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Volume 18 Issue 1
February 2003
    Citation: ZHANG Dong—wei, LIANG Bu—feng, LI Wei-dong, QI Zi—bai, LING Shi—gan. HCV E2 Prodaryotic system Gene expression Purification .VIROLOGICA SINICA, 2003, 18(1) : 1-4.
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    HCV E2 Prodaryotic system Gene expression Purification

    • 1.

      1．Wuhan Institute of Virology，Academia Sinica，Wuhan 430071，．China

    • 2.

      Department ofHepatitis，National Institute for the Control ofPharmaceutical and Biology Products，Beijing 100050，China

    • 3.

      Military Medical Academia Sinica， Beijing 100850，China

    • Corresponding author: LIANG Bu—feng, 
    • Available online: 02 February 2003
    • Rceomdinant plasmid pET-E2 was transformed into BL21 (DE3) competent cell, then induced the amplified culture with IPTG. There was a 34 kDa band in SDS-PAGE gel. The molecular weight of the expressed protein was comsistent to that of deduced E2 protein. Tested by western-blot, It showed that the product was HCV E2 protein. It contained a six-Histidine tag and aggregated into the inclusion body. Computer scan analysis showed the protein interested took the percentage more than 36 of the total bacterial proteins and the rate of purification was higher than 95% after purified by Ni-column. The activity of pruified E2 protein was tested by ELISA. The result showed that the 5/15 HCVpositive sera had anti-E2 antibody, and the other 5 negative HCV sera hadn’t anti-E2 antibody
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      HCV E2 Prodaryotic system Gene expression Purification

        Corresponding author: LIANG Bu—feng,
      • 1. 1．Wuhan Institute of Virology，Academia Sinica，Wuhan 430071，．China
      • 2. Department ofHepatitis，National Institute for the Control ofPharmaceutical and Biology Products，Beijing 100050，China
      • 3. Military Medical Academia Sinica， Beijing 100850，China

      Abstract: Rceomdinant plasmid pET-E2 was transformed into BL21 (DE3) competent cell, then induced the amplified culture with IPTG. There was a 34 kDa band in SDS-PAGE gel. The molecular weight of the expressed protein was comsistent to that of deduced E2 protein. Tested by western-blot, It showed that the product was HCV E2 protein. It contained a six-Histidine tag and aggregated into the inclusion body. Computer scan analysis showed the protein interested took the percentage more than 36 of the total bacterial proteins and the rate of purification was higher than 95% after purified by Ni-column. The activity of pruified E2 protein was tested by ELISA. The result showed that the 5/15 HCVpositive sera had anti-E2 antibody, and the other 5 negative HCV sera hadn’t anti-E2 antibody

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