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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.author | Hsin-Wei Wang | en |
| dc.contributor.author | 汪心薇 | zh_TW |
| dc.date.accessioned | 2021-07-01T08:21:05Z | - |
| dc.date.available | 2021-07-01T08:21:05Z | - |
| dc.date.issued | 1998 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/76397 | - |
| dc.description.abstract | FTZ-Fl是首先在果蠅中發現，能夠活化homeodomain蛋白質Fushitarazu (Ftz)表現的一個轉錄因數，因此被稱為Fushi-tarazu Factor l。許多生物如斑馬魚體內的同系物(zFF1)已經被發現，由於FTZ-Fl是核受體蛋白質中的一員，而這類的蛋白質通常在生物體內有著重要的功能，例如老鼠SF1(steroidogenic factor l)就是在哺乳動物體內對腎上腺和性腺發育有重要的影響；而大鼠FTF (fetoprotein transcrption factor)則對肝等內臟形成有影響。因此為瞭解zFF1在斑馬魚胚胎中所扮演的角色，我們想要研究zFF1的表現型態及調控機制。從RT-PCR的結果，我們發現zFF1在成魚許多的組織中都有表現，顯示其在斑馬魚體內有重要的功能。另外。目前已知zFF1有兩個分開利用的起始外顯子，即外顯子l和外顯子2，且各自接到外顯子3。在胚胎發育過程中，zFF1在早期胚層發育的時候表現較少，真正大量產生則是在Segmentation即體節發育的時期之後。在表現位置上，zFF1在胚胎的許多地方都有表現，例如頭部腦下垂體、背部及頭部神經脊細胞和緊靠卵黃囊的內胚層部位及肝胰臟，其中zFF1A的表現位置和含量都大於zFF1B。另外，在成魚的腦部靠近下視丘的部位有zFF1的表現。在細胞株中的實驗證明，zFF1的蛋白質可以辨認mSF1的binding site，並可活化報導基因的表現。對zFF1外顯子2啟動子所做的分析發現，有一可能的調控區域位在-79bp到-126bp之間。從這些研究結果中我們發現，zFF1兩個外顯子的表現型態並無太大差異，但 zFF1A在胚胎發育過程中較zFF1B多。zFF1的蛋白質具有轉錄活化的功能。而zFF1在胚胎發育過程中的多重表現也顯示其對斑馬魚胚胎某些重要組織的發育有重要的功能，但與FTF的作用較相似。 | zh_TW |
| dc.description.abstract | EXPRESSION PATTERN AND FUNCTIONAL ANALYSIS OF ZEBRAFISH FTZ-F1 GENE
Zebrafish FTZ-F1 (zFF1) is a homolog of Drosophila FTZ-Fl that activates the homeodomain protein Fushi-tarazu and belongs to the nuclear hormone receptor superfamily. This superfamily includes many important members such as steroidogenic factor l(SF1), a mammalian homolog of FTZ-Fl, which is a key regulator in the synthesis of steroidogenic enzymes and the development of adrenal and gonads ; and fetoprotein-transcription factor (FTF). To find out the role of zFF1 in the development of zebrafish, we examined the expression pattern and function of zFF1. zFF1 mRNA is widely distributed in many tissues, as shown in our RT-PCR results, indicating that zFF1 has essential function. zFF1 has two leader exons, 1 and 2 , which are differentially utilized and are spliced into exon 3. During the process of development, zFF1 starts to be expressed from the segmentation stage when somites start to form. The in situ hybridization shows zFF1 can be expressed in the hypothalamus of adult brain and many different areas of the embryo, including pituitary, neural crest cell and endoderm in the early segmentation stage and mandibular arch, liver and pancreas in the late stage. Transfection studies in