Construction of Recombinant Canine Adenovirus Type-2 Expressing Rabies Virus Glycoprotein
Abstract: 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. ˎ̥,Verdana,Arial; mso-font-kerning: 1.0pt; mso-bidi-font-family: 'Times New Roman'; mso-fareast-font-family: 宋体; mso-ansi-language: EN-US; mso-fareast-language: ZH-CN; mso-bidi-language: AR-SA">Preliminary immunization trial in dogs showed that the recombinant virus could stimulate specific immune response to both vector virus and Rabies virus .