Serving the Society Since 1986
Advanced Search
Volume 19 Issue 4
August 2004
    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.
    shu

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

    • 1.

      Institute of Virology, Wuhan University, Wuhan 430072, China

    • 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.
    • 加载中
    • 加载中
    PDF

    Article Metrics

    Article views(3814) PDF downloads(878) Cited by()

    Related
    Proportional views

      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.

        HTML

      Relative (20)

      目录

      This journal is a member of, and subscribes to the principles of, the Committee on Publication Ethics (COPE)

      鄂ICP备05001977号

         

      鄂公网安备 42010602003674号

      Copyright © 2019 Virologica Sinica. All rights reserved

      /

      DownLoad:  Full-Size Img  PowerPoint
      Return
      Return