Establishment and Characterization of a Vero Cell Line Stably Expressing N Protein of Porcine Deltacoronavirus
To obtain a African green monkey kidney cell(Vero)line stably expressing nucleocapsid(N)protein of Porcine deltacoronavirus(PDCoV),the PDCoV N gene was cloned into a lentiviral vector to obtain the recombinant plasmid PLVX-PDCoV-N.Then,the recombinant plasmid PLVX-PDCoV-N was transfected into 293T cells by lentivirus packaging system and packaged into lentivirus particles for N protein expression.Vero cells were infected then with the lentivirus and the target cells were screened by puromycin.The results of RT-PCR and sequencing of PCR product showed the genome of the resulting Vero cell line containing N gene.The N protein that was stably expressed in the Vero cell line was confirmed by Western blot and IFA.The Vero cell line was used to detect clinical PDCoV positive serum samples and the coincidence with the results of ELISA was 100％.The study successfully established a Vero cell line stably expressing PDCoV N protein,which laid a foundation for further research of biological characteristics of N protein and clinical detection and epidemiological investigation of PDCoV.
Porcine deltacoronavirusnucleocapsid proteinLentiviruscell line
万方数据
维普
