Establishment of C genotype hepatitis B virus genome transgenic mice

AIM:To generate HBV (C) genome transgenic mice to provide suitable animal models for the related researches in China. METHODS:The HBV transgenic mice were generated by microinjection of 1.3 copies HBV genome into the pronucleus of FVB/N zygotes. The PCR assay,ELISA,RT-PCR and immunohistochemical methods were used to detect the HBV integration,replication and expression in the transgenic mice. RESULTS:In this study,2840 zygotes were injected,the survived 2256 zygotes after microinjection were reimplanted into the oviducts of 85 psudopregnant KM female mice. After birth,42 female mice were pregnant and 168 F0 offsprings were born. PCR analysis indicated that 14 out of 168 F0 were integrated with HBV genome. With ELISA,5 of them expressed HBsAg in serum. And two mice were positive for HBV DNA replication in serum with 3.12×105copies/L and 1.98×105 copies/L respectively. Then the transgenic mice were mated to normal mice to establish transgenic mouse strain. 61 F1 offsprings were born. Further study demonstrated that 5 of them were integrated with HBV genome by PCR,4 of them were serum HBsAg positive by ELISA and 2 of them with the intracellular distribution of HBsAg in hepatocytes by immunohistochemistry. Both serum HBeAg and liver HBcAg immunohistochemistry were negative. CONCLUSION:The established HBV (C) genome could be not only replicated and expressed in transgenic mice but also inherited to the next generations.