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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 張俊哲(Chun-che Chang) | |
dc.contributor.author | Nai-Wei Chiang | en |
dc.contributor.author | 姜乃瑋 | zh_TW |
dc.date.accessioned | 2021-05-16T16:18:08Z | - |
dc.date.available | 2018-08-01 | |
dc.date.available | 2021-05-16T16:18:08Z | - |
dc.date.copyright | 2013-08-27 | |
dc.date.issued | 2013 | |
dc.date.submitted | 2013-08-15 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/5896 | - |
dc.description.abstract | 黑腹果蠅 (Drosophila melanogaster) 的體軸取決於卵的不對稱性,而卵的不對稱性又源於表皮生長因子訊號 (EGF signaling) 的影響。目前已知在赤擬穀盜 (Tribolium castaneum)、寄生蜂 (Nasonia vitripennis) 和黃斑黑蟋蟀 (Gryllus bimaculatus) 的卵發育過程中發現 EGF 訊號的表現,但卵的極性究竟是如何影響體軸決定的仍是未知。在行無性生殖胎生豌豆蚜蟲 (Acyrthosiphon pisum) 的微卵管 (ovariole) 中,卵發育後緊接著是胚胎發育 (embryogenesis),這提供絕佳的機會來觀察 EGF 訊號在卵和胚胎發育中的表現。藉由分析 EGF 訊號的相關基因和 par-1 基因的表現,期望能找出這些基因是否會在卵或胚胎發育過程中有不對稱的分布情形。另一方面,因為不完全變態昆蟲的眼部發育所知甚少,用無性生殖豌豆蚜蟲來作研究,希望能了解不完全變態昆蟲眼部是如何在胚胎發育中特化的,故選殖了在其他昆蟲中數個已知調控眼部發育的工具基因 (toolkit genes) 如 twin of eyeless (toy)、eyes absent (eya) 和 eyegone (eyg),藉由原位雜合實驗去分析基因的表現,能初步了解蚜蟲在胚胎發育中的眼部特化以及體軸決定機制是否類似於其他昆蟲。研究結果顯示 EGF 訊號可能與卵母細胞分化有關,但卻無法觀察決定背腹極性的現象,而 Par-1 蛋白質在卵母細胞中的分佈,可能與微管極性有關,進一步決定前後體軸。眼部發育基因的表現位置和比較,顯示不完全變態昆蟲眼部發育的過程,與已知完全變態昆蟲眼部發育上許多共同之處。 | zh_TW |
dc.description.abstract | Body axis determination in the fruit fly Drosophila melanogaster depends on the establishment of egg asymmetry through EGF (epidermal growth factor receptor) signaling between germline and somatic cells. EGF signaling has been identified in the oocytes of beetle Tribolium castaneum, wasp Nasonia vitripennis, and cricket Gryllus bimaculatus. However, whether the formation of egg polarity can drive the determination of body axes remains unclear. Because in the asexual and viviparous pea aphid Acyrthosiphon pisum oogenesis and embryogenesis occur within the ovariole, this provides an excellent opportunity to observe how EGF signaling is patterned from oogenesis to embryogenesis consecutively. In order to understand how body axes are determined in the asexual pea aphid, we analyzed the developmental expression of genes involving EGF signaling and par-1 gene, aiming to find whether these genes are asymmetrically localized in oocytes and early embryos. In order to understand how eyes are specified in hemimetabolous insects, we selected to study twin of eyeless (toy), eyes absent (eya), and eyegone (eyg), all of which are eye toolkit genes in insects. Through the analysis of in situ hybridization results, a preliminary scheme about how eyes are specified in the asexual pea aphid has been established and whether body axis determination in other model insect is different. EGF signaling may