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目的:构建HBV表面抗原preS2 S 基因表达质粒,并探讨其在小鼠体内的表达及诱导体液免疫应答的能力。方法:采用PCR方法,以PBR322 HBV2.0(adr亚型)质粒DNA为模板获得HBV preS2 S 基因,并将其重组进入pcDNA3.0表达载体中,转染7721细胞系进行稳定表达;以此重组质粒免疫小鼠,ELISA方法检测免疫小鼠抗HBs抗体浓度。结果:构建了HBV preS2 S 基因的表达质粒pcDNAS2 S,该表达质粒可在7721 细胞中稳定高效表达;免疫接种小鼠2 周后抗HBs抗体浓度明显升高,接种后第4周布比卡因处理组小鼠抗HBs抗体的浓度达到峰值161.4 mIU/ml,布比卡因非处理组第5周抗HBs抗体的浓度可达133.7 mIU/ml。结论:所构建的pcDNAS2 S表达质粒能有效表达并诱导小鼠体液免疫应答。
OBJECTIVE: To construct the HBV preS2 S gene expression plasmid and investigate its expression in mice and its ability to induce humoral immune response. Methods: HBV preS2 S gene was obtained by PCR using the plasmid DNA of PBR322 HBV 2.0 (adr subtype) as a template and recombined into the pcDNA3.0 expression vector and transfected into the 7721 cell line for stable expression. Recombinant Mice were immunized with plasmid and the concentration of anti-HBs antibody in immunized mice was detected by ELISA. Results: The expression plasmid pcDNAS2 S of HBV preS2 S gene was constructed and expressed stably and efficiently in 7721 cells. After 2 weeks of immunization, the concentration of anti-HBs antibody was significantly increased. After 4 weeks of vaccination, The concentration of anti-HBs antibody peaked at 161.4 mIU / ml, and the anti-HBs antibody concentration at week 5 in non-treated bupivacaine group reached 133.7 mIU / ml. Conclusion: The constructed pcDNAS2 S expression plasmid can effectively express and induce humoral immune response in mice.