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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 韓玉山(Yu-San Han) | |
| dc.contributor.author | Ping-Fu Liu | en |
| dc.contributor.author | 劉秉賦 | zh_TW |
| dc.date.accessioned | 2021-05-20T21:29:01Z | - |
| dc.date.available | 2012-08-20 | |
| dc.date.available | 2021-05-20T21:29:01Z | - |
| dc.date.copyright | 2010-08-20 | |
| dc.date.issued | 2010 | |
| dc.date.submitted | 2010-08-18 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/10431 | - |
| dc.description.abstract | 日本鰻為一降海洄游性魚類，擁有複雜的生活史。在亞洲，日本鰻是重要的經濟養殖魚種之一，然而其養殖產業所需的魚苗(玻璃鰻)，卻只能依靠捕撈天然的玻璃鰻供應之。但氣候變遷、環境汙染與日漸嚴重之過漁的影響，日本鰻苗的自然資源量逐漸稀少，導致養鰻產業的發展受到阻礙，而日本鰻人工繁殖成為解決此瓶頸的重要關鍵。由於現存之鰻魚人工繁殖技術無法成功應用在產業上，我們需要更進一步了解其生殖調控機制，以找出日本鰻自然性成熟所需的條件。鰻魚與其他硬骨魚的生殖週期同樣是由下視丘¬¬-腦下垂體-生殖腺所組成的神經內分泌軸線所控制，能夠整合外部環境之刺激以誘發青春期的活化，而促性腺激素釋放素 (GnRH) 即為此軸線之關鍵調控者。因此本研究針對 GnRH 基因進行啟動子位置的分析，將其基因轉錄起始位 (start condon, +1) 上游可能為啟動子的區域，以 PCR 增幅放大後，構築到pGL3-basic載體中，再轉殖進GT1-7細胞株中，以冷光強度分析這些區域轉錄表現情況，確定GnRH基因啟動子的關鍵區域以及其轉錄調控情形。研究結果顯示，GnRH基因轉錄起始位上游 -388 至 -499 為此啟動子重要的區域；當移除此位置，啟動子之活性會受到明顯的抑制。針對此啟動子區域進行轉錄因子分析，在 -488 與 -432 位置上分別發現有 Oct-1 和 Pbx-1a 轉錄因子的結合位，且此兩種轉錄因子已被證實與GnRH轉錄之啟動有極大的關係，顯示此區域 ( -388 ~ -499 ) 為具有調控GnRH基因表現的關鍵位置。而在外源性荷爾蒙刺激GnRH表現的實驗中，發現雄性素與雌性素的刺激，同樣會受到啟動子中此位置 ( -388 ~ -499 ) 的調控，此外不論是雄性素或雌性素，在濃度的變化下對GnRH表現均無顯著作用，但若縮短處理的時間，則會失去促進GnRH轉錄的能力，表示性類固醇在GnRH轉錄的作用裡，可能以間接的方式調控其基因表現。 | zh_TW |
| dc.description.abstract | Anguilla japonica is one of important species in Asia aquaculture. However, glass eels production is continuously declining, resulting in great impact on the development of eel aquaculture industry. In order to reach a sustainable eel production, we should develop complete artificial propagation. For this reason we need to know the mechanisms of eel nature maturation. Like other teleost, the center of eel reproduction endocrine control system is hypothalamus-pituitary-gonad (HPG) axis, and GnRH is the major regulator in HPG axis. For this reason, this study focuses on promoter analysis of eel GnRH gene. In the promoter assay, the upstream sequences before GnRH start coding will be available from 5’-Genome Walking. Difference of GnRH promoter area is zonated by specific primers and PCR. After that, PCR product is constructed into pGL3-basic vector then transfect in GT1-7 cell line. Promoter region of GnRH is determinated by assaying the firefly luciferase activity. The results indicate that the critical region for GnRH transcription locates in -388 to -499 that contain the important transcription factors Oct-1 and Pbx-1a for GnRH expression. Moreover, sex-steroids maybe indirectly activate GnRH promoter via this region from -388 to -499. | en |
| dc.description.provenance | Made available in DSpace on 2021-05-20T21:29:01Z (GMT). No. of bitstreams: 1 ntu-99-R97b45016-1.pdf: 1944802 bytes, checksum: 4e4a586db5136eff746dd2d7aed0d728 (MD5) Previous issue date: 2010 | en |
| dc.description.tableofcontents | Abstract----------------------------------------i
Introduction------------------------------------1 Materials and methods---------------------------9 Biological Materials----------------------------9 Reaction reagents and Mediums-------------------9 Experimental methods---------------------------11 Results----------------------------------------21 GnRH Genome Walking in Japanese eel------------21 Basal GnRH promoter activity assay-------------21 Effects of Sex-steroids on GnRH expression-----23 Discussion-------------------------------------26 GnRH Genome Walking in Japanese eel------------26 Basal GnRH promoter activity assay-------------26 Effects of Sex-steroids on GnRH expression-----29 References-------------------------------------33 Tables-----------------------------------------39 Figures----------------------------------------44 | |
| dc.language.iso | en | |
| dc.title | 日本鰻之促性腺激素釋放素啟動子分析 | zh_TW |
| dc.title | Gonadotropin-releasing hormone (GnRH) promoter analysis of Anguilla japonica | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 98-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 鄭達智(Ta-Chih Cheng),蕭仁傑(Jen-Chieh Shiao) | |
| dc.subject.keyword | 促性腺激素釋放素,啟動子,轉錄因子,性類固醇刺激, | zh_TW |
| dc.subject.keyword | Gonadotropin-releasing hormone,Promoter,Transcription factors,Stimulation of sex-steroids, | en |
| dc.relation.page | 62 | |
| dc.rights.note | 同意授權(全球公開) | |
| dc.date.accepted | 2010-08-19 | |
| dc.contributor.author-college | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 漁業科學研究所 | zh_TW |
| 顯示於系所單位： | 漁業科學研究所 | |
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