Milman G, Herzberg M
Somatic Cell Genet. 1981 Mar;7(2):161-70. doi: 10.1007/BF01567655.
We describe DEAE dextran-mediated DNA transfection in suspension which routinely gives transient gene expression in 0.1--1% of the transfected cells. We have used normal diploid human skin fibroblasts, monkey BSC cells, and mouse L or 3T6 cells with almost equal efficiency. Gene expression is detected 1--3 days after addition of the DNA. SV40 and polyoma T and V antigen are detected by in situ immunofluorescence and thymidine kinase gene expression is detected by in situ autoradiography. The high efficiency of transfection and the speed of detection together provide a means to study transfecting gene functions that does not rely on selection to obtain stable transformants. It should be possible to screen for the expression of any gene product which can be assayed by in situ immunofluorescence or autoradiographic techniques.
我们描述了在悬浮液中通过二乙氨基乙基葡聚糖介导的DNA转染方法,该方法通常能使0.1%至1%的转染细胞实现瞬时基因表达。我们使用正常二倍体人皮肤成纤维细胞、猴BSC细胞以及小鼠L或3T6细胞,转染效率几乎相同。在加入DNA后1至3天可检测到基因表达。通过原位免疫荧光检测SV40、多瘤病毒T和V抗原,通过原位放射自显影检测胸苷激酶基因表达。转染的高效率和检测速度共同提供了一种研究转染基因功能的方法,该方法不依赖于选择来获得稳定的转化体。应该可以筛选出任何能够通过原位免疫荧光或放射自显影技术进行检测的基因产物的表达。