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題 名 | Protamine Enhances the Efficiency of Liposome-Mediated Gene Transfer in a Cultured Human Hepatoma Cell Line=魚精蛋白可增強微脂體攜帶基因進入肝癌細胞株之效能 |
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作 者 | 倪衍玄; 許宏遠; 陳培哲; 張美惠; | 書刊名 | 臺灣醫學會雜誌 |
卷 期 | 98:8 1999.08[民88.08] |
頁 次 | 頁562-566 |
分類號 | 418.1 |
關鍵詞 | 魚精蛋白; 微脂體攜帶基因; 肝癌; 細胞株; Gene transfection; Protamine; Liposome; Green fluorescent protein; |
語 文 | 英文(English) |
英文摘要 | Protamine, used clinically as an antidote for heparin, is a small protein with high arginine content and is potent in folding DNA. Protamine and DNA can form a compact structure, protecting DNA from digestion by intracellular enzymes. Protamine may, therefore, enhance the efficiency of gene transfer. In this study, we tested the ability of protamine to improve liposome-mediated gene transfer efficiency in a human hepatoma cell line. The results of a preliminary gel retardation assay indicated that 10 μg was the minimal amount of protamine sulfate needed to completely bind 5 μg of a plasmid containing a reporter gene, green fluorescent protein (GFP). For transfection assays, protamine (0, 10, 50, 100 and 500 μg) was added to a DNA-liposome mixture (5 μg DNA and 20 μg of a mixed formulation of 2, 3-dioleyloxy-N-[2(sperminecarboxamido)ethyl]-N, N-dimethyl-1-propanaminium trifluoroacetate and dioleoylphosphatidyletha-nolamine) to transfect cultured Huh7 cells. Transfected cells (those expressing GFP) were counted by using flow cytometry. The expression index (EI) was calculated as the transfection efficiency (% of transfected cells) with protamine divided by the transfection efficiency with DNA and liposome only. Our results show that protamine sulfate (in a range of 10-100, 10 μg being most efficient) addition to the liposome-DNA mixture significantly increases the EI, and transfection efficiency of GFP in Huh7 cells. |
本系統中英文摘要資訊取自各篇刊載內容。