Ratl cells showed that zFF1 can recognize the consensus sequence of SF1 binding site and transactivate the expression of the reporter gene. Deletion analysis of the zFF1 promoter 2 in Ratl cells shows that a potent regulatory element may exist between 79bp and 126bp upstream from the transcription start site. Our studies indicate that the utilization of zFF1 exon 1 and 2 is very similar and zFF1 protein has the transactivation ability. Also, the expression of zFF1 in the zebrafish embryos implies that it may have important function in the development, and is probably more related to FTF. | en |
| dc.description.provenance | Made available in DSpace on 2021-07-01T08:21:05Z (GMT). No. of bitstreams: 0 Previous issue date: 1998 | en |
| dc.description.tableofcontents | 中文摘要 英文摘要 前言 1 一、新興的模式生物(Model Organism)-斑馬魚(Zebrafish) 1 二、斑馬魚的胚胎發育過程 1 三、核受體家族(nuclear receptor superfamily) 2 四、Ftz-F1基因 4 五、其他生物體內的FTZ-Fl基因 5 六、斑馬魚FTZ-Fl(zFF1)的基因結構 6 七、zFF1的功能 8 八、Steroidogenic factor l (SF1)的功能 8 九、P450scc酵素 10 十、pax-b基因在斑馬魚胚胎中的表現 11 材料與方法 12 一、反轉錄聚合?連鎖反應(RT-PCR) 12 1.斑馬魚Total RNA的萃取 12 2.寡核糖酸的合成 12 3.南氏墨染法 13 二、暫時轉染法 13 1.細胞培養(cell culture) 15 2.質體之構築 15 A. zFFl蛋白質功能的測試：zFF1 cDNA表現質體 15 B. zFF1基因與mSF1基因5'端啟動子接報導基因質體 16 C. zFF1基因5'端調節區連續刪除質體 16 3.β-Gal、Luciferase及CAT assay 18 三、原位雜交法(In situ hybridization) 20 1.探針(probe)的製備 20 A.質體線性化 20 B.聚合脢連鎖反應片段 21 2.斑馬魚組織及胚胎的準備 22 3.雜交反應 22 4.Antibody及呈色反應 23 5.Sample之保存及照相 23 四、雙原位雜交反應(Double in situ hybridization) 24 五、顯微注射(microinjection) 25 1.斑馬魚卵的收集與注射 25 2.報導基因表現之分析 25 結果 27 一、zFF1在成魚的許多組織中均有表現 27 二、zFF1從Segmentation時期開始大量表現 27 三、zFF1在胚胎發育過程中的表現位置 28 A. zFF1外顯子2全長探針 29 B.刪除掉DNA結合區域的zFF1 30 C.刪除DNA結合區探針與pax-b做雙原位雜交法 31 D. zFF1A專一性探針 31 E. zFF1B專一性探針 32 四、zFF1蛋白質具有轉錄活化(transactivation)的功能 32 五、zFF1基因5'端在細胞株中的分析 33 六、mSF1基因5'端部份接LacZ在基因轉殖班馬魚中的表現 34 討論 36 一、zFF1的生理意義 36 二、zFF1外顯子1與外顯子2的比較 37 三、zFF1A和zFF1B的比較 38 四、全長與刪除DNA結合區域的探針表現型態的比較 39 五、zFF1A和zFFIB探針的比較 40 六、zFF1的功能探討 41 七、zFF1與mSF1的關聯及在生理上的意義 42 八、zFF1外顯子2啟動子分析 43 九、基因轉殖斑馬魚(transgenic zebrafish)的研究 43 參考文獻 45 表格 52 圖片 54 | |
| dc.language.iso | zh-TW | |
| dc.title | 斑馬魚 FTZ-Fl 基因的表現型態及功能分析 | zh_TW |
| dc.title | Expression Pattern and Functional Analysis of Zebrafish FTZ-F1 gene | en |
| dc.date.schoolyear | 86-2 | |
| dc.description.degree | 碩士 | |
| dc.relation.page | 83 | |
| dc.rights.note | 未授權 | |
| dc.contributor.author-dept | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 動物學研究所 | zh_TW |
| 顯示於系所單位： | 動物學研究所 | |
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