Expression of VP2 Hydrophilic Coding-region of Canine Parvovirus and Analysis of Immunogenicity
Yong Wang
Abstract
Yong Wang
Abstract
Objective:To investigate the Immunogenicity of VP2 hydrophilic coding-region of canine parvovirus and lay foundation for further study of Subunitvaccine.Method:Protein analysis software-PROTEAN was used to find VP2 major antigenic epitopes according to hydrophile and antigen character of VP2 amino acids and then the corresponding region of nucleotide was amplified by PCR and cloned into prokaryotic expression vector pQE-31.After transformation,the recombinant protein was analyzed by isopropy1β-D-1 thiogalactopyranoside(ITPG) induction.Finally,mice were immunized with VP2S and the antibody were measured by hemagglutination inhibition(HI) assay.Result:The hydrophilic and antigenic region from 293 to 520 amino acids of the VP2 protein of canine parvovirus,called VP2S,was located.After amplification the prokaryotic expression vector pQE-31-VP2S was obtained.The protein with molecular weight of 29kD,were detected in the lysates of the recombinant E.coli by IPTG induction.Western-blotting assay showed that the recombinant proteins were recognized by CPV antiserum.And VP2S-immunized mice appeared hemagglutination inhibition(HI) antibody compared to the control.which demonstrated the VP2S had good immunogenicity.Conclusion:These results indicated that the VP2S had good immunogenicity.
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Objective:To investigate the Immunogenicity of VP2 hydrophilic coding-region of canine parvovirus and lay foundation for further study of Subunitvaccine.Method:Protein analysis software-PROTEAN was used to find VP2 major antigenic epitopes according to hydrophile and antigen character of VP2 amino acids and then the corresponding region of nucleotide was amplified by PCR and cloned into prokaryotic expression vector pQE-31.After transformation,the recombinant protein was analyzed by isopropy1β-D-1 thiogalactopyranoside(ITPG) induction.Finally,mice were immunized with VP2S and the antibody were measured by hemagglutination inhibition(HI) assay.Result:The hydrophilic and antigenic region from 293 to 520 amino acids of the VP2 protein of canine parvovirus,called VP2S,was located.After amplification the prokaryotic expression vector pQE-31-VP2S was obtained.The protein with molecular weight of 29kD,were detected in the lysates of the recombinant E.coli by IPTG induction.Western-blotting assay showed that the recombinant proteins were recognized by CPV antiserum.And VP2S-immunized mice appeared hemagglutination inhibition(HI) antibody compared to the control.which demonstrated the VP2S had good immunogenicity.Conclusion:These results indicated that the VP2S had good immunogenicity.
Key concepts: Immunogenicity, Recombinant DNA, Hemagglutination assay, Molecular biology, Canine parvovirus, Virology, Antiserum, Porcine parvovirus