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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.author | Wen-Hai Tsai | en |
| dc.contributor.author | 蔡文海 | zh_TW |
| dc.date.accessioned | 2021-07-01T08:16:22Z | - |
| dc.date.available | 2021-07-01T08:16:22Z | - |
| dc.date.issued | 1992 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/75911 | - |
| dc.description.abstract | 近幾年來本實驗室研究老鼠肝臟的α l－Acid Glycoprotein ( AGP )基因表現之調控機制，此基因啟動子 ( promoter )的鹼基序列 －180/+10 之間，包括五個主要序列，分別為A，B，C，D及E。同時也鑑定出幾 個轉錄因數，其中之一AGP/EBP可以結合到A，C，D，E四個序列，加強AGP基因的表現，而另一方面B序列經 特異位置突變( site-directed mutagenesis )及功能性分析 ( functional assays )已確定其為一負向序 列( negative element )。 本篇論文主要就是應用傳統的生物化學方法，將肝臟細胞的核萃取液通過離子交換層析管柱及DNA親和性層 析管柱，藉以純化和B序列相結合，且具有負向調節效果的轉錄因數AGP/NTF。此因數對熱大致穩定，經 south–western blot的分析，發現至少有六個多勝?，其中分子量45一66kDa之間，具有主要結合B序列的活性。 | zh_TW |
| dc.description.abstract | α 1-acid glycoprotein (AGP) is an acute-responsive, 1iver-specific gene. Its expression is regulated by both positive and negative trans-acting factors. At least five cis-e1ements have been identified within the 180 bp region of AGP promoter. Four of these sites, A, C, D and E are recognized by AGP/EBP or AGP/EBP-like factors in liver nuclear extracts. Site-directed mutagenesis and trasfection analysis indicate that A, C, D and E regions are positive regulatory elements enhancing the expression of AGP gene while B motif is a negative regu1atory element repressing the expression of AGP gene. We have purified several proteins that bind to the B region in the AGP gene promoter. These proteins were purified from rat liver nuclear extracts by chromatography on heparin-agarose and oligonucleotide affinity columns. Analysis of purified protein fractions by southwestern blot reveals more than six polypeptides having binding activity to B motif. The purified protein from each fraction was recovered from SDS-PAGE, and those of the molecular size ranging from 45 to 66 kDa were demonstrated to have major binding activity toward B region of the AGP promoter. | en |
| dc.description.provenance | Made available in DSpace on 2021-07-01T08:16:22Z (GMT). No. of bitstreams: 0 Previous issue date: 1992 | en |
| dc.description.tableofcontents | 論文縮寫(Abbrcviations)…………………………………………………………………………Ⅰ 英文摘要(English Abstract)………………………………………………………………………Ⅱ 中文摘要(Chinese Abstract)……………………………………………………………………Ⅲ 第一章 緒言( Introduction )……………………………………………………………………1 第二章 材料與方法(Materials and Methods)……………………………………………………4 一、老鼠肝臟細胞核萃取液之製備………………………………………………………………4 二、蛋白質濃度之測定……………………………………………………………………………6 三、Heparin - Agarose chromatography……………………………………………………………7 四、Oligonucleotide 親和性管柱層析……………………………………………………………8 五、DNase I Footprinting Assay…………………………………………………………………11 六、Gel Retardation Assay…………………………………………………………………………14 七、Southwestern Blot Assay……………………………………………………………………16 八、Denaturation / Renaturation Assay……………………………………………………………18 第三章 實驗結果( Results )……………………………………………………………………20 第四章 討論(Discussion）………………………………………………………………………37 第五章 參考文獻(References)…………………………………………………………………41 | |
| dc.language.iso | zh-TW | |
| dc.title | 純化及鑑定負向調節AGP基因表現的轉錄因數 | zh_TW |
| dc.title | Purification And Characterization Of A Negative Transcription Factor For α 1-Acid Glycoprotein Gene Expression | en |
| dc.date.schoolyear | 80-2 | |
| dc.description.degree | 碩士 | |
| dc.relation.page | 52 | |
| dc.rights.note | 未授權 | |
| dc.contributor.author-dept | 生命科學院 | zh_TW |
| dc.contributor.author-dept | 生化科學研究所 | zh_TW |
| 顯示於系所單位： | 生化科學研究所 | |
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