Construction and identification of a rabies virus nucleoprotein-canine adenovirus recombinant

Objective To construct a rabies virus nucleoprotein containing canine adenovirus type 2 (CAV-2) recombinant, and to evaluate the biological and immunological properties of the recombinant virus. Methods The expression cassette of nucleoprotein consisted of the human cytomegalovirus (CMV) immediate early promoter, rabies virus strain SRV_9 nucleoprotein cDNA, and polyadenylation signal of the bovine growth hormone. By using two SspⅠ sites in the E3 region of CAV-2 strain YCA18, a DNA fragment of 859 bp was replaced by the 2 392 bp expression cassette of the nucleoprotein. The recombinant genome was released by AscⅠand PvuⅠ double digestion,then and was transfected into MDCK cells by Lipofectamine TM reagent. Results Typical cytopathogenic effect was observed 10 d post-transfection. SDS-PAGE and Western blotting analysis showed that the nucleoprotein was expressed. Conclusion The recombinant virus could stimulate specific immune responses to CAV-2 and rabies virus.