Establishment of stable avian cell line expressing V protein of Newcastle disease virus

In order to obtain the stable DF-1 cell lines expressing Newcastle disease virus(NDV) V protein,NDV V gene from the recombinant plasmid pEGFT-N1-V was amplified,the restriction sites of EcoRⅠ and BamHⅠ were inducted into the amplicon.The amplicon were then connected into the piggyBac transposon cloning plasmid pTKL1-CMV-Puro-EGFP vector,which was checked by enzyme digestion and sequence analysis.The recombinant plasmids were co-transfected with the helper plasmid mPB into DF-1 cells using LipofectamineTM2000.The cell line was selected with puromycin and detected with green fluorescent protein observation,and V protein expression was detected by Western-blot.Results showed that the stable monoclonal cell lines expressing V protein was constructed successfully.The results revealed that piggyBac transposon system can be used for the construction of avian stable cell lines and the DF-1 cell line(DF-1-V) expressing NDV V protein was established successfully.The present study provi-ded reference materials for application of the piggyBac transposon system in avian cell and research of NDV V protein.