In Europe, Puumala virus and Dobrava virus are the major hantaviruses that cause hemorrhagic fever with renal syndrome in humans. As hantaviruses can cause diseases with high morbidity and mortality rates, and as to date there is no specific treatment, efforts are concentrated on the development of vaccines. In this study we characterized the immunogenicity of recombinant nucleocapsid proteins of Puumala virus (PUUV) linked to a carrier protein corresponding to the outer membrane protein A from Klebsiella pneumoniae (rP40). The rP40 molecule is a novel carrier protein that facilitates exogenous antigen uptake by dendritic cells. We cloned and expressed the recombinant PUUV proteins in the E. coli mutant ICONE 200 using the tryptophan promoter-controlled pTEXmp18 expression vector. All recombinant PUUV proteins were found to be highly immunogenic in NMRI mice after three immunizations of 10 mu5g each of the protein. Only the truncated construct, P40-Puu118, gave high antibody titers after two vaccinations of 0.2 mug each. Likewise in the challenge experiments in NMRI mice, only the truncated construct P40-Puu118 resulted in 100% protection after three immunizations of 10 mug each. The results suggest that P40-Puu118 in particular is a good candidate for a recombinant vaccine against Puumala virus. All recombinant proteins linked to rP40 induced high antibody responses, indicating that rP40 is a carrier protein with potential for use in other vaccines.