Establishment of P815 cell line stably expressing porcine circovirus type 2 capsid protein

Recombinant plasmids pcDNA3.1-orf2 expressing PCV2 capsid protein were transfected into P815 line cells mediated by liposomes.The positive anti-G418 monoclonal cells were screened and obtained in the medium with G418,and identified by PCR,RT-PCR and indirect immunofluorenscence assay.Subsequently,the stability of the positive cells expressing aimed protein was detected.The results showed that orf2 gene integrated into the genome of P815 cells,the capsid protein coded by PCV2 ORF2 was expressed in the acquired positive P815 monoclonal cells,and the P815 cell line with a stable expression of PCV2 capsid protein was obtained.The cell line will be necessary in the study of PCV2 new type vaccines,and the experiment method may be of referential value in other virus vaccines development.