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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 張鑫(Shin C. Chang) | |
| dc.contributor.author | Chia-Yi Chou | en |
| dc.contributor.author | 周佳儀 | zh_TW |
| dc.date.accessioned | 2021-06-16T10:19:19Z | - |
| dc.date.available | 2018-09-24 | |
| dc.date.copyright | 2013-09-24 | |
| dc.date.issued | 2013 | |
| dc.date.submitted | 2013-08-16 | |
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Gerechet, H. Greif, L. Bachmatove, Y. Birk, A. Golan-Goldhirsh, M. Kunin, Y. Berdichevsky, I. Benhar and R. Tur-Kaspa (2001). 'Cell transformation induced by hepatitis C virus NS3 serine protease.' J Viral Hepat 8(2): 96-102. 寇怡衡，2007，非結構性蛋白質NS3和NS4A參與C型肝炎病毒致病之分子機構。國立臺灣大學醫學院微生物學研究所博士論文。 | |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/60480 | - |
| dc.description.abstract | C型肝炎病毒具有一個單股正向的RNA基因體，感染宿主細胞時，會利用此RNA轉譯出一多蛋白質前驅物，再利用宿主細胞及病毒自身的蛋白酶進行切割形成有功能性的結構與非結構性蛋白質。非結構性蛋白質NS4A作為NS3 蛋白酶的輔助因子，幫助NS3進行病毒多蛋白質前驅物的切割，此外，NS4A存在下，亦能幫助NS3進行內部截切，提高NS3的細胞轉型能力。在本論文中，首先在C型肝炎病毒sub-genomic replicon系統，發現C型肝炎病毒非結構性蛋白質表現時會造成DNA雙股斷裂訊號γ-H2AX的增加，以及參與DNA修復的WRN蛋白質表現量降低，顯示C型肝炎病毒非結構性蛋白質可能會增加細胞DNA受損的情形。進一步表現NS3-4A蛋白質進行分析，結果顯示NS3-4A蛋白質亦能使γ-H2AX增加，表示細胞DNA有受損的情況。分析DNA修復系統，發現當病毒蛋白質NS3-4A存在下，會使修復DNA雙股斷裂的non-homologous end-joining修復能力降低。由這些結果推測，增加細胞DNA受損的情況以及降低DNA修復系統的修復能力可能是NS3-4A蛋白質造成細胞轉型的一種機制。 | zh_TW |
| dc.description.abstract | Hepatitis C virus (HCV) possesses a positive single-stranded RNA genome. Upon infection, the RNA genome is used as a template to encode a polyprotein precursor which is then processed by host and the viral proteases to form functional proteins. Nonstructural protein (NS) 4A is a cofactor of the NS3 protease involved in the polyprotein processing. In addition, NS4A facilitates the internal NS3 cleavage that produces products with higher transforming activity than the full-length NS3. In this study, an increased level of the DNA damage marker γ-H2AX and a reduced expression level of WRN which participates in DNA repair were detected in HCV sub-genomic replicon cells. The increased DNA damage was also detected in NS3-4A expressing cells. In addition, expression of NS3-4A protein reduced the activity of non-homologous end-joining involved in the repair of DNA double-strand breaks. The results suggest that an increased DNA damage coupled with a reduced DNA repair ability could be one of the mechanisms involved in the transforming activity of the HCV NS3-4A protein. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-16T10:19:19Z (GMT). No. of bitstreams: 1 ntu-102-R00445123-1.pdf: 1596629 bytes, checksum: a67c2ba5234e19f89a3d95b172ab75a5 (MD5) Previous issue date: 2013 | en |
| dc.description.tableofcontents | 摘要 i
Abstract ii 目錄 iii 圖表目錄 v 緒論 1 一、C型肝炎病毒的歷史 1 二、C型肝炎病毒相關的肝病與治療 1 三、C型肝炎病毒的基因體結構及功能 2 (一) 結構性蛋白質 2 (二) 非結構性蛋白質 3 四、C型肝炎病毒非結構性蛋白質NS3的特性 4 (一) NS3的內部截切 5 (二) NS3對於細胞轉型的作用 5 (三) NS3對於細胞週期及細胞凋亡機制的影響 6 五、C型肝炎病毒非結構性蛋白質NS4A的特性 6 六、C型肝炎病毒與肝細胞癌 (hepatocellular carcinoma) 7 (一) C型肝炎病毒蛋白質與DNA損傷 7 (二) C型肝炎病毒蛋白質與DNA修復 7 七、Werner syndrome protein (WRN) 8 八、研究目的 9 材料與方法 10 一、藥品 10 三、抗體 11 四、細胞培養液及轉染試劑 11 六、其他 12 七、細胞株 12 八、實驗室提供的質體 14 九、DNA轉染 (DNA transfection) 15 十、流氏細胞儀分析 (Flow cytometry analysis) 16 十一、收取細胞蛋白質 16 十二、蛋白質定量 17 十三、正十二烷硫酸鈉-聚丙烯醯胺板電泳 (SDS-polyacrylamide gel electrophoresis， SDS-PAGE) 17 十四、西方墨點法 (Western blot analysis) 18 十五、Cell fractionation 19 十六、Non-homologous end-joining (NHEJ) repair 分析 20 實驗結果 21 一、HCV非結構性蛋白質對細胞g-H2AX及WRN表現量的影響 21 二、NS3-4A蛋白質在細胞中的分布 21 三、NS3-4A蛋白質對細胞g-H2AX及WRN表現量的影響 22 四、IR刺激下，NS3-4A蛋白質對DNA損傷及修復的影響 22 五、NS3-4A蛋白質對DNA修復系統的影響 24 討論 26 一、C型肝炎病毒蛋白質對於DNA損傷的影響 26 (一) 蛋白酶活性是否會影響NO及ROS的產生 26 (二) NS3或NS3-4A是否透過蛋白酶活性影響DNA修復系統中的蛋白質 26 二、在外力IR造成DNA損傷的情況下，病毒蛋白質NS3-4A對於DNA修復的影響 27 三、NS3、NS3-4A、NS3pd-4A對於NHEJ的影響 27 圖表 29 參考文獻 36 圖表目錄 圖一、HCV非結構性蛋白質對細胞內g-H2AX及WRN表現量的影響 29 圖二、NS3-4A蛋白質在細胞中的分布 30 圖三、NS3-4A蛋白質對細胞內g-H2AX及WRN表現量的影響 31 圖四、IR刺激下，NS3-4A蛋白質對DNA損傷及修復的影響 32 圖五、NS3-4A蛋白質對受到IR照射的S phase細胞在DNA損傷上的影響 34 圖六、NS3-4A蛋白質對DNA修復系統的影響 35 | |
| dc.language.iso | zh-TW | |
| dc.subject | NS3-4A蛋白質 | zh_TW |
| dc.subject | C型肝炎病毒 | zh_TW |
| dc.subject | DNA修復系統 | zh_TW |
| dc.subject | HCV | en |
| dc.subject | DNA repair | en |
| dc.subject | NS3-4A protein | en |
| dc.title | C型肝炎病毒NS3-4A蛋白質對DNA損傷及修復的影響 | zh_TW |
| dc.title | The Effects of Hepatitis C Virus NS3-4A Protein on DNA Damage and Repair | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 101-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 陳美如(Mei-Ru Chen),許金玉(Jin-Yuh Shew),詹世鵬(Shih-Peng Chan) | |
| dc.subject.keyword | C型肝炎病毒,NS3-4A蛋白質,DNA修復系統, | zh_TW |
| dc.subject.keyword | HCV,NS3-4A protein,DNA repair, | en |
| dc.relation.page | 43 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2013-08-16 | |
| 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-102-1.pdf 未授權公開取用 | 1.56 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。