Ray J, Gage F H
Department of Neurosciences, University of California, San Diego La Jolla 92093-0627.
Biotechniques. 1992 Oct;13(4):598-603.
The stable transfection of immortalized Rat-1 and rat skin primary fibroblast cell lines by calcium phosphate precipitation, lipofection and electroporation methods have been examined. The lipofection method was found to be better than the other methods in terms of higher transfection efficiency and convenient use. Expression of beta-galactosidase from two different viral promoters showed that the level of transgene expression depends on the promoter strength in a particular cell type. The results presented here show that the transgene expression is extremely variable among different colonies generated from individually transfected cells. Therefore, it is necessary to examine individual colonies of cells for the production of reporter gene to obtain cell lines expressing high amounts of gene products.
已通过磷酸钙沉淀法、脂质体转染法和电穿孔法检测了永生化大鼠-1细胞系和大鼠皮肤原代成纤维细胞系的稳定转染情况。结果发现，脂质体转染法在转染效率较高且使用方便方面优于其他方法。来自两种不同病毒启动子的β-半乳糖苷酶表达表明，转基因表达水平取决于特定细胞类型中的启动子强度。此处呈现的结果表明，在由单独转染的细胞产生的不同克隆中，转基因表达差异极大。因此，有必要检测单个细胞克隆以产生报告基因，从而获得能够表达大量基因产物的细胞系。
