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頁籤選單縮合
題 名 | 動物細胞基因轉染方法之比較=A Comparison of Gene Transfection Methods for Mammalian Cells |
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作 者 | 倪志文; 謝學真; | 書刊名 | 界面科學會誌 |
卷 期 | 21:3 1998.09[民87.09] |
頁 次 | 頁121-130 |
分類號 | 362.6 |
關鍵詞 | 基因轉染; 微脂體轉染法; 中國大頰鼠卵巢細胞; Gene transfection; Lipofection; Chinese hamster ovary cells; CHO; |
語 文 | 中文(Chinese) |
中文摘要 | 基因轉染(gene transfection)技術廣泛應用於研究基因調控機制、改質細胞以生產高附加價值的醫藥與複雜的生物分子、以及臨床上的基因治療(gene therapy)等。此項技術係將經過重組後的基因送入細胞之中,但是由於細胞膜表面與DNA分子皆帶有大量的負電荷,使得DNA分子無法有效穿透細胞膜而進入細胞中。因此研究上若採用非病毒的基因轉染方法時,必須在轉染的過程中加入界面活性分子以改質DNA分子的界面特性,達成轉染的目的。目前常採用的技術包括有:磷酸鈣沉澱法、微脂體轉染法、DEAE-Dextran轉染法等等。本研究採用幾種常用的轉染技術對中國大頰鼠卵巢細胞(CHO-K1)進行基因轉染,比較各轉染方式的效率,以作為日後研究的參考。實驗的結果發現,微脂體轉染法的轉染效率約為25%~30%,優於磷酸鈣沉澱法(<1%以及DEAE-Dextran轉染法(∼2%),也優於電擊穿孔法(∼3%)。 |
英文摘要 | Transfection is the technique of introducing a foreign DNA molecule into a eucaryotic cell. This technique has been widely utilized to study the function and regulation of gene/protein, as well as to produce recombinant products in mammalian cells. Moreover, it plays an important role in gene therapy. However, the negative charges on the surface of cell membrane form a barrier for DNA molecule to across cell membrane. Therefore, the use of surfactants to alter the properties of interface between cell membrane and DNA molecule is a way leading to effective transfections. This study chose several methods including surfactant-mediated transfection to transfect CHO-K1 cells and compared the efficiencies of these methods. The results indicated that transfection of CHO-K1 cells by the method of lipofection had the highest efficiency (about 20~30%). The transfection efficiencies of calcium phosphate coprecipitation (<1%), DEAE-Dextran mediated tansfection (~2%) and electroporation (~3%) were significantly lower than that of lipofection. |
本系統中英文摘要資訊取自各篇刊載內容。