%0 Journal Article
%T Study on a Novel Hepatitis B Virus Multi-epitope DNA Vaccine with siRNA,hIL-12 Coexpressing<br>共表达siRNA和hIL-12的新型乙肝多表位DNA疫苗的研究
%A HUANG Jing-xian
%A CAO Yi-cheng
%A DU Zheng-ping
%A TAO Chang-li
%A YANG Hua-qiang
%A <br>黄镜贤
%A 曹以诚
%A 杜正平
%A 陶嫦立
%A 杨化强
%J 中国生物工程杂志
%D 2008
%I 
%X Objective To construct a DNA vaccine co-expressing the HBV compound multi-epitope antigen gene, the hIL-12 and the anti-HBV siRNA genes, and to express this DNA vaccine in HepG2 cells. Methods The HBV multi-epitope antigen gene was designed and synthesized before it was fused with enhanced green fluorescent protein(EGFP) gene, and cloned into the multi-clone site(MCS) of the eukaryotic expression vector pVAX1. The expressinig units of hIL-12 and siRNA were cloned into the BspH I and Mlu I site of pVAX1 respectively. Then the recombinant plasmid pVAX1-siHBV-HB-EGFP-hIL12 was transiently transfected HepG2 cells. The expression of HBV compound multi-epitope gene was observed through EGFP report gene. The expression of hIL-12 was analyzed by ELISA and the effects of anti-HBV siRNA was confirmed with rtPCR . Results The analysis of enzyme digestion and sequencing both demonstrated that the trible-expressing HBV DNA vaccine has been constructed successfully. The green fluorescent image was detected in the transfected cells which could confirm the expression of the multi-epitope antigen gene. The amount of hIL-12 secretion was 1289pg/ml in supernatant at 48h after transfection and 1712pg/ml at 72h after transfection. The mRNA amount of HBV S gene, which was the siRNA target, had been obviously knockdown. Conclusion The DNA vaccine co-expressing the HBV compound multi-epitope antigen gene, the hIL-12 and the siRNA genes was constructed and transiently expressed in HepG2 cells, and siRNA had shown us a good anti-HBV effect. It laid a foundation of further study on anti-HBV effect of the new DNA vaccine.
%K Hepatitis B virus
%K DNA vaccine
%K multi-epitope gene
%K hIL-12
%K siRNA<br>乙型肝炎
%K DNA疫苗
%K 多表位抗原
%K hIL-12
%K siRNA
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=951380788B355DEA7D75520FA3B199C9&aid=49FEF2EB6EB0F2A0FA79FCFAE386F425&yid=67289AFF6305E306&vid=D3E34374A0D77D7F&iid=5D311CA918CA9A03&sid=933658645952ED9F&eid=ECE8E54D6034F642&journal_id=1671-8135&journal_name=中国生物工程杂志&referenced_num=0&reference_num=18