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目的制备高表达乙型肝炎病毒(HBV)转基因小鼠,为乙型肝炎研究提供较理想动物模型。方法构建、筛选高表达HBV质粒载体,用显微注射法导人昆明鼠受精卵雄性原核,用PCR、Southern杂交、放射免疫、免疫组织化学等方法分析HBV基因在小鼠体内的整合、表达情况。结果显微注射1456枚卵,产仔118只,整合阳性25只,11只血清HBV DNA、HBsAg阳性,其中2只血清中HBsAg含量高于1000ng/ml,且子代鼠中达到900ng/ml。肝组织免疫组化分析发现有HBsAg、HBcAg表达,未见明显病理学改变。结论已初步获得高表达HBV转基因小鼠。
Objective To prepare highly expressed hepatitis B virus (HBV) transgenic mice and provide an ideal animal model for the study of hepatitis B. Methods The HBV vector with high level of expression was screened, and the prokaryotic nuclei of Kunming mouse fertilized eggs were induced by microinjection. The integration and expression of HBV gene in mice were analyzed by PCR, Southern blot, radioimmunoassay and immunohistochemistry . RESULTS: A total of 1456 eggs were injected microinjected, and 118 were born and 25 were positive for integration. Serum HBV DNA and HBsAg were positive in 11 serum samples. Serum HBsAg levels were higher than 1000ng / ml in 2 serums and 900ng / mL in offspring rats. Immunohistochemical analysis of liver tissue found HBsAg, HBcAg expression, no significant pathological changes. Conclusion Highly expressed HBV transgenic mice have been obtained initially.