Study on Transfecting the Anti-Swine Fever Virus Plasmid DNA into the Piglet Fibroblasts by Electroporation

The aim of this research is to transfect the anti-swine fever virus plasmid into the piglet fibroblasts by electroporation and optimize the conditions of electroporation.The anti-swine fever virus plasmid was transfected into the P6 piglet fibroblasts cultured in our lab by electroporation,and transfection factors which includes voltage,pulse time,the number of pulse,the amount of plasmid DNA,the temperature after electric shock,electroporation buffer,the types of plasmid were explored and optimized.The results showed that the transfection efficiency is the highest,reaching 47.49%,when transfecting the fibroblasts with 220V voltage,25ms pulse time,1pulse,12μg annular plasmid DNA,the limited electroporation fluid as the buffer of electric shock and the transfected cells kept in 4℃ for 10min.These results indicated when transfecting the anti-swine fever virus plasmid into the piglet fibroblasts by electroporation,an ideal transfection efficiency could be obtained.