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Abstract After the eukaryotic expression vector pcDNA3-NDV-F was constructed successfully, the recombinant was transfected into P815 cell by LipofectamineTM 2000, and the cells were screened with G41 8 to obtain stable clone. The total RNA was prepared from transfected cells according to the instructions of the Trizol reagent and the 1600bp fragment was obtained by RT-PCR. Westem blotting and immunofluorescence were used to detect the expression of NDV-F protein. In order to detect the stability of the transfected cell line, the cell line was freezed in the liquild nitrogen for six months. The experiments proved NDV–F gene were expressed well. These results suggested that the stable transfected P815 cell line was successfully established by Lipofectin, which provides solid foundation for further experimental studies on the function NDV nucleotide vaccine.
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Received: 14 March 2008
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