Abstract：Objective To construct a recombinant prokaryotic expression vector of influenza virus M2 gene(M2e)and Qβ,and to conduct immunological evaluation of Qβ-M2e virus-like particles(VLPs),to make a foundation for the development of preventive vaccines.Methods By means of molecular biology,M2e and Qβ plasmid vector were connected to construct a recombinant prokaryotic expression vector,which was identified by restriction enzyme digestion and sequencing.Then,transformed the recombinant plasmids into Escherichia coliBL21,the fusion protein Qβ-M2e was recovered and purified by density gradient centrifugation.After the animals were immunized,the proliferation of germinal center(GC)B cells was detected by immunohistochemistry and flow cytometry.Results The enzyme digestion and sequencing results were consistent with the expected.The fusion protein was observed under the electron microscope as a particle-like protein.After immunizing animals,GC B cells proliferated significantly.Conclusion The recombinant Qβ-M2e vector has been constructed successfully.And the expressed Qβ-M2e virus particles were evaluated by preliminary cytology.The vector might be able to serve as a candidate vector vaccine for influenza virus VLP vaccine,which provides information for further study of general influenza vaccine.
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