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Virologica Sinica, 20 (2) : 155-158, 2005
Research Article
Construction of Recombinant Canine Adenovirus Type-2 Expressing Rabies Virus Glycoprotein
To construct a new type of vaccine for dog Rabies prevention, Canine adenovirus type-2 was developed for Rabies virus glycoprotein expression. A 1044bp fragment of the E3 region of canine adenovirus type-2 in plasmid pVAX-E3(7.86kb) was deleted by Dra Ⅲ/SspⅠdouble digestion, and an expression cassette of 2424bp in length, composed of the cytomegalovirus immediately early promoter, Rabies virus strain SRV_9 glycoprotein cDNA and SV40 polyadenylation signal, was cloned into the deleted site. The NruⅠ-SalⅠfragment of pPolyⅡ-CAV-2 excluding the E3 region and the NruⅠ-SalⅠfragment of pVAX-E3 containing the expression cassette of the Rabies virus glycoprotein were recombined into pPolyⅡ-CAV2-CGS. The recombinant CAV-2 genome was released by AscⅠ/ClaⅠdigestion and was transfected into MDCK cells by Lipofectamine~(TM) 2000. Canine adenovirus typical CPE and the recombinant virus, CAV-2-CGS, were observed 8 d after transfection. Rabies virus glycoprotein expression was confirmed by Western blotting analysis. Preliminary immunization trial in dogs showed that the recombinant virus could stimulate specific immune response to both vector virus and Rabies virus .
  Published online: 20 Apr 2005
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