play a role in oocyte differentitaion but not in determination of body axes in asexual pea aphid. Distribution of Par-1 protein indicates the polarity of microtubules, which determine localization of anterior-posterior determinant genes. The expression patterns and comparisons of eye specific genes in hemimetabolous insect indicate the conserved patterns of eye development with holometabolous insect. | en |
dc.description.provenance | Made available in DSpace on 2021-05-16T16:18:08Z (GMT). No. of bitstreams: 1 ntu-102-R00632004-1.pdf: 66284530 bytes, checksum: 6af451ce5c840ab539c882bd647390cd (MD5) Previous issue date: 2013 | en |
dc.description.tableofcontents | 目錄
口試委員會審定書 2 致謝 3 圖目錄 6 中文摘要 7 ABSTRACT 8 第一章 前言 10 1.1 體軸決定 10 1.1.1 模式生物的體軸決定機制 10 1.1.2 以豌豆蚜蟲 tgfα、EGFR 和 par-1 基因來研究其體軸決定機制 14 1.2 眼部特化 15 1.2.1 模式生物的眼部特化機制 15 1.2.2 以豌豆蚜蟲 toy、eya 和 eyg 基因來研究其眼部特化 18 第二章 實驗材料與方法 20 2.1 豌豆蚜蟲的飼養 20 2.2 合成探針 20 2.2.1 萃取 total RNA 和反轉錄 20 2.2.2 設計專一性引子 22 2.2.3 PCR 操作和產物回收 23 2.2.4 接合反應和轉型 24 2.2.5 養菌和質體萃取 26 2.2.7 DNA 沈澱 27 2.2.8 胞外轉錄 (in vitro transcription) 28 2.2.9 RNA 沈澱 28 2.3. 原位雜合反應 29 2.3.1 解剖豌豆蚜蟲 29 2.3.2 原位雜合反應 29 2.4 蛋白質表現 32 2.4.1 設計引子與接合反應 32 2.4.2 C41 勝任細胞處理和轉型 33 2.4.3 蛋白質小量表現和大量表現 34 2.5 蛋白質純化 36 2.5.1 可溶性測試 36 2.5.2 親和層析法 (affinity chromatography) 37 2.5.3 SDS-PAGE 膠體蛋白質電泳 39 2.5.4 抗體製備與純化 40 2.6 西方墨點法 42 2.6.1 豌豆蚜蟲的蛋白質萃取 42 2.6.2 西方墨點法 (immersion version) 42 2.7 抗體免疫染色 44 2.7.1 酵素受質染色 44 2.7.2 螢光受質染色 46 第三章 實驗結果 48 3.1 基因選殖 48 3.1.1 體軸決定基因 tgfα、EGFR 和 par-1 選殖 48 3.1.2 眼部特化基因 toy、eya 和 eyg 選殖 50 3.2 原位雜合表現結果 52 3.2.1 體軸決定基因 tgfα、EGFR 和 par-1 原位雜合結果 52 3.2.2 眼部特化基因 toy、eya 和 eyg 原位雜合結果 54 3.3 豌豆蚜蟲 Par-1 蛋白質表現與純化 56 3.4 豌豆蚜蟲 Par-1 西方墨點法之結果 57 3.5 豌豆蚜蟲 Par-1 和 dpERK 免疫染色結果 57 第四章 討論 59 4.1 各基因在豌豆蚜蟲發育過程的表現 59 4.1.1 體軸決定基因 tgfα、EGFR 和 par-1 59 4.1.2 眼部特化基因 toy、eya 和 eyg 64 4.2 豌豆蚜蟲的 RNAi 技術應用 69 參考文獻 71 附圖 89 附錄 115 口試問答和建議 116 | |
dc.language.iso | zh-TW | |
dc.title | 體軸決定基因和眼部發育基因在孤雌胎生豌豆蚜之表現分析 | zh_TW |
dc.title | Developmental Analysis of Genes Involved in Axis Determination and Eye Development in the Parthenogenetic and Viviparous Pea Aphid Acyrthosiphon pisum | en |
dc.type | Thesis | |
dc.date.schoolyear | 101-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 孫以瀚(Y. Henry Sun),蔡玉真(Yu-Chen Tsai) | |
dc.subject.keyword | 體軸決定,EGF 訊號,par-1 基因,昆蟲眼部發育,眼部特化基因, | zh_TW |
dc.subject.keyword | Body axes determination,EGF signaling,par-1 gene,Insect eye development,Eye specific genes, | en |
dc.relation.page | 123 | |
dc.rights.note | 同意授權(全球公開) | |
dc.date.accepted | 2013-08-15 | |
dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
dc.contributor.author-dept | 昆蟲學研究所 | zh_TW |
顯示於系所單位: | 昆蟲學系 |
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