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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 ,这一品系的建立为中蛋白的相关研究提供了技术平台
Objective: To establish a transgenic mouse strain that expresses the envelope protein of hepatitis B virus (adr subtype). Methods: Transgenic mice with Hepatitis B virus pread - S gene were prepared by pronuclear microinjection. The founder mice of hepatitis B virus preS2 - S gene were screened by PCR. The expression of hepatitis B virus protein in these mice was analyzed by immunohistochemistry. The expression of hepatitis B virus Positive transgenic mice were crossed with normal male mice for subculturing and their offspring mice were detected by PCR. Results: A total of 69 fertilized eggs were injected. Fifty surviving fertilized eggs were selected and implanted into four pseudopregnant mice, respectively, and survived 23. Five founder S2 cells pre S2 - S gene were screened by tail PCR, and HBs Ag was expressed in the liver of 3 of them by immunohistochemical staining, and then the positive mice with stronger expression were conserved. Conclusion: The established transgenic strain of pre S2 - S gene of hepatitis B virus (adr subtype) has the biological characteristics of expressing the protein of hepatitis B virus. The establishment of this line provides a technical platform for the related research of mesoderm