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[Transgenic mice model of human hepatitis B virus x gene]
1National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, CAMS, PUMC, Beijing 100005, China.
Objective:
To set up a human hepatitis B virus x (HBx) gene transgenic model.
Methods:
The model was set up by microinjection, the integration and expression of the founders were identified by molecular hybridization.
Results:
A 1,800 bp DNA fragment containing the human HBx promoter and genomic DNA was microinjected into the male pronucleus of the fertilized eggs of mice. Three hundred and thirty-eight eggs survived from microinjection were transplanted into the oviduct of 16 pseudopregnant female, 12 pseudopregnant mice got pregnant and gave birth to 74 pups. Genomic DNA were extracted from the tails of the pups, Southern blot hybridization was used for determination of the integration of human HBx gene. The 1,800 bp microinjected fragment was used as the probe, 17 mice (founders) were identified of the integration of human HBx gene. Total RNA were extracted from the livers of the founders, all 17 founders were identified of the expression of human HBx gene by using Northern blot hybridization.
Conclusions:
These results may provide animal models for studying on the correlation between chronic HBV infection and the development of hepatocellular carcinoma.