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Published on: June 19, 2013
[Construction of eukaryotic expression vector containing foot-and-mouth disease virus VP1 gene and its expression in
1Department of Immunology and Cell Biology, College of Veterinary Medicine, Agricultural University of Hebei, Baoding 071000, China. lijie2006698@yahoo.com.cn
Aim:
To construct the eukaryotic expression vectors for the FMDV VP1 gene and transfect dendritic cells(DC) for the expression of VP1.
Methods:
The plasmid pMD-18-VP1 was subcloned into pcDNA3.1(+) to construct recombinant eukaryotic expression plasmids named as pcDNA3.1-VP1. The recombinant plasmids were transfected into DC by lipofectamine method and the positive cell clones were screened with G418. The expression of FMDV VP1 gene in DC was determined by Western blot.
Results:
The correct construction of pcDNA3.1-VP1 was identified by means of restriction enzyme analysis and nucleotide sequence determination. It was showed by SDS-PAGE and Western blot that transfected cells, DC, expressed FMDV VP1 gene constantly.
Conclusion:
The pcDNA3.1-VP1 eukaryotic expression plasmids are successfully constructed and VP1 gene can be sustainly expressed in DC.
