人乙型肝炎病毒x基因转基因小鼠模型的建立

Transgenic Mice Model of Human Hepatitis B Virus x Gene

目的 建立人乙型肝炎病毒 x(HBx)基因转基因小鼠模型。方法 用显微注射法制备转基因小鼠 ,用分子杂交法鉴定转基因小鼠 HBx的整合与表达。结果 建立了人 HBx基因转基因小鼠模型。经 Southern杂交鉴定得到 17只转基因首建鼠。自肝脏提取总 RNA进行 Northern杂交 ,结果显示 17只首建鼠肝脏中均有 HBx表达。结论 为从整体水平研究慢性乙型肝炎患者诱发肝癌的作用机制及 HBx的反式激活作用机制提供了理想的动物模型。

To set up a human hepatitis B virus x(HBx) gene transgenic model. Methods The model was set up by microinjection,the integration and expression of the founders were identified by molecular hybridization. Results A 1 800 bp DNA fragment containing the human HBx promoter and genomic DNA was microinjected into the male pronucleus of the fertilized eggs of mice.Three hundred and thirty-eight eggs survived from microinjection were transplanted into the oviduct of 16 pseudopregnant female,12 pseudopregnant mice got pregnant and gave birth to 74 pups.Genomic DNA were extracted from the tails of the pups, Southern blot hybridization was used for determination of the integration of human HBx gene.The 1 800 bp microinjected fragment was used as the probe,17 mice(founders)were identified of the integration of human HBx gene.Total RNA were extracted from the livers of the founders,all 17 founders were identified of the expression of human HBx gene by using Northern blot hybridization. Conclusions These results may provide animal models for studying on the correlation between chronic HBV infection and the development of hepatocellular carcinoma.