Study on establishment of cellular model transcripting HCV structural genes

Objective In order to study the effect of Chinese herbs' on the transcription of hepatitis C virus (HCV) structural genes,the cell model being able to transcript the HCV structural genes was established.Methods Using lipofectin,human HeLa cells were transfected with the eukaryotic expression vector pBK-CMV which contained HCV structural gene fragments 1.73-kb.The selective medium containing G418(600-mg·L -1) was used to select the correctly trasfected HeLa cells that was named as HeLaD.Then the total RNA of the transfected HeLa cells was extracted by Trizol reagent and RT-PCR assays were used to further verify the HCV structural gene fragments in mRNA level.Results The RT-PCR results showed that a specific product of 144-bp DNA fragment could be seen in HeLa-D cells transfected with the recombinant plasmid pBK-CMV indicating that the HeLa-D cells could transcript the HCV structural genes successfully.Conclusion Human HeLa cells can be transfected with the HCV structural genes.Further more,the HCV genes can be expressed and transcripted successfully in the HeLa-D cells.