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目的建立一种水动力注射将外源基因导入树鼩肝脏的方法,并用此方法建立HBV树鼩模型。方法用萤火虫荧光素酶(firefly luciferase,Fluc)和β-半乳糖苷酶(β-galactosidase,β-Gal)作为报告基因,通过树鼩大隐静脉将报告基因用水动力注射方法导入树鼩肝脏,活体成像和β-Gal染色观察水动力转染效果。将pHBV1.2质粒用水动力注射方法转入树鼩肝脏,检测树鼩血清中ALT、实时荧光定量检测血清中HBV DNA,ELISA检测血清中HBsAg、HBsAb。结果通过大隐静脉水动力注射,报告基因能够在肝脏特异性表达,转染HBV1.2质粒后树鼩血清中能够检测到相关阳性指标。结论大隐静脉水动力注射法能够作为树鼩肝脏外源基因导入的一种方法,并用此方法初步探索了树鼩的HBV模型。
Objective To establish a method of hydrodynamic injection of exogenous genes into the liver of tree shrews, and use this method to establish a model of HBV tree shrews. Methods Firefly luciferase (Fluc) and β-galactosidase (β-Gal) were used as reporter genes and the reporter genes were introduced into the liver of tree shrews by means of hydrodynamic injection through the saphenous vein of tree shrews. Live imaging and β-Gal staining were used to observe the effect of hydrodynamic transfection. The pHBV1.2 plasmid was transferred into the liver of tree shrew by hydrodynamic injection, the ALT in serum of tree shrews was detected, the serum HBV DNA was detected by real-time fluorescence quantitative detection, and the serum HBsAg and HBsAb were detected by ELISA. Results Through the hydrodynamic injection of saphenous vein, the reporter gene was specifically expressed in the liver. After the transfection of HBV1.2 plasmid, the positive correlation could be detected in the tree shrew serum. Conclusions Hydrodynamic injection of saphenous vein can be used as a method of introduction of exogenous genes in tree shrew liver. The HBV model of tree shrew was initially explored by this method.