Department of Cardiology, Children's Hospital Boston, Enders 1261, 320 Longwood Avenue, Boston, MA, 02115, USA.
Cytotechnology. 2006 Jun;51(2):105-10. doi: 10.1007/s10616-006-9018-3. Epub 2006 Sep 21.
Although various gene delivery techniques are available, their application in zebrafish cell cultures has not been extensively studied. Here, we report that nucleofection of zebrafish primary embryonic fibroblasts results in higher transfection efficiency in comparison to other non-viral gene delivery methods. The transfection was performed using green fluorescent protein (GFP) gene constructs of a different size. Greatest DNA uptake was obtained with 4.9-kb plasmid, resulting in 43% GFP positive cells. Nucleofection with 7.4-kb pH2B-GFP plasmid followed by geneticin (G418) selection was successfully used to establish a cell line expressing nuclear histone 2B-GFP fusion protein. Efficient transfection of zebrafish fibroblasts by nucleofection offers a non-viral technique of plasmid delivery and can be used to overexpress genes of interest in these cells.
虽然有各种基因传递技术，但它们在斑马鱼细胞培养中的应用尚未得到广泛研究。在这里，我们报告说，与其他非病毒基因传递方法相比，斑马鱼原代胚胎成纤维细胞的电穿孔可导致更高的转染效率。转染使用了不同大小的绿色荧光蛋白（GFP）基因构建体。用 4.9kb 的质粒获得了最大的 DNA 摄取量，导致 43%的 GFP 阳性细胞。用 7.4kb pH2B-GFP 质粒进行电穿孔，然后进行遗传霉素（G418）选择，成功地建立了表达核组蛋白 2B-GFP 融合蛋白的细胞系。电穿孔可有效地转染斑马鱼成纤维细胞，提供了一种非病毒的质粒传递技术，并可用于在这些细胞中转染感兴趣的基因。
