Abstract: Objective To construct a multi?gene recombinant pcDNA3?HBsAg?p30?ROP2 expression vector and identify it preliminarily. Methods According to recombinant pcDNA3?p30?ROP2 restriction sites，HBV HBsAg gene sequences of primers were designed and synthesized to amplify target fragment，and then cloned into pcDNA3?HbsAg?p30?ROP2 expression vector. Af? ter sequencing，it was identified finally by restriction enzyme digestion and other molecular biology techniques. Results HBV HBsAg gene segment was amplified by PCR and the multi?gene recombinant pcDNA3?HBsAg?p30?ROP2 expression vector was constructed and identified to be correct as theoretical values. The PCR and restriction enzyme digestion results showed that HBsAg and p30?ROP2 gene in recombinant plasmid were confirmed by DNA sequencing. Conclusion The multi?gene recombinant pcD? NA3?HBsAg?p30?ROP2 expression vector is successfully constructed.

Key words: Toxoplasma gondii；Surface antigen 1（p30）； Rhoptry protein 2（ROP2）； HBsAg；Gene recombination

摘要： 目的 目的 构建pcDNA3?HBsAg?p30?ROP2多基因重组表达载体， 并对其进行初步鉴定。方法 方法 根据重组体pcDNA3? p30?ROP2酶切位点和乙型肝炎表面抗原 （HBsAg） 基因序列等因素设计合成引物， 扩增HBsAg目的基因片段， 再应用酶切、 连接等分子生物学技术将HBsAg目的基因克隆至pcDNA3?p30?ROP2表达载体中。应用聚合酶链反应 （PCR） 初筛， 再采用 酶切、 测序等技术对构建的重组表达载体pcDNA3?HBsAg?p30?ROP2进行鉴定。 结果 结果 PCR扩增出HBsAg基因片段， 构建 了pcDNA3?HBsAg?p30?ROP2多基因真核表达载体。PCR与酶切结果显示， 该基因片段大小均与理论值相符； 测序结果显 示该重组表达载体包含了p30?ROP2和HBsAg目的基因的完整序列。 结论 结论 成功构建了多基因重组表达载体pcDNA3? HBsAg?p30?ROP2， 为进一步研究多基因核酸疫苗奠定了基础。

关键词: 弓形虫； 表面抗原1 （p30）； 棒状体分泌抗原2 （ROP2）； 乙肝表面抗原； 基因重组