[Construction of HCV-producing cell model based on self-cleaving ribozyme].
Zhonghua Gan Zang Bing Za Zhi
; 18(6): 437-9, 2010 Jun.
Article
en Zh
| MEDLINE
| ID: mdl-20587314
ABSTRACT
OBJECTIVES:
To construct a stable HCV-producing cell model for anti-HCV drug research.METHODS:
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.RESULTS:
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.CONCLUSION:
We constructed a stable HCV-producing cell model which can be used for anti-HCV drug research.
Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
Proteínas del Núcleo Viral
/
ARN Catalítico
/
Hepacivirus
Límite:
Humans
Idioma:
Zh
Revista:
Zhonghua Gan Zang Bing Za Zhi
Asunto de la revista:
GASTROENTEROLOGIA
Año:
2010
Tipo del documento:
Article
País de afiliación:
China