Expression of Grass carp reovirus RNA Polymerase Gene and Purification of its Product

Abstract：Grass carp reovirus(GCRV)is a disaster agent to aquatic animals，which belongs to genus Aquareovirus．family Reoviridae．Sequences an alysis revealed GCRV S2 was 3877 nucleotides long encoding a 1 38kDa protein VP2，which is deduced as virus RNA polymerase．To understan d the properties of its RN A polym erase，here we constructed 2 expression recombinan ts as pR／RRpN an d pR／RRpC，that covered the gene sequences of N terminal an d C terminal region of RN A polymerase． The 2 recombinan ts were demonstrated in frame expression by SDS—PAGE，an d their molecular weight are about 98kDa an d 103kDa，which were interest fusion proteins．It showed the fusion proteins were able to bind to rabbit serum an ti GCRV—VP2 by using W estern Blot an alysis．In addition，6XHis—tagged GCRV RN A polymerase products were purified by afinity chromatography an d got around 90％ purification of the interest proteins．Th is data provided the evidence for further GCRV RN A polymerase characterization．