Effect of viral core protein mutant on HBV encapsidation
- VernacularTitle:乙型肝炎病毒核心蛋白突变体干扰HBV包装的实验研究
- Author:
Wenbin WU
;
Wei PAN
;
Xinyu WEN
- Publication Type:Journal Article
- Keywords:
Hepatitis B virus;
Dominant negative mutant;
Gene therapy
- From:
Chinese Journal of Infectious Diseases
2001;0(05):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To observe a recombinant mutant of HBV core protein for dominant negative gene therapy against HBV encapsidation in vitro. Methods C gene and S gene of HBV were acquired through PCR and subcloned into pGEM T to construct pGEM T C and pGEM T S respectively. After digestion and ligation of these two plasmids, pGEM T CS was constructed. The cloned gene was inserted into pcDNA3.1 + to construct pcDNA3.1 + CS, which was identified by DNA sequencing. The recombinant plasmids were transformed into HepG2 cells, and screened with G418. The resistant HepG2 cell clones were chosen to test the expression of core surface protein by RT PCR, and the expressing HepG2 clones were cultured with 10% HBV DNA positive human serum for 72 hours. The intracellular HBV particles were extracted and the DNA was subjected to dot hybridization. Results The analysis showed that the HepG2 cells expressing mutant C protein had capabilities to resist HBV invasion in varied degrees. The mutant C protein had a dominant negative role in the encapsidation of HBV compared with the naive part of core protein. Conclusions The production of recombinant mutant core protein has a potential value for gene therapy against HBV infection.
