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Volume 19 Issue 4
August 2004
    Citation: WU Dong, DENG Fei, HU Zhi-hong, YUAN Li, SUN Xiu-lian. Molecular Cloning and Expression of the fp25k Gene of HaSNPV in E. coli and Preparation of Its Antiserum .VIROLOGICA SINICA, 2004, 19(4) : 380-384.
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    Molecular Cloning and Expression of the fp25k Gene of HaSNPV in E. coli and Preparation of Its Antiserum

    • 1.

      1. Laboratory of Molecular Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China

    • 2.

      Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China

    • 3.

      Graduate School of the Chinese Academy of Sciences, Beijing 100039, China

    • Corresponding author: SUN Xiu-lian, 
    • Available online: 25 August 2004
    • The PCR product of Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus HaSNPV fp25k gene was cloned into a prokaryotic expression vector pProEXHTb. After inducing with IPTG, the fp25k gene was successfully expressed in E.coli DH5α. The expression product was 32 kDa. This protein was purified and injected into rabbit to raise polyclonal antiserum. Western blot analysis using this antiserum showed the specific reaction to FP25K protein expressed in both prokaryotic with GST fusion and viral infected insect cells. The antibody made it possible to analysis the function this protein in infected insect cells.
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      Molecular Cloning and Expression of the fp25k Gene of HaSNPV in E. coli and Preparation of Its Antiserum

        Corresponding author: SUN Xiu-lian,
      • 1. 1. Laboratory of Molecular Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China
      • 2. Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China
      • 3. Graduate School of the Chinese Academy of Sciences, Beijing 100039, China

      Abstract: The PCR product of Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus HaSNPV fp25k gene was cloned into a prokaryotic expression vector pProEXHTb. After inducing with IPTG, the fp25k gene was successfully expressed in E.coli DH5α. The expression product was 32 kDa. This protein was purified and injected into rabbit to raise polyclonal antiserum. Western blot analysis using this antiserum showed the specific reaction to FP25K protein expressed in both prokaryotic with GST fusion and viral infected insect cells. The antibody made it possible to analysis the function this protein in infected insect cells.

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