乙型肝炎病毒(adr亚型)preS2-S基因转基因小鼠的建立  

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

作　　者：苏小平[1] 姚玉成[1] 訾晓渊[1] 赵书民[2] 熊俊[1] 李建秀[1] 王新民[1] 丁佳[1] 许燕[1] 余宏宇[3] 胡以平[1] 

机构地区：[1]第二军医大学基础医学部细胞生物学教研室,上海200433 [2]第二军医大学长征医院感染科 [3]长征医院病理科

出　　处：《第二军医大学学报》2003年第2期168-171,共4页Academic Journal of Second Military Medical University

基　　金：国家九五攻关项目 (TJ99-L A0 1) ;国家自然科学基金(3 9670 811) ;上海市科学技术发展基金 (9949190 3 3 )

摘　　要：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系。 方法 :通过原核显微注射法制备带有乙型肝炎病毒 (adr亚型 ) pre S2 - S基因的转基因小鼠。应用 PCR方法筛选乙型肝炎病毒 pre S2 - 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 6 9枚。选取存活受精卵 5 8枚分别植入 4只假孕小鼠 ,产仔并存活 2 3只。经尾组织 DNA PCR筛选得到 5只整合 pre S2 - 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 HBs Ag,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 ) pre S2 - 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 。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.

关 键 词：乙型肝炎病毒 MHBS 转基因小鼠 preS2-S基因 

分 类 号：R373.21[医药卫生—病原生物学]