Basic & Clinical Medicine ›› 2009, Vol. 29 ›› Issue (6): 570-574.

• 研究论文 • Previous Articles     Next Articles

Establishment of a cell model supporting HCV1b subgenomic replication

Hui-yi CHEN, Jie LU, Rui LI, Zhong-shuai XIN, Bin YIN, Yan-hua GONG, Xiao-zhong PENG   

  1. Institute of Basic Medical Sciences, CAMS Institut Pasteur of Shanghai, CAS National Laboratory of Medicine Molecular Biology, IBMS, CAMS,School of Basic Medicine PUMC National Lab of Medical Molecular Biology, Institute of Basic Medical Science& School of Basic Medicine, CAMS & PUMC
  • Received:2009-04-15 Revised:2009-04-20 Online:2009-06-25 Published:2009-06-25
  • Contact: Xiao-zhong PENG

Abstract: Objective To establish a cell model for supporting HCV 1b subgenomic replicon of Chinese population replication in vitro. Methods Plasmid containing HCV 1b subgenomic replicon of Chinese population was constructed. HCV subgenomic replicon RNA was obtained by in vitro transcription, and then delivered into Huh-7.5.1 cell by electroporation. After screening the cells in the medium containing G418, we detected the HCV RNA in the obtained cell colonies by RT-PCR and the HCV NS5B protein by western blot. The cell colonies which contain HCV RNA were treated by IFN in order to observe the diminished level of HCV RNA. Results In 4-6 weeks after transfection, visible colonies were stained by crystal violet. The HCV subgenomic replicon RNA NS4B was detected by RT-PCR and the HCV NS5B protein was detected by western blot. After treatment by IFN, HCV RNA diminished obviously. Conclusion We have successfully established a cell model for supporting HCV 1b subgenomic replicon of Chinese population replication in vitro. This cell model is a useful tool for the study on HCV pathogenesis, the screening of antiviral drugs and the development of vaccines.