- 學位論文
經電擊處理後之微脂體對轉染效率的影響之研究
The effects of electric-treated DNA-liposome complex on transfection efficiency
摘要
目前已有許多動物細胞的轉染技術，包括利用病毒性載體（例如：腺病毒或反轉錄病毒等）、物理性基因傳遞的方法（例如：電擊、基因槍或微泡等）或使用商業上常用的化學轉染試劑（例如：聚合物或陽離子型的微脂體等）。而在轉染的過程中有許多影響轉染效率的因子，若以微脂體為例，以其粒徑大小和轉染後的培養時間等最被廣為研究。 在此，我們以在哺乳動物細胞能表現綠色螢光蛋白的pIRES2-EGFP作為表現質體，並使用Lipofectamine 2000作為攜帶此質體的轉染試劑。本實驗研究中，我們證明了電擊後的DNA-liposome complex粒徑大小會增加；不過，轉染效率反而下降。從許多結果顯示出不同於發表的文獻所提出的DNA-liposome complex粒徑大小增加會提高轉染效率的論點。
並列摘要
There are many transfection technologies of animal cell, including the use of viral vectors, such as adenovirus and retrovirus, physical gene delivery method, such as electroporation, gene gun and microbubbles, and commercial chemical transfection reagents, such as polymer and cationic liposome. Many factors affect transfection efficiency in the transfection process. For example, the size of liposome particles and incubation time after transfection are most widely studied. Here we used pIRES2-EGFP as an expression plasmid that could produce green fluorescent protein in mammalian cells, and Lipofectamine 2000 as the transfection reagents. In this study, we demonstrate that the size of DNA-liposome complex increased after electric shock; however, the transfection efficiency decreased. These results are different from the researchs which reported that the size of DNA-liposome complex increased, the transfection efficiency increased.