乙型肝炎病毒(adr亚型)preS2-S基因转基因小鼠的建立

目的：建立可表达乙型肝炎病毒（adr亚型）包膜中蛋白的转基因小鼠品质。方法：通过原核显微注射法制备带有乙型肝炎病毒（adr亚型）preS2-S基因的转基因小鼠。应用PCR方法筛选乙型肝炎病毒preS2-S基因转基因首建者（founder）小鼠，再以免疫组织化学法分析乙型肝炎病毒蛋白在这些小鼠中的表达特性，PCR和免疫组化均阳性的转基因小鼠与正常同系异性小鼠交配用于传代培育，并用PCR法检测其子代小鼠。结果：共注射受精卵69枚。选取存活受精卵58枚分别植入4只假孕小鼠，产仔并存活23只。经尾组织DNA PCR筛选得到5只整合preS2-S基因的founder小鼠，免疫组织化学法发现其中3只小鼠的肝脏中表达HBsAg，进而对其中表达较强的阳性鼠进行保种。结论：所建立的乙型肝炎病毒（adr亚型）preS2-S基因转基因小鼠品系具有表达乙肝病毒中蛋白的生物学特性，这一品系的建立为中蛋的相关研究提供了技术平台。

Generation of transgenic mice harbouring hepatitis B virus(adr subtype) preS2-S gene

Objective:To establish the transgenic mice model expressing the middle surface protein of hepatitis B virus (HBV) (adr subtype). Methods: The expression vectors pcDNA3 S2 S containing CMV promoter and HBV preS2 S gene were microinjected into the pronucleus of fertilized eggs to generate transgenic mice, the pups were evaluated by polymerase chain reaction (PCR) at genomic DNA level. Expression of preS2 S gene in transgenic mice were confirmed by immunohistochemistry. The transgenic mice were mated with normal mice to establish transgenic mice strains. Results:Totally 58 effective eggs of 69 fertilized eggs were implanted into oviducts of 4 pseudopregnant recipients. There were 23 pups were born and survived. Five of them were identified to have the preS2 S gene in their genomic DNA by PCR using a set of specific primers. Three were detected to express of the HBsAg in their liver by immunohistochemistry. One transgenic mouse was mated with normal mouse to establish transgenic mice strains. Conclusion:The transgene integrated in the founder can be transmitted to the subsequent generations. This transgenic mouse line is useful to study the function of the MHBs protein.