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乙型肝炎病毒(adr亚型)转基因小鼠的研究 |
胡以平,王肖鹏,郝光荣,孙伟,王新民,戴德顺,余宏宇,李建秀,胡卫江 |
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摘要: |
目的:制备含有2.0拷贝乙型肝炎病毒(HBV,adr亚型)基因组的转基因小鼠,为HBV相关医学问题的研究准备实验动物模型。方法:应用受精卵原核显微注射的方法,产生HBV转基因小鼠。用PCR, Southern-blotting杂交、放射免疫、免疫组织化学以及亚显微结构分析等方法,研究外源基因在转基因小鼠体内的整合、表达以及复制。应用常规病理切片及血清生物化学方法分析转基因小鼠的病理学改变。结果:得到了4只整合有HBV基因组的建立者小鼠,并证明HBV基因在转基因小鼠体内可以稳定遗传,在肝、肾组织内可特异性表达、复制,并在肝细胞内发现了Dane颗粒。转基因小鼠的肝、肾组织未见明显的病理学改变。结论:获得了HBV的转基因小鼠,其病毒基因可以表达,并有病毒颗粒形成。该小鼠对于HBV的各个基因产物是免疫耐受的,其基本生物学特性与人类的HBV携带者特征相似。 |
关键词: 乙型肝炎病毒 转基因小鼠 Dane颗粒 |
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Study of transgenic mice harbouring hepatitis B virus genomes (adr subtype) |
胡以平,王肖鹏,郝光荣,孙伟,王新民,戴德顺,余宏宇,李建秀,胡卫江 |
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Abstract: |
Objective:To prepare the transgenic mice harbouring 2 copies genome of hepatitis B virus(HBV) as a useful model for the studies of medical issues related to HBV. Methods:The HBV(adr subtype)transgenic mice were prepared by microinjection of complate 2.0 copy HBV genome into pronuclei fertilized eggs. The PCR assay, Southern-blotting, radioimmunity, immunohischemistry and ELISA methods were used to detect the HBV intergration,expression and replication in the transgenic mice. The dane particles were observed in the liver. The pathological change was tested with ALT,AST in the serum and the normal histological slides. Results:The 4 transgenic mice were obtained and the HBV DNA can be expressed and replicated in the transgenic mice. HBV genomic DNA in mouse line could be transmitted to next generations. No evidence suggested the existence of pathogenesis. Conclusion:The HBV transgenic mice were obtained and the virus genes can be expressed, the Dane particles can be formed in the liver.The mice are tolerant to HBV gene products. The transgenic mice are similar to human chronic HBV carrier. The structure and the function of the transgenic mice are stable. |
Key words: hepatitis B virus transgenic mouse Dane particle |