摘要
Grass carp reovirus （GCRV）, a double stranded RNA virus that infects aquatic animals, often with disastrous effects, belongs to the genus Aquareovirus and family Reoviridea. Similar to other reoviruses, genome replication of GCRV in infected cells occurs in cytoplasmic inclusion bodies, also called viral factories. Sequences analysis revealed the nonstmctural protein NS80, encoded by GCRV segment 4, has a high similarity with μNS in MRV（Mammalian orthoreovimses）, which may be associated with viral factory formation. To understand the function of the μNS80 protein in virus replication, the initial expression and identification of the immunogenicity of the GCRV NS80 protein inclusion forming-related region （335-742） was investigated in this study. It is shown that the over-expressed fusion protein was produced by inducing with IPTG at 28℃. In addition, serum specific rabbit antibody was obtained by using super purified recombinant NS80（335-742） protein as antigen. Moreover, the expressed protein was able to bind to anti-his-tag monoclonal antibody （mouse） and NS80〈335.742） specific rabbit antibody. Further western blot analysis indicates that the antiserum could detect NS80 or NS80C protein expression in GCRV infected cells. This data provides a foundation for further investigation of the role of NS80 in viral inclusion formation and virion assembly.
Grass carp reovirus(GCRV),a double stranded RNA virus that infects aquatic animals,often with disastrous effects,belongs to the genus Aquareovirus and family Reoviridea. Similar to other reoviruses,genome replication of GCRV in infected cells occurs in cytoplasmic inclusion bodies,also called viral factories. Sequences analysis revealed the nonstructural protein NS80,encoded by GCRV segment 4,has a high similarity with μNS in MRV(Mammalian orthoreoviruses) ,which may be associated with viral factory formation. To understand the function of the μNS80 protein in virus replication,the initial expression and identification of the immunogenicity of the GCRV NS80 protein inclusion forming-related region(335-742) was investigated in this study. It is shown that the over-expressed fusion protein was produced by inducing with IPTG at 28oC. In addition,serum specific rabbit antibody was obtained by using super purified recombinant NS80(335-742) protein as antigen. Moreover,the expressed protein was able to bind to anti-his-tag monoclonal antibody(mouse) and NS80(335-742) specific rabbit antibody. Further western blot analysis indicates that the antiserum could detect NS80 or NS80C protein expression in GCRV infected cells. This data provides a foundation for further investigation of the role of NS80 in viral inclusion formation and virion assembly.
基金
National Basic Research Program of China (973 Program, Grant No. 2009CB118701)
National Natural Scientific Foundation of China (Grant Nos. 30671615, 30871940)
Innovation project of the Chinese Academy of Sciences (Grant No.KSCX2-YW-N-021)
