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• By utilizing bioinformatics techniques, we have identified conserved regions within the m genes of HTNV and SEOV. These regions have been utilized to design a universal subunit protein vaccine for HFRS. • The transmembrane domain of Gn and Gc in the envelope glycoproteins of hantaan and seoul viruses was eliminated to enhance the solubility of protein and augment the level of vaccine expression. • The S2 drosophila expression system was utilized to acquire the universal subunit vaccines for hantaan and seoul viruses. • The neutralization induced by HTNV-Gc protein exhibited a greater protective effect compared to inactivated vaccines, as observed both in vitro and in vivo. Due to the global resurgence of hemorrhagic fever with renal syndrome (HFRS), more attention is being focused on this dangerous illness. In China and Korea, the only vaccines available are the virus-inactivated vaccine against Hantaan virus (HTNV) or Seoul virus (SEOV), but their efficacy and safety are inadequate. Therefore, it is important to develop new vaccines that are safer and more efficient to neutralize and regulate areas with a high prevalence of HFRS. We employed bioinformatics methods to design a recombinant protein vaccine based on conserved regions of protein consensus sequences in HTNV and SEOV membranes. The S2 Drosophila expression system was utilized to enhance protein expression, solubility and immunogenicity. After the Gn and Gc proteins of HTNV and SEOV were successfully expressed, mice were immunized, and the humoral immunity, cellular immunity, and in vivo protection of the HFRS universal subunit vaccine were systematically evaluated in mouse models. These results indicated that the HFRS subunit vaccine generated elevated levels of binding and neutralizing antibodies, particularly IgG1, compared to that of the traditional inactivated HFRS vaccine. Additionally, the spleen cells of immunized mice secreted IFN-r and IL-4 cytokines effectively. Moreover, the HTNV-Gc protein vaccine successfully protected suckling mice from HTNV infection and stimulated GC responses. In this research, a new scientific approach is investigated to develop a universal HFRS subunit protein vaccine that is capable of producing effective humoral and cellular immunity in mice. The results suggest that this vaccine could be a promising candidate for preventing HFRS in humans. [ABSTRACT FROM AUTHOR]