Evaluation on expression of the recombinant S gene of human hepatitis B virus in vitro and in vivo.
OBJECTIVE
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.


CONCLUSIONS
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.
Yang
Rui-Yi
RY
Tropical Medicine Institute, Guangzhou University of Traditional Chines e Medicine, Guangzhou 510405, China. yangruiyi@21cn.com
Zeng
Qing-Ping
QP
Fu
Lin-Chun
LC
Chen
Zheng-Tu
ZT
D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D045744:Cell Line, Tumor; D015342:DNA Probes; D004797:Enzyme-Linked Immunosorbent Assay; D006508:Hepatitis Antibodies / Q000097:blood; D006509:Hepatitis B / Q000276:immunology / Q000517:prevention & control*; D006514:Hepatitis B Surface Antigens / Q000032:analysis / Q000235:genetics* / Q000276:immunology; D017325:Hepatitis B Vaccines / Q000235:genetics*; D006515:Hepatitis B virus / Q000235:genetics* / Q000276:immunology; D018197:Hepatoblastoma; D006801:Humans; D066298:In Vitro Techniques; D008113:Liver Neoplasms; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D010957:Plasmids / Q000235:genetics; D051381:Rats; D011994:Recombinant Proteins / Q000235:genetics
D015342:DNA Probes; D006508:Hepatitis Antibodies; D006514:Hepatitis B Surface Antigens; D017325:Hepatitis B Vaccines; D011994:Recombinant Proteins