高表达乙型肝炎病毒转基因小鼠的制备

目的 制备高表达乙型肝炎病毒（ＨＢＶ）转基因小鼠，为乙型肝炎研究提供较理想动物模型。方法 构建、筛选高表达ＨＢＶ质粒载体。用显微注射法导入昆明鼠受精卵雄性原核，用ＰＣＲ、Ｓｏｕｔｈｅｒｎ杂交、放射免疫、免疫组织化学等方法分析ＨＢＶ基因在小鼠体内的整合、表达情况。结果 显微注射１４５６枚卵，产仔１１８只，整合阳性２５只，１１只血清ＨＢＶ ＤＮＡ、ＨＢｓＡｇ阳性，其中２只血清中ＨＢｓＡｇ含量高于１０００ｎｇ／ｍｌ，且子代鼠中达到９００ｎｇ／ｍｌ。肝组织免疫组化分析发现有ＨＢｓＡｇ、ＨＢｃＡｇ表达，未见明显病理学改变。结论 已初步获得高表达ＨＢＶ转基因小鼠。

High-level hepatitis B virus expression in transgenic mice

Objective To prepare hepatitis B virus (HBV) transgenic mice expressing high level HBV that can be used as a model for study of human hepatitis B. Methods HBV plasmid of high-level expression was prepared and selected, introduced into the zygote pronuclei by microinjection. The PCR assay, southern-blotting, radioimmunoassay and im-munohistochemistry were used to detect the HBV intergration and expression. Results A total of 1456 zygotes were injected, 118 resultant mice were obtained. Twenty-five mice were HBV DNA positive in the tail tissue and 11 were HBV DNA and HBsAg positive in the serum. Two mice had over 1000ng/ml. HBsAg and their G1mice had 900ng/ml HBsAg. HBsAg and HBcAg expression were observed in liver tissue by immunohistochemistry but no evidence suggested the existence of pathogenesis. Conclusions HBV transgenic mice of high-level expression had been preliminarily established.