Elsevier

Virology

Volume 304, Issue 2, 20 December 2002, Pages 197-210
Virology

Regular Article
Subgenomic Hepatitis C Virus Replicons Inducing Expression of a Secreted Enzymatic Reporter Protein

https://doi.org/10.1006/viro.2002.1652Get rights and content
Under an Elsevier user license
open archive

Abstract

We constructed dicistronic, subgenomic hepatitis C virus (HCV) replicons in which the sequence encoding the human immunodeficiency virus (HIV) tat protein was placed in the upstream cistron, between the HCV 5′NTR and a picornaviral 2A proteinase sequence fused to the selectable marker Neo. Stably transformed Huh7 cells expressing secreted alkaline phosphatase (SEAP) under transcriptional control of the HIV LTR promoter actively secreted SEAP following transfection with these replicon RNAs. Extracellular SEAP activity correlated closely with intracellular HCV RNA levels, as determined by Northern blotting and real-time RT-PCR analysis. These RNAs replicated efficiently despite the absence of core-protein-coding sequence downstream of the HCV IRES. The replication efficiency of replicons derived from the HCV-N strain of HCV was significantly greater than those derived from Con1 in transiently transfected cells. Using this reporter system, we have demonstrated significant differences in the response to interferon α-2b in cell lines containing replicons derived from these two strains of HCV.

Keywords

hepatitis C virus
replication
internal ribosome entry site
core protein
replicon
interferon
tat
secreted alkaline phosphatase

Cited by (0)

1

To whom correspondence and reprint requests should be addressed. Fax: 1-409-772-9598. E-mail: [email protected].