Braga Daniel, Laizé Vincent, Tiago Daniel M, Cancela M Leonor
Centre of Marine Sciences (CCMAR), University of Algarve, Campus de Gambelas, 8005-139 Faro, Portugal.
Mol Biotechnol. 2006 Sep;34(1):51-4. doi: 10.1385/MB:34:1:51.
The use of in vitro cell culture systems to assess gene function largely depends on the successful transfer of DNA into target cells. Well developed in mammals, transfection methods are still to be optimized for non-mammalian cell culture systems, like fish. Here we describe a rapid, cost-efficient, and successful method to transfer DNA into a fish bone-derived cell line using polyethylenimine (PEI) as the DNA carrier. Using this method, DNA transfer was remarkably enhanced in comparison with commercially available reagents, as demonstrated by the increased activity of both luciferase and green fluorescent protein observed in the transfected cells. Its efficiency in transferring DNA into a wide range of cell types, including non-mammalian and hard-to-transfect cells, in addition to a low cost, show that PEI is a reagent of choice for nonviral vector transfection.
利用体外细胞培养系统评估基因功能很大程度上取决于将DNA成功导入靶细胞。转染方法在哺乳动物中已得到充分发展,但对于非哺乳动物细胞培养系统(如鱼类)仍有待优化。在此,我们描述了一种快速、经济高效且成功的方法,即使用聚乙烯亚胺(PEI)作为DNA载体将DNA导入一种源自鱼骨的细胞系。使用该方法,与市售试剂相比,DNA转染显著增强,转染细胞中观察到的荧光素酶和绿色荧光蛋白活性增加证明了这一点。其将DNA导入多种细胞类型(包括非哺乳动物细胞和难以转染的细胞)的效率,以及低成本,表明PEI是用于非病毒载体转染的首选试剂。