The Establishment Of Mdbk Stable Cell To Express The Capsid Protein Of Bovine Virus Diarrhea Virus

Bovine viral diarrhea-mucosal virus (BVDV) is a virus that causes bovine viral diarrhea-mucosal disease, which poses a serious threat to animal husbandry and aquaculture. To change this serious situation, the research of BVDV vaccine is of great urgency. In this paper,we studied how to construct stable cell lines that expressed the capsid protein of bovine viral diarrhea virus. It is of great important to provide a basis for the study of the vaccine of the virus in the next step, so that we are able to form a guarantee for the defense of food-borne hazards.In this study, recombinant plasmids expressing capsid proteins were obtained by plasmid construction and transfected into cells by electroporation. Through screening of stable cell lines and identification of Western blotting. Finally we obtained a Madin-Darby Bovine Kidney cell line, which can stably express the capsin protein of BVDV. The main results are as follows (1)with BVDV strain SD-1 acting as a template gene, the target gene Npro-C-Erns was obtained by PCR;(2) On the basis of the eukaryotic expression vector pcDNA3.1 (+), we constructed recombinant plasmid pc-Npro-C-Erns which were able to express the viral proteins Npro-C-Erns in cells; (3) by the method of electroporation, the plasmid pc-Npro-C- Erns were transfected into MDBK cells; (4) Via Western blotting, we verified whether the cells can stably express viral proteins.After the all steps, the present experiment finally obtained a stable cell line pc-Npro-C-Erns-457. The successful construction of this cell line will provide a supplementary material for the further development of bovine viral diarrhea-mucosal virus vaccine, laying a solid foundation for research...