Effect of structural domain II of HCV 5' noncoding region on its translation initiation activity.
- Author:
Shui-Ping LIU
1
;
Jun-Qin ZHAO
;
Hong-Tao LI
;
Tao YANG
Author Information
- Publication Type:Journal Article
- MeSH: 5' Untranslated Regions; Hep G2 Cells; Hepacivirus; chemistry; genetics; metabolism; Hepatitis C; Humans; Nucleic Acid Conformation; Peptide Chain Initiation, Translational; RNA, Viral; chemistry; genetics
- From: Chinese Journal of Experimental and Clinical Virology 2009;23(3):200-203
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo examine the role of domain II of hepatitis C virus (HCV) 5' noncoding region (5' NCR) in its translation initiation activity.
METHODSThe fragment of HCV 5' NCR with deletions of 5'-proximal 118 nucleotides were amplified with PCR, then was used to substitute for the full length HCV 5' NCR of plasmid pCMVNCRluc, a firefly luciferase (Fluc) eukaryotic expression plasmid regulated by HCV 5' NCR, to generate recombinant plasmid pCNl-d3. pCMVNCRluc, pCNl-d2 (modified pCMVNCRluc with deletions of HCV 5' NCR ntl-43) and pCNl-d3 were transfected into HepG2 cells with liposome transfection protocol, respectively, and the relative luciferase activity was measured to analyze the regulatory effect of the truncated 5'NCR on Flue gene expression. Meanwhile the Flue mRNA levels were detected by RT-PCR.
RESULTSThe recombinant plasmid was successfully constructed. The Flue mRNA levels of the 3 plasmids were not significantly different (P > 0.05), and there were also no significant difference between the relative luciferase activity of plasmids pCNl-d2 and pCMVNCRluc (P > 0.05). However, that of pCNl-d3 was decreased significantly (P < 0.01, compared with pCNl-d2 or pCMVNCRluc).
CONCLUSIONStructural domain II of HCV 5' NCR plays an important role in its translation initiation activity.
