微环载体结合水动力转染技术制备乙肝病毒受体小鼠模型  

Establishment of Hepatitis B Virus Receptor(NTCP) Mouse Model Using Minicircle DNA with Hydrodynamic Injection

作　　者：李秀梅[1] 刘光泽[1] 谢勇 佟明华[1] 孔祥平[1] 

机构地区：[1]解放军第458医院传染病防治中心全军转基因动物重点实验室,广州510600

出　　处：《实验动物科学》2017年第6期26-29,33,共5页Laboratory Animal Science

基　　金：广东省科技基础条件建设领域项目(No.2014A030304054);广东省科技计划项目(No.2014A020212639);广州市科技计划项目(No.201510010004),广州市科技计划项目(No.201604046020)

摘　　要：目的制备乙肝病毒受体钠离子/牛磺胆酸共转运多肽(Na+/taurocholate co-transporting polypeptide,NTCP)小鼠基因转染模型,为乙肝的防治研究提供动物模型。方法制备携带人NTCP真核表达框的亲本质粒ZY781-CMV-NTCP及微环DNA p MC-CMV-h NTCP,采用水动力转染技术将微环DNA p MC-CMV-h NTCP经尾静脉注入ICR小鼠体内,采用PCR和免疫组化方法检测NTCP基因在小鼠体内的表达情况。结果成功构建了携带人NTCP基因的微环载体。PCR检测表明人NTCP基因成功转入小鼠肝组织内,同时肝组织免疫组化检测表明h NTCP可以在小鼠的肝脏内表达。结论成功制备出乙肝病毒受体(NTCP)基因转染小鼠模型。Objective The aim of this study was develop a hNTCP-transfected mouse model. Method Parental plasmid harboring human NTCP eukaryotic expression cassette ZY781-CMV-NTCP and minicircle DNA pMC-CMV- hNTCP were prepared using molecular cloning technique. Then the minicircl DNA pMC-CMV-hNTCP was injected into ICR mice via tail vein by hydrodynamic Injection. The expression of NTCP gene in mice was detected by PCR and immunohistochemistry. Result The minicircle DNA carrying human NTCP gene pMC-CMV-hNTCP was successfully constructed. PCR analysis showed that human NTCP gene was successfully transferred into mouse liver tissue. And Immunohistochemistry of liver tissue suggested that hNTCP could be expressed in the liver of the gene- transfected mice. Conclusion The mouse model of hepatitis B virus receptor （NTCP） gene transfection was successfully established.

关 键 词：钠离子/牛磺胆酸共转运多肽 微环载体DNA 水动力转染技术 

分 类 号：R-332[医药卫生]