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  1. NTU Theses and Dissertations Repository
  2. 生命科學院
  3. 動物學研究所
請用此 Handle URI 來引用此文件: http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/10631
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dc.contributor.advisor郭光雄(Guang-Hsiung Kou)
dc.contributor.authorYing-Ru Linen
dc.contributor.author林映如zh_TW
dc.date.accessioned2021-05-20T21:45:17Z-
dc.date.available2010-08-09
dc.date.available2021-05-20T21:45:17Z-
dc.date.copyright2010-08-09
dc.date.issued2010
dc.date.submitted2010-08-06
dc.identifier.citation參考文獻
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dc.identifier.urihttp://tdr.lib.ntu.edu.tw/jspui/handle/123456789/10631-
dc.description.abstract中文摘要
水溫升高至 32℃ 時蝦類因白點症病毒 (White spot syndrome virus) 感染的死亡率可大幅降低,但其機制尚未完全明瞭,惟推論可能與高溫下抑制白點症病毒的關鍵調控因子有關。按本實驗室先前的研究指出,草蝦於高溫感染白點症病毒後,醛脫氫酶(NAD-dependent aldehyde dehydrogenase;ALDH)與蛋白酶體第四 α 亞基(Proteasome alpha 4 subunit)的蛋白質表現量會明顯上升。另外,目前的研究亦發現,蝦內熱休克蛋白 70 (Heat shock protein 70;HSP70) 的 mRNA 在高溫病毒感染後其表現量亦會增高,為了探討上述蛋白質是否為高溫抑制白點症病毒的關鍵調控因子,本實驗利用RNA靜默化技術 (RNA interference, RNAi),使上述特定基因的轉錄表現量下降,並於注射雙股 RNA 48 小時後,注射感染白點症病毒。檢測病毒量之結果發現:注射 aldh 及 hsp70 雙股 RNA 的高水溫 (32℃) 感染組,其病毒量在感染 48 小時後相較於控制組有明顯上升;注射proteasome雙股RNA之高水溫感染組,其病毒量相對於控制組則無明顯變化。反轉錄聚合酶鏈鎖反應 (RT-PCR) 的分析結果也確認實驗組基因在高溫狀況下確實可被靜默化,因此推測 ALDH 與 HSP 70 可能為宿主於高溫時抑制白點病毒複製之關鍵因子,但其詳細機制仍待深入探討。
zh_TW
dc.description.abstractAbstract
Water temperature is one of the most important factors of environment. High temperature (32-33℃) has been proven to reduce the mortality in WSSV infected shrimps, but the mechanism still remains unclear. In previous researches, two-dimensional gel electrophoresis analysis revealed two proteins, NAD-dependent aldehyde dehydrogenases (ALDH) and proteasome alpha 4 subunit that were markedly up-regulated after WSSV infected shrimps at 32℃. Moreover, as referring to heat treatments, it has to be mentioned the heat shock protein family. Heat shock protein family has been considered as one of the most important indicators for heat shock treatments. We then used gene-specific (aldh, proteasome, and heat shock protein 70) dsRNA to silence these genes and found that the high temperature-induced suppression of WSSV was prevented when aldh and hsp70 were silenced. Our results therefore suggest that HSP70 and ALDH might be critical for the suppression of WSSV replication at 32℃.
en
dc.description.provenanceMade available in DSpace on 2021-05-20T21:45:17Z (GMT). No. of bitstreams: 1
ntu-99-R97b41013-1.pdf: 3872631 bytes, checksum: 38b152ad4b755ae265247780cd2dc56a (MD5)
Previous issue date: 2010
en
dc.description.tableofcontents目錄
誌謝............................... ii
中文摘要..............................iii
英文摘要..............................iv
緒論................................1
材料與方法.............................8
1. 白點症病毒液之製備 8
2. 檢體RNA之萃取 8
3. 反轉錄酶—聚合酶鏈反應(Reverse transcription PCR;RT-PCR) 9
4. 雙股核醣核酸 (Double stranded RNA, dsRNA) 之製備 10
5. RNA靜默化試驗 (RNA interference) 10
6. 養殖及實驗設計 11
7. 人工注射感染試驗與蝦體組織檢體之收取 12
8. 檢體之DNA萃取與白點症病毒檢測 12
結果.................................14
1. 溫度對熱休克蛋白質家族基因表現量的影響 14
2. RNA靜默化試驗 15
3. 特定基因靜默化於常溫 (25℃) 對白點症病毒複製之影響 15
4. 特定基因靜默化於高溫 (32℃) 對白點症病毒複製之影響 16
討論.................................17
參考文獻...............................22
圖表................................ 31
dc.language.isozh-TW
dc.subject白點症病毒zh_TW
dc.subject高溫zh_TW
dc.subjectRNA靜默化試驗zh_TW
dc.subject醛脫氫&#37238zh_TW
dc.subject熱休克蛋白質zh_TW
dc.title探討高溫(32℃)抑制白點症病毒於白蝦之複製分子機制zh_TW
dc.titleStudy of the molecular mechanisms inhibiting White Spot Syndrome Virus (WSSV) replication at high temperature (32℃) in Litopenaeus vannameien
dc.typeThesis
dc.date.schoolyear98-2
dc.description.degree碩士
dc.contributor.coadvisor羅竹芳(Chu-Fang Lo)
dc.contributor.oralexamcommittee周信佑(Hsin-Yiu Chou),王涵青(Han-Ching Wang)
dc.subject.keyword白點症病毒,高溫,RNA靜默化試驗,醛脫氫&#37238,熱休克蛋白質,zh_TW
dc.subject.keywordWSSV,high temperature,RNAi,ALDH,HSP,en
dc.relation.page39
dc.rights.note同意授權(全球公開)
dc.date.accepted2010-08-08
dc.contributor.author-college生命科學院zh_TW
dc.contributor.author-dept動物學研究所zh_TW
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