Subgenomic hepatitis C virus replicons inducing expression of a secreted enzymatic reporter protein

Min Kyung Yi, Francis Bodola, Stanley M. Lemon

Research output: Contribution to journalArticle

89 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)197-210
Number of pages14
JournalVirology
Volume304
Issue number2
DOIs
StatePublished - 2002

Fingerprint

Replicon
Hepacivirus
Proteins
Alkaline Phosphatase
RNA
HIV Long Terminal Repeat
tat Gene Products
Human Immunodeficiency Virus Proteins
Northern Blotting
Interferons
Transfection
Real-Time Polymerase Chain Reaction
Peptide Hydrolases
Cell Line
Genes

Keywords

  • Core protein
  • Hepatitis C virus
  • Interferon
  • Internal ribosome entry site
  • Replication
  • Replicon
  • Secreted alkaline phosphatase
  • Tat

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases

Cite this

Subgenomic hepatitis C virus replicons inducing expression of a secreted enzymatic reporter protein. / Yi, Min Kyung; Bodola, Francis; Lemon, Stanley M.

In: Virology, Vol. 304, No. 2, 2002, p. 197-210.

Research output: Contribution to journalArticle

Yi, Min Kyung ; Bodola, Francis ; Lemon, Stanley M. / Subgenomic hepatitis C virus replicons inducing expression of a secreted enzymatic reporter protein. In: Virology. 2002 ; Vol. 304, No. 2. pp. 197-210.
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