Citation: FANG Qin, DING Qing—quan, WAN G Ya-ping, ZHU Zuo—yan. Expression of Grass carp reovirus RNA Polymerase Gene and Purification of its Product .VIROLOGICA SINICA, 2003, 18(2) : 169-173.

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

  • Available online: 15 April 2003
  • 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.

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

    • 1. Wuhan Institute of Virology,Chinese Academy of Sciences,Wuhan 430071,China
    • 2. State Key Laboratory of Freshwater Ecology and Biotechnology,Institute ofHydrobiology,Chinese Academy ofSciences,Wuhan 430072,China

    Abstract: 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.

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