Optimization of Parameters of Anti-Classical Swine Fever virus Gene Transfection of porcine fetus fibroblasts Mediated by Liposome

Take large white pig as the study materials,porcine fetus fibroblasts was obtained by the method of tissue culture.Plasmid pGPU6/GFP/Neo of eGFP as report gene and Anti-Swine fever virus gene as aim gene was transfected into porcine fetus fibroblasts by lipofectamineTM 2000.By adjusting transfection time,concentrations of plasmid DNA and liposome to improve transfection rate.Perfect cell transfection efficiency(32.5%) was obtained on the condition of the complex of 1.2μg Plasmid and 2.5uL lipofectamine TM 2000 transfecting PFF in 6 hours.Both high concentration of LipofectamineTM 2000 and longer incubation time(6h) will cause cell shrinkage and death.The present study would provide donor cell for the production of transgenic pig which can resist classicial swine fever vires infection.