Co-expression of rabies virus nucleoprotein and glycoprotein using Bac-to-Bac baculovirus expression system

Objective To co-express rabies virus CTN-1V strain nucleoprotein (NP) and glycoprotein(GP) using Bac-to-Bac baculovirus expression system.Methods The ORF of GP and NP were amplified from CTN-1V strain by RT-PCR and cloned into plasmid vector pFastBacDual PP10 and PPH,respectively.The constructed recombinant donor plasmid P-NG was transformed to DH10Bac E.coli and obtained the recombinant bacmid shuttle plasmid A-NG.Then the plasmid A-NG was transfected into Sf9 cells to construct recombinant baculovirus Bac-NG.The co-expressed protein was identified by Western blot.Resuits It was proved by PCR that recombinant bacmid shuttle plasmid A-NG was constructed correctly.The recombinant protein NP and GP,with a relative molecular mass of about 51 × 103 and 59 × 103,were co-expressed in Sf9 cells.Both expressed NP and GP showed the specific binding reactivity to murine sera against RV.Conclusion The NP and GP of rabies virus CTN-1V strain were successfully co-expressed and showed good immunogenicity,which could serve as a technical basis for new form of vaccine and diagnostic reagents for rabies virus. Key words: Rabies virus; Nucleoprotein; Glycoprotein; Bac-to-Bac baculovirus expression system