【摘要】 目的 研究C57BL/6J-HBV转基因小鼠的繁育规律，并从DNA、RNA、蛋白质、病毒学角度对该品系转基因小鼠进行综合分析。方法 C57BL/6J-HBV转基因小鼠以两种不同的交配方式进行繁殖，利用PCR法对每一世代仔鼠检测其阳性率。利用RT-PCR和real-time PCR对转基因小鼠肝组织中的HBV DNA进行定性和半定量检测并对HBV基因在转基因小鼠中的转录特征进行分析。利用血清ELISA和Western Blotting方法，分析HBV病毒抗原在转基因小鼠中的表达和分泌特征。结果 两种交配方式窝产仔数和离乳数存在显著性差异(P<0.05)，互交后代小鼠的阳性率高于回交后代。4只乙型肝炎病毒转基因小鼠分别与正常C57BL/6J鼠杂交进行传代，其阳性率始终保持在40%～50%，表明C57BL/6J-Tg(A1b1HBV)44Bri/J型HBV已稳定整合表达，并能可靠地遗传给下一代。转基因小鼠的肝组织可稳定表达的病毒抗原并可分泌入血。结论 该转基因小鼠可长期稳定高表达HBV病毒抗原，可应用于乙型肝炎病毒相关研究。更多还原

【Abstract】 Objective To explore the breeding rule of C57BL/6J-HBV transgenic mice and to conduct a comprehensive study on biological characteristics of the transgenic mice from the DNA, RNA, proteins and virological level. Method C57BL/6J-HBV transgenic mice were breeded in two different ways of mating and breeding, PCR was used to detect the positive rate of transgenic in each generation. RT-PCR and real-time PCR were used to semi-qualitatively or quantitatively detect the expression of HBV DNA in transgenic mice and to explore the transcription characteristics of mRNA coded by HBV. ELISA and Western Blotting were used to HBV virus antigen expression and secretion. Results There were significant differences(P<0.05) of both litter size and weaning between two mating mode. The positive rate of reciprocal cross progeny was higher than the rate of backcross progeny. Four founder HBV positive transgenic mice were crossed with normal C57 BL/6J mice, and real-time PCR was performed to detect the HBV genome of the offspring during every passage. Analysis showed that the positive rate was always around 40-50%, indicating the HBV genome has integrated into the C57BL/6J-Tg mice’s genome, expressed stably and can transmit into the next generation reliably. Viral antigen can be stablely expressed in the transgenic mice liver tissue and secreted into the blood. Conclusion The transgenic mouse model can be of high expression model of HBV viral antigen with long-term stability and be applicted as the animal model for hepatitis B virus-related research.更多还原