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Citation: LIU Chao-hong, JIAO Chen-song, CHEN Xin-wen. Cloning of HCV ns2 Gene and its Expression in Prokaryotic and Mammalian cells [J].VIROLOGICA SINICA, 2006, 21(3) : 217-220.

Cloning of HCV ns2 Gene and its Expression in Prokaryotic and Mammalian cells

  • Corresponding author: CHEN Xin-wen, 
  • The full-length and truncated ns2 gene (2881-3078 bp) were gained through PCR using p90-HCV as template. The truncated ns2 gene was cloned into the prokaryotic expression vector PET32a. The fusion protein Trx-His-NS2C expressed in a soluble form in E.coli, and then was purified by His resin. Highly specific and efficient antibody was produced by immunizing the rabbit with purified fusion protein. In addition to this, recombinant adenovirus containing the full-length ns2 gene was constructed. The NS2 protein was expressed successfully in a size of 23.2kDa by infecting 293 and HepG2 cells with recombinant adenovirus. These results had set a base for the further stulies on the structure and function of the NS2 protein.

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    Cloning of HCV ns2 Gene and its Expression in Prokaryotic and Mammalian cells

      Corresponding author: CHEN Xin-wen,
    • 1. 1.State Key Laboratory of Virology, Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China
    • 2. Wuhan General Hospital of Guangzhou Command. Wuhan 430071,China

    Abstract: The full-length and truncated ns2 gene (2881-3078 bp) were gained through PCR using p90-HCV as template. The truncated ns2 gene was cloned into the prokaryotic expression vector PET32a. The fusion protein Trx-His-NS2C expressed in a soluble form in E.coli, and then was purified by His resin. Highly specific and efficient antibody was produced by immunizing the rabbit with purified fusion protein. In addition to this, recombinant adenovirus containing the full-length ns2 gene was constructed. The NS2 protein was expressed successfully in a size of 23.2kDa by infecting 293 and HepG2 cells with recombinant adenovirus. These results had set a base for the further stulies on the structure and function of the NS2 protein.

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