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, 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.
Kong
L B
LB
State Key Laboratory of Virology, College of Life Science, Wuhan University, Wuhan, P.R. China.
Ye
L B
LB
Ye
L
L
Timani
K A
KA
Zheng
Y
Y
Liao
Q J
QJ
Li
B Z
BZ
Gao
B
B
D015870:Gene Expression; D006367:HeLa Cells; D016174:Hepacivirus / Q000201:enzymology*; D006801:Humans; D012324:RNA-Dependent RNA Polymerase / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism*; D017361:Viral Nonstructural Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism*
D017361:Viral Nonstructural Proteins; C087036:NS-5 protein, hepatitis C virus; D012324:RNA-Dependent RNA Polymerase