Construction and Preliminary Identification of Vero Cell Line Expressing N Protein of Porcine Epidemic Diarrhea Virus
[Objective]The construction of Vero cell line expressing porcine epidemic diarrhea virus(PEDV)N protein will contribute to the further study of PEDV.[Methods]N gene was cloned into lentiviral vector to construct recombinant plasmid pLenti-CMV-N,which was transfected into 293T cells by lentiviral vector packaging system in this study.The lentivirus particles expressing N protein were packaged and transferred to Vero cells.The positive cells were initially screened by hygromycin B.The Vero cell lines expressing PEDVN protein were initially screened by end point dilution method,and finally identified.[Results]The result of RT-PCR showed that there was a N gene sequence in the genome of the constructed cell line.Western blot and IFA tests confirmed the expression of N protein in the cell line.[Conclusion]A Vero cell line expressing PEDVN protein was successfully constructed by our research.
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