鄭登貴2006-07-262018-06-292006-07-262018-06-292003http://ntur.lib.ntu.edu.tw//handle/246246/16456近年來，已有多種可於生物體外具備可誘導性質之基因表現系統陸續發展而出，以進一步突破傳統產製轉基因動物之諸多限制，尤有更甚者，係成功應用於生物體內之四環黴素調控系統表現系統。我們嚐試使用四環黴素來控制rtTA或tTA 的表現;而PDGF-C 則結合在rtTA 或tTA 相對應的啟動子之後，故可用四環黴素去控制PDGF-C 基因之表現 (tet operator sequence; tet O)，以條件化控制目標蛋白質 (PDGF-C);另為能夠進一步讓四環黴素辨識及調節轉C 型血小板生長因子基因轉殖鼠之表現與否和量上之表現情形，我們使內源性基因(PDGF-C)受特定活化啟動子(promoter)控制，此活化啟動子可接受四環黴素控制支配的轉錄活化子(transactivator) tTA 及rtTA 所調控，故理論上同時具備此二種共同構築的轉基因鼠，可使用四環黴素來調控目標基因的生理作用;且亦有研究指出四環黴素衍生物-Doxycycline，此化合物對於上述雙基因轉基因鼠具更高敏感性之控制能力。針對於此，吾等已成功構築四種表現載體(Expression vector)及二種反應載體(Response vector)，前者攜帶有特定組織專一性表現之啟動子諸如:四環黴素反應啟動子(Tet-responsive promoter)或者是4311 基因上之特定胎盤組織調控區域( trophoblast-specific promoter)，後者則攜帶有報導基因(綠螢光蛋白質)及有興趣之轉殖基因(PDGF-C)，我們樂觀地預期上述數種構築之轉殖基因所產製的轉基因小鼠，更能提供吾等針對PDGF-C基因於活體動物內更具真實性的biologicalfunction 之資料收集及探索，提供日後進行有關該基因之生理學功能及各項應用之可能性等探討時的依據。Several inducible gene expression systems have been developed in vitro in recent years to overcome limitations with traditional transgenic mice. One of these, the tetracycline-regulated system, has been used successfully in vivo. The expression of the targeted protein are controlled by a promoter containing seven DNA operator sequences from the Escherichia coli transposon 10 ( Tn10). To control the expression of PDGF-C in transgenic (Tg) mice, we used a tetracycline-controlled transactivator (tTA or rtTA) and a tTA/rtTA-dependent promoter linked to a PDGF-C gene. Doxycycline (Dox)-sensitive co-regulation of two transcriptionally coupled transgenes will be investigated in the mouse. For this, we generated four independent mouse line clone carrying coding regions for mouse PDGF-C gene and green fluorescent protein (GFP) in a bicistronic, bidirectional module under the control of the Tet-responsive promoter and/or 4311 regulatory regions of trophoblast- specific promoter. It is anticipated that the rtTA andor tTA-sensitive bi-directional expression module is well suited to express genes of interest in a regulated manner and that GFP can be used to track transcriptional activity of the module in the living mouse.application/pdf554656 bytesapplication/pdfzh-TW國立臺灣大學動物科學技術學系暨研究所C 型血小板生長因子四環黴素調控系統綠螢光蛋白質轉基因小鼠PDGF-CTetracycline-regulated systemgreen fluorescent proteintransgenic micereporthttp://ntur.lib.ntu.edu.tw/bitstream/246246/16456/1/912313B002384.pdf