Abstract:

Objective To construct DNA vaccine encoding ubiquitinated hepatitis B virus core antigen (HbcAg) against hepatitis B virus. Methods Gene encoding mutant ubiquitin (Ub) was amplified by RT-PCR from the liver of BALB/c mice, and HBcAg gene fragment was amplified from the plasmid pADR containing the genome of hepatitis B virus by PCR. Ub-HBcAg fusion gene was spliced by recombinant PCR. After purification and collection, Ub-HBcAg was connected with pUCm-T vector to construct the recombinant plasmid pUCm-Ub-HBcAg. After identification of pUCm-Ub-HBcAg by enzyme digestion analysis, Ub-HBcAg was inserted into eukaryotic expression plasmid pcDNA3.1(-) to construct the recombinant eukaryotic expression plasmid Ub-HBcAg-pcDNA3.1(-), which was identified through sequencing. Results Gene encoding HbcAg and gene encoding mutant Ub were amplified. Ub-HBcAg fusion gene eukaryotic expression plasmid Ub-HBcAg-pcDNA3.1(-) was confirmed through sequencing. Fusion gene Ub-HBcAg was inserted into the right direction of plasmid pcDNA3.1(-), the mutant gene fragment was confirmed to accord to expectation, and there was no other difference between the target gene and published gene in GenBank. Conclusion DNA vaccine encoding ubiquitinated HBcAg against hepatitis B virus has been successfully constructed.

Key words: hepatitis B virus core antigen, DNA vaccine, ubiquitin, fusion gene