Aujame L, Seguin D, Droy C, Hessler C
Biotechniques. 2000 Jun;28(6):1202-6, 1208, 1210 passim. doi: 10.2144/00286rr01.
A previous study showed that filamentous phage could be efficiently transfected into mammalian cells in the presence of the cationic lipid Transfectam. In the present study, we used an experimental plan based on a uniform network (Doehlert) matrix to estimate optimal transfection conditions in two different cell lines, CHO and Cos-7. Using the cationic lipid RPR120535b as a model, we show that optimal conditions can be determined much more readily than with standard response curves. Under optimal conditions as analyzed by FACS, up to 60% of Cos-7 and 50% of CHO cells can be transfected. Furthermore, a comparison of different lipids (Transfectam, RPR120535b, TC1-12 and GAP-DLRIE/DOPE) suggests that lipids with multiple amine groups are more efficient for the transfection of filamentous phage.
先前的一项研究表明，在阳离子脂质转染胺存在的情况下，丝状噬菌体能够高效转染进入哺乳动物细胞。在本研究中，我们使用基于均匀网络（Doehlert）矩阵的实验方案来评估在两种不同细胞系（CHO和Cos-7）中的最佳转染条件。以阳离子脂质RPR120535b作为模型，我们发现与标准响应曲线相比，确定最佳条件要容易得多。通过荧光激活细胞分选术（FACS）分析，在最佳条件下，高达60%的Cos-7细胞和50%的CHO细胞能够被转染。此外，对不同脂质（转染胺、RPR120535b、TC1-12和GAP-DLRIE/DOPE）的比较表明，具有多个胺基的脂质对丝状噬菌体的转染更有效。
