Vaccine potential analysis of Grass carp reovirus capsid protein VP7 through in vitro microneutralization assay

Yang Qian; Lu Liqun

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Vaccine potential analysis of Grass carp reovirus capsid protein VP7 through in vitro microneutralization assay

Yang Qian,Lu Liqun * #

College of Fisheries and Life Science, Shanghai Ocean University

*Correspondence author

#Submitted by

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Funding: Specialized Research Fund for the Doctoral Program of Higher Education of China（No.No.20093104120003）

Opened online: 2 April 2011

Accepted by: none

Citation: Yang Qian,Lu Liqun.Vaccine potential analysis of Grass carp reovirus capsid protein VP7 through in vitro microneutralization assay[OL]. [ 2 April 2011] http://en.paper.edu.cn/en_releasepaper/content/4418350

In this paper, the Glutathione S-transferase (GST) fusion protein expression vector pGEX-4T-3 was employed to clone and expression of GCRV outer capsid gene vp7, which was amplified by RT - PCR from infected Grass carp. The recombinant GST-fusion protein rVP7 was induced by 1mM IPTG in Dh5αand confirmed by SDS-PAGE and Western blot assays using both anti-GST-tag and anti-VP7 monoclonal antisera. An expected 52-kDa rVP7 was highly expressed, and was mainly exhibited in the formation of the inclusion body. After purification, rVP7 was intraperitoneally injected to the experimental mice to produce anti-rVP7 polyclonal serum. In vitro microneutralization assay indicated that polyclonal antibody against rVP7 could neutralize GCRV, and suggested that rVP7 had the potential to be used as subunit vaccine against GCRV infection. The present study paved the way for further characterization of the immunogenicity of viral outer capsid protein VP7 in grass carp Ctenopharyngodon idellus and could be based to develop antibody or antigen detection assays for GCRV pathogen.

Keywords:Grass carp reovirus;VP7 protein;microneutralization

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