Establishment of stable HeLa cell lines expressing enzymatically active hepatitis C virus RNA polymerase

The hepatitis C virus RNA polymerase (NS5B) is strictly required for viral replication and thus represents an attractive target for antiviral drug development. In this study, stable HeLa cell lines with an integrated NS5B gene were selected by G418 and then confirmed by genome PCR. Subsequently, tra...

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Bibliographic Details
Published in:Archives of virology 2006-02, Vol.151 (2), p.361-367
Main Authors: KONG, L. B, YE, L. B, YE, L, TIMANI, K. A, ZHENG, Y, LIAO, Q. J, LI, B. Z, GAO, B
Format: Article
Language:English
Subjects:
Biological and medical sciences
Cloning
Cytomegalovirus
Drug resistance
Fundamental and applied biological sciences. Psychology
Gene Expression
Genes
HeLa Cells
Hepacivirus - enzymology
Hepatitis C
Hepatitis C virus
Humans
Microbiology
Miscellaneous
Proteins
RNA polymerase
RNA-Dependent RNA Polymerase - biosynthesis
RNA-Dependent RNA Polymerase - genetics
RNA-Dependent RNA Polymerase - metabolism
Viral Nonstructural Proteins - biosynthesis
Viral Nonstructural Proteins - genetics
Viral Nonstructural Proteins - metabolism
Virology
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Summary:The hepatitis C virus RNA polymerase (NS5B) is strictly required for viral replication and thus represents an attractive target for antiviral drug development. In this study, stable HeLa cell lines with an integrated NS5B gene were selected by G418 and then confirmed by genome PCR. Subsequently, transcription and expression of the integrated NS5B genes were demonstrated by RT-PCR and Western blot analysis. Further analysis demonstrated enzymatic activity of the expressed NS5B polymerase. The stable HeLa cell lines should be useful for the identification of NS5B inhibitors and for studying the mechanisms of HCV replication.
ISSN:0304-8608
1432-8798
DOI:10.1007/s00705-005-0656-0