Expression Of GE Protein Of PRV And Nucleocapsid N Protein Of Prrsv Using Baculovirus Expression System

Pseudorabies virus (PRV) and Porcine reproductive and respiratory syndrome virus (PRRSV) are two kinds of infectious pathogen that greatly endangers swine industry. gE protein gene of PRV and nucleocapsid N protein gene of PRRSV are target genes for developing subunit vaccines against PRV and PRRSV respectively.By using baculovisus expression system, gE protein gene of PRV and nucleocapsid N protein gene of PRRSV were subcloned into a bacterial plasmid vector pMAGIC-HIS to yield the recombinant plasmids pMAGIC-HIS-gE、 pMAGIC-HIS-N respectively. Then transformed E.coliDH10β obtained recombinant strain E.coliDH10β/MAGIC-HIS-gE and E.coliDH10β/MAGIC-HIS-N, and as a donor strain and contains a plasmids PML300and AcNPV bacmid-Phes recipient strain E.coliBUN21through MAGIC method (mating-assisted geneticaly integrated cloning) obtained recombinant AcNPV Bacmid-gE、AcNPV Bacmid-N. Recombinant baculoviruses AcNPV-gE and AcNPV-N, were obtained after the two bacmids were used to transfected by liposome mediated cultured Spodoptera frugiperda (Sf9) cell lines respectively. Western blotting detected gE and N proteins with molecular weight of approximately43kD and20kD in Sf9cell lines respectively, after about2-3days later infected by recombinant baculoviruses that by means of two rounds virions amplication, indicating that gE protein and N protein had been successfully expressed inside Sf9cells. The above results established a precious basis for developing subunit vaccines and diagnostic agents against PRV and PRRSV by using baculovirus expression system.