Abstract: In order to establish highly efficient system of transfected sheep fibroblast, cationic liposome method was used to transfect subcultured 3 to 5 passage ovine fibroblasts by plasmids with green fluorescence protein (GFP) report gene, with the view of confirming the best transfection method and its optimization. Results showed that transfection efficiency could get maximal with 0.2 μg DNA and 2.0 μL LipofectamineTM 2000 in 12 hours under the circumstance of using plasmid DNA taken by Plasmid Purification Mini Kit-spin column. Green fluorescence near the nucleus became dimmer and dimmer with the lapse of time after the initial strong expression, on the contrary, it enhanced gradually in the cytoplasm.

Key words: plasmid DNA; sheep fibroblasts; transfection; cationic liposome method