流体动力学法表达乙肝病毒X蛋白小鼠模型的建立

目的建立表达乙肝病毒X蛋白的小鼠模型。方法实验动物分成模型组和对照组,模型组利用已构建的含有HBX基因的真核表达质粒pcDNA3.1（＋）-HBX,对照组以等量生理盐水代替,采用流体动力学法将质粒经尾静脉高压注入小鼠体内,24h后取小鼠肝组织,行免疫荧光、RT-PCR和Western blot法从不同水平检测HBX在小鼠肝组织内的表达情况。结果模型组小鼠RT-PCR显示肝组织内有HBX mRNA的存在,免疫荧光和Western blot检测均有HBX蛋白的表达;对照组小鼠则无HBX表达。结论成功建立了表达乙肝病毒X蛋白小鼠模型,为进一步探讨HBX蛋白在动物体内的生物学作用提供实验基础。

Establishment of Mouse Model Harboring Hepatitis B Virus X Gene by the Means of Hydrodynamic Injection

Objective To establish a mouse model haboring hepatitis B virus X gene. Methods The immunofluoreseency, Western blotting and RT-PCR were performed on mouse liver tissue obtained from 4 experimens（ pcDNA3.1 （ ＋ ）-HBX） and 1 normal control（NS）, 24 h after delivery the HBX gene eukaryon expression vector peDNA3, 1（ ＋ ）-HBX into mouse by the means of hydrodynamic injection. Results HBX mRNA was detected in transfected mouse liver tissue by RT-PCR, expression of HBX protein was identified through immunofluoresceney and Western blot, while no expression of HBX was detected in the normal control. Conclusion The mouse model haboring hepatitis B virus X gene is established successfully, and the result provide a base for studying the biological ruction of HBX gene advanced.