Construction and Identification of a Recombinant Canine Adenovirus Expressing Nucleocapsid Protein of Canine Distemper Virus
Abstract: The nucleocapsid protein gene (N) located 108 to 1679 situation of Canine distemper virus (CDV) genome and was immunogenicity protein with strong conservatism, so N gene was choosed as aim gene. A shuttle plasmid pVAXΔE3LPN containing the gene encoding the nucleocapsid protein of CDV was constructed by restriction endonuclease digestion and ligation. A recombinant plasmid pCAV-2-CDVLPN was constructed with pPoly2-CAV-2 vector containing CAV-2 SY genome. The plasmid pCAV-2-CDVLPN was transfected into MDCK cells by Lipofectamine and after three transfections, a typical CAV-2 cytopathic effect (CPE) was observed. We demonstrated that a recombinant canine adenovirus type-2 expressing nucleocapsid protein of canine distemper virus was successfully constructed and observed by electron microscopy, enzyme digestion, PCR and sequencing. The molecular mass of nucleocapsid protein was 58 kDa.