Construction Of Recombinant Canine Adenovirus Type 2 Expressing Rabies Virus Protective Antigens And Immunization In Canis

Rabies continues to be a serious problem in China due to the reservoirof rabies virus(RV) in dogs. The control and eradication of rabiesdepends on the development of safe, effective and economical vaccine thatmight be used in preexposure vaccination programs for dogs. Fiveproteins are encoded by the rabies virus genome. The glycoprotein isgenerally regarded as the only protein that can stimulate neutralizingantibody. Several studies have also shown that the nucleoprotein protectsmice and dongs against lethal rabies virus infections. To construct arecombinant canine adenovirus against Rabies, the N gene of RV virus andcanine adenovirus type 2(CAV-2) were chosen as foreign gene and thevector, respectively. The expression cassette of nucleoprotein consists of thehuman cytomegalovirus(CMV) immediate promoter, rabies virus strainSRV9 nucleoprotein cDNA and polyadenylation signal of the bovine growthhormone. Using two Ssp Ιsites in the E3 region of canine adenovirustype 2 strain YCA18, a 859 bp DNA fragment was replaced by the 2 392 bpexpression cassette of the nucleoprotein. The recombinant genome wasreleased by AscΙand PvuΙdouble digestion and was transfected intoMDCK cells by Lipofectamine ? . Typical cytopathogenic effect wasobserved 5 passages after transfection. Using the restriction endonucleasedigestion and Southern blot, we confirmed that the recombinant canineadenovirus type 2 expressing rabies nucleoprotein (CAV-2-RN) had been...