HBV转基因在C57-TgN(HBVadr2.0)SMMU3小鼠基因组中的整合分析

目的:分析HBV转基因在乙肝转基因小鼠C57 TgN(HBVadr2.0)SMMU"3号"品系基因组中的整合特征.方法:以F6～F17代乙肝转基因小鼠C57 TgN(HBVadr2.0)SMMU为研究对象,采用基因组DNA PCR、Southern印迹和DNA测序的方法,分析HBV转基因在该品系转基因小鼠基因组中的整合特征.结果:PCR分析显示,F6～F17代乙肝转基因小鼠基因组中均已稳定整合了相同拷贝数的HBV转基因,整合的HBV DNA可通过生殖系在世代间稳定遗传.整合在转基因小鼠基因组中的HBV DNA含有HBVpres、s、c、x基因,Southern印迹证实HBV转基因含有全长HBV基因组DNA,DNA测序结果表明,该品系转基因小鼠所整合的转基因为adr亚型的HBV DNA.结论:C57-TgN(HBVadr2.0)SMMU"3号"品系转基因小鼠基因组中均整合有adr亚型的全长HBV基因组DNA,从DNA水平证实该转基因小鼠品系已具备了乙肝实验动物模型的基本条件.

Integration analysis of HBV transgene in genome of C57-TgN(HBVadr2.0)SMMU3 transgenic mice

Objective:To analyze the characters of HBV DNA integration in the transgenic mice genome of C57-TgN(HBVadr2.0)SMMU3 strain. Methods: F6-F17 C57-TgN(HBVadr2.0) SMMU3 transgenic mice were used.PCR, Southern blotting and DNA sequencing were used to analyze the characters of hepatitis B virus transgene integrated in F6-F17 transgenic mice genomic DNA. Results: The same copies of hepatitis B virus genomic DNA was stably integrated into the genome of F6-F17 C57-TgN(HBVadr2.0)SMMU3 transgenic mice and genetically transmitted from parents to offspring. The integrated transgene contained the complete HBV genomic DNA including HBV pres, s, c and x gene. DNA sequencing indicated that the HBV transgene was adr subtype HBV DNA. Conclusion:The full-long HBV genomic DNA of adr subtype has been integrated in the genome of the C57-TgN(HBVadr2.0)SMMU3 line transgenic mice.