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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 郭光雄,羅竹芳 | |
| dc.contributor.author | Yu-Chen Kuo | en |
| dc.contributor.author | 郭昱辰 | zh_TW |
| dc.date.accessioned | 2021-06-13T08:03:56Z | - |
| dc.date.available | 2006-07-26 | |
| dc.date.copyright | 2005-07-26 | |
| dc.date.issued | 2005 | |
| dc.date.submitted | 2005-07-21 | |
| dc.identifier.citation | 參考文獻
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/36519 | - |
| dc.description.abstract | 本研究由實驗室建構之草蝦5’EST資料庫和5’RACE選殖出草蝦IAP基因完整序列，稱為草蝦inhibitor of apoptosis protein (IAP)基因，草蝦IAP包含三個桿狀病毒IAP重複區域(BIR)以及一個RING區域，且進行序列上的分析發現草蝦IAP蛋白之BIR1、BIR2、BIR3和RING finger區域與人類cIAP1、cIAP2、XIAP及果蠅DIAP2相似。此基因cDNA全長為4769 bps ，轉錄區域為2097bps ，轉譯長度為698個氨基酸，分子量為76.9 kDa。以RTPCR得知IAP基因在類淋巴器官相對於其他器官如：肝胰腺、胃、血球、鰓、神經組織、心臟和肌肉組織有最高表現量，在感染白點症病毒後也發現在類淋巴器官與胃器官表現量並未隨感染時序而改變，進一步將草蝦IAP基因暫時性轉染進Sf-9昆蟲細胞株，經由免疫螢光染色得知此基因產物主要是位於細胞質。對於草蝦IAP拮抗細胞凋亡測試，以轉染果蠅Reaper使細胞產生細胞凋亡，發現共同轉染草蝦IAP與果蠅Reaper時，草蝦IAP具有抑制細胞凋亡之能力。 | zh_TW |
| dc.description.abstract | We cloned a novel inhibitor of apoptosis protein (IAP) family member, PmIAP, from Penaeus Monodon. PmIAP contains three baculoviral IAP repeat (BIR) domains followed by a RING finger domain. The BIR and RING finger domains of PmIAP share striking amino acid sequence similarity with those of cIAP1, cIAP2, XIAP, and DIAP2, suggesting evolutionary conservation. Pm-IAP was 4.7 kb long, including a coding region of 2097 bp preceded by 633 bp (5' untranslated region) and followed by 2039 bp (3' untranslated region). Pm-IAP encoded a protein (Pm-IAP) of 698 amino acids with a predicted molecular mass of 76.9 kDa. RT-PCR analysis revealed that the steady-state level of PmIAP mRNA is highest in lymphoid organ among examined tissues, including hepatopancrease, stomach, hemocyte, gill, nerve, heart and muscle. Furthermore, the PmIAP RNA level in LO cells was not changed significantly after WSSV infection, although LO cells undergo apoptosis upon viral infection. The immunofluorescence staining of Spodoptera frugiperda Sf-9 cells transiently expressing PmIAP revealed that the PmIAP was mainly localized at cytoplasm of Sf-9 cells. To demonstrate the anti-apoptosis activity of Pm-IAP, we cotransfected the Drosophila reaper and Pm-IAP gene into Sf-9 cells. The result showed that Pm-IAP can block apoptosis in Sf-9 cells induced by Drosophila Reaper. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-13T08:03:56Z (GMT). No. of bitstreams: 1 ntu-94-R92b41021-1.pdf: 1355801 bytes, checksum: 9a82c77ea9d1e6bfcf4b1a22675ef4c0 (MD5) Previous issue date: 2005 | en |
| dc.description.tableofcontents | 前言................................................1
材料與方法......................................................................................................12 1. 草蝦 Inhibitor of Apoptosis Protein (IAP)基因之篩選..................................12 2. 人工感染試驗..................................................................................................12 3. 檢體RNA收取................................................................................................13 4. 轉型作用 (Transformation).............................................................................13 5. 草蝦IAP基因之構築......................................................................................14 6. AcMNPV-IE1基因、AcMNPV-P35基因之構築..........................................14 7. 反轉錄酶-聚合酶鏈反應.................................................................................15 8. 淋巴器官與胃器官經WSSV病毒感染後之IAP表現量.............................17 9. 抗草蝦IAP重組蛋白抗體之製備..................................................................17 10. 轉染作用 (Transfection)..................................................................................20 11. 免疫螢光分析法(Immunofluorescent assay)...................................................20 12. 西方轉印法 (Western blot)..............................................................................21 13. 功能分析...........................................................................................................22 結果...........................................................................................................25 草蝦IAP基因核苷酸序列與胺基酸序列分析....................................................25 草蝦IAP同源性分析...........................................................................................26 草蝦IAP基因轉錄特性分析與白點症病毒感染之影響...................................26 草蝦IAP基因抗體的生產...................................................................................27 草蝦IAP於Sf-9細胞之表現定位......................................................................28 草蝦IAP基因之功能分析...................................................................................28 討論..............................................................................................31 結論..............................................................................................37 參考文獻........................................................................ ...........................................38 圖表及附錄................................................................................................................51 | |
| dc.language.iso | zh-TW | |
| dc.subject | 草蝦 | zh_TW |
| dc.subject | 細胞凋亡 | zh_TW |
| dc.subject | Penaeus monodon | en |
| dc.subject | Inhibitor of apoptosis protein(IAP) | en |
| dc.title | 草蝦細胞凋亡抑制蛋白基因之表現及其功能分析 | zh_TW |
| dc.title | Expression and Functional analysis of the Inhibitor of
Apoptosis Protein of Penaeus monodon | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 93-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 廖一久,王重雄,陳歷歷 | |
| dc.subject.keyword | 草蝦,細胞凋亡, | zh_TW |
| dc.subject.keyword | Penaeus monodon,Inhibitor of apoptosis protein(IAP), | en |
| dc.relation.page | 64 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2005-07-21 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 動物學研究研究所 | zh_TW |
| 顯示於系所單位： | 動物學研究所 | |
文件中的檔案：
| 檔案 | 大小 | 格式 | |
|---|---|---|---|
| ntu-94-1.pdf 未授權公開取用 | 1.32 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。