含CpG motifs的乙肝表面抗原载体的构建及表达(英文)

Construction and Expression of Vector Containing Hepatitis B Virus Surface Antigen Gene and CpG Motifs

构建了编码乙肝表面抗原中蛋白的卡那霉素抗性真核表达质粒 pVAX_CpGS2 S ,其中通过PCR克隆了若干个CpGmotifs到质粒载体中 .经酶切鉴定和测序正确后将重组质粒体外转染COS7细胞 ,ELISA法检测上清液中的HBsAg呈阳性 ,说明重组质粒 pVAX_CpGS2 S在体外能够有效表达HBsAg ,为探讨CpGmotifs克隆入DNA疫苗载体后对体内免疫反应的影响打下基础 .

Recombinant eukaryotic expression plasmid pVAX_CpGS 2S with kanamycin resistance was constructed containing HBV surface antigen M protein encoding gene, with CpG motifs being cloned into its bone structure by PCR method. After identification by restriction enzyme digestion analysis and DNA sequencing, the plasmid was then used to transfect COS7 cells, ELISA assay for HBsAg in the supernatant was positive. The results show that recombinant plasmid can produce HBsAg effectively in vitro. This work forms the foundation for further immune responses research in vivo.