Construction of HCV-producing cell model based on self-cleaving ribozyme / 中华肝脏病杂志
Chinese Journal of Hepatology
;
(12): 437-439, 2010.
Article
in Chinese
| WPRIM
| ID: wpr-326340
ABSTRACT
<p><b>OBJECTIVES</b>To construct a stable HCV-producing cell model for anti-HCV drug research.</p><p><b>METHODS</b>The HCV-ribozyme recombinant plasmid pJFH1-Rbz was constructed to generate the exact 5' and 3' ends of HCV genomic RNA by placing two self-cleaving ribozymes at both ends of the HCV JFH-1 cDNA. The plasmid was then transfected into HepG2 cells and the resultant clones were screened with G418. Subsequently, immunofluorescence and Western blot were performed to detect the expression of HCV core protein, HCV RNA level was quantitated by TaqMan real-time PCR method and HCV particles was detected by electron microscopy.</p><p><b>RESULTS</b>HCV core protein was detected in the screened cell clone, and the level of HCV RNA was up to 1000,0000 copies/ml in the culture medium. Electron microscopy showed the viral particles in the culture suspension were approximately 55 nm in diameter. IFN-treating experiment demonstrated that the HCV RNA level decreased with the increasing concentration of IFN alpha.</p><p><b>CONCLUSION</b>We constructed a stable HCV-producing cell model which can be used for anti-HCV drug research.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Plasmids
/
Virion
/
Virus Replication
/
Transfection
/
Viral Core Proteins
/
RNA, Catalytic
/
Genome, Viral
/
DNA, Complementary
/
Hepacivirus
/
Hep G2 Cells
Limits:
Humans
Language:
Chinese
Journal:
Chinese Journal of Hepatology
Year:
2010
Type:
Article
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