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研究生:張曉瑩
研究生(外文):Shiao-Ying Chang
論文名稱:核醣毒蛋白α-sarcin及重組之核酸脢RNaseT-tat進入細胞之機制
論文名稱(外文):The cell entry of ribotoxin α-sarcin and engineering RNase T-tat
指導教授:林茂榮 老師
指導教授(外文):Alan Lin
學位類別:碩士
校院名稱:國立陽明大學
系所名稱:遺傳學研究所
學門:生命科學學門
學類:生物訊息學類
論文種類:學術論文
論文出版年:2001
畢業學年度:89
語文別:中文
論文頁數:47
中文關鍵詞:蛋白藥物遞送細胞通透愛滋病毒
外文關鍵詞:α-sarcinribotoxinT-tatHIVprotein drug deliverycell entry
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本實驗的主旨在於研究專一性對抗病毒的蛋白藥物其進入細胞之機制,其中包含從真菌類純化出來的蛋白質毒素α-sarcin,及由本實驗室構築的重組核醣核酸脢T-tat。
α-sarcin為一核醣毒蛋白,因其具有專一性水解真核細胞中28S rRNA的特性,使得蛋白質無法合成,具致死細胞之能力。α-sarcin在正常狀況下無法進入細胞,但在細胞受到病毒感染之狀況下可進入細胞,並執行水解細胞28S rRNA的作用。因此希望藉由此研究結果,發展專一性對抗受病毒感染的細胞之蛋白藥物。我們推測α-sarcin進入細胞的動作與序列有關,因此使用多個變異株試驗其是否也能進到細胞中,希望能從中找到一些序列與進入細胞之關連。
T-tat是由核醣核酸脢T1改造而來,經in vitro測試,證實它對愛滋病毒HIV 的TAR RNA具專一性辨認及水解能力,並且T-tat具進入細胞之能力。此論文試驗T-tat是否對細胞具有毒性,及T-tat在細胞中是否有辨認HIV之TAR RNA之能力。結果發現在低劑量、短時程內T-tat對細胞並無毒性,並且在細胞內具有專一性辨認及水解TAR RNA能力。此實驗結果將增進T-tat在將來用作治療愛滋病的潛力。

In this study, we test how α-sarcin enters into cell. α-sarcin is purified from the fungus Aspergillus giganteus. It cleaves 28S rRNA of eukaryotic cells at a specific site. It is a useful tool to investigate ribosome complex. Whetherα-sarcin makes the cell entry is the object of this study. Recently, reports have pointed out that cell suffers with viral infection, becoming α-sarcin transportable. In this study, we have confirmed such phenomenon. We also assume that the phenomenon is sequence dependent. By using a series of deletion mutants, we attempt to find out the exact sequence that governs the cell entry.
In this study we also examine the cell entry of RNase T-tat. The T-tat is an engineering protein, which specifically binds to the TAR RNA of HIV. Because of the ribonuclease activity of T-tat, the protein is potential to be useful as an anti-HIV drug. In this thesis, we have confirmed that T-tat is capable of entering cells. The toxicity of T-tat to mammalian cells and its biological activity in cells is also measured.

英 文 摘 要‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧ 1
中 文 摘 要‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧ 2
緒 論‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧ 3
材 料 及 方 法‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧10
結 果‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧20
討 論‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧24
圖 與 附 錄‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧28
參 考 文 獻‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧44

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謝孟迪 設計專一性之核醣核酸脢具辨認及水解HIV-Ⅰ特定之核酸序列 國立陽明大學遺傳學研究所碩士論文 民國八十八年七月

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