT assay的結果也發現， integrin β1存在會抑制細胞生長，而integrin β1與NS3(1-402) 同時存在時的影響更為明顯。因此推測，NS3可能藉由影響integrin β1活化的訊息傳遞，進而影響細胞生長、貼附的能力，這個結果提供細胞轉型的可能機制。 | zh_TW |
| dc.description.provenance | Made available in DSpace on 2021-06-15T05:59:18Z (GMT). No. of bitstreams: 1 ntu-99-R97445123-1.pdf: 2482246 bytes, checksum: 2950b452239ee0f7a5e438ca082958d8 (MD5) Previous issue date: 2010 | en |
| dc.description.tableofcontents | 口試委員會審定書
摘要 i Abstract ii 目錄 iii 圖表目錄 vi 緒論 1 一、C型肝炎病毒的歷史及現況 1 二、C型肝炎病毒的基因體構造及功能 2 三、非結構性蛋白質NS3的特性 4 四、Focal adhesion訊息傳遞 6 研究目的 11 材料與方法 12 一、藥品 12 二、酵素 14 三、抗體 14 四、細胞培養液及轉染試劑 15 五、套組試劑 15 六、染劑 15 七、其它材料 15 八、細胞株 16 十、質體的建構 18 十一、DNA 接合 (Ligation) 19 十二、細菌轉型 (Transformation) 19 十三、DNA 轉染 (DNA transfection) 20 十四、細胞全蛋白質收取 21 十五、蛋白質定量 21 十六、正十二烷硫酸鈉–聚丙醯胺板膠電泳 (SDS-polyacrylamide gel electrophoresis, SDS-PAGE) 21 十七、西方點墨法 22 十八、共同免疫沉澱法 22 十九、Glutathione S-transferase (GST)融合蛋白質的製備與GST pull-down 分析 23 二十、MTT assay 24 二十一、免疫螢光染色 (Immuno-fluorescence staining) 24 二十二、細胞黏著度測試 24 實驗結果 26 一、焦點黏著因子與NS3蛋白質的表現 26 二、焦點黏著因子與NS3蛋白質的交互作用 26 三、NS3 蛋白質次區域與integrin β1的交互作用 27 四、NS3和integrin β1蛋白質在細胞中的分布情形 28 五、NS3蛋白質對於integrin β1及細胞週期調控蛋白質表現量的影響 28 六、NS3蛋白質對於細胞貼附能力的影響 29 七、NS3及integrin β1存在之下對於細胞生長情形的影響 29 討論 30 一、焦點黏著因子的表現情形 30 二、焦點黏著因子與NS3蛋白質之間的交互作用 30 三、Integrin β1與NS3蛋白質在細胞中分布的位置 31 四、NS3蛋白質可能參與focal adhesion訊息傳遞活化機制 31 五、NS3蛋白質對於integrin β1及細胞週期相關蛋白質的調控 32 六、NS3蛋白質對於細胞貼附能力的影響 32 七、NS3與integrin β1蛋白質同時存在下對於細胞生長的影響 33 圖表 34 附圖 41 參考文獻 42 圖表目錄 圖一、焦點訊息分子zyxin、parvin、vinculin、integrin β1與NS3在293細胞的表現情形 34 圖二、焦點黏著因子與NS3蛋白質的交互作用 35 圖三、利用GST pull-down assay分析，NS3-GST融合蛋白質和integrin β1的交互作用 36 圖四、NS3和integrin β1蛋白質在Huh7細胞的分布情形 37 圖五、NS3蛋白質表現對於integrin β1及細胞週期調控蛋白質表現量的影響 38 圖六、NS3及NS3(1-402)片段對於細胞貼附能力的影響 39 圖七、NS3及integrin β1對於細胞生長情形的影響 40 | |
| dc.language.iso | zh-TW | |
| dc.subject | 整合蛋白訊息傳遞路徑 | zh_TW |
| dc.subject | C型肝炎病毒非結構性蛋白質NS3 | zh_TW |
| dc.subject | HCV NS3 | en |
| dc.subject | integrin signaling pathway | en |
| dc.title | C型肝炎病毒非結構性蛋白質NS3調節integrin訊息傳遞路徑 | zh_TW |
| dc.title | Hepatitis C virus NS3 protein regulates integrin signaling pathway | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 98-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 陳美如,董馨蓮 | |
| dc.subject.keyword | C型肝炎病毒非結構性蛋白質NS3,整合蛋白訊息傳遞路徑, | zh_TW |
| dc.subject.keyword | HCV NS3,integrin signaling pathway, | en |
| dc.relation.page | 47 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2010-08-17 | |
| dc.contributor.author-college | 醫學院 | zh_TW |
| dc.contributor.author-dept | 微生物學研究所 | zh_TW |
| 顯示於系所單位： | 微生物學科所 | |
文件中的檔案：
| 檔案 | 大小 | 格式 | |
|---|---|---|---|
| ntu-99-1.pdf 未授權公開取用 | 2.42 MB | Adobe PDF |
系統中的文件，除了特別指名其著作權條款之外，均受到著作權保護，並且保留所有的權利。