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目的:评价乙肝转基因小鼠品系C57-TgN(HBV adr2.O)SMMU生物学特征的稳定性。方法:以F5代乙肝转基因小鼠C57-TgN(HBV adr2.O)SMMU为研究对象,采用基因组DNA PCR、血清ELISA检测、Western印迹分析、免疫组织化学、血清DNA PCR、透射电镜和H-E染色的方法分析HBV基因在转基因小鼠中的整合、表达、复制和组织学变化。结果:F1代乙肝转基因小鼠基因组中稳定整合有HBV基因,肝组织中可检测到HBsAg、HBcAg和X蛋白3种病毒蛋白,血清中HB-sAg和HBeAg的表达率分别为19.54％和3.39％,且在血清和肝组织中存在病毒DNA和病毒样颗粒;长期的病毒DNA整合、表达和复制可以引起转基因小鼠肝、肺等组织的病理性损伤。结论:乙肝转基因小鼠品系C57-TgN(HBV adr2.O:)SMMU具有基因组中稳定整合病毒DNA、血清和肝组织中有病毒蛋白表达和病毒复制的特征,并具有一定的组织学变化,作为生物医药研究的实验动物模型具有推广应用价值。
Objective: To evaluate the stability of the biological characteristics of C57-TgN (HBV adr2.O) SMMU in hepatitis B transgenic mice. Methods: The C57-TgN (HBV adr2.O) SMMU of F5 hepatitis B transgenic mice was used as the research object. Genomic DNA PCR, serum ELISA test, Western blot analysis, immunohistochemistry, serum DNA PCR, transmission electron microscopy and HE staining Methods The integration, expression, replication and histological changes of HBV gene in transgenic mice were analyzed. Results: The HBV gene was stably integrated into the genome of F1 generation hepatitis B virus, and three kinds of viral proteins of HBsAg, HBcAg and X protein were detected in liver tissues. The expression rates of HBsAg and HBeAg in serum were 19.54% and 3.39% , And there is virus DNA and virus-like particles in serum and liver tissue. Long-term integration, expression and replication of viral DNA can cause pathological damage in the liver, lung and other tissues of transgenic mice. CONCLUSION: The C57-TgN (HBV adr2.O:) SMMU strain of hepatitis B transgenic mice has the characteristics of stable integration of viral DNA in the genome, expression of viral proteins and virus replication in serum and liver tissues, and has certain histological changes as Experimental animal model of biomedical research has the promotion and application value.