Effect of structural domain Ⅱ of HCV 5' noncoding region on its translation initiation activity
10.3760/cma.j.issn.1003-9279.2009.03.014
- VernacularTitle:丙型肝炎病毒5'端非编码区结构域Ⅱ在其翻译启动活性中的作用
- Author:
Shui-Ping LIU
1
;
Jun-Qin ZHAO
;
Hong-Tao LI
;
Tao YANG
Author Information
1. 中南大学湘雅医学院
- Keywords:
Hepatitis C;
Hepacivirs;
Luciferase;
Viral structural proteins;
Translation,genetic
- From:
Chinese Journal of Experimental and Clinical Virology
2009;23(3):200-203
- CountryChina
- Language:Chinese
-
Abstract:
Objective To examine the role of domain Ⅱ of hepatitis C virus(HCV) 5'noncoding region (5'NCR) in its translation initiation activity. Methods The 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 pCMVNCRIuc, a firefly luciferase(Flue) eukaryotic expression plasmid regulated by HCV 5' NCR, to generate recombinant plasmid pCN1-d3, pCMVNCRIuc, pCN1-d2 (modified pCMVNCRluc with deletions of HCV 5' NCR nt1-43) and pCN1-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 Fluc gene expression. Meanwhile the Fluc mRNA levels were detected by RT-PCR. Results The recombinant plasmid was successfully constructed. The Fluc 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 pCN1-d2 and pCMVNCRluc(P 0.05). However, that of pCN1-d3 was decreased significantly(P < 0.01, compared with pCN1-d2 or pCMVNCRluc). Conclusion Structural domain Ⅱ of HCV 5' NCR plays an important role in itstranslation initiation activity.
