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目的 :构建乙肝表面抗原 DNA疫苗 ,并探讨其在实验动物体内整合到宿主细胞基因组上的可能性。 方法 :采用PCR法扩增乙肝病毒的 S基因片段后装入 p VAX载体 ,并以此为候选疫苗免疫 (BAL B/c)小鼠 ,在不同时间、不同部位取其组织抽提基因组 DNA,用 PCR检测 DNA整合情况。结果 :构建了乙肝核酸疫苗 p VAX- HBs Ag。免疫接种该疫苗第 3周时在小鼠肌肉、肝脏、脾脏、胸腺均有不同量的游离质粒存在 ;第 6周除在股四头肌可检测到外 ,其余组织中均为阴性 ;第 12、18周时检测所有组织中均为阴性。 结论 :构建的 p VAX- HBs Ag DNA疫苗在小鼠体内无染色体整合。
Objective: To construct hepatitis B surface antigen DNA vaccine and explore the possibility of its integration into the genome of host cells in experimental animals. Methods: The S gene fragment of hepatitis B virus was amplified by PCR and inserted into p VAX vector. BALB / c mice were immunized with different doses of genomic DNA at different time and place. DNA integration was detected by PCR. Results: HBV DNA vaccine p VAX-HBs Ag was constructed. Immunization The vaccines had different amounts of free plasmids in the muscle, liver, spleen and thymus in the 3rd week of the vaccine. In the 6th week, except the quadriceps, all the other tissues were negative; , All tissues tested negative at 18 weeks. CONCLUSION: The constructed p VAX-HBs Ag DNA vaccine has no chromosomal integration in mice.