Expression Of Hbv Virus Receptor Sccai Annexinv Hbv Susceptible Mouse Model To Establish

The host of HBV coronavirus is restricted to humanbeing and a few kinds of primates, the lack of appropriate animal model impeded a more thorough understanding of HBV and HBV infection associated disease.Some HBV receptors already been reported,Among them,the human annexin V and HBV-BP protein ever been transfected into HepG2 and CHO cells, which resulted the replication of HBV particles in the above cells, the titers of HBV showed positive relation with the expressed protein levels of above two receptors in HepG2 cell, more interestingly, the CHO cell,which already known can not be infected by HBV, become susceptible because of the exogenous expression of human Annexin V and HBV-BP..In order to get the cheap and useful HBV animal model, the transgenic mice which express human Annexin V and HBV-BP were established.Firstly, the cDNA coding for AnnexinV and BP was amplified by PCR and then cloned into a eukaryotic expression vector PCAGGS, under the control of the CAG promoter, a composite promoter consisting of the cytomegalovirus immediate-early enhancer, which resulted the two kinds expression vector PCAGGS-AnnexinV and PCAGGS-BP.For the establishment of transgenic mice, The purified DNA fragment generated by SalI and DrdI double digestion of PCAGGS-AnnexinV and PCAGGS-BP were used for co-microinjection. transgenic founders were established and identified by PCR and Southern blot analysis, the transgenic positive rate were 27.2%. These founders were backcrossed with wild-type KM mice and set lineages. The expression of exogenous AnnexinV and BP in these transgenic mice were identified, the total RNA of mice livers were extracted and then be subjected to RT-PCR analysis. Our results indicated that the two exogenous human receptor genes were actively transcribed.. Then the total protein of liver tissue for PCAGGS transgenic mice was extracted for the western blot analysis, results showed that AnnexinV and BP had been strongly expressed in the liver tissue of transgenic mice.Until now We get the transgenic mice that express HBV functional receptor AnnexinV and BP. It will provide an useful animal model for the research of HBV infection.