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Virologica Sinica, 21 (5) : 421-425, 2006
Research Article
The Effect of Various Domains of HCV 5’UTR on the Promoter Activity
Institution of Viral Hepatitis, Key Laboratory of Molecular Biology on Infectious Disease, Chongqing University of Medical Science,Chongqing 400016, China
Based on a previous study, a pGL3-5’UTR plasmid was constructed by substitution of the SV40 promoter with HCV 5’UTR cDNA. A set of deletion mutant plamids: pGL3-5’UTR1 comprising the full sequence of 5’UTR; pGL3-5’UTR2 containing domains II, III and IV; pGL3-5’UTR3 containing domains I, II and III; pGl3-5’UTR4 with domains III and IV; pGl3-5’UTR5 with domains I and II were constructed. HepG2 cells were transfected with the above plasmids. The plasmid carrying the full-length of 5’UTR was transfected into hepG2, Hela, HEK293 and Hela cells. The luciferase mRNA and the enzymatic activity were then detected. The relative luciferase activity associated with the pGL3-5’UTR1, pGL3-5’UTR2, pGL3-5’UTR3, pGL3-5’UTR4 and pGL3-5’UTR5 were 5.91±0.65, 9.52±0.32, 2.72±0.45, 2.64±0.25 and.32±0.09, respectively. The luciferase activity was evident in all four cell lines. The data suggested that the DNA sequence of stem-loop III was the core structure and the domain I, II and IV had a regulating effect. It also showed that cells originated from various tissues could provide efficient accessory factors for HCV 5’UTR sequence than strictly act as a promoter and that the original cells may be the most suitable.
Key Words: HCV;  5’UTR;  Domains;  Promoter activity
  Published online: 20 Sep 2006
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