| Hapatitis core antigen (HBcAg) can be used as a vector vaccine platform because of its ability to form virus-like particles (VLP) activating humoral immunity and cellular immunity. In this study, ZsGreen contains 231AA macromolecules protein, as an antigen, was inserted into major epitope of HBcAg which is capable to present heterologous protein at the surface of the VLP. The gene of ZsGreen was fused with HBcAg and cloned into pET28a to generate a recombinant plasmid. The recombinant plasmid was transformed into E.coli BL21(DE3), after optimized the fermentation of expression conditions, recombinant fusion protein (Zscore) expression in E.coli is more than 20%. After cell homogeneous,centrifugal clarification,ammonium sulfate precipitation,cation column and molecular sieve chromatography, the purity of recombinant fusion protein is greater than 80%. Structural analysis and immunological experiments of the fusion protein were performed systemically. The relevant results confirmed that (1) the fusion protein was assembled into VLP structure, and (2) the fused ZsGreen presenting at the surface of the particle of recombinant protein could activating a higher immunogenicity. All of the results demonstrate that HBcAg can be a foreign protein expression vector vaccine platform. This study set up the foundation platform for further investigation and development of vaccine.
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