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Construction of Canine parvovirus DNA Vaccine and Detection of Its Inoculated Mice |
XIE Zhi-jing;XIA Xian-zhu;HU Rong-liang;YANG Song-tao;ZOU Xiao-huan;HUANG Geng |
Institute of Military Veterinary, Academy of Medical Sciences of PLA, Changchun 130062, China |
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Abstract The VP2 gene of Canine parvovirus (CPV) isolated from a raccoon dog was amplified by PCR and cloned into the vector pMD18-T. The recombinant, named pTCPV, was sequenced and analyzed. As a result, a G→S substitution at amino acid residue 32 was observed besides of 300 G→S. Then, the VP2 gene was sub-cloned into the vector pVAX1 at the downstream of CMV. The recombinant, named pVCPV, was used as CPV DNA vaccine. BHK-21 cells transfected with pVCPV did express CPV VP2 protein specifically reacting to anti-CPV sera. The mice vaccinated with pVCPV developed both anti-, CPV ELISA antibody and SN antibody, but was not detected anti-CPV HI antibody. Lympho-spleencytes of the inoculated mice was proliferated with CPV and ConA stimulation, respectively. The results indicate that the mice inoculated with pVCPV develops the humoral immunity and cellular immunity against CPV.
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Received: 13 July 2005
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