Construction of recombinant adenovirus hepatitis B virus1.3 and its expression

Abstract:

Objective：To construct the recombinant adenovirus vector of hepatitis B virus（HBV） and to provide a more effective and convenient method for HBV-relative research. Methods：HBV 1.3 fold genome was subcloned to shuttle vector pAdTrack-TO4，then pAdTrack-TO4-HBV1.3 was linearized by PmeⅠ and transfected into BJ5183 cells containing pAd-Easy1. The confirmed recombi－nant plasmid with PacⅠ restriction endonuclease was linearized and transfected into HEK-293 cells to generate recombinant aden－oviruses containing HBV1.3 fold genome. Efficiency of the infection was detected by fluorescence microscope and expression of HBV proteins was tested by ELISA and Western blot. Intravenous recombinant adenovirus HBV1.3 was injected into C57BL mouse and changes of HBs expression in mouse were observed. Results：Cellular efficiency of recombinant adenovirus HBV1.3 infection could reach above 95%. Expression of HBs and HBe in experimental group was much higher than that in control group according to ELISA test. High-level HBs expression was detected by Western blot after recombinant adenovirus HBV1.3 infection. Animal experiment showed that after being injected with the recombinant adenovirus HBV1.3，HBs began to express on the 3rd d，peaked on the 5th d and declined gradually on the 7th d. Conclusion：Recombinant adenovirus with high infective efficiency is constructed and can express HBV easily，providing a much more efficiency method for HBV-relative study.