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Volume 19 Issue 4
August 2004
    Citation: LI Ti-yuan, XU Yi-ying, Kevin Philips. Purification and Enzymatic Analysis of the Recombinant Hepatitis C Virus NonStructure Protein 3 with Protease and Helicase Activity .VIROLOGICA SINICA, 2004, 19(4) : 325-328.
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    Purification and Enzymatic Analysis of the Recombinant Hepatitis C Virus NonStructure Protein 3 with Protease and Helicase Activity

    • 1.

      1. Shenzhen People’s Hospital, 2nd Affiliated Hospital of Medical College, Jinan University, Shezhen 518020, China

    • 2.

      Department of Biochemistry, Ohio State University, Columbus, Ohio 43210，USA

    • Corresponding author: LI Ti-yuan, 
    • Available online: 25 August 2004
    • To study the enzymatic activity，the recombinant HCV NS3 protein with both protease and helicase activity was expressed and purified. The nonstructure gene 3 (NS3) of HCV was amplified and inserted into plasmid pPIC9 and the recombinant plasmid pPIC-NS3 was transformed into Pichia pastoris strain GS115. Recombinant protein was produced by induction of the AOX1 promoter with methanol. The recombinant protein was first purified by Hitrap SP cation exchange and then by Mono S HR column. In vitro cleavage assay showed that the recombinant protein has protease and helicase activity.
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      Purification and Enzymatic Analysis of the Recombinant Hepatitis C Virus NonStructure Protein 3 with Protease and Helicase Activity

        Corresponding author: LI Ti-yuan,
      • 1. 1. Shenzhen People’s Hospital, 2nd Affiliated Hospital of Medical College, Jinan University, Shezhen 518020, China
      • 2. Department of Biochemistry, Ohio State University, Columbus, Ohio 43210，USA

      Abstract: To study the enzymatic activity，the recombinant HCV NS3 protein with both protease and helicase activity was expressed and purified. The nonstructure gene 3 (NS3) of HCV was amplified and inserted into plasmid pPIC9 and the recombinant plasmid pPIC-NS3 was transformed into Pichia pastoris strain GS115. Recombinant protein was produced by induction of the AOX1 promoter with methanol. The recombinant protein was first purified by Hitrap SP cation exchange and then by Mono S HR column. In vitro cleavage assay showed that the recombinant protein has protease and helicase activity.

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