Citation: YANG Xiao-jun, YE Lin-bai*, GAO Jin-rong, LIU Jing, YANG Fan, YE Li, SHE Ying-long, LIAO Qing-jiao, WU Zheng-hui, ZHENG Yi. Stable Expression of NS5A Gene of HCV in Hela Cells and Inhibition to the Growth of Hela Cells .VIROLOGICA SINICA, 2004, 19(4) : 340-344.

Stable Expression of NS5A Gene of HCV in Hela Cells and Inhibition to the Growth of Hela Cells

  • Corresponding author: YE Li, 
  • Available online: 25 August 2004
  • Full-length NS5A gene of Hepatitis C virus(HCV) was amplified by PCR, using the plasmid pBRTM/HCV 1-3011 containing HCV full-length open reading frame(ORF)as template, and cloned into the eukaryotic expressing plasmid pcDNA3.1(-)by DNA recombination technique. The recombin- ant vector was identified by digestion with restriction enzymes and polymerase chain reaction and by directly sequencing. Then both the recombinant vector pcDNA3.1(-)-NS5A and the control vector pcDNA3.1(-)were transfected Hela cells using LipoVecTM. The cells expressing NS5A stablely were selected by G-418 and further proved by RT-PCR and Western blot analysis. We found the growth of Hela cells expressing NS5A was slower than the cells transfected by pcDNA3.1(-)in the same culture condition, and the population doubling time of Hela cells expressing NS5A gene is increased about 50%(about 35-35 hours). There was no significant difference between the control cells and the cells transfected with pcDNA3.1(-) (about 23-24 hours). The results indicate that NS5A can inhibit the proliferation of Hela cells.

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    Stable Expression of NS5A Gene of HCV in Hela Cells and Inhibition to the Growth of Hela Cells

      Corresponding author: YE Li,
    • 1. Institute of Virology, Wuhan University, Wuhan 430072, China

    Abstract: Full-length NS5A gene of Hepatitis C virus(HCV) was amplified by PCR, using the plasmid pBRTM/HCV 1-3011 containing HCV full-length open reading frame(ORF)as template, and cloned into the eukaryotic expressing plasmid pcDNA3.1(-)by DNA recombination technique. The recombin- ant vector was identified by digestion with restriction enzymes and polymerase chain reaction and by directly sequencing. Then both the recombinant vector pcDNA3.1(-)-NS5A and the control vector pcDNA3.1(-)were transfected Hela cells using LipoVecTM. The cells expressing NS5A stablely were selected by G-418 and further proved by RT-PCR and Western blot analysis. We found the growth of Hela cells expressing NS5A was slower than the cells transfected by pcDNA3.1(-)in the same culture condition, and the population doubling time of Hela cells expressing NS5A gene is increased about 50%(about 35-35 hours). There was no significant difference between the control cells and the cells transfected with pcDNA3.1(-) (about 23-24 hours). The results indicate that NS5A can inhibit the proliferation of Hela cells.

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