Evaluation on expression of the recombinant S gene of human hepatitis B virus in vitro and in vivo.
Hepatobiliary Pancreat Dis Int
; 2(2): 259-64, 2003 May.
Article
em En
| MEDLINE
| ID: mdl-14599980
ABSTRACT
OBJECTIVES:
To construct a DNA vaccine capable of expressing the S gene of hepatitis B virus (HBV) and evaluate the expression of the recombinant S gene in vitro and in vivo.METHODS:
A cloned S-X gene fragment was inserted into an eukaryote expression vector to construct a recombinant expressing plasmid pCMV-SX. The recombinant plasmid was transcribed in vitro with a T7 promoter transcription system and transfected into a human hepatoblastoma cell line HepG2. The expression of the S gene was detected by Northern blot hybridization, Western blot hybridization, and enzyme-linked immunosorbent assay (ELISA), respectively. BALB/c mice were inoculated with the recombinant plasmid, and the efficiency of DNA-based immunization in eliciting anti-HBs was evaluated by ELISA.RESULTS:
In vitro transcription of the subcloned HBV S gene was confirmed by Northern blot hybridization. The results of Western blot hybridization and ELISA showed that the S gene was expressed exactly in HepG2. In immune experiment, 2 of 10 immunized mice were shown to induce antibody against HBsAg.CONCLUSION:
The recombination and expression of the S gene can be achieved successfully in vitro. And the recombinant plasmid is able to elicit humoral immune response in mice.
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Bases de dados:
MEDLINE
Assunto principal:
Vírus da Hepatite B
/
Vacinas contra Hepatite B
/
Hepatite B
/
Antígenos de Superfície da Hepatite B
Limite:
Animals
/
Humans
/
Male
Idioma:
En
Revista:
Hepatobiliary Pancreat Dis Int
Assunto da revista:
GASTROENTEROLOGIA
Ano de publicação:
2003
Tipo de documento:
Article
País de afiliação:
China