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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 蕭信宏(Shin-Hong Shiao) | |
dc.contributor.author | Tsun Wang | en |
dc.contributor.author | 王尊 | zh_TW |
dc.date.accessioned | 2021-06-15T16:18:56Z | - |
dc.date.available | 2015-09-25 | |
dc.date.copyright | 2015-09-25 | |
dc.date.issued | 2015 | |
dc.date.submitted | 2015-08-17 | |
dc.identifier.citation | Anders, R.F., Adda, C.G., Foley, M., and Norton, R.S. (2010). Recombinant protein vaccines against the asexual blood stages of Plasmodium falciparum. Human vaccines 6, 39-53.
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/52570 | - |
dc.description.abstract | 瘧疾、登革熱、西尼羅等病媒傳染病,至今仍對人類造成重大的危害。這些疾病每年造成全球超過三億人感染,以及至少一百萬人死亡,而臺灣在2014年也爆發嚴重的登革熱疫情,病例數總計超過15000例,對我國經濟及國民健康影響甚鉅。而多數病媒蚊傳染病並無有效藥物及疫苗,因此控制病媒蚊數量將是可行的替代策略之一,欲達到此目標,詳細探討病媒蚊生殖調控機制是第一要務。含硫脂蛋白 (thioester-containing protein, TEP) 為人類補體 C3 的同源蛋白,過去的研究發現其參與果蠅體內對抗細菌的吞噬作用 (phagocytosis),在甘比亞瘧蚊 (Anopheles gambiae) 腸道中也扮演毒殺瘧原蟲的重要角色。此外,近期的研究也發現抑制兩個營養運輸蛋白 Lipophorin和Vitellogenin後會影響Anopheles gambiae TEP1之免疫功能,但 TEP1如何參與臺灣的重要登革病媒蚊─埃及斑蚊的生殖調控還未被深入探討。本研究利用 RNAi 的方式抑制埃及斑蚊 (Aedes aegypti) 體內TEP1的表現,發現TEP1被抑制的母蚊平均產卵量有明顯的減少,若以另一組dsRNA進行實驗結果亦然;此外,我們再利用回補實驗表現TEP1蛋白質,也發現其產卵量上升,証明此一產卵減少現象為TEP1 的專一作用。我們利用 qPCR 以及 Western blot觀察埃及斑蚊各組織中 TEP1 於吸血後的表現趨勢,結果顯示 TEP1 不論是 mRNA 或 protein 都在吸血後48-72小時的ovary 有最高表現量。有趣的是,TEP1被抑制後會影響Vitellogenin蛋白質的表現,我們發現TEP1可藉由調控Akt的磷酸化來影響Vitellogenin蛋白質的表現,另外,我們也發現TEP1的抑制會影響Vitellogenin receptor的表現。因此,綜合我們的研究結果,我們認為TEP1會藉由調控Akt來影響埃及斑蚊的產卵能力。 | zh_TW |
dc.description.abstract | Mosquito-borne diseases are the most devastating agents for human beings, such as malaria, dengue fever, West Nile fever…etc. The WHO reported that some 2.5 billion people are now at risk and more than a million people are killed by mosquito-borne diseases annually. More than 15,000 dengue cases were reported in Taiwan at 2014. Therefore, any possible avenue for developing novel control strategies against mosquito-borne diseases is urgently needed. Detailed investigation of mosquito reproduction should benefit the development of novel vector control strategies. TEP1 is a human complement-like protein. It plays a decisive role in fighting against invading pathogens in the mosquito. This well-defined macroglobulin-family protein has been reported to be participated in the regulation of mosquito reproduction. However, the mechanism of TEP1 in the regulation of mosquito reproduction remains unknown. In this study, silencing of TEP1 through RNA interference revealed a significant reduction of egg production in the mosquito Aedes aegypti. We conducted a rescue experiment to elucidate the effect of TEP1 in the egg production. RNA expression profile showed abundant mRNA expression of TEP1 in the ovary post blood meal. Interestingly, silencing of TEP1 leads to a reduction of vitellogenin production and this is independent of S6K phosphorylation. In addition, our data showed that silencing of TEP1 inhibited the phosphorylation of AKT, indicating that TEP1 may regulate vitellogenin production through a TOR signaling-independent pathway. Therefore, our results shed light on the essential role of TEP1 in the regulation of reproduction in the mosquito Aedes aegypti. | en |
dc.description.provenance | Made available in DSpace on 2021-06-15T16:18:56Z (GMT). No. of bitstreams: 1 ntu-104-R02445203-1.pdf: 2999325 bytes, checksum: 8c891ce184361c707d2c11ee7328c46b (MD5) Previous issue date: 2015 | en |
dc.description.tableofcontents | 中文摘要 i
Abstract ii 目錄 iii 圖目錄 vi 表目錄 vii 第一章 緒論 1 1.1 病媒蚊傳播疾病 1 1.1.1. 瘧疾 (Malaria) 2 1.1.2. 登革熱 (Dengue fever) 3 1.2 蚊子的先天性免疫反應 (innate immunity) 4 1.2.1 含硫酯蛋白 (Thioester-containing protein, TEP) 4 1.2.2 蚊子含硫酯蛋白 5 1.3 埃及斑蚊之生活史 6 1.4 埃及斑蚊卵黃生成機制 (vitellogenesis) 6 1.5 TOR訊息傳遞路徑介紹 (TOR signaling pathway) 7 1.6 Akt磷酸酶 (Akt kinase) 8 1.7 蚊子免疫系統與生殖調控之交互關係 8 1.8 實驗目的與研究動機 9 第二章 實驗材料與方法 10 2.1 實驗步驟與流程 10 2.1.1. 埃及斑蚊之飼養與繼代 10 2.1.2. 雙股RNA(Double-stranded RNA, dsRNA)製備 10 2.1.2.1. 質體建構 (Plasmid construction) 10 2.1.2.2. 雙股RNA合成 (dsRNA synthesis) 11 2.1.3. RNAi-mediated silencing與knockdown效率測試 12 2.1.4. RNA萃取(RNA extraction) 12 2.1.5. 反轉錄作用 (Reverse Transcription, RT) 13 2.1.6. 聚合酶連鎖反應 (Polymerase Chain Reaction, PCR) 13 2.1.7. 即時定量聚合酶連鎖反應 (Real-time Quantitative PCR, qPCR) 14 2.1.8. 產卵實驗與計數 (Egg production assay) 14 2.1.9. 埃及斑蚊TEP1抗體之製備 15 2.1.10. 西方點墨法 (Western blot analysis) 15 2.1.11. TEP1回補實驗 (TEP1 rescue assay) 16 2.1.11.1. 質體建構 (Plasmid construction) 16 2.1.11.2. 質體與dsRNA注射 16 2.1.12. 抑制劑注射 (Inhibitor injection) 17 2.2 實驗試劑之製備 17 第三章 結果 20 3.1 埃及斑蚊TEP1蛋白質區域 (protein domain) 的預測 20 3.2 演化樹 (phylogenetic tree) 分析 20 3.3 RNAi抑制TEP1對埃及斑蚊產卵之影響 20 3.4 TEP1抑制後補回對產卵數之影響 21 3.5 埃及斑蚊吸血後各組織不同時期TEP1 mRNA表現情形 22 3.6 埃及斑蚊吸血後各組織不同時期TEP1 蛋白質表現情形 22 3.7 抑制TEP1對埃及斑蚊卵巢與卵泡發育之影響 23 3.8 抑制TEP1對埃及斑蚊Vg蛋白質表現量之影響 24 3.9 抑制TEP1對埃及斑蚊TOR signaling pathway之影響 24 3.10 抑制TEP1後Vg進入卵巢中之情形 25 3.11 抑制TEP1對Akt之影響與Akt對產卵之影響 25 3.12 抑制TEP1對埃及斑蚊Vitellogenin receptor (VgR) 之影響 26 3.13 抑制Akt對埃及斑蚊VgR之影響 26 3.14 抑制硫酯鍵斷裂 (thioester bond cleavage) 對埃及斑蚊產卵之影響 27 3.15 抑制LRIM1對埃及斑蚊產卵之影響 27 第四章 討論 29 附圖 34 附表 60 附錄 65 參考文獻 68 | |
dc.language.iso | zh-TW | |
dc.title | 類補體蛋白TEP1對埃及斑蚊生殖調控之決定性影響 | zh_TW |
dc.title | Complement-like Protein TEP1 is a Determinant for Regulating Reproduction in the Mosquito Aedes aegypti | en |
dc.type | Thesis | |
dc.date.schoolyear | 103-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 陳俊宏(Chun-Hong Chen),張俊哲(Chun-che Chang) | |
dc.subject.keyword | 埃及斑蚊,含硫脂蛋白,生殖,RNA干擾,卵黃蛋白, | zh_TW |
dc.subject.keyword | Aedes aegypti,thioester-containg protein,reproduction,RNA interference,Vitellogenin, | en |
dc.relation.page | 73 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2015-08-17 | |
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 |
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