臺灣博碩士論文加值系統

雙剪接核醣核酸(double-spliced RNA)是B型肝炎病毒(HBV)在自然狀態下的產物之一(11)，過去的研究中證明，此雙剪接核醣核酸會抑制B型肝炎病毒基因的表現，而且此抑制的作用點是在轉錄的起始步驟(transcription initiation)(33)。在本論文中，我們嘗試瞭解雙剪接核醣核酸抑制基因表現的機制，實驗的結果如下：首先，pCMV/DS/3081轉錄產物的5’端有一段162鹼基對(base pair)的載體序列(vector sequence)，對於抑制作用而言此序列並非是必需的。再者，五個截短選殖株(deletion clone)用來探求雙剪接核醣核酸分子對抑制作用的決定區域，實驗結果指出雙剪接核醣核酸的5’端對於抑制現象是重要的。蟲螢光酶基因(luciferase gene)分別由TATA區域(TATA box)或是含完整上游活化序列(up-stream activating sequence)的巨細胞病毒起動子(CMV promoter)所控制，我們觀察到雙剪接核醣核酸對此兩個不同長度起動子有相似的抑制比例，推論雙剪接核醣核酸是藉由影響與TATA區域作用的共同轉錄因子(general transcription factor)達成抑制的作用。關於B型肝癌雙剪接核醣核酸的作用機制與生物意義需要更進一步的研究來探討。

The double-spliced RNA (dsp-RNA) is a naturally produced RNA by HBV (10). From previous studies, it has been demonstrated that the double-spliced RNA, pCMV/DS/3081, played an inhibitory role in the expression of HBV genes and this inhibition was regulated at the level of the transcription initiation (29). In this thesis, we would like to study further on the inhibitory role of the double-spliced RNA in HBV. The results are as follows: First, it is demonstrated that the 5’ end of transcript from pCMV/DS/3081, in which 162 nucleotides was transcribed from the putative transcription start site of CMV promoter, is not essential for the inhibitory effect. Also the inhibitory effect of the dsp-RNA could be observed by 24 hours after transfection. Second, five deletion clones were used to map the important regions for the inhibition. The data indicate that the 5’ region of the double-spliced RNA is crucial to the inhibitory activity. Finally, luciferase gene that was controlled either by single transcription element (TATA box) or complex up-regulatory cis-elements (authentic cytomealovirus immediate early promoter). We found that the dsp-RNA exhibited similar inhibitory activity in both systems. This results suggest that dsp-RNA inhibits the gene expression through its interaction with common trancriptional factors of TATA box.

中文摘要-------------------------------------------------1
英文摘要-------------------------------------------------2
壹. 緒論-------------------------------------------------3
貳. 材料與方法-------------------------------------------8
參. 結果------------------------------------------------16
肆. 討論------------------------------------------------20
伍. 附圖------------------------------------------------24
陸. 附表------------------------------------------------45
柒. 參考文獻--------------------------------------------46